PINK BOOK 3

Sebacic Acid/Dicarboxylic Acids

CIR EXPERT PANEL MEETING AUGUST 30-31, 2010

Memorandum

To: CIR Expert Panel Members and Liaisons

From: Monice M. Fiume MMF Senior Scientific Analyst/Writer Bart A. Heldreth, Ph.D. BAH Chemist

Date: July 30, 2010

Subject: Draft Report on Dicarboxylic Acids (previously called Diisopropyl Sebacate)

The draft report on dicarboxylic acids was last reviewed in December 2009, under the title Diisopropyl Sebacate. At that time, the report was tabled for reorganization. Also at that time, it was determined that oxalic acid would not be part of the safety assessment.

The entire report has been reorganized and rewritten. It will be obvious that the Chemistry section is now prepared in a way that allows you to view the dicarboxylic acids in order of increasing chain length. Also included are charts demonstrating the relationship between molecular weight and the log octanol – water partitioning coefficient.

Additionally, the report, from the General Biology section on, is now divided into two sections, as was suggested by the Panel in December. The first section addresses dicarboxylic acids and their salts, while the second part addresses esters of dicarboxylic acids. These two subsets of ingredients have different functions in cosmetics, and this will allow the Panel to view the data on each subset separately.

In addition to the reorganization, the report has change greatly in the extent of data included. A complete new search of the literature has been performed, and a substantial amount of new data has been added. A great deal of the information included in this report has come from summary documents, such as HPV robust summaries, that cite unpublished sources. Whatever details were available have been included, but often the summaries were brief. Additional unpublished data received from Council, including concentration of use data, have been added.

Also included are summaries of information from the 1984 Final Assessment of Dioctyl Adipate and Diisopropyl Adipate and from the 2006 Amended Final Report on the Safety Assessment of Dibutyl Adipate as Used in Cosmetics. (Dioctyl adipate is now correctly named diethylhexyl adipate.) These reports have been included for your use.

The diesters have been shown to metabolize. Accordingly, data on esterase metabolites have been included as support information. For your ease in use of this data, an Appendix to the report, that

CIR Panel Book Page 1 summarizes these data, immediately follows the reference section. When applicable, these data are also included in the appropriate tables.

There are many aspects for discussion in this report, and we have tried to present it to you as cohesively as possible.

The following are included as paper copy:

1. Final report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate; 2. Amended Final Report on the Safety Assessment of Dibutyl Adipate as Used in Cosmetics; 3. Updated concentration of use data – dated February 25, 2010; 4. Unpublished data on Diethylhexyl Succinate, Diisobutyl Adipate, Diisocetyl Adipate, Diisodecyl Adipate, Dioctyldodecyl Adipate, Diethylhexyl Sebacate and Diisopropyl Sebacate; submitted January 27, 2010. (One in French has not yet been translated, so is not included in text.); 5. Unpublished data on Diisostearyl Adipate, Diisocetyl Dodecanedioate, and Dioctyldodecyl Dodecanedioate; submitted January 15, 2010; and 6. Unpublished data - Physical/Chemical Properties on Dibutyl Octyl Sebacate, Dihexyldecyl Sebacate, and Dioctyldodecyl Sebacate; dated February 15, 2010

CIR Panel Book Page 2

CIR Panel Book Page 3 REPORT HISTORY: DICARBOXYLIC ACID AND THEIR SALTS AND ESTERS

July 10, 2009: SLR was issued

September 2009 Panel meeting Initial review of the draft report. The recommendation was made to add 5 additional dicarboxylic acids – oxalic, malonic, succinic, glutaric, and adipic – and their salts and esters

December 2009 Panel meeting The report was tabled for reorganization into (1) acids and salts and (2) esters. It was also agreed that oxalic acid should be removed. It was also felt that more data should be available.

August 2010 Panel meeting The report has been completely reorganized. A new search was done and the text was updated with much new data.

CIR Panel Book Page 4 Sebacates Search Update June 2010 Toxline NTP HPV EU/SCCP EPA IARC OTC SIDS Sebacic Acid 0 0 NR/NS 0 0 Dibutyl Sebacate 0 0 NR/NS 0 0 Dibutyloctyl Sebacate 0 0 NR/NS 0 0 Dicapryl/Capryl Sebacate 0 0 NR/NS 0 0 Diethyl Sebacate 0 x NR/NS 0 0 Diethylhexyl Sebacate G 0 NR/NS 0 0 Dihexyldecyl Sebacate 0 0 NR/NS 0 0 Diisooctyl Sebacate 0 0 NR/NS 0 0 Diisopropyl Sebacate 0 0 NR/NS 0 0 Diisostearyl Sebacate 0 0 NR/NS 0 0 Dioctyldodecyl Sebacate 0 0 NR/NS 0 0 Disodium Sebacate 0 0 NR/NS 0 0 Isostearyl Sebacate 0 0 NR/NS 0 0 Malonic Acid G 0 NR/NS 0 0 Diethyl Malonate 0 0 NR/NS 0 0 Succinic Acid 0 x NR/NS 0 0 Sodium Succinate 0 0 NR/NS 0 0 Disodium Succinate 0 0 NR/NS 0 0 x Dimethyl Succinate G x NR/NS 0 0 Diethyl Succinate 0 0 NR/NS 0 0 Diethylhexyl Succinate 0 0 NR/NS 0 0 Decyl Succinate 0 0 NR/NS 0 0 Dicapryl Succinate 0 0 NR/NS 0 0 Dicetearyl Succinate 0 0 NR/NS 0 0 Diisobutyl Succinate 0 0 NR/NS 0 0 Glutaric Acid G x NR/NS 0 0 Dimethyl Glutarate G x NR/NS 0 0 Diisostearyl Glutarate 0 0 NR/NS 0 0 Diisobutyl Glutarate 0 0 NR/NS 0 0 Adipic Acid G x NR/NS 0 0 x Dimethyl Adipate G x NR/NS 0 0 Diethyl Adipate 0 0 NR/NS 0 0 Diethylhexyl Adipate C,G x NR/NS x 2 0 x Dipropyl Adipate 0 0 NR/NS 0 0 Dibutyl Adipate 0 x NR/NS 0 0 x Di-C12-15 Alkyl Adipate 0 0 NR/NS 0 0 Dicapryl Adipate 0 0 NR/NS 0 0 Dicetyl Adipate 0 0 NR/NS 0 0 Diheptylundecyl Adipate 0 0 NR/NS 0 0 Dihexyl Adipate 0 0 NR/NS 0 0 Dihexyldecyl Adipate 0 0 NR/NS 0 0 Diisobutyl Adipate 0 0 NR/NS 0 0 Diisocetyl Adipate 0 0 NR/NS 0 0 Diisodecyl Adipate 0 x NR/NS x 0 0 Diisononyl Adipate 0 x NR/NS x 0 0 Diisooctyl Adipate 0 x NR/NS x 0 0 Diisopropyl Adipate 0 0 NR/NS 0 0 Diisostearyl Adipate 0 0 NR/NS 0 0 Dioctyldodecyl Adipate 0 0 NR/NS 0 0 Ditridecyl Adipate 0 x NR/NS x 0 0 Azelaic Acid 0 0 NR/NS 0 0 Dipotassium Azelate 0 0 NR/NS 0 0 Disodium Azelate 0 0 NR/NS 0 0 Dodecanedioic Acid 0 x NR/NS 0 0 x Diisocetyl Dodecanedioate 0 0 NR/NS 0 0 Dioctyldodecyl Dodecanedioate 0 0 NR/NS 0 0 Totals 2744

CIR Panel Book Page 5

Toxline - searched 6-10-10; included Medline NTP - searched 6-20-10; G - genotoxicity, C - carcinogenicity HPV - searched 6-20&21-10; x - listed in HPV database EU - searched 6-20-10; NR - no restrictions; NS - no SCCP opinion IARC - searched 6-21-10 OTC - searched 6-22-10 (April 7, 2010 update) EPA - searched 7-8-10 found updated Robust Summaries for HPV items

TOXNET Search Strategy:

Sebacic 06-11-10 306 hits 111-20-6 OR 184706-97-6 OR 109-43-3 OR 110-40-7 OR 122-62-3 OR 359073-59-9 OR 10340-41-7 OR 7491-02- 3 OR 69275-01-0 OR 17265-14-4 OR 478273-24-4 OR (DICAPRYL AND CAPRYL AND SEBACATE) OR (DIISOSTEARYL AND SEBACATE)

Malonic-Succinic-Glutaric 06-10-10 2511 hits 141-82-2 OR 105-53-3 OR 110-15-6 OR 2922-54-5 OR 150-90-3 OR 106-65-0 OR 123-25-1 OR 2915-57-3 OR 2530-33-8 OR 14491-66-8 OR 93280-98-9 OR 925-06-4 OR 110-94-1 OR 1119-40-0 OR 71195-64-7 OR (DIISOSTEARYL AND GLUTARATE)

Adipic 06-10-10 1167 hits 124-04-9 OR 627-93-0 OR 141-28-6 OR 103-23-1 OR 106-19-4 OR 105-99-7 OR 105-97-5 OR 26720-21-8 OR 155613-91-5 OR 110-33-8 OR 141-04-8 OR 57533-90-1 OR 58262-41-2 OR 59686-69-0 OR 27178-16-1 OR 33703-08-1 OR 108-63-4 OR 6938-94-9 OR 62479-36-1 OR 155613-91-5 OR 85117-94-8 OR 16958-92-2 OR (ALKYL AND ADIPATE) OR (DIHEXYLDECYL AND ADIPATE)

Azelaic-Dodecanedioic 06-10-10 208 hits 123-99-9 OR 9619-43-3 OR 52457-54-2 OR 17265-13-3 OR 27825-99-6 OR 132499-85-5 OR 693-23-2 OR 131252-83-0 OR 129423-55-8

Combined files minus existing references – 2744

June 21, 2010 – 47 published papers ordered July 8, 2010 - 20 additional papers ordered July 15, 2010 – 10 additional papers ordered

*Metabolite alcohols and monoesters were searched for supplemental information in TOXLINE and RTECS, via STN. CAS Registry files were used to search in each of the databases. 55 published papers were ordered. Numerous other references on metabolite alcohols were available from prior safety assessments.

CIR Panel Book Page 6

Transcripts/ Minutes

PANEL - DEC 2009 CIR Meeting day 2 Page: 40 CIR Meeting day 2 Page: 41

1 isononanoates and all that -- those parts. And, 1 the aim of splitting the acids and salts into one 2 hopefully, we can have our -- you know, at the 2 group -- they are used as pH adjusters -- and then 3 next meeting, have a good discussion on whether 3 the esters into another group, rather than having 4 these are long shots or not, in other words, if 4 them all mixed in as is in the present document. 5 they're lipophilic, probably really concerned and 5 That could either be, as we discussed, 6 so forth, because I'd hate to pull -- 6 whether this should be two separate documents or 7 unnecessarily pull ingredients off the list when 7 one document and just have the document organized 8 there's a good reason to believe that it's not a 8 so these ingredients were split, and we felt that 9 problem. 9 we could proceed with one document. And then we 10 DR. BERGFELD: Well, thank you for that 10 also felt that within this list, that we should 11 clarification of who does what. 11 reopen three of the esters that have already been 12 All right. Well, then we move on. 12 determined to be safe by the CIR panel, and that's 13 We're moving on to the second ingredient under the 13 the diethylhexyl adipate, the dibutyl adipate, and 14 reports advancing and that's sebacates by Dr. 14 the diisopropyl adipate. So, we felt some more 15 Marks. 15 work needed to be done on the document and, 16 DR. MARKS: So, this is the second time 16 therefore, recommend to table it. 17 we've looked at the sebacates. In the September 17 DR. BERGFELD: Is there a second to 18 2009 meeting we decided to table these ingredients 18 table? 19 so that we could expand the number and I have a 19 SPEAKER: Second. 20 feeling we're going to have a discussion again 20 DR. BERGFELD: Second. There's no 21 procedurally how to go. Our team felt that we 21 discussion on the table. Motion, all those in 22 should continue to table these ingredients with 22 favor of tabling, please indicate by raising your

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net

CIR Meeting day 2 Page: 42 CIR Meeting day 2 Page: 43

1 hand? 1 into two or split into two pieces in one document. 2 DR. BELSITO: Any discussion? 2 We have lots of experience with both approaches. 3 DR. BERGFELD: No discussion on the 3 It's simply a question of approach. It doesn't 4 table. It's four against four? So, the chair has 4 address the question of any real additional data 5 to vote. I'll vote to table. So, this particular 5 needs. So, I'm not sure there's a benefit to 6 ingredient has been tabled and let's go over why 6 tabling from that standpoint. 7 it's being tabled again. 7 DR. BERGFELD: How about the addition of 8 DR. MARKS: Well, the first is that we 8 the other previously approved ingredients? 9 felt the document could be reorganized and that 9 DR. ANDERSON: I think -- 10 really by uses, so the acids and the salts are 10 DR. BERGFELD: Does that fit with the 11 used as pH adjusters and then the esters had other 11 first comment? 12 uses. And so the document would be split to 12 DR. ANDERSON: There is -- there are 13 discuss those two groups together, and that we, as 13 data that are available in the other safety 14 part of this document, reopened three esters which 14 assessments that previously the panel has 15 had previously been determined by the CIR as all 15 completed. To the extent that you want those data 16 are safe to be included in this new amended. 16 incorporated, tabling it to do that step is, I 17 DR. BERGFELD: May I ask, Alan, is this 17 think, a valid thing to do because clearly they're 18 really editorial or is this truly -- does it truly 18 not in there now. 19 have a need for being tabled? 19 DR. BERGFELD: Don? Comment? 20 DR. ANDERSON: Yes, I would have said 20 DR. BELSITO: Well, I mean, we obviously 21 that was an editorial change. It's just a matter 21 didn't vote to table, but some comments. First, 22 of reorganizing the document, whether it is split 22 yes, I mean, I think the other three should be

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net CIR Panel Book Page 7 CIR Meeting day 2 Page: 44 CIR Meeting day 2 Page: 45

1 included. Jim, I think you said dibutyl, you mean 1 review and grapple with. 2 dioctyl. 2 DR. LIEBLER: If I could just elaborate 3 DR. MARKS: Okay. Thank you, Don. 3 on that. 4 DR. BELSITO: The biggest discussion we 4 DR. BERGFELD: Don? 5 had revolving around this was oxalic acid, which 5 DR. LIEBLER: Yeah, I think the idea of 6 did not -- I mean, all of the safety data on that 6 including the smaller diacids and their esters and 7 specific acid, particularly in terms of causing 7 salts was mainly to expand the chemical space to 8 renal calculi, was not in this document. And we 8 things that are possibly relevant. But in getting 9 actually thought since we didn't see it listed as 9 down to oxalic acid, you run into some unique 10 having any cosmetic uses, that unless there were 10 toxicology that's very well documented, very 11 cosmetic uses for oxalic acid or its esters that 11 prominent, and quite problematic, and we don't 12 it be dropped from this family because of those 12 even know if this is used as a cosmetic 13 issues with renal toxicity. 13 ingredient. So, rather than get tangled up in all 14 DR. BERGFELD: So, your recommendation 14 of that, it's worth considering deleting the 15 under the conditions that we were tabling this is 15 oxalic acid from this report. 16 to reconsider the inclusion of oxalic acid? 16 DR. BERGFELD: John Bailey? 17 DR. BELSITO: Well, to first of all 17 MR. BAILEY: Yeah, I think we didn't 18 confirm whether, in fact, it's used -- currently 18 have time in getting these additional ingredients 19 being used, and, if not, then our recommendation 19 to survey -- do our usual survey of use and use 20 would be to remove it from this family because of 20 levels, so we can't answer definitively whether 21 those issues that were being countered and because 21 that's the situation or not, so I think we need 22 of the huge amount of data that we'd have to 22 some more information. I would point out that in

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net

CIR Meeting day 2 Page: 46 CIR Meeting day 2 Page: 47

1 Europe, oxalic acid is on annex 3 for hair dyes, 1 better simply to keep all the ingredients, which 2 so presumably, you know, that may be a use. And 2 is a large number of ingredients we have right 3 we haven't done the research necessary to get more 3 now, get rid of oxalic acid and, in fact, if it's 4 information about how it got there and use levels 4 used, we can always look at oxalic acid 5 and so forth. So, I would say, certainly oxalic 5 specifically rather than as a member of this 6 acid is in the textbooks. We all know about it, 6 group? Because I just think we're going to get 7 but there may be some additional information that 7 hugely bogged down even if it's used in trying to 8 would be relevant that we simply, you know, didn't 8 explain why it'd say brief discontinuous use on 9 have time to get. 9 the hair, no absorption, yadda, yadda, yadda. But 10 DR. BERGFELD: So, that's a promise to 10 we're going to have to look at all the data 11 get? 11 anyway. 12 MR. BAILEY: Certainly, yeah. We'll do 12 MR. BAILEY: Yeah, I concur. 13 our usual survey of -- 13 DR. BERGFELD: So, we might entertain a 14 DR. BERGFELD: Okay. Any further 14 motion at this time to delete oxalic acid from the 15 discussion? Don? 15 list. 16 DR. BELSITO: Yeah, I guess, just to 16 DR. BELSITO: I would like to make that 17 follow up on Dan's point, there is such a 17 motion. 18 voluminous amount of literature on oxalic acid 18 DR. BERGFELD: Is there a second? 19 that even if it's used in cosmetics, do we want to 19 Second. Is there a discussion of that 20 get bogged down dealing with why that is going to 20 recommendation? 21 be safe in a cosmetic product and reviewing all of 21 Curt? 22 that literature on oxalic acid? Or would it be 22 DR. KLAASSEN: No problem.

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net CIR Panel Book Page 8 CIR Meeting day 2 Page: 48 CIR Meeting day 2 Page: 49

1 DR. BERGFELD: Paul? Dan? Ron? 1 here. You'd have to pull out that list, Ron, that 2 That's okay. 2 you had where we would seem to be depending on 3 DR. HILL: I have no problem with that. 3 read-across data for things like genotoxicity, 4 DR. BERGFELD: Okay, we'll call for the 4 tumor-promoting potential, and even sensitization. 5 vote to delete oxalic acid from the list of 5 Am I mistaken there? We have direct data on the 6 ingredients. All those in favor? Unanimous, with 6 sebacates, some of the adipates, and a number of 7 the intent of taking it up as a separate 7 the others we don't. Isn't that correct? 8 ingredient at some time in the future when it's 8 DR. SHANK: That's correct. 9 been declared how it's used and how frequent it's 9 DR. HILL: So, we're looking at a pretty 10 used. 10 high number of read-across, which was another 11 MR. BAILEY: To that end, we'll include 11 reason, I think, basis, for why we had suggested 12 that in our survey just so we'll have that 12 the tabling idea, so that we had the time to 13 information. 13 really better assess those gaps. 14 DR. BERGFELD: Thank you. Ron? 14 DR. BERGFELD: We need to record your 15 DR. HILL: Yeah, in our discussions 15 response, Ron. 16 yesterday our starting point was really a 16 DR. SHANK: I agree with Dr. Hill that 17 suggestion that Ron Shank made to separate into 17 we have some information on the esters, but the 18 two documents, and then we talked about two sort 18 esters are used quite differently from the acids 19 of separate sections of the main document because 19 and salts. And that's why I had suggested that we 20 in the consideration of the diacids, they're 20 split them, either two reports or within the same 21 pretty much -- their uses are as pH adjusters and 21 report, make it very clear that the acids and 22 then similar like that. There are a lot of esters 22 their salts are -- can be handled one way and the

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net

CIR Meeting day 2 Page: 50 CIR Meeting day 2 Page: 51

1 esters will have to be handled a different way. 1 DR. HILL: Well, I'm relatively new, 2 DR. ANDERSON: Yeah, I really don't 2 but, you know, I mean, I still have the same 3 think that that's a problem in terms of overall 3 issues. There's some of the alcohols that we just 4 strategy. It provides the opportunity to be a lot 4 talked about in the pelargonate section that show 5 clearer to the reader, that we understand the 5 up again here. And if there's no reason to 6 fundamental difference between how these chemicals 6 believe that there's any significant generation of 7 are used in cosmetics. So I think we'll simply do 7 those alcohols in the skin, if there's no reason 8 that. 8 to believe that those alcohols would lead to 9 Now, I am concerned about the suggestion 9 sensitization in the skin, then there's probably 10 that read-across may be problematic. I would ask 10 -- and there's no reason to believe that either 11 the panel to reserve that, to focus on the data, 11 the alcohols or, in fact, these esters that we 12 where the data gaps are, and what your comfort 12 have no data on could promote tumor development in 13 level is going to be with the ability to use data 13 skin, then we don't have a problem. But I'd like 14 on one chain length ester of dicarboxylic acid to 14 to know, when I make a final vote, that I'm 15 inform your decision about another chain length, 15 comfortable whether that is valid scientifically, 16 all other things being equal. So, having all data 16 and that's really the only concern I have is that 17 on all ingredients is not the norm. So, I think 17 it's scientifically valid to make that conclusion. 18 there needs to be a comfort level with some degree 18 DR. ANDERSON: Point well taken. 19 of read-across as you evaluate these data. Only 19 DR. BERGFELD: Thank you. So we're 20 you folks on the panel can reach that level of 20 going to be tabling the sebacates and we've had 21 comfort, but I would ask you not to veto it a 21 the discussion and we're going to reorganize the 22 priori. 22 document, and we're going to get some more

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net CIR Panel Book Page 9 CIR Meeting day 2 Page: 52

1 information on some of the ingredients. Is that 2 correct? 3 DR. ANDERSON: Mm-hmm. 4 DR. BERGFELD: All right. 5 DR. ANDERSON: And we're deleting oxalic 6 acid -- 7 DR. BERGFELD: And we're deleting oxalic 8 acid, thank you. 9 DR. ANDERSON: Deleting oxalic acid. 10 Nobody has specifically said it, but let me say 11 it, we're also going to delete the oxalic acid 12 esters. 13 DR. BERGFELD: Correct. All right. 14 It's time to move on to the next ingredient, the 15 PEGs. Dr. Belsito. 16 DR. BELSITO: Yes. At our September 17 meeting, we reviewed results from industry 18 essentially done on tape- stripped skin as a 19 method of looking at PEG toxicity on damaged skin 20 because previously we had a conclusion that 21 restricted the use of PEGs on damaged skin. We 22 had a chance to look at what would be absorbed.

ANDERSON COURT REPORTING 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net

CIR Panel Book Page 10 Cosmetic Ingredient Review Panel December 8, 2009 Page: 143 Cosmetic Ingredient Review Panel December 8, 2009 Page: 144

1 Sebacate? Sebacate? 1 this morning on dermal penetration. We don't have 2 SPEAKER: You asked that last time. 2 a lot of genotox and carcinogenicity here. But we 3 DR. BELSITO: Sebacates. Sebaceous. 3 do for the azelaic acid, which is used as a 4 Okay, so we're going to go to the -- 4 topical, have some information. So if there is a 5 SPEAKER: Sebacates. 5 concern about that, I guess we still have Bob here 6 DR. BELSITO: Dicarboxylic Acid Salts 6 and he can tell us what he would think about the 7 and Ester Report. That, I know how to pronounce. 7 absorption of these. 8 Okay. And I guess as part of the 8 So with that as an intro, the question 9 add-ins now, we've been provided with a final 9 is do we have enough here for this group of 10 report that we did on dioctyl and diisopropyl 10 dicarboxylic acids, which now will contain azelaic 11 adipate. And we're back in December. We tabled 11 acid, malonic, succinic, glutaric, adipic, and 12 this to look at a whole bunch of related 12 sebacic acid and their salts and esters that are 13 dicarboxylic acids that we agreed to incorporate 13 listed in the Cosmetic Dictionary to go with a 14 into this report. And we've gone out and got 14 safe as used conclusion? 15 information on azelaic acid as used in topical 15 SPEAKER: We have, to date, not received 16 drugs. And we've got some new unpublished data 16 use data. 17 that we saw briefly at the September meeting but 17 MS. ROBINSON: It's forthcoming. Carol 18 didn't really have time to digest. And so now 18 (inaudible) by the next -- 19 we've got this tentative safety assessment and all 19 SPEAKER: Mic, please. 20 of the information in it is in here, short of a 20 MS. ROBINSON: The use information is 21 discussion and a conclusion, so. 21 forthcoming. Carol has said that it may be here 22 Then we've heard a little bit from Bob 22 by the next meeting.

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net

Cosmetic Ingredient Review Panel December 8, 2009 Page: 145 Cosmetic Ingredient Review Panel December 8, 2009 Page: 146

1 DR. BELSITO: But how can we -- 1 DR. BELSITO: Okay. And one comment I 2 SPEAKER: By the next meeting? April? 2 have is are the impurities sufficient? We have 3 DR. ANDERSON: We don't have it. 3 impurities only for dimethyl malonate and 4 DR. LIEBLER: Can I get a clarification 4 diethylmalonate and diethylhexyl adipate. We 5 of that? Were these the data that were from the 5 don't have impurities for the other compounds 6 American Chemistry Council? 6 we're reviewing. 7 MR. ANSELL: That data has been 7 So I just, you know, sort of throw that 8 included. This is the use concentration on all 8 out as an aside, to get the sense of chemists 9 the add-ons, which came to us in too short a time 9 looking at the way these are manufactured as to 10 to turnaround between these. 10 whether they would be concerned about that. 11 DR. LIEBLER: Okay, so we did -- last 11 DR. LIEBLER: I'd like to see data that 12 time we did talk about the American Chemistry 12 describes the impurities, at least the types of 13 Council data that we thought would be useful. And 13 compounds that would be present as impurities and 14 there was a question of whether or not we joined 14 their approximate ranges of concentrations. 15 -- or CIR joined in or somehow got access to that. 15 DR. BELSITO: And then in terms of 16 So I see some of the data in this report. And 16 concentrations of use, I mean, we do have some 17 just, I'm curious, how do we get that? 17 information here if we go to Table 4, particularly 18 MS. ROBINSON: Well, to date, we haven't 18 for the sebacic acid groups we have. We have some 19 received any additional data from the last 19 for the succinate. We have some for adipic acid 20 meeting, from the reports. So what you see in the 20 and its groups, and then we have azelaic acid. So 21 report is essentially the same that we saw at the 21 we have some concentrations of use. And we have 22 last meeting. 22 that standard, you know, asterisk boilerplate that

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net CIR Panel Book Page 11 Cosmetic Ingredient Review Panel December 8, 2009 Page: 147 Cosmetic Ingredient Review Panel December 8, 2009 Page: 148

1 if where we're not giving a concentration of use, 1 aware of any sort of pharmacodynamic reasons why 2 we assume it's being used in the same 2 different chain length dicarboxylates would have 3 concentration as the other groups. 3 different toxicological effects. 4 So I'm not sure -- I mean, it would be 4 DR. BELSITO: Curt? 5 nice to get updated information, and there are 5 DR. KLAASSEN: I agree. 6 lots and lots of blanks here, I agree. But I 6 DR. SNYDER: And the overall toxicity is 7 think the issue that bothered me the most was the 7 relatively low. Pretty high OB50s. 8 lack of impurities in all these groups that we're 8 SPEAKER: Okay. 9 adding in. 9 DR. BRONAUGH: I guess there was an 10 And then, of course, we're relying on 10 absorption study in guinea pig skin of the diethyl 11 azelaic acid and the data from the Finacea and the 11 -- what is it? -- the diethylhexyl sebacate, which 12 Azelex cream to support a lot of our systemic or 12 the molecular weight looks to be under 500. You 13 lack thereof of systemic toxicity. So again, I 13 know, sometimes, if you have a large family of 14 just throw out is there any concern here about 14 ingredients, it might be useful if we could know 15 absorption of the other molecules? Because for -- 15 what the molecular weight is, what the logP is, so 16 and again, it's back to Dan whether you can. You 16 you could look through and have a better sense. 17 know, is the systemic toxicity going to be 17 But, you know, this data is in a 18 generalizable across all these dicarboxylic acids 18 hairless guinea pig -- or the guinea pig, anyway, 19 or are there any that you would be concerned 19 that was given to us, where they said it was 20 about? 20 readily absorbed. But I would imagine some of 21 DR. LIEBLER: Aside from parameters that 21 these molecules are absorbed. But, you know, it's 22 would affect absorption and distribution, I'm not 22 really kind of hard to say without, you know, how

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net

Cosmetic Ingredient Review Panel December 8, 2009 Page: 149 Cosmetic Ingredient Review Panel December 8, 2009 Page: 150

1 to compare them without knowing a little bit more 1 you can calculate logP and water solubility just 2 about their chemistry. 2 knowing the SMILES code of any ingredient. And 3 DR. KLAASSEN: One would think that 3 again, the SMILES codes are available on the NIH 4 these would be extremely water soluble and not be 4 site, for many compounds. 5 absorbed so well, right? 5 So it might be useful to have that 6 SPEAKER: (inaudible) the mic. 6 sometimes. 7 DR. KLAASSEN: Yeah. 7 DR. KLAASSEN: Yeah, I agree 100 8 SPEAKER: Sometimes I -- if it -- 8 percent. 9 DR. KLAASSEN: You know, in regard to 9 DR. BELSITO: But if the toxicity is the 10 these dicarboxylic acids, I guess it'd be my 10 same, at least systemic toxicity is, that was the 11 impression that they'd be so water soluble that 11 reason I asked the question. And we have use that 12 they probably wouldn't be absorbed. I think it 12 looks like it's not going to above 10 percent in 13 would be nice to see some partition coefficients, 13 cosmetics. And then we have Azelex cream, which 14 etcetera. 14 has been looked at by FDA, that's 20 percent for 15 DR. BRONAUGH: It would take a little 15 treatment of acne. Then I think I'm okay as long 16 extra time, but maybe it'd be worthwhile if -- I 16 as I don't see cosmetic products coming out above 17 mean, this data could be calculated in a family 17 10 percent or certainly over the 20 percent in 18 like this and we'd have a better idea. 18 terms of, you know, okay, you know, is there 19 DR. KLAASSEN: And in your talk this 19 absorption, you know, and what are slight 20 morning didn't you say that there was some EPA or 20 differences in absorption. 21 FDA website that had a lot of that data on there. 21 But I agree in principle with what Bob 22 DR. BRONAUGH: It has the software where 22 is saying. As we create these superfamilies, it

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net CIR Panel Book Page 12 Cosmetic Ingredient Review Panel December 8, 2009 Page: 151 Cosmetic Ingredient Review Panel December 8, 2009 Page: 152

1 would be nice if CIR would go to that website and 1 report, in fact, did use the EPA program to 2 provide us with a list of molecular weights and 2 calculate logPs, and these are included. So for 3 logPs. You know, particularly when it comes to 3 dimethyl sebacate, it's 3.4 for a logP; for 4 picking, you know, if there is an issue about 4 diethylhexyl sebacate, it's 3.74 there. Just 5 absorption, what ingredient do we want as the 5 outside, if you will, of the -1 to 3 range that 6 model ingredient to assure the safety for the 6 would suggest penetration, but not far enough 7 entire family. Well, it would be the one that we 7 outside so that I'd be comfortable for giving them 8 would predict from these various parameters would 8 a free ride. I think you'd have to presume there 9 be most likely to be absorbed. So I think it 9 is some absorption of these things. Then the 10 would be helpful. 10 question becomes what's the toxicity, which is 11 MR. ANSELL: To the discussion on 11 rather low. And I wouldn't kiss them off on the 12 systemic safety, we would like to point out that 12 basis of dermal penetration. 13 CIR has already conducted and found safe as used 13 SPEAKER: Yeah. 14 three of the adipates, including diethylhexyl, 14 DR. ANDERSON: But I take your point 15 dibutyl, and diisopropyl. And that the 15 that a table that presents just molecular weight 16 diethylhexyl has already gone through a complete 16 and logP, if those data are available, paints an 17 NTP screen through carcinogenicity. 17 interesting picture that you'd like to see. 18 DR. ANDERSON: I think the, excuse me, 18 DR. BRONAUGH: Yeah, I forgot about that 19 Table 2 already includes many of the data you're 19 data. I don't have a copy. Linda has our copy of 20 talking about. Admittedly they are buried in 20 the book. I forgot that that was done for this 21 amongst all of the other chemical and physical 21 series of compounds. And I think it was very 22 properties. But the American Chemistry Council 22 useful. There were some gaps in there that maybe

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net

Cosmetic Ingredient Review Panel December 8, 2009 Page: 153 Cosmetic Ingredient Review Panel December 8, 2009 Page: 154

1 could have been filled in, but I think that's 1 vapor pressure, density even, we have all of these 2 useful. 2 other things. It's kind of like the important 3 SPEAKER: That's good to know. 3 data gets lost in the minutiae. 4 DR. LIEBLER: You could represent the 4 SPEAKER: Yep, the -- 5 calculated logP values in a plot, like the plot 5 DR. LIEBLER: With the Henry's Law 6 for the phenyls that Bob showed during his talk, 6 constant, you're saying. 7 in which you'd number, instead of just using round 7 DR. ANDERSON: I take your point. Just, 8 dots as the points, you actually use numbers as 8 we need a good chemist on staff. 9 the points. And the numbers could refer back to 9 DR. BELSITO: Now, were the data from -- 10 the compounds in a table. And what that would 10 all of the data from this dioctyl and diisopropyl 11 allow us to do is see much better which compounds 11 adipate report incorporated into this report? 12 are within the range or close to the range that's 12 Because it seems like they have not been. 13 typically associated with the potential for 13 And did I understand, Jay, from your 14 absorption. 14 comment, that there is another adipate that we've 15 DR. ANDERSON: That raises an 15 also previously reviewed out there? 16 interesting idea of what would that plot look like 16 MR. ANSELL: Yeah, we have three 17 since we have calculated logP values on one axis 17 adipates that were CIR reviewed, which have not 18 as a function of molecular weight on another. 18 been included. These were diethylhexyl, dibutyl, 19 DR. KLAASSEN: Well, those are two of 19 and diisopropyl adipate. 20 the more important characteristics of chemicals 20 DR. BELSITO: Okay. So then there's the 21 that are important to us. And, you know, a lot of 21 diethylhexyl has also not been incorporated into 22 these that we have, you know, like boiling point, 22 this at this point?

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net CIR Panel Book Page 13 Cosmetic Ingredient Review Panel December 8, 2009 Page: 155 Cosmetic Ingredient Review Panel December 8, 2009 Page: 156

1 MS. ROBINSON: If it was from the 1 Dan's question. And we did find robust summaries, 2 original report, it has not been incorporated. 2 and I do believe they have been incorporated. 3 But an updated search was performed, so anything 3 DR. LIEBLER: I couldn't tell what was 4 outside of that is in the report. 4 different. I guess I didn't have the previous 5 DR. BELSITO: Okay. So we have one 5 book, so. I saw some references to the American 6 other report, other than this one, that needs to 6 Chemistry Council HPV summaries. Is that what 7 be incorporated into this document, as well? 7 you're referring to, Jay? 8 SPEAKER: (inaudible) 8 MR. ANSELL: Yeah. 9 DR. BELSITO: Okay. 9 MS. ROBINSON: Yes, those are new. 10 MR. ANSELL: We also -- the NTP study, 10 DR. LIEBLER: Those are new. 11 we don't believe has been incorporated on a whole 11 MS. ROBINSON: Yes. 12 series of materials. I'm also unclear as to 12 DR. LIEBLER: Since September. 13 whether the -- we did obtain from ACC robust 13 DR. ANDERSON: That's correct. Those 14 summaries that we were requested to -- and the 14 were not in the document. 15 cover letter suggests they were included, but 15 DR. LIEBLER: Okay. 16 Valerie's suggesting they're not. So I think 16 DR. ANDERSON: And the only issue from 17 there's some data that needs to be included. 17 the panel's perspective is that now that you 18 MS. ROBINSON: The robust summaries from 18 understand dicarboxylic acids were in the American 19 ACC were incorporated, but we haven't received any 19 Chemistry Council report, there is a couple of 20 additional information that we asked for prior. 20 points of overlap with this safety assessment, and 21 MR. ANSELL: The data which was 21 is that enough? 22 requested of us in September, though, I think was 22 DR. BELSITO: Okay. Where do we get the

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net

Cosmetic Ingredient Review Panel December 8, 2009 Page: 157 Cosmetic Ingredient Review Panel December 8, 2009 Page: 158

1 information as to what was in the American 1 impurities? Are we going insufficient for more 2 Chemical report? How do we divine that from this 2 concentration of use? Are we going safe as used 3 document? 3 when we add in the NTP summaries and the 4 DR. ANDERSON: Well, if you go to page 4 information from the dioctyl, diisopropyl, 5 34, for example, for the question, really, of the 5 diethylhexyl adipate documents that were 6 genotoxicity data, then there's specific mention, 6 previously published? 7 you know: Mammalian cell gene mutation assay. 7 Paul, where are you? 8 Two ingredients in this assessment were included 8 DR. SNYDER: Sounding an awful lot like 9 in the ACC report and were negative in the mouse 9 insufficient. I mean, Dan's comment to the 10 lymphoma assay. 10 impurities issue, he's not comfortable with the 11 DR. BELSITO: Okay, so you're talking 11 absence of additional impurities data. 12 about the mutagenicity studies, in particular. 12 DR. LIEBLER: So I'd be happy with 13 DR. ANDERSON: Yes. 13 representative data for some of these compounds 14 DR. BELSITO: Okay. 14 for impurities. I mean, we have a wide range of 15 DR. ANDERSON: Yeah, that was the -- 15 things from small dicarboxylates to big, 16 DR. BELSITO: Wasn't clear where you 16 long-chain esters, which are going to result from 17 were. 17 different processes and are going to have 18 DR. ANDERSON: Yeah, I'm sorry. That 18 different impurities. We may not have impurity 19 was the major gap. Without those data, the 19 data for every compound in the table, but I'd like 20 genotoxicity section was really quite bleak. 20 to see representative impurity data from compounds 21 DR. BELSITO: Okay. So where are we 21 that represent the processes that go into these 22 with this? Are we going insufficient for 22 products.

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net CIR Panel Book Page 14 Cosmetic Ingredient Review Panel December 8, 2009 Page: 159 Cosmetic Ingredient Review Panel December 8, 2009 Page: 160

1 And I was struck, in concentrations and 1 council and industry for some representative 2 frequency of use, that it seemed like they were 2 impurity data? 3 inversely related. The compounds with recorded 3 DR. LIEBLER: I'd like to see impurity 4 significant frequencies of use had almost no 4 data on the dicarboxylates. 5 concentration data and the compounds with bigger 5 DR. BELSITO: But which ones? Any 6 numbers for frequency of use tended to have no 6 specific one that you're concerned about, or? 7 concentration data. 7 DR. LIEBLER: How about sebacate. 8 Talking about Table 4. 8 DR. BELSITO: Okay. 9 DR. ANDERSON: Yeah, and that's very 9 DR. LIEBLER: As representative. And 10 likely an artifact of the late addition of many of 10 then how about succinate as -- or malonate as 11 the compounds. 11 representative? So you get a longer one and a 12 DR. LIEBLER: Do you think the 12 shorter one. 13 information's available and we just haven't gotten 13 DR. BELSITO: Okay. 14 it into a table yet? 14 DR. LIEBLER: And then how about the 15 DR. ANDERSON: I think that's the case. 15 impurity data on short esters of both of those and 16 DR. BELSITO: That's what we're told, 16 long esters of both of those? I think that 17 that Carol -- 17 represents the chemical space reasonably well. 18 DR. LIEBLER: I see, okay. 18 DR. BELSITO: Okay. So then what we're 19 DR. BELSITO: So then what we're looking 19 suggesting is that we are going insufficient for 20 at is Dan would like some representative impurity 20 impurities. And we would like to see something on 21 data. Would you like to mention any specific ones 21 the sebacates or sebacic acid, and either malonic 22 you'd like or just leave it open-ended back to the 22 or succinic acid, and then whatever two you

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net

Cosmetic Ingredient Review Panel December 8, 2009 Page: 161 Cosmetic Ingredient Review Panel December 8, 2009 Page: 162

1 choose, some impurity data on short- and a 1 mean, it's not like it's going out for a final. 2 long-chain ester of each of those acids. And we 2 And if we were going out for a final, it was going 3 would appreciate getting updated concentration of 3 to be, you know, we're saying it's insufficient, 4 use, particularly for those ingredients that have 4 and you're going to get the data for us before the 5 considerable use. And assuming that that data 5 next meeting. But, I mean, if anything, I mean, 6 looks clear, I am guessing that we would be going 6 PC has been -- 7 safe as used for these dicarboxylic acids. 7 DR. BERGFELD: Are you asking for an 8 DR. BERGFELD: So are you going out 8 announcement? 9 insufficient or are you tabling it? 9 DR. BELSITO: -- after us to expedite 10 DR. BELSITO: No, insufficient. I mean, 10 the process. Well, this expedites the process. 11 it's only -- it's at a pink stage. 11 It moves it up to the next stage and it tells you 12 MR. ANSELL: But there's so much data 12 where we're at. 13 which was not included in the report, and so many 13 DR. BERGFELD: Are you going out for 14 added after, it would be possible for us to have 14 insufficient data announcement or -- 15 provided the data. We would prefer it be tabled 15 DR. BELSITO: Yeah. 16 to include CIR's reports, to include the NTP 16 DR. BERGFELD: -- insufficient final 17 study, to include the actual concentrations of 17 announcement? 18 use. 18 DR. BELSITO: No, this is not even a 19 DR. BELSITO: But I guess, Jay, I don't 19 final. This is pink, Wilma. 20 see how tabling it versus us telling you what we 20 DR. BERGFELD: No, no, I know, but I'm 21 find is insufficient in the current report really 21 -- 22 changes things since it's at a pink stage. I 22 DR. BELSITO: We haven't even gotten --

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net CIR Panel Book Page 15 Cosmetic Ingredient Review Panel December 8, 2009 Page: 163 Cosmetic Ingredient Review Panel December 8, 2009 Page: 164

1 DR. BERGFELD: I know, but we had jumped 1 flagged these very long list of additions too late 2 over the announcement part in our new progression. 2 for the council to in any way be expected to 3 But you're going to input in this one 3 respond with use concentration data. So those are 4 announcement, insufficient data announcement. 4 -- there's some serious gaps in those data. 5 DR. BELSITO: Right. 5 But it is at a stage when you would be 6 DR. BERGFELD: It's different than a 6 expected to do one of two things: Issue an 7 final. 7 insufficient data announcement or issue a 8 DR. BELSITO: It's not a final, it's 8 tentative report. And you flag that there are 9 just a -- 9 clearly data that you want. In our expedited 10 DR. BERGFELD: No, no, I know that. But 10 process of keep it moving, this is ready for an 11 we had done away with announcements. We've done a 11 insufficient data announcement. That's, 12 few of them, but we've done away with that process 12 processwise, that's where it is. 13 a little bit. Haven't we? Alan? We were trying 13 There is the issue of the couple of 14 to expediate the flow. 14 oopses. And I wouldn't be upset if you used that 15 DR. ANDERSON: Yeah. I think in terms 15 as a basis for saying, look, we got to fix these 16 of process, at the last meeting the panel agreed 16 gaps. We've got the reports from previous safety 17 to move forward to prepare a draft report which in 17 assessments that weren't captured. You need to do 18 our judgment, given your comments, would be ready 18 that. And arguably that would allow enough time 19 to be issued as a tentative report. There were a 19 for the use data to come in. And I don't see any 20 couple of oopses in that process in that a couple 20 difficulty in tabling it. I would write in the 21 of background reports prepared by the panel didn't 21 post-meeting announcement that part of tabling it 22 get captured. The other oops is that the panel 22 is the expectation for the impurities data that

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net

Cosmetic Ingredient Review Panel December 8, 2009 Page: 165 Cosmetic Ingredient Review Panel December 8, 2009 Page: 166

1 Dan has outlined. 1 So, I mean, it's -- we go insufficient, we list 2 The risk of that is that we get to April 2 what we need, clean up the document, update 3 and those data don't show up. 3 concentration of use, give us some impurities, 4 DR. BELSITO: Exactly. 4 then I think it's very clear what we're looking at 5 DR. ANDERSON: And that's -- 5 for safety. And that's what I would -- I mean, 6 DR. BERGFELD: However, the document is 6 that's my argument. In the spirit of trying to 7 cleaned up and all of the -- 7 expedite getting these documents done, I think the 8 DR. ANDERSON: Yeah, but you just 8 best way to expedite it is to be very upfront as 9 delayed it -- 9 to where we are. 10 DR. BELSITO: But cleaning up the 10 SPEAKER: Well said. 11 document is not -- I mean, if all of the 11 DR. KLAASSEN: There's one additional 12 information we needed was in the oops documents 12 comment I'd like to make. And that is one major 13 that didn't get in and an update in the 13 group of chemicals in this larger class is oxalic 14 concentration of use, then I would be very happy 14 acid and its various compounds. And the toxicity 15 tabling it. But I'm just concerned that if we 15 of oxalic acid is well known, if it gets absorbed, 16 want the impurities data and, you know, Dan is the 16 in that it's quite -- produces kidney injury. 17 man I go to to tell me if we need that, and we 17 And, in fact, that's the toxicity from ethylene 18 just table it, even if Alan says, oh, by the way, 18 glycol, is that it's metabolized oxalic acid and 19 you know, I don't think, based upon past 19 it causes severe kidney injury. And while I don't 20 performance of industry, we're going to get it. 20 think we're probably getting enough absorbed here 21 And then we're going to be sitting at the April 21 to be a problem, there's really not much data in 22 meeting where we were at the December meeting. 22 this entire document on oxalic acid. And I just

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net CIR Panel Book Page 16 Cosmetic Ingredient Review Panel December 8, 2009 Page: 167 Cosmetic Ingredient Review Panel December 8, 2009 Page: 168

1 want to make sure people are aware of that. 1 September, this category was expanded at Ron 2 DR. BELSITO: There's a lot of 2 Shank's suggestion based on incorporating the 3 literature on it. 3 greater range of chemistries. But I think the 4 DR. KLAASSEN: There's a lot of 4 rationale was going across the chemical space as 5 literature, but it's not in here. 5 opposed to the cosmetic ingredient space, and we 6 DR. BELSITO: Yeah, I know. 6 might have picked up some unnecessary compounds in 7 DR. KLAASSEN: And I think somehow we 7 doing so; oxalic acid and its esters being 8 need to at least address oxalic acid in a 8 representative of that. 9 discussion or something, if there is good reasons 9 DR. BELSITO: But they're in the 10 why we haven't kind of paid special attention to 10 dictionary. And we thought we could cover them in 11 it. And, in fact, it's -- yes, there's a lot of 11 this report. One of the things that we get 12 information out there. In fact, it's -- also, a 12 criticized for by groups such as the Environmental 13 lot of plants contain oxalic acid and -- so, 13 Working Group is that there are X-number of 14 anyhow, it needs to be somehow addressed. 14 thousands of chemicals in the dictionary and we 15 DR. LIEBLER: I didn't see oxalic acid 15 have looked at only a certain small percentage of 16 listed in Table 4, or any of its derivatives 16 them. And so that was the whole purpose for 17 listed in Table 4. Is it used in cosmetic 17 grouping these. And so if oxalic acid is in the 18 products? Oxalic acid or oxalate esters? 18 dictionary and the functions are similar to the 19 MR. HAVERY: We don't have any reported 19 other dicarboxylic acids, and we feel that the 20 uses for oxalic acid. 20 safety can adequately be reviewed, you know, we do 21 DR. LIEBLER: So I think that -- my 21 it. And then there's that footnote at the end 22 recollection is that last time we met, in 22 saying, well, you know, oxalic acid and its esters

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net

Cosmetic Ingredient Review Panel December 8, 2009 Page: 169 Cosmetic Ingredient Review Panel December 8, 2009 Page: 170

1 aren't being used, but if they were to be used, 1 level of absorption from a cosmetic product, I 2 we're assuming that they would have the same 2 mean, if it's a true no-brainer, then we should 3 function and the same concentration as everything 3 add it. If it's not a true no- brainer, then we 4 else in this report. 4 shouldn't. And I guess that's a point that, I 5 SPEAKER: (inaudible) review. 5 mean, if you want to raise tomorrow, we can 6 DR. LIEBLER: They may just -- and 6 certainly do that. 7 oxalic acid may be somewhat unrepresentative of 7 DR. LIEBLER: Yeah, I just don't know 8 this class of compounds. I mean, in the case of 8 what the data are on absorption of oxalic acid in 9 oxalic acid in the kidney, it forms these very 9 a use that would be consistent with a cosmetic 10 insoluble crystals with calcium, calcium oxalate, 10 product, oxalic acid or oxalic acid derived from 11 very insoluble. So it's kind of the -- 11 oxalate esters. 12 mechanistically, it's an outlier. 12 So if it's insignificant, yeah, then I'm 13 DR. KLAASSEN: And it is at the end of 13 not worried. If it's likely to be more 14 the chain that we're talking about. I mean, it is 14 significant, then I would worry. 15 the smallest one. 15 DR. BELSITO: Well, 10 percent would the 16 DR. BELSITO: Well, if mechanistically 16 highest concentration, at least from the limited 17 it's an outlier, and if that's the issue, and then 17 use data we have right now, in a cosmetic product. 18 if you're concerned about absorption, then we 18 Bob, what is your sense for absorption 19 could easily delete it by saying that it is an 19 of oxalic acid versus a shorter chain acid? 20 outlier. You know, that we cannot group this 20 DR. BRONAUGH: I don't know. It's 21 specific dicarboxylic acid with the others. On 21 really hard to say. Oh, excuse me. It's really 22 the other hand, if you're not concerned about the 22 hard to say. I guess, and not being a chemist, I

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net CIR Panel Book Page 17 Cosmetic Ingredient Review Panel December 8, 2009 Page: 171 Cosmetic Ingredient Review Panel December 8, 2009 Page: 172

1 don't automatically know solubility properties. I 1 here, or at least in my mind. 2 mean, it's certainly an acid; it's going to be 2 DR. SNYDER: I guess my question goes to 3 water soluble. I don't know to the extent that it 3 why didn't the scientific literature review then 4 would penetrate through the stratum corneum if it 4 identify some of this information regarding the 5 has no lipid solubility. But it depends. I don't 5 renal toxicity? So did you search under the term, 6 know the pKa of oxalic acid. I'm not sure what 6 all of those terms? 7 that is. 7 MS. ROBINSON: Mm-hmm. Yes, I did. 8 DR. BELSITO: Well, we have in, let's 8 Yes, I did the search on the terms. And a couple 9 see, the table, the solubility is 1 gram per 7 mls 9 of the studies Halyna and I took out because there 10 of water. And -- 10 wasn't enough information in the published report. 11 SPEAKER: That's very soluble. 11 I can actually bring those back in. 12 DR. BELSITO: -- that's it. 12 DR. BELSITO: Well, what I'm hearing is 13 DR. KLAASSEN: But then you have the -- 13 that, at least from both Dan and Curt, is that 14 DR. BELSITO: We don't have a -- 14 they have enough concern that, again, the add-ons 15 DR. KLAASSEN: That's the dibutyl 15 are supposed to be no- brainers, and we've now 16 oxalate. 16 spent at least 10 minutes discussing whether 17 DR. BRONAUGH: Right. Now there you 17 oxalic acid actually fits into this report, and 18 would start expecting some penetration. 18 we're not seeing any evident cosmetic use of it. 19 DR. KLAASSEN: Yeah. 19 So the no- brainer, you know, swore it would say 20 DR. BRONAUGH: In lipophilicity. 20 that we should eliminate oxalic acid and its 21 DR. KLAASSEN: I guess the point is this 21 esters from this report. 22 is not a no-brainer. I mean, there's enough doubt 22 DR. KLAASSEN: I agree. And just to put

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net

Cosmetic Ingredient Review Panel December 8, 2009 Page: 173 Cosmetic Ingredient Review Panel December 8, 2009 Page: 174

1 this in perspective, some people, like myself, and 1 So we'll follow up on that and see whether that 2 maybe us around this table have a job because of 2 would have any bearing on that conclusion. 3 the toxicity of oxalic acid. So in the 1930s, to 3 DR. BELSITO: So, to recap. We are 4 get a drug on the market what you had to do was to 4 suggesting that the addition of oxalic acid to 5 show that it was effective. But then in the 5 this group is not a no-brainer and that we would 6 1930s, when the sulphonamides came out, they 6 recommend that oxalic acid and the esters be 7 dissolved the sulphonamides in ethylene glycol. 7 removed. Once that is done, if it's done, we find 8 And then we had a number of people that died, 8 this group to be insufficient for impurities, and 9 especially children, from kidney injury. And the 9 we would like impurities on a representative short 10 toxicity really was due to the ethylene glycol 10 and long chain. The suggested ones were sebacate 11 being metabolized oxalic acid. And as a result of 11 and malonic or succinic acid. And then 12 that, the FDA, Congress changed the laws for FDA 12 subdividing those, we would like short- and long- 13 to get a drug on the market. You not only had to 13 chain esters of a short- and long-chain acid, 14 show that it was effective, but that it was not 14 impurities for those representative ones. And 15 toxic. So this whole business around oxalic acid 15 then concentration of use. And the assumption, 16 is kind of, like, no small story in the history of 16 also, is that when this document comes back to us, 17 toxicology, for whatever that's worth. 17 that the NTP summary and that the information on 18 MR. ANSELL: And, you know, I certainly 18 the dioctyl, diisopropyl, and diethylhexyl adipate 19 agree with these comments. I just want to throw 19 studies will be included in this report. 20 out that I can't actually find the reference for 20 Is that where we're at? 21 it, but apparently it's used as a pH adjuster to a 21 SPEAKER: Mm-hmm. 22 concentration of about 5 percent in hair products. 22 DR. BERGFELD: Well, and use, because

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net CIR Panel Book Page 18 Cosmetic Ingredient Review Panel December 8, 2009 Page: 175 Cosmetic Ingredient Review Panel December 8, 2009 Page: 176

1 they promised you use. 1 at.001 percent in a hairspray and it's not going 2 DR. BELSITO: I said that. 2 to get absorbed, then, you know, we might change 3 DR. BERGFELD: Did you say that? 3 our minds. So I guess that we're considering 4 DR. BELSITO: Yeah. 4 deleting oxalic acid unless we get information on 5 DR. BERGFELD: Okay. 5 absorption and concentration of use in products 6 DR. SNYDER: Unless we're provided data 6 that it would be used in. But if it's open-ended 7 on oxalic acid regarding dermal absorption 7 and we assume that it's going to be used in all 8 toxicity. 8 the same type of products as these other 9 DR. BELSITO: Yeah, I suppose that could 9 dicarboxylic acids in concentrations up to 10 10 be done. But again, the boilerplate that we've 10 percent, we don't have any information, we'll 11 had for the decision to go ahead with additions 11 probably just remove it from the report. 12 has been that it be a no-brainer. And that we 12 SPEAKER: Lunchtime. 13 wouldn't need that additional data; that the data 13 DR. BELSITO: It is 12:10. Be back at 14 was already there. I don't know. 14 1:10. 15 DR. ANDERSON: I think the option would 15 (Recess) 16 always exist for any interested party to provide 16 DR. BELSITO: Okay, welcome back. So, 17 data. 17 we're back here at the PEGS. And at the last 18 DR. BELSITO: Yeah, if there was data 18 meeting, as you know, the issue here is to reopen 19 provided on lack of absorption, then I guess that 19 it to get rid of this damaged skin restriction in 20 the no-brainer part of it is if it gets absorbed, 20 the conclusion. At the last meeting we had a 21 how much and what would it do to the kidney. If 21 wonderful presentation on transepidermal water 22 we see information that, you know, it's used 22 loss and various skin diseases, including atopic

ANDERSON COURT REPORTING ANDERSON COURT REPORTING 706 Duke Street, Suite 100 706 Duke Street, Suite 100

Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net Anderson Court Reporting -- 703-519-7180 -- www.andersonreporting.net

CIR Panel Book Page 19          1    hiiv‡† T‚Dt‚‡‡‚ur r‡uh‡h†qr vtˆrˆ  tr‚‡‚‘hqph pv‚trvpv‡’qh‡hU‚€’‚ˆrrq‡‚   ‡uh‡’‚ˆqvq‡r‰r v‡r h‡v‡u‚ˆ‡ƒˆ‡‡vt p‚€€r‡‚ 6q‡ur‚i‰v‚ˆ†y’‚‡ur v††ˆr†‚    ‡ur†ƒrpvr†vs ‚‡‚sv‡r††r‡vhyy’r‰r  p‚pr † T‚†u‚ˆyqrt‚v‡u‡urvt rqvr‡   !9S H6SFT'Pxh’ T‚D‡uvxr r !yv†‡4   *svv†urqv‡u‡ur†rt ‚ˆƒ†‚svt rqvr‡† Xrph *9S TC6IF'Duh‰rh†‡ htr†ˆttr†‡v‚    ,v††ˆrhsvhyh€rqrq†hsr‡’h††r††€r‡v‡u†hsr ,Dh†p‚†vqr vt//‚ €h’irrphp‚†vqr    .h†pˆ r‡y’ˆ†rqv‚/p‚y‚ vtuhv ph r .†ƒyv‡‡vt‡uv†h††r††€r‡v‡‚‡‚ƒh ‡†'Prs‚    0ƒ ‚qˆp‡†v‡u‡urrqv‡‚ vhyp‚€€r‡†‡uh‡r 0‡urhpvq†hq‡ur†hy‡†hq‚rs‚ ‡urr†‡r †    qv†pˆ††rq  Uurhpvq†hyy†rr€‡‚irˆ†rqh†ƒChqwˆ†‡r ††‚   1Ir‘‡†rihph‡r† D‡†v‡urQvx Gr‡ 1‡ur†hy‡†h r‰r ’†v€vyh //‚ˆyqir†v€vyh ‡‚   €r†rr 7‚‚xˆqr qvv†‚ƒ ‚ƒ’y†rihph‡r  iruh‰r‡ur†h€rv‡r €†‚sƒ‚††viyrurhy‡u   Pxh’ T‚h‡‡urTrƒ‡r€ir 1€rr‡vt rssrp‡† Uurr†‡r †‚ˆyqirqvssr r‡ 6qDh€   9 Tuhx rp‚€€rqrq‡uh‡‡ur rƒ‚ ‡irr‘ƒhqrq wˆ†‡†ˆttr†‡vt‡uh‡r€vtu‡p‚†vqr //‡uv†v†   !‡‚sv‰rhqqv‡v‚hyvt rqvr‡†hqhy†‚‡uh‡r !huˆtrq‚pˆ€r‡//†ƒyv‡‡vtv‡s‚ ‡urhpvq†h†   * r‚ thv“rv‡hq†‚v‡h†‡hiyrq 6q†‚‚ *ƒChqwˆ†‡r †hq‡urv †hy‡†hq‡urh†rƒh h‡r   ,r rur ry‚‚xvth‡‡ur r‰v†rq rƒ‚ ‡‡uh‡† ,h††r††€r‡s‚ ‡ur‚ thvpr†‡r †    .irr r‚ thv“rqv‡ur‡hiyr† 6qS‚’‚ˆ .9S TG6B6'Dyvxr‡uh‡    0uh‰r‡ur†ƒ rhq†urr‡v‡u‡uru‚yr‡uvtq‚ 09S TC6IF'A‚ ‡urr†‡r †    ‡ur r  9S CDGG'`rhu€r‡‚‚    1T‚yr‡†//D‡uvxruh‰r†r‰r hy 19S TC6IF'Pxh’    ‡uvt† Prv†//h r‡urvt rqvr‡†‡uh‡h r 9S TG6B6'Duhqp‚pr ‡uh‡‡uh‡†   vpyˆqrq Xuvpu‚r†//h r‡ur’‚xh’4Xruh‰r ‡‚‚€ˆpuur r 6q‡ur r†‚q‚ˆi‡vsv‡†hƒC       6I9@STPI8PVSUS@QPSUDIB 6I9@STPI8PVSUS@QPSUDIB .1,9ˆxrT‡ rr‡Tˆv‡r11 .1,9ˆxrT‡ rr‡Tˆv‡r11 6yr‘hq vhW6! 6yr‘hq vhW6! Qu‚r?.1@*/.01Ah‘?.1@*/.1 Qu‚r?.1@*/.01Ah‘?.1@*/.1  hqr †‚ rƒ‚ ‡vt r‡  hqr †‚ rƒ‚ ‡vt r‡

            hqwˆ†‡r //  r/ r‰vrDtˆr††‡‚hqq€‚ r†ˆi†‡hpr†    9S TC6IF'Uuh‡†‚r‡uvt  9S TC6IF'Uur’‚ˆp‚ˆyq‡hxr‡uh‡   9S TG6B6'Uuh‡†‚r‡uvt 6qr q‚pˆ€r‡‚ƒrv‡s‚  r/ r‰vrhqhqq‡ur†r   !x‚‡uh‡†hr‘‡ r€ry’†€hyyp‚pr‡ h‡v‚  !‚‡ur r†‡r † 9‚v‡ihpxh q†    *6’h’t‚‚q  *9S 76DG@`'Pxh’ D‡uvxr rt‚vt   ,9S TC6IF'T‚hs‡r ’‚ˆ‰rq‚rhyy ,‡‚uh‰r‡‚‡uvxhi‚ˆ‡‡uv†irphˆ†r//   .‡uv†‚ x  .9S TC6IF'Duh‰r‡‚†rr‡uh‡qh‡hih†r    0HT SP7DITPI'6y†‚‡ur rh r†‚€r 07ˆ‡vsr‰rhy rhq’ r‰vrrq    ‡uvt†€v††vth†ryyirphˆ†rqvr‡u’yur‘’y TQ@6F@S'I‚Dht rrv‡u‡uh‡iˆ‡//   1hqvƒh‡rh†ƒ r‰v‚ˆ†y’ r‰vrrqi’8DS†‚‡uh‡ 1TQ@6F@S'Xruhq‡u rr    †‡ˆssrrq†‡‚irvp‚ ƒ‚ h‡rqh†ryy T‚v‡† TQ@6F@S'Uu rr 6yy vtu‡    t‚vt‡‚irhr‰ryh tr q‚pˆ€r‡  TQ@6F@S'6q‡ur r†‡‚ rƒ‚ ‡†    9S H6SFT'Ds‡uh‡†irrƒ r‰v‚ˆ†y’ HT SP7DITPI'D‡†qvr‡u’yur‘’y   ! r‰vrrqruh‰r‡‚ r‚ƒrv‡  !hqvƒh‡rqviˆ‡’yhqvƒh‡rhqqvv†‚ƒ ‚ƒ’yhqvƒh‡r    *HT SP7DITPI'Dth‰r’‚ˆ‡ur‚ vtvhy  *9S 76DG@`'T‚‡u‚†rh ry‚tvphy‡‚   ,9S H6SFT'`r†  ,t ‚ˆƒD‡uvxhq‡ur’‚ˆhqq‡ur‚‡ur r†‡r †   .HT SP7DITPI'Pxh’  .v‡‚v‡ 6q‡ur’‚ˆ‰rt‚‡‡urhpvq†hq‡ur   09S H6SFT'7ˆ‡D€rhv‡r €†‚s 0†hy‡†h†h†rƒh h‡rq‚pˆ€r‡ Uur’ rpur€vphyy’   ƒ ‚‡‚p‚y  †‚€ruh‡qvssr r‡ T‚//   19S TC6IF'`rhu  19S H6SFT'Xuh‡r r‡u‚†rhthv   9S 76DG@`'Dsv‡†t‚vt‡‚irhqqrq Whyr vr49vur‘’y//   ‡urr//ih†vphyy’’‚ˆ r r‚ƒrvthr‘v†‡vt HT SP7DITPI'9vr‡u’yur‘’yhqvƒh‡r       6I9@STPI8PVSUS@QPSUDIB 6I9@STPI8PVSUS@QPSUDIB .1,9ˆxrT‡ rr‡Tˆv‡r11 .1,9ˆxrT‡ rr‡Tˆv‡r11 6yr‘hq vhW6! 6yr‘hq vhW6! Qu‚r?.1@*/.01Ah‘?.1@*/.1 Qu‚r?.1@*/.01Ah‘?.1@*/.1  hqr †‚ rƒ‚ ‡vt r‡  hqr †‚ rƒ‚ ‡vt r‡

CIR Panel Book Page 20          !   qviˆ‡’yhqvƒh‡rhqqvv†‚ƒ ‚ƒ’yhqvƒh‡r  HT @DT@IH6II'6pp‚ qvt‡‚‡urIUQ   9S H6SFT'8‚€€r‡†hi‚ˆ‡//v‡ ‚‘hyvp    pr ‡hvy’†rr€†y‚tvphyS‚  HT SP7DITPI'P‘hyvp    !9S 76DG@`'Xh†‡ur‚y’€v††vt !HT @DT@IH6II'Qˆiyv†urqvChy‚ ‡u   *vs‚ €h‡v‚‡urtr‚‡‚‘‡uh‡h†h†xrqs‚ v *r‡hy 0 D‡†‚‡v‡ur r’r‡    ,Trƒ‡r€ir 47rphˆ†rD‡uvx‡uh‡h†ƒ ‚‰vqrq ,9S TC6IF'Pxh’    .h†‡v‡4 .9S 76DG@`'Pxh’ T‚ruh‰r€‚ r   09S TC6IF'Xruh‰rtr‚‡‚‘‚‡‚‚s 0vs‚ €h‡v‚‡uh‡†t‚vt‡‚irrpr††h ’4   ‡ur†rihph‡rr†‡r †‚‡€ˆ‡htrvp Xruh‰rtr‚ HT @DT@IH6II'`rhu Uur r†hy‚‡‚s   1//v€€ˆ‚trrpv‡’qh‡h‚qvr‡u’y€hy‚h‡r 1vs‚ €h‡v‚‡uh‡††‡vyy‚‡v‡uv† rƒ‚ ‡   rth‡v‰rE†ˆppvvphpvqrth‡v‰rEtyˆ‡h vphq vpyˆqvt‡ur8DS rƒ‚ ‡†irphˆ†r‡ur r†hIUQ   hqvƒvphpvq†rth‡v‰rEqvr‡u’yur‘’yhqvƒh‡r iv‚h††h’‚‡urqvr‡u’yur‘’yhqvƒh‡r    rth‡v‰rEqv‚‘’/‚’y/hqvƒh‡r//hqvƒh‡rr†‡r  9S TG6B6'Tu‚ˆyqr‡hiyrv‡4   !h“ryhvphpvqrth‡v‰r 6yy‚s‡ur€ˆ‡htrvpv‡’ !9S H6SFT'Xryy‡uh‡†‡urr‘‡   *qh‡hh rrth‡v‰r 7ˆ‡‡ur r†‚‡// *†ˆttr†‡v‚    ,TQ@6F@S'D‡††xv€ƒ’ph pv‚trvp ,9S TG6B6'D€rhr r‡hiyvtv‡v   .?vhˆqviyr@  . rhyv‡’h’h’    09S TC6IF'Uur rh ry‚‡†‚s€v††vt// 0HT @DT@IH6II'Xryyhq’‚ˆq‚‡uh‰r   ‚‡uvt‚‚‘hyvphpvq‚ ‡ur‚‘hyh‡r†  ‡urp‚pr‡ h‡v‚‚sˆ†rvs‚ €h‡v‚s‚ h’‚s   1HT @DT@IH6II'6qhp‡ˆhyy’IUQuh†// 1‡urrvt rqvr‡†rv‡ur Uuh‡uh†‡irrq‚r   v‡†rth‡v‰rv†hy€‚ryyh  ’r‡irphˆ†rDqvq‡x‚‡ur’r rt‚vt‡‚ir   9S TC6IF'Xuvpu4 hqqrq        6I9@STPI8PVSUS@QPSUDIB 6I9@STPI8PVSUS@QPSUDIB .1,9ˆxrT‡ rr‡Tˆv‡r11 .1,9ˆxrT‡ rr‡Tˆv‡r11 6yr‘hq vhW6! 6yr‘hq vhW6! Qu‚r?.1@*/.01Ah‘?.1@*/.1 Qu‚r?.1@*/.01Ah‘?.1@*/.1  hqr †‚ rƒ‚ ‡vt r‡  hqr †‚ rƒ‚ ‡vt r‡

        * ,   9S TC6IF'Xryyvsrp‚ˆyqqv†pˆ†† ur rr††r‡vhyy’v‡†‡‚t ‚ˆƒ†4   sv †‡†u‚ˆyqr†ƒyv‡v‡ˆƒ‚ // 9S 76DG@`'Svtu‡ Dtˆr††h‡‡urrq   9S H6SFT'XryyD‡uvx‡uh‡†v‡ D ‚s‡urqh’‡urvs‚ €h‡v‚‡uh‡†t‚vt‡‚ir   !‡uvxr†u‚ˆyq//Dtˆr††v‡yyp‚€rq‚‡‚ !vs‚ €vt‡ur†hsr‡’ur rvyyirs ‚€‡urhpvq†   *‡hiyrv‡uuh‡v†‡urhp‡v‚rh‡‡‚‡hxr  *hq‡urr†‡r † vtu‡4D€rhv‡†‚‡t‚vt‡‚   ,6qv‡†rr€†yvxr†ƒyv‡‡vtv‡v†h‰r ’ ,irh†rƒh h‡r†r‡‚sqh‡hs‚ ‡urhpvq†‰r †ˆ†h   . rh†‚hiyrh’‡‚hƒƒ ‚hpu‡uv†  .†rƒh h‡r†r‡‚sqh‡hs‚ ‡urr†‡r †4D€rhD€   09S TG6B6'6qƒˆ‡‡vt‡urhƒƒ ‚ƒ vh‡r 0wˆ†‡uh‰vthuh q‡v€r‰v†ˆhyv“vtu‚‡uv†‚ˆyq   qh‡hv‡‚uh‡r‰r v††ƒyv‡  ir†ƒyv‡hqu‚8DS†‡hss’‚ˆx‚‡uv†v†h   19S CDGG'Uuv†v†u‚†r rƒ‚ ‡4 1†vtvsvph‡iˆ qr‡‚‚ T‚D€wˆ†‡‡ ’vt‡‚   HT SP7DITPI'D‡†€’ rƒ‚ ‡  ‡uvx‚srssvpvrpvr†ur r‡uh‡€vtu‡‚ x    9S TG6B6'Whyr vr  Xuh‡q‚’‚ˆtˆ’†‡uvx4   9S H6SFT'9vqE‚u8h ‚yS‚//U‚€ 9S TG6B6'8‚ˆyqruh‰rv‡h†‡‚   !Dt‚‡‡ur†r†r’‚ˆyvxr‡urvqrh‚s†ƒyv‡‡vtv‡  !ƒh ‡†‡urur r’‚ˆqrhyv‡u‡urƒChqwˆ†‡r †   *D‡pr ‡hvy’€hxr††r†r  *‡urhpvq†hq‡urs‚yy‚v‡i’‡urr†‡r †v‡ur   ,9S 76DG@`'Uur?vhˆqviyr@‡uvtD€ ,†h€rq‚pˆ€r‡4   .‡uvxvt‚sv†‡uh‡v‡u‡urty’p‚yvphpvqrqvq .9S TC6IF'Uurˆ†r†s‚ ‡urhpvq†h r   0‡urty’p‚yvphpvqhq†hy‡†hqr†‡r † T‚‡ur r 0„ˆv‡rqvssr r‡s ‚€‡urˆ†r†‚s‡urr†‡r † 6q   rxrƒ‡v‡hy‡‚tr‡ur Dph†rr‡ury‚tvp ‡uh‡†ur rD‚ˆyqyvxr‡‚†ƒyv‡‡ur€ 7ˆ‡r   1ir‡rr†ƒyv‡‡vtEDwˆ†‡‚qr uh‡v†‡ur€‚†‡ 1q‚‡uh‰rhu‚yry‚‡‚sˆ†rvs‚ €h‡v‚rv‡ur    rssvpvr‡h’‡‚q‚‡uv†  TQ@6F@S'`rhu    9S H6SFT'P q‚ruh‰r‚rq‚pˆ€r‡ 9S TC6IF'Xruh‰r‡‚‡hxr‡uru‚yr       6I9@STPI8PVSUS@QPSUDIB 6I9@STPI8PVSUS@QPSUDIB .1,9ˆxrT‡ rr‡Tˆv‡r11 .1,9ˆxrT‡ rr‡Tˆv‡r11 6yr‘hq vhW6! 6yr‘hq vhW6! Qu‚r?.1@*/.01Ah‘?.1@*/.1 Qu‚r?.1@*/.01Ah‘?.1@*/.1  hqr †‚ rƒ‚ ‡vt r‡  hqr †‚ rƒ‚ ‡vt r‡

CIR Panel Book Page 21         . 0   ‡uvtrvtu‡‚sr‰vqrpr‡‚€hxrhqrpv†v‚‚ 9S 76DG@`'//‡ur’‚ˆ rih†vphyy’   ‡ur†r 7ˆ‡vs’‚ˆy‚‚xh‡uh‡vs‚ €h‡v‚r t‚vt‡‚uh‰r‡‚qˆƒyvph‡rv‡’‚ˆx‚ir‡rr   uh‰rh‡yrh†‡uh‡†v‡uv† rƒ‚ ‡‡ur r†‚‡ ‡ur‡‚ rƒ‚ ‡†uvpuv†‚xh’ 7ˆ‡hthv   !‰r ’€ˆpu‚h’‚r‚s‡ur€ 6q‚‡uvt‚‡ur !‡uvxvt‚srssvpvrp’hq//   *yrhqvt rqvr‡  *9S TC6IF'Dwˆ†‡q‚‡†rrˆ†vtr†‡r    ,9S 76DG@`'7ˆ‡D‡uvxv‡†h ,qh‡h‡‚†ˆƒƒ‚ ‡‡ur†hsr‡’‚s‡urhpvq†    . rh†‚hiyrƒ ‚ƒ‚†hy Hh’irrph//’‚ˆtˆ’†ph .9S CDGG'Hrrv‡ur    0ƒ r†r‡v‡‡‚€‚ ‚hq‡uruh‰rhqv†pˆ††v‚ 09S 76DG@`'Xryy‡ur€r‡hi‚yv†€‚s   v‡u‡ur‚‡ur ‡rh€hq†rr  r†‡r †vyytrr h‡r‡urhpvq†    19S H6SFT'6 rrt‚vt‡‚ˆ†rqh‡hq‚ 19S TC6IF'`rhuiˆ‡‡urhpvq†h rˆ†rq   ’‚ˆ‡uvxU‚€hqS‚s ‚€‡urhpvq†‡uh‡ qvssr r‡y’ Uur’ rˆ†rqh†ƒChqwˆ†‡r †v‡ur   †ˆƒƒ‚ ‡‡ur†hsr‡’‚s‡urr†‡r †hq‰vpr‰r †h4 s‚ €ˆyh‡v‚ƒ ‚pr†† Uuh‡†qvssr r‡s ‚€uh‰vt   Dsr rt‚vt‡‚q‚‡uh‡‡urv‡†rr€†yvxrv‡ €r‡hi‚yvptrr h‡v‚‚s‡urhpvq†    !‚ˆyq€hxr†r†r‡‚uh‰rv‡hyy‡‚tr‡ur iˆ‡ !9S 76DG@`'I‚Dph?vhˆqviyr@‡uh‡    *†ƒyv‡ Xr‰rq‚r‡uh‡irs‚ rur rr‰r‡hxrh *9S TC6IF'`rhuDphuhqyrv‡vs   ,t ‚ˆƒ‚svt rqvr‡†hq†ƒyv‡‡ur€v‡ur†h€r ,v‡†hyyv‚riˆ‡D‡uvxv‡†//D‡uvxv‡†   . rƒ‚ ‡  .rh†vr ‡‚†ƒyv‡    09S 76DG@`'7rphˆ†rv‡‚ˆyqir 09S CDGG'Dsˆyy’†ˆƒƒ‚ ‡’‚ˆ    qˆƒyvph‡vthy‚‡‚s€h‡r vhy‡‚t‚v‡‚‡ur ƒ ‚ƒ‚†hy    1 rƒ‚ ‡ Ds’‚ˆ rt‚vt‡‚ˆ†rr†‡r qh‡h‡‚ 19S TC6IF'Pxh’ Uuhx’‚ˆ    †ˆƒƒ‚ ‡‡urhpvq†rp‡v‚hq‰vpr‰r †h// HT X@DIUS6V7'D†‡uri‚q’‚sr‰vqrpr   9S H6SFT'Svtu‡  ‡uh‡qvssr r‡‚ v†‡v‡†‚€r‚‡ur ?vhˆqviyr@4       6I9@STPI8PVSUS@QPSUDIB 6I9@STPI8PVSUS@QPSUDIB .1,9ˆxrT‡ rr‡Tˆv‡r11 .1,9ˆxrT‡ rr‡Tˆv‡r11 6yr‘hq vhW6! 6yr‘hq vhW6! Qu‚r?.1@*/.01Ah‘?.1@*/.1 Qu‚r?.1@*/.01Ah‘?.1@*/.1  hqr †‚ rƒ‚ ‡vt r‡  hqr †‚ rƒ‚ ‡vt r‡

         1   9S CDGG'`‚ˆ‚ˆyqirˆ†vthpvqqh‡h 8h ‚ytr‡†‡urp‚pr‡ h‡v‚†‚sˆ†r‡uh‡€vtu‡   ‡‚†ˆƒƒ‚ ‡‡urr†‡r qh‡hiˆ‡ƒ ‚ihiy’‚‡‡ur uryƒˆ†‡‚‚†‚ ‡‡u‚†r‚ˆ‡vh€‚ r//vh   ‚‡ur h’h ‚ˆq Dph‡†rrh’ƒyhprur r pyrhr h’s‚ ‚r rƒ‚ ‡ 7rphˆ†r‡ur’h r   !‡uh‡‚ˆyqir‡urph†r T‚’‚ˆ‚ˆyquh‰r‡‚ !t‚vt‡‚‡hyxihpxhqs‚ ‡u    * rsr rpr‡urhpvq rƒ‚ ‡†ƒ ‚ihiy’ rƒrh‡rqy’ *9S H6SFT'Uur r†‚„ˆr†‡v‚‡ur’ r   ,iˆ‡Dq‚‡†rr‡urq‚†vqr‚s‡uh‡  ,t‚vt‡‚ir†ƒyv‡ D‡uvx‡ur‚y’v††ˆrv†h r   .9S H6SFT'Xryy‡‚€rv‡‚ˆyqirvpr .‡ur’t‚vt‡‚†ƒyv‡//ir†ƒyv‡v‡‚‡‚q‚pˆ€r‡†   0‡ur‡‚uh‰rh rƒ‚ ‡ur r’‚ˆ‚ˆyq†‡h ‡‚ss 0‚ h r‡ur’t‚vt‡‚irv‚rq‚pˆ€r‡hq†ƒyv‡   hq†h’r rt‚vt‡‚qv†pˆ††‡urhpvq†sv †‡  v‡uv‡ur‚rq‚pˆ€r‡46qDtˆr††‡ur„ˆr†‡v‚   1Uursv †‡ƒh ‡‚s‡ur rƒ‚ ‡‚ˆyqirhyy‡ur 1v†u‚ˆvryq’‡uh‡v†iˆ‡vs’‚ˆ rt‚vt‡‚   hpvq† 6q‡ur rh†‚v‡††ƒyv‡v†irphˆ†r‚s ˆ†rh’r†‡r q‚pˆ€r‡hy‚‡‚s rsr rpr†‡‚   ˆ†rhq‡urv †hy‡† 6q‡ur‡ur†rp‚qƒh ‡ ‡uvt†v‡urhpvqq‚pˆ€r‡v‡€vtu‡irryy‡‚   ‡urr†‡r †ir‰r ’pyrh ‡ur’uh‰rqvssr r‡ xrrƒv‡hyy‡‚tr‡ur    !ˆ†r†iˆ‡r rt‚vt‡‚ˆ†r†‚€r‚s‡urhpvq !9S 76DG@`'D‡uvxvthi‚ˆ‡‡ur   *‡‚‘vp‚y‚t’‡‚†ˆƒƒ‚ ‡‡ur†hsr‡’r†‡r † 6q *ƒˆ ƒ‚†r‚s‡ur†r rƒ‚ ‡†‡‚‚v†v€ƒ‚ ‡h‡   ,hthvv‡†hyyv‚r rƒ‚ ‡†‚’‚ˆq‚‡uh‰r‡‚ ,irphˆ†r‡uv†p‚€€ˆvph‡r†‡‚ˆ†r †‡urvqˆ†‡ ’   .t‚ihpx  .hq‚‡ur †uh‡†r‘ƒrp‡rq’‚ˆx‚‡urih†v†   09S 76DG@`'Uuh‡†xvq‚suh‡D€ 0s‚ †hsr‡’hqh’p‚qv‡v‚†‡uh‡€vtu‡ir   ‡uvxvt  v€ƒ‚†rqhq‡uh‡‡u‚†rh rƒ ‚ihiy’€‚ r   19S H6SFT'`rhu  1rssvpvr‡y’p‚€€ˆvph‡rqv‚rq‚pˆ€r‡‡uh‡‚    9S 76DG@`'9‚’‚ˆx‚u‚‡uv†v† 9S H6SFT'`rhu‡uh‡†//   hp‡ˆhyy’t‚vt‡‚y‚‚x46q8h ‚y//D‡uvx‚pr 9S CDGG'D‚ˆyqwˆ†‡rpu‚U‚€†‰vr       6I9@STPI8PVSUS@QPSUDIB 6I9@STPI8PVSUS@QPSUDIB .1,9ˆxrT‡ rr‡Tˆv‡r11 .1,9ˆxrT‡ rr‡Tˆv‡r11 6yr‘hq vhW6! 6yr‘hq vhW6! Qu‚r?.1@*/.01Ah‘?.1@*/.1 Qu‚r?.1@*/.01Ah‘?.1@*/.1  hqr †‚ rƒ‚ ‡vt r‡  hqr †‚ rƒ‚ ‡vt r‡

CIR Panel Book Page 22             ‡uh‡‡urƒˆ ƒ‚†r†‚sˆ†rh r rhyy’„ˆv‡r uh‡‡ur‚‡ur ‡rh€//‡uh‡†‚‡huˆtrv††ˆr   qvssr r‡ Uuriv‚uhqyvt‚s‡urp‚€ƒ‚ˆq† ‚r‰r †ˆ†‡‚q‚pˆ€r‡† Uur€‚†‡v€ƒ‚ ‡h‡v†   †u‚ˆyqirr‘ƒrp‡rq‡‚ir„ˆv‡rqvssr r‡ @‰r †ƒyv‡‡vtv†‡v‡4   !‡u‚ˆturr‘ƒrp‡‡urr†‡r †‡‚iriv‚‡ h†s‚ €rq !9S TC6IF'Xryyvs’‚ˆ†ƒyv‡‡ur€//   *v‡‚hpvq††’†‡r€vph‰hvyhivyv‡’†v‡ˆh‡v‚† *ƒˆ‡‡ur€v‚rq‚pˆ€r‡‚ ‡‚//Dq‚‡ph r    ,†u‚ˆyqir„ˆv‡rqvssr r‡ir‡rr‡urhpvq†hq ,D‡€h’ir‡uh‡9 Cvyy†hi†‚yˆ‡ry’sh †vtu‡rq   .†hy‡†h†‚ƒƒ‚†rq‡‚‡urr†‡r † 6q†‚D€ .r‚ˆtu‡uh‡urrp‚€r‡‚‡urp‚pyˆ†v‚‡ur   0‡uvxvt‡urp‚pyˆ†v‚h‡‡urrqp‚ˆyqtr‡„ˆv‡r 0p‚pyˆ†v‚vyyir†‚p‚€ƒyr‘‡uh‡ryyqrpvqr‡‚   ˆvryq’ih†rq‚‡uh‡‰h†‡qvssr rprv‡urh’ ‡hxr‡ur‡‚ƒh ‡†hqƒˆ‡‡ur€v‡‚‡‚qvssr r‡   1‡ur†r‡uvt†h rˆ†rq 6q†‚D‡uvxv‡‚ˆyq 1q‚pˆ€r‡†    rqˆƒ//vs’‚ˆuhq‡‚q‚pˆ€r‡†D€rh‡ur’ 9S TG6B6'Xryyiˆ‡y‚‚xh‡uh‡r   p‚ˆyqirƒˆiyv†urqihpx/‡‚/ihpx 7ˆ‡vs’‚ˆuhq uh‰rhquh‡xvq‚sqh‡h‡ur rv† `‚ˆx‚   ‡‚q‚pˆ€r‡†’‚ˆ‚ˆyquh‰rD‡uvxpyrhr  ‡ur r† rhyy’ƒ ‚ihiy’‚‡t‚vt‡‚ir€ˆpu‚sh   !p‚pyˆ†v‚†h‡‡urrq  !ƒ ‚iyr€ Uurp‚pyˆ†v‚v†t‚vt‡‚irƒ ‚ihiy’   *9S H6SFT'Dtˆr††‡uh‡†h††ˆ€vt‡uh‡ *‰r ’†v€ƒyr D€rh‡uh‡†uh‡D‚ˆyqƒ rqvp‡    ,’‚ˆ rt‚vt//vs’‚ˆtr‡hyy‡urqh‡hv‡uh‡ ,9S 76DG@`'D‚ˆyqr‘ƒrp‡‡uh‡ `‚ˆ   .’‚ˆphƒˆiyv†u‡ur€ihpx/‡‚/ihpx  .p‚ˆyq†h’u’ƒ‚‡ur‡vphyy’‡uh‡‡urhpvq†hq†hy‡†   09S CDGG'Qˆiyv†u‡ur€ihpx/‡‚/ihpxhq 0h r†hsrs‚ ˆ†r’‚ˆx‚h†hƒC/hqwˆ†‡vt   hv‡‚‡ur‚r  htr‡ Uuh‡p‚pr‡ h‡v‚v†ˆƒ‡‚ˆ†r//   19S H6SFT'`‚ˆphƒˆ‡‚ˆ‡h 1†ˆpu/hq/†ˆpuhˆ†rhq‡urr†‡r †‚ˆyqir†‚€r   v†ˆssvpvr‡hq†‚ ‡‚sƒˆ†uv‡‡‚tr‡v‡hyy ‰h vh‡v‚‚‡uh‡ T‚D‚ˆyq‡r‰v†v‚h   ‡‚tr‡ur 7ˆ‡hthvD‡uvx‡uh‡†//ryy†rr p‚€ƒyr‘//iˆ‡’‚ˆx‚vsrtr‡‡‚‡uh‡ƒ‚v‡       6I9@STPI8PVSUS@QPSUDIB 6I9@STPI8PVSUS@QPSUDIB .1,9ˆxrT‡ rr‡Tˆv‡r11 .1,9ˆxrT‡ rr‡Tˆv‡r11 6yr‘hq vhW6! 6yr‘hq vhW6! Qu‚r?.1@*/.01Ah‘?.1@*/.1 Qu‚r?.1@*/.01Ah‘?.1@*/.1  hqr †‚ rƒ‚ ‡vt r‡  hqr †‚ rƒ‚ ‡vt r‡

         !   hq‡ur’h rp‚€ƒyr‘‡urrph‡uvxhi‚ˆ‡ 9S TC6IF'6q‡u‚†rh rhyyr†‡r †‚s   †ƒyv‡‡vtv‡ˆƒ‡ur 6qv‡ƒ ‚ihiy’‚ˆyqir hqvƒvphpvq4   shv y’rh†’‡‚q‚ 9‚‡’‚ˆ‡uvx4 HT SP7DITPI'`r†    !9S TG6B6'Pxh’ Xr‚ˆyqhv‡ˆ‡vy !9S TC6IF'Uur‡u rrvt rqvr‡†4   *r†rrhyy‡urqh‡h  *9S TG6B6'Uu‚†rr rhyy†hsrr r‡   ,9S 76DG@`'Svtu‡  ,‡ur’4   .9S H6SFT'T‚Dyy€‚‰r‡uh‡r‡hiyr .HT SP7DITPI'`r†    0‡uv† 9‚r†‡uh‡†‚ˆqt‚‚q4S‚U‚€S‚4 09S H6SFT'6qv‡uh‡y‚tyv†‡v‡ur   9S TC6IF'Xrq‚‡h‡‡‚t‚‡u ‚ˆtu irtvvt‡urqvur‘’y†ur rˆqr ‡ur†ˆiwrp‡4   1‡ur rƒ‚ ‡v‡†rys4Xhv‡‡vyyrtr‡hr 18h’‚ˆ//hqvƒh‡r†u‚ˆyqirq‚‡‚h q†‡ur   ‰r †v‚4 €vqqyr‡‚‡urrq D†‡uh‡ vtu‡Whyr vr4Xr   9S H6SFT'6q†‚ryyƒ ‚prrqs‚ h q uhq‡urqviˆ‡’yhqqvv†‚ƒ ‚ƒ’y4   v‡u‡urvqrh‡uh‡r rt‚vt‡‚†ƒyv‡‡urhpvq† T‚v‡†qvur‘’yhqvƒh‡r4D†‡uh‡//   !hq‡urv †hy‡†s ‚€‡urr†‡r † Xr r‚‡†ˆ r !HT SP7DITPI'9vr‡u’yur‘’yhqvƒh‡r    *’r‡ur‡ur ‡uv†v†t‚vt‡‚ir‚r‚ ‡‚ *D‡†‡ur‡uv qs ‚€hqvƒvphpvq    ,q‚pˆ€r‡† Xr‡rq‡‚iryrhvt‡‚h q†‚r ,9S H6SFT'6qvƒvphpvq Dyy‡ryy’‚ˆ   .q‚pˆ€r‡h‡yrh†‡‡‚irtvv‡u  .uh‡ Dyytr‡‡uh‡s ‚€’‚ˆyh‡r    09S 76DG@`'Xv‡u‡urƒ‚††vivyv‡’‚s 09S TC6IF'9vr‡u’yur‘’y4   t‚vt‡‚‡‚  9S H6SFT'9vr‡u’yur‘’y    19S H6SFT'6qv‡uv†ƒ ‚pr††r‰r 19S TC6IF'Xuh‡r r‡ur‚‡ur ‡‚4   t‚‡‡‚ r‚ƒr‡u rrvt rqvr‡† D†‡uh‡‚‡ 9S H6SFT'6q‡ur‡ur rh†qviˆ‡’y    p‚ rp‡4 9S TC6IF'9viˆ‡’yhqvƒh‡r4       6I9@STPI8PVSUS@QPSUDIB 6I9@STPI8PVSUS@QPSUDIB .1,9ˆxrT‡ rr‡Tˆv‡r11 .1,9ˆxrT‡ rr‡Tˆv‡r11 6yr‘hq vhW6! 6yr‘hq vhW6! Qu‚r?.1@*/.01Ah‘?.1@*/.1 Qu‚r?.1@*/.01Ah‘?.1@*/.1  hqr †‚ rƒ‚ ‡vt r‡  hqr †‚ rƒ‚ ‡vt r‡

CIR Panel Book Page 23         * ,   9S H6SFT'6qvƒh‡r Uuh‡†‚‡urr‘‡ ‡uh‡vs‚ €h‡v‚ 6qrh ry‚‚xvth‡hy‚‡‚s   yvr 6q‡urqvv†‚ƒ ‚ƒ’y D†‡uh‡p‚ rp‡4 vt rqvr‡†//h‡yrh†‡uh‡†vur r//r r   HT SP7DITPI'`r†‡uh‡†p‚ rp‡  t‚vt‡‚irv‡r ƒ‚yh‡vthqv†‚€rph†r†   !9S H6SFT'9vv†‚ƒ ‚ƒ’y Xur rv†‡uh‡4 !r‘‡ hƒ‚yh‡vt 6q‡uh‡€vtu‡iruvtuy’ ryr‰h‡    *9vv†‚ƒ ‚ƒ’y Pxh’ Uur rrt‚ 6qvƒh‡r T‚ *HT SP7DITPI'Uuhx’‚ˆ    ,‡ur†r‡u rr Pxh’  ,9S H6SFT'D†‡ur rh’‡uvtv‡uv†   .6’‡uvtry†r4 .y‚tyv†‡ur r‚//U‚€‚ S‚//‡uh‡rrq‡‚ir   09S CDGG'6†’‚ˆ†‡h ‡rq‡‚†h’r 0qryr‡rqh‡‡uv†ƒ‚v‡47rphˆ†rrqvq‡hp‡ˆhyy’   uh‰rh†‡h‡r€r‡//†‚ ’D€ƒhtvth ‚ˆq// t‚‚‰r ‡urvqv‰vqˆhyvt rqvr‡†yvxrr‰rq‚r   1hi‚ˆ‡‡urˆh‰hvyhivyv‡’‚sƒu’†vphyhqpur€vphy 1v‡uryh†‡ Xr‰rt‚rq‚ur r//v†‡ur r   ƒ ‚ƒr ‡vr† 6qDh†‚qr vtvs‡ur//‡uv† h’‡uvt‚h’‚s‡ur†r‡uh‡wˆ†‡q‚‡€hxr   €vtu‡//Fr‰v†‚‡vur rv†ur4 †r†r4   HT SP7DITPI'I‚  9S TC6IF'Xryyˆqr €hy‚vphpvq   !9S CDGG'Ds‡ur7rvy†‡rvqh‡hih†ruhq !qvr‡u’y€hy‚h‡r‡‚‘vpv‡’qh‡hh rtv‰rv‡ur   *irr†rh purqs‚ ‡uh‡  * rƒ‚ ‡iˆ‡v‡†‚‡‚r‚s‡urvt rqvr‡†v‡ur   ,HT SP7DITPI'Uur7rvy†‡rv4 ,‡v‡yr T‚r†u‚ˆyqqv†pˆ†††u‚ˆyqryrh‰r‡uh‡   .9S CDGG'7rvy†‡rv7/@/D/G/T/U/@/D/I  .virphˆ†rv‡v†€hy‚vphpvqr†‡r hq‡ur rh r   09S 76DG@`'6yy‚sˆ†‚yqpur€v†‡†x‚ 0qh‡h47ˆ‡v‡†‚‡hvt rqvr‡ 7ˆ‡v‡€vtu‡   uh‡‡uh‡v†  uryƒˆ†€hxrqrpv†v‚†‚‡ur‚‡ur //‡ur‚r   19S CDGG'D‡ˆ†rq‡‚irhyyqˆ†‡’ 1r†‡r ‡uh‡v†‡ur rv†‡urqvr‡u’y    †ury‰r†vyvi h vr† Tury‰r†‡uh‡r‡‚hq‚ 9S CDGG'Xryyv‡p‚ˆyqirvpyˆqrq   hq‚hq‚ 7ˆ‡’rhu 7rphˆ†r’‚ˆ€vtu‡svq s‚ p‚†vqr h‡v‚v‡u‚ˆ‡uh‰vtv‡ir‚r‚s‡ur       6I9@STPI8PVSUS@QPSUDIB 6I9@STPI8PVSUS@QPSUDIB .1,9ˆxrT‡ rr‡Tˆv‡r11 .1,9ˆxrT‡ rr‡Tˆv‡r11 6yr‘hq vhW6! 6yr‘hq vhW6! Qu‚r?.1@*/.01Ah‘?.1@*/.1 Qu‚r?.1@*/.01Ah‘?.1@*/.1  hqr †‚ rƒ‚ ‡vt r‡  hqr †‚ rƒ‚ ‡vt r‡

        . 0   yv†‡‚svt rqvr‡†p‚ˆyq‡v‡4D€rh// 7ˆ‡‡uh‡h†‡‡ur„ˆr†‡v‚Duhq Pƒhtr!   9S TC6IF'7ˆ‡D‡uvxr†u‚ˆyq ruh‰rhqvv†‚‚p‡’y†rihph‡r 6q‡uh‡†   vqr‡vs’v‡h†v‡†ur riˆ‡v‡†‚‡h hp‡ˆhyy’hqvssr r‡qvv†‚‚p‡’yt ‚ˆƒv‡r €†‚s   !vt rqvr‡ 7ˆ‡r rˆ†vt‡u‚†rqh‡h  !ur r‡uru’q ‚‘’yv†h‡‡hpurq    *9S CDGG'6q†vpr9 7hvyr’v†ur r *9S 76DG@`'Uuh‡†.C‡uh‡’‚ˆ r//   ,‡uv†v†xvq‚shqvp‡v‚h ’„ˆr†‡v‚ Dy‚‚xvt ,9S CDGG'`rhu Xryyhp‡ˆhyy’’r†   .h‡‡ur†‡ ˆp‡ˆ r†uruh†Uur rh r‡‚qvssr r‡ ..C‡urv†‚‚p‡’yt ‚ˆƒv†qvssr r‡‡uhv‡v†//   0syh‰‚ †‚sqvv†‚‚p‡’y Prv†hhqvƒh‡rhq‚r 0‡ur’ ri‚‡uv†‚‚p‡’yiˆ‡‡urt ‚ˆƒv†qvssr r‡   ‚s‡ur‚‡ur †// ‚hqvƒh‡r‰r †ˆ††rihph‡r 6q‡uh‡†ƒ ‚ihiy’   19S 76DG@`'Xuh‡ƒhtrh r’‚ˆ‚4 1wˆ†‡hh ‡vshp‡‚s‡urqvp‡v‚h ’hq‡ur   9S CDGG'Xryy‚xh’vs’‚ˆt‚‡‚‡ur vt rqvr‡†‡uh‡r rhp‡ˆhyy’irvtˆ†rq 7ˆ‡D   ‡hiyr†‡uh‡†//‚‚‡‡ur‡hiyr†‡ursvtˆ r†  h†ƒˆ““yrqi’‡uh‡irphˆ†rv‡ r€hv†‡ur†h€r   Avtˆ r†p‚€rhs‡r ‡ur‡hiyr† Uur r†hƒvx iˆ‡‡ur†‡ ˆp‡ˆ rv†„ˆv‡rqvssr r‡    !ƒhtr  !9S 76DG@`'Xryyy‚‚x‡uh‡ˆƒ    *9S 76DG@`'Xuh‡ƒhtr4 *9S CDGG'Pxh’    ,9S CDGG'T‚D€y‚‚xvth‡‡ur‰r ’ ,9S TC6IF'6†y‚th†r r‚‡ur   .i‚‡‡‚€‚sƒhtrqvv†‚‚p‡’yhqvƒh‡r 6p‡ˆhyy’ .svtˆ r†Avtˆ r@//D‡uvxv‡†@‚ƒhtr!   0u’v†v‡‚‡ur r‡vpr4*Ghq*Q†rr€‡‚ir 0//v‡†h’†qvr‡u’yur‘’yhqvƒh‡r D†‡‡uh‡   ‡ur†h€ruˆu4 qvr‡u’yur‘’y†ˆppvh‡r4   19S 76DG@`'*Ghq*Q†rr€‡‚ir‡ur 19S 76DG@`'D‡xvq‚sy‚‚x†yvxrv‡    †h€ruˆu4 HT SP7DITPI'T‚ ’9 Tuhxuvpu   9S CDGG'Uur’†rr€‡‚ir‡ur†h€r  //       6I9@STPI8PVSUS@QPSUDIB 6I9@STPI8PVSUS@QPSUDIB .1,9ˆxrT‡ rr‡Tˆv‡r11 .1,9ˆxrT‡ rr‡Tˆv‡r11 6yr‘hq vhW6! 6yr‘hq vhW6! Qu‚r?.1@*/.01Ah‘?.1@*/.1 Qu‚r?.1@*/.01Ah‘?.1@*/.1  hqr †‚ rƒ‚ ‡vt r‡  hqr †‚ rƒ‚ ‡vt r‡

CIR Panel Book Page 24          !1   9S TC6IF'D‡†ƒhtr!‚s‡ursvtˆ r† ‡‚ˆ†rphƒv‡hyv“rqh€r† T‚rvyyˆ†r‡ur   Avtˆ r@ Uur‡v‡yr†h’†//‡urh€r‚s‡ur phƒv‡hyv“rqh€r†urr r rsr vt   †‡ ˆp‡ˆ r†h’†qvr‡u’yur‘’yhqvƒh‡r Dshp‡ †ƒrpvsvphyy’‡‚‡urDI8Dh€riˆ‡ruh‰r‡‚t‚   !v‡†qvr‡u’yur‘’y†ˆppvh‡r  !ihpxhq rpuhtr‡ur€‡‚y‚r phƒh‡‡urƒ‚v‡   *9S CDGG'6quvyrr r‡ur r‡uv† *‚sƒˆiyvph‡v‚ T‚r r‡ ’vt‡‚h‰‚vq   ,v†hs‚ €h‡‡vt„ˆr†‡v‚iˆ‡‚rDrp‚ˆ‡r  ,phƒv‡hyv“vt‡ur€urrph 9‚r†‡uh‡€hxr//   .trr hyy’ Ds’‚ˆuh‰rhpur€vphyh€ryvxr .9S TC6IF'Xuvyrr r‚‡uh‡   0qvr‡u’y†ˆppvh‡r‡ur r†‚ rh†‚‡‚ˆ†r 0w‚ˆ hy†s‚ €h‡‡vtuh‡r‰r uhƒƒrrq‡‚‡uv†   ˆƒƒr ph†ryr‡‡r † 7ˆ‡D€h††ˆ€vt‡uh‡ v‡ ‚qˆp‡‚ ’†ˆ€€h ’‡uh‡Duhqhqhƒ‡rq‡‚hq   1ˆƒƒr ph†ryr‡‡r †h rs r„ˆr‡y’ˆ†rqv‡ur 1r‰r ’i‚q’ry†r†hvq‡ur’ rt rh‡vhyy‡ur    rƒ‚ ‡†irphˆ†r‡ur’ rqvp‡v‚h ’vt rqvr‡†   rƒ‚ ‡† D‡uvxx‚wvphpvqv†‡ur‚y’‚r‡uh‡   Uuh‡†ur rv‡p‚€r†s ‚€ T‚vs‡ur’ r uh†‡uv† Xur rh rr‚‡uh‡46 rrt‚vt‡‚   qvp‡v‚h ’vt rqvr‡†D€tˆr††vtv‡ur uh‰r‡ur€‚ ‚‡4   !svtˆ r†‡ur’‚ˆtu‡‡‚xrrƒ‡uh‡†h€rs‚ €h‡‡vt !9S 7S@TG6X@8'Xryvxr‡urhƒƒ ‚hpu    *ˆ†vtˆƒƒr ph†r†  *Xru‚ƒr‡‚v‡ ‚qˆpr‡urhƒƒ ‚hpu Hh’‚s‡ur   ,9S CPX6S9'D‡uvxr€hqr‡urpuhtr ,q‚pˆ€r‡†‡uh‡’‚ˆ†rr‚uh‰rirrv   .hqChy’hph†ƒrhx‡‚‡uv†h†ryy‡uh‡v‡uv .ƒ rƒh h‡v‚s‚ huvyrhq†‚v‡†‚‡irvt   0‡ur rƒ‚ ‡rr r‡t‚vt‡‚phƒv‡hyv“r 0v€ƒyr€r‡rqxvq‚shp ‚††‡uri‚h qs ‚€‡uv†   vt rqvr‡h€r†h’€‚ r  ƒ‚v‡‚ D‡vyyirv€ƒyr€r‡rqh†rq‚pˆ€r‡†   19S 7S@TG6X@8'`rhu Xr rtr‡‡vth 1irp‚€rh‰hvyhiyr    y‚‡‚sp‚€€r‡†s ‚€‡urD‡r h‡v‚hyE‚ˆ hy‚s 9S TC6IF'Pxh’ Uur†h€r‡uvtv‡u   U‚‘vp‚y‚t’urr rƒˆiyv†uvt‚ˆ  rƒ‚ ‡†‚‡ ‡ur rsr rpr//pv‡vt rsr rpr†v‡ur‡r‘‡4       6I9@STPI8PVSUS@QPSUDIB 6I9@STPI8PVSUS@QPSUDIB .1,9ˆxrT‡ rr‡Tˆv‡r11 .1,9ˆxrT‡ rr‡Tˆv‡r11 6yr‘hq vhW6! 6yr‘hq vhW6! Qu‚r?.1@*/.01Ah‘?.1@*/.1 Qu‚r?.1@*/.01Ah‘?.1@*/.1  hqr †‚ rƒ‚ ‡vt r‡  hqr †‚ rƒ‚ ‡vt r‡

        ! !   9S 7S@TG6X@8'Vs‚ ‡ˆh‡ry’’r†  Xr rt‚vt‡‚ r‚ƒr‡ur‡u rrr†‡r †‡uh‡h rv   9S TC6IF'Uuh‡†‡ur†h€r4Uuh‡†v ‡uv†q‚pˆ€r‡‡uh‡uh‰rhy rhq’irr r‰vrrqhq   ƒ ‚t r††4 ‡ur@‘ƒr ‡Qhrys‚ˆq‡‚ir†hsr'Uur   !9S 7S@TG6X@8'Uuh‡v†vƒ ‚t r††  !qvr‡u’yur‘’yhqvƒh‡r‡urqviˆ‡’yhqvƒh‡rhq‡ur   *9S TG6B6'6q‡ur†h€rh’v‡u‡ur *qvv†‚ƒ ‚ƒ’yhqvƒh‡r    ,hi†‡ hp‡?vhˆqviyr@‡‚‚4 ,D†‡uh‡v‡4T‚ˆqt‚‚q4   .9S 7S@TG6X@8'Uuh‡hy†‚v†v .9S TC6IF'H€/u€€    0ƒ ‚t r†† Xrwˆ†‡uh‰rqvssr r‡q‚pˆ€r‡†‡uh‡ 09S H6SFT'Pxh’    h rqvssr r‡ƒuh†r†‚sƒ rƒh h‡v‚ 6qr€hqr 9S 76DG@`'Pr‡uvt rth qvt‡ur   1hqrpv†v‚‡‚v€ƒyr€r‡hyy‡urpuhtr†‚r 1qvp‡v‚h ’hqh€vtp‚‰r‡v‚† Uurr‡u’yur‘’y   q‚pˆ€r‡†h†‡ur’irtv h‡ur ‡uh†h’vt‚xh’ hqv†‚‚p‡’y’‚ˆx‚D‡uvxh r//D‡uvxs‚    r‰r ’i‚q’puhtr‡urs‚ €h‡‚s‡ur‡uvt†‡uh‡ huvyr‡ur’r rh€rqh†v†‚‚p‡’yhqD‡uvx   ’‚ˆ r‚ xvt‚ T‚v‡vyyir//v‡vyy ‡ur’ rphyyrqr‡u’yur‘’y‚ 7ˆ‡ryys‚yy‚   !p‚‡vˆr‡‚irp‚sˆ†vts‚ h‡yrh†‡†v‘€‚ r !ˆƒ‚‡uh‡hqsvq‚ˆ‡uh‡‡urp‚‰r‡v‚v†v   *€‚‡u†vs‚‡y‚tr s‚ ‚‡ur  rh†‚†  *‡urqvp‡v‚h ’wˆ†‡s‚ p‚†v†‡rp’r‰r‡u‚ˆtu   ,9S H6SFT'6’‚‡ur p‚€€r‡†4T‚ ,v‡€h’‚‡ir vt‚ ‚ˆ†VQ68‚ ‚‡ur h€vt   .r rt‚vt‡‚‡hiyr‡uv†yv†‡‚svt rqvr‡†s‚  .p‚‰r‡v‚† Xrq‚uh‰rh†u‚ ‡uhq‡uh‡rˆ†r   0€‚ rqh‡h Xr rt‚vt‡‚ rp‚€€rq‡uh‡‡ur 0v‡urqvp‡v‚h ’hqD‡uvx†vpr‚ˆ hˆqvrpr   hpvq†hq†hy‡†ir†ƒyv‡v‡‚‚rt ‚ˆƒ‡ur v†‡urp‚†€r‡vp†‡hxru‚yqr †vqˆ†‡ ’hq   1r†‡r †vh‚‡ur t ‚ˆƒuvpuƒ r†ˆ€hiy’h‡‡uv† 1‚‡ur †‡urD‡uvxrrrq‡‚€hxr‡uh‡h†   ƒ‚v‡vyyir‚rq‚pˆ€r‡v‡u‡ur‚ƒ‡v‚‚s p‚†v†‡r‡h†rph‡u ‚ˆtu‚ˆ‡    irp‚€vt‡‚q‚pˆ€r‡†h†rƒ ‚prrq‡u ‚ˆtu  9S CDGG'`rhu 6qD€‚‡†ˆttr†‡vt       6I9@STPI8PVSUS@QPSUDIB 6I9@STPI8PVSUS@QPSUDIB .1,9ˆxrT‡ rr‡Tˆv‡r11 .1,9ˆxrT‡ rr‡Tˆv‡r11 6yr‘hq vhW6! 6yr‘hq vhW6! Qu‚r?.1@*/.01Ah‘?.1@*/.1 Qu‚r?.1@*/.01Ah‘?.1@*/.1  hqr †‚ rƒ‚ ‡vt r‡  hqr †‚ rƒ‚ ‡vt r‡

CIR Panel Book Page 25     !  ‡uh‡vs‡uh‡†irr‡ur†‡h‡r‚s‡urh ‡hq  ‡uh‡†uh‡ƒr‚ƒyruh‰r‚‡urv yhiry†‡uh‡h’  puhtr‚ˆyqrpr††h vy’irrrqrq 7ˆ‡‡urr  !€h’irwˆ†‡rrq‡‚ƒ‚v‡‡uh‡‚ˆ‡‡uh‡‡ur rv†h  *qvssr rprv‡urq‚pˆ€r‡uvpuD‡uvxrqir  ,q‚vth’h’irphˆ†rvsu’q ‚y’†v†v†‚ppˆ vt  .r rtrr h‡vtqvssr r‡hyp‚u‚y†   09S 76DG@`'Svtu‡ I‚Dsˆyy’ht rr   9S CDGG'Uur†u‚ ‡uhqv†‡ur  1†u‚ ‡uhq   9S 76DG@`'Svtu‡   9S H6SFT'Ir‘‡r rt‚vt‚‡‚‡ur  ƒ‚y’r‡u’yrrty’p‚y†‡urQ@B† D‡†vQvx7‚‚x  ! Xruh‰r†‚€rrvs‚ €h‡v‚   *T‚h†’‚ˆ rphyyv9rpr€ir ‚s110  ,r r‚ƒrrq‡uv††hsr‡’h††r††€r‡hqr‡ur  .ƒ ‚prrqrq‡‚v††ˆrh‡r‡h‡v‰rh€rqrq†hsr‡’  0h††r††€r‡‚sQ@B† D‡urTrƒ‡r€ir €rr‡vt‡uv†  ’rh rurh qhƒ r†r‡h‡v‚p‚pr vt‡ ’vt‡‚  1qrhyv‡u‡urv††ˆr‚sqh€htrq†xvhqqr‰ry‚ƒvt  h€h tv‚s†hsr‡’†‚‡uh‡rp‚ˆyqsrry  p‚€s‚ ‡hiyr‡uh‡Q@B†‚ˆyqir†hsrur r‡ur rv†    6I9@STPI8PVSUS@QPSUDIB .1,9ˆxrT‡ rr‡Tˆv‡r11 6yr‘hq vhW6! Qu‚r?.1@*/.01Ah‘?.1@*/.1  hqr †‚ rƒ‚ ‡vt r‡

CIR Panel Book Page 26      PANEL - SEPT 2009     7                                    E    6 F                 E,A                     !"# $%#&'%(")*   +, + !   E,A           -" +#+.+/   -            0"# 12#34)56    0>                  "# 12#34)=         7   7           8"# $%#&'%(")9:  2  8                 +# +* ;  2 .      >                  2 6   <   . : 5       "         =  >            !            !            -?4 ? " $   - ;  !    #,.   0"# $%(4.9)@ =:   0               .     :        F      7  =           7"# $%#&'%(")4       8 : :,*,*       8 +        "# 12#34)@    =       "# $%#&'%(").           25"%#495*9<#.#%,9#.5& 25"%#495*9<#.#%,9#.5& 08-":4 4 88 08-":4 4 88 2>  A2 2>  A2 ,B08C!7D08' >B08C!7D078 ,B08C!7D08' >B08C!7D078

        8        2       "# $%#&'%(")     >      . : 6 :       D     .            4             >               !*   >"   DD  !        :    -"   0+ -   :           0"# 25"%#4%5)=    0                      +   7         7"# 25"%#4%5)@    8    8"# $%#&'%(")9:     "# $%#&'%(")9:   "# 25"%#4%5)@      "# 25"%#4%5)=       :       : :   :  /                " $  " 1 :      :    !        !        -        -"# $%#&'%(")9:     0      D 6  0"# 25"%#4%5)4  >        4                7          7        8 > >  .  8=      DD                                          ;        25"%#495*9<#.#%,9#.5& 25"%#495*9<#.#%,9#.5& 08-":4 4 88 08-":4 4 88 2>  A2 2>  A2 ,B08C!7D08' >B08C!7D078 ,B08C!7D08' >B08C!7D078

CIR Panel Book Page 27                :    =   E,A                 D     >  4             ;      DD :        :         "# $%#&'%(")/      !??      : !       2 6    -            -          0         0       =                +=   >+   7        7"# 254%(()   :   8           8  : *#              ,             :               $             :   E,A       G   4 :       :     $       !          !      4   -      4   :   -;  )E       0          0         +2              : 2        7         7     8"# E(()26    8"# $%#&'%("). ;            :     :                 D                      25"%#495*9<#.#%,9#.5& 25"%#495*9<#.#%,9#.5& 08-":4 4 88 08-":4 4 88 2>  A2 2>  A2 ,B08C!7D08' >B08C!7D078 ,B08C!7D08' >B08C!7D078

       :88DD  2        DD   +"              +  !"# 254%(()= *#  -4   4 *    :   0  :  =     6   :        7     DD   8                   4    :   *#     "# $%#&'%(")9:    "# 254%(()       !      :    -#          0       "# $%#&'%("). : . :   7"# 25"%#4%5)&    8"# $%#&'%(")2      >       ,%&" 1 :     25"%#495*9<#.#%,9#.5& 08-":4 4 88 2>  A2 ,B08C!7D08' >B08C!7D078

CIR Panel Book Page 28 TEAMMEETINGMINUTESONSEBACATESͲDR.BELSITO’STEAM 198

7 DR. BELSITO: If you could do that, let 1 SPEAKER: Green one.

8 us know because then it would help. I mean, I 2 DR. BELSITO: Okay. Okay, so this is a

9 don't think we're going to reopen the report. The 3 first for us. A SLR, so it's in green along with

10 question is how to wordsmith the discussion 4 kojic acid.

11 regarding this. 5 And we've gotten some new data. And

12 Okay. Anything else on the bromates? 6 again, this was one of the reports, like kojic

13 Okay. So we're moving to the sebacates. How do 7 acid, that was sort of formulated in, hopefully,

14 you pronounce that? 8 the new CIR approach to doing documents, which I

15 DR. ANDERSEN: We had a short discussion 9 think we already discussed with kojic acid that we

16 on the other team and nobody knew how to pronounce 10 liked. And so now we're just looking at this data

17 it. 11 and deciding what we need.

18 SPEAKERS: Sebacates. 12 On Table 1 on page 42 and 43, there were

19 DR. BELSITO: Sebacates. 13 just a couple that I thought needed to be

20 SPEAKER: What color? 14 amplified like dioctyl sebacate is an organic

21 DR. BELSITO: Green. It's with the 15 compound. That's the definition. See Structure.

22 kojic acid. 16 DR. LIEBLER: The middle of page 32

ANDERSON COURT REPORTING 17 you're talking about?

706 Duke Street, Suite 100 18 DR. BELSITO: No, top.

Alexandria, VA 22314 19 MS. BECKER: Top of page.

Phone (703) 519-7180 Fax (703) 519-7190 20 DR. BELSITO: And then on page 33, the 21 same thing with dodecanedioic acid is an organic

22 compound.

ANDERSON COURT REPORTING

706 Duke Street, Suite 100

Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190

199 200

1 DR. ANDERSEN: Coagula extract is an 1 DR. LIEBLER: Well, you've got the

2 extract of coagulum. 2 world's tallest carbonyls.

3 DR. BELSITO: Right. 3 MS. BECKER: I don't have steady hands.

4 DR. ANDERSEN: This is the dictionary 4 DR. LIEBLER: There's a bug, so if you

5 we're talking about. 5 did it in ChemDraw and it looked right, then

6 DR. BELSITO: Okay. 6 there's a problem with cutting and pasting or

7 DR. ANDERSEN: It is what it is. 7 something. But that just needs to be fixed

8 DR. BELSITO: So straight out of the 8 because they're unreadable.

9 dictionary? 9 MS. BECKER: Okay. Okay.

10 DR. ANDERSEN: Yes. 10 DR. ANDERSEN: All right. And this is a

11 DR. BELSITO: Because all the others say 11 perfect opportunity to get some feedback from you

12 it's the diester of isostearyl alcohol and sebacic 12 on -- flip back to the kojic acid report, since

13 acid. 13 it's in the same document, real quick. My own

14 Okay. Just questioning. 14 personal preference, which hasn't succeeding in

15 DR. LIEBLER: You know, the structures 15 posing yet, my own personal preference is that you

16 in this table, I realize it's tough, but the 16 get more out of showing the stick figures.

17 structures are just about unreadable. And I think 17 DR. SNYDER: Yes.

18 -- I don't exactly know how you do these, but if 18 DR. ANDERSEN: A better understanding of

19 you can perhaps paste in better quality structures 19 what the heck this molecule really looks like in

20 from something like ChemDraw or ChemOffice. 20 its length, especially if your eyes aren't good

21 MS. BECKER: Yeah, these were done in 21 enough to figure out if that's an 11 or a 17 in

22 ChemDraw. 22 the little side script in the "formula." So

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

CIR Panel Book Page 29 201 202

1 that's the direction that I would like to go. 1 DR. BELSITO: Yes.

2 And frankly, since that looks hugely 2 SPEAKER: Okay.

3 awkward when you try to put it in tables, I'd pull 3 DR. SEIDMAN: Which, I might point out,

4 it out of the table. 4 is how Valerie had it in the first place.

5 DR. BELSITO: Right, as in figure 7. 5 SPEAKER: (off mike) acid.

6 Right. 6 DR. SNYDER: What are we going to title

7 DR. LIEBLER: The figure is fine and 7 it?

8 these figures for the kojic acid compounds look 8 DR. BELSITO: Basic acids and salts and

9 just fine. 9 esters as used in cosmetic products. So basic

10 SPEAKER: What page is that on? 10 acids and other related dicarboxylic acids and

11 DR. LIEBLER: 55. 11 their salts and esters as used in cosmetic

12 MS. BECKER: 55. 12 products. You don't like that sound?

13 SPEAKER: (off mike) the kojic ones. 13 DR. SNYDER: No, I'm just curious.

14 DR. SEIDMAN: Alan, what are you 14 MR. RE: If you would like to see the

15 suggesting gets pulled out of the table? 15 stick figures, just turn to the HPB.

16 DR. ANDERSEN: The figures. 16 DR. BELSITO: Yeah. Okay. On page 3,

17 SPEAKER: So all the figures. 17 this West Germany reference. First of all,

18 DR. BELSITO: The figures. Do it as a 18 there's not even a reference. It just says

19 figure. 19 amongst Germany that's got to be at least 20 years

20 SPEAKER: They don't fit. 20 old. So I would say that unless we have some

21 SPEAKER: So do it as a separate (off 21 update from the EU or from the current German

22 mike)? 22 state, we delete that.

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

203 204

1 Page 4, the azelaic acid, 20 percent is 1 DR. SNYDER: Oh, okay. I was looking at

2 approved by the FDA. It's a topical cream for the 2 (off mike).

3 treatment of mild to moderate acne. There is also 3 DR. BELSITO: No.

4 a 15 percent gel for the treatment of rosacea. 4 DR. SNYDER: Okay. Never mind.

5 DR. SNYDER: And along those lines, 5 DR. BELSITO: Page 7, under Acute

6 that's not -- transillic acid is not listed in the 6 Toxicity, the dibutyl and sebacate and azelaic

7 use table. 7 acid. I thought that the last couple of sentences

8 SPEAKER: I thought it was. 8 in both of those paragraphs -- while dibutyl is

9 MS. ROBINSON: It is. It should be. 9 not currently used, it would unlikely cause acute

10 DR. SNYDER: I thought it was at the 10 toxicity, and the rest of that paragraph. And

11 very end. 11 then these data indicate that azelaic acid is not

12 MS. ROBINSON: Yes. It's on page 39, 12 highly toxic (off mike) oral. Shouldn't those be

13 all the way at the bottom it starts. 13 in the discussion rather than here? Or are we

14 DR. SNYDER: I'm looking at the new use 14 making -- again, I guess we're changing the format

15 (off mike). 15 so this -- is it likely renal?

16 MS. ROBINSON: Oh, it's in the actual 16 MS. ROBINSON: I can move it to the

17 tables. 17 discussion if it's an easier read. But what do

18 DR. BELSITO: Page 39. 18 you think, Brenda?

19 MS. ROBINSON: Yeah, at the end of the 19 DR. ANDERSEN: I think what we've been

20 actual document. 20 doing is trying to calibrate how it is we handle

21 SPEAKER: At the bottom. 21 giving the reader a sense of the import of the

22 DR. BELSITO: Azelaic acid shaving care. 22 section.

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

CIR Panel Book Page 30 205 206

1 DR. BELSITO: Okay. 1 so you might want to just consider this. Revisit

2 DR. ANDERSEN: This goes much further 2 it in just a few minutes when you look at the

3 than we did in the examples for kojic acid. So 3 summary tables because you might want to think

4 the question is what's your level of comfort with 4 about putting summary tables in the individual

5 that? Kojic acid was more of an attempt simply to 5 sections instead of a summary statement or you

6 summarize what data you'll find in the following 6 might want a combination. So you can discuss this

7 paragraphs. This says these data are gone because 7 further, of course, now. But you might just want

8 they are. But if you want to reserve any such 8 to think of shelving it for just a few minutes

9 statements for the discussion, we can do that. We 9 until you see the tables in 5 (off mike).

10 wanted to push the envelope a little bit and see 10 DR. BELSITO: Okay. We can do that.

11 what your reaction was. 11 Page 10.

12 DR. SNYDER: I like it better in the old 12 DR. SNYDER: Well, in that same area, I

13 report where it was italicized right up front 13 prefer to say the studies that reported NOEL are

14 rather than here it's kind of -- 14 less than 4,000 milligrams rather than "CIR

15 DR. BELSITO: Buried. 15 concluded." We had a couple things CIR concluded,

16 DR. SNYDER: -- buried at the summary 16 CIR determinate. I think we (off mike).

17 statement or discussion-like statement instead of 17 MS. ROBINSON: What paragraph?

18 being a summary-like statement there. 18 DR. SNYDER: On page 7, (off mike). The

19 DR. SEIDMAN: May I suggest something? 19 third line in the first paragraph there where it

20 I think revisit this in just a few minutes when

21 you get to the tables. The new tables we're 20 says, "CIR concluded that the study supported (off

22 throwing out there as floaters or trial balloons, 21 mike)." Say, "The study supported NOEL."

ANDERSON COURT REPORTING 22 MS. ROBINSON: Okay.

706 Duke Street, Suite 100 ANDERSON COURT REPORTING

Alexandria, VA 22314 706 Duke Street, Suite 100

Phone (703) 519-7180 Fax (703) 519-7190 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190

207 208

1 DR. SNYDER: And then down at the next 1 maybe like to communicate with you about what

2 one under azelaic acid with rat and rabbit, CIR 2 details are actually available in these tox of

3 determined the maximum tolerated dose. It should 3 studies. Now, maybe the acute tox aren't the most

4 just say, "Based on the reported endpoints of 4 important studies on earth, but in getting ready

5 animal deaths, the maximum tolerated dose (off 5 for this meeting that's what we had time to play

6 mike)." 6 with.

7 DR. BELSITO: On page 10 on the Mingrone 7 And I think that there is a prevailing

8 Study 1083, were the ends given? There were no 8 sentiment that those tables have more information

9 ends. 9 content than any of the verbiage that we can

10 DR. ANDERSEN: A perfect segue to let's 10 write. At a glance you can see what it is you

11 talk tables. What Brenda was referring to earlier 11 see.

12 was that there had originally been captured Table 12 DR. BELSITO: So we would get rid of all

13 5 that summarized at least one study. And not to 13 the verbiage. These paragraphs would disappear

14 put words in Brenda's mouth, but the idea of 14 and under Acute Toxicity it would just say --

15 having a full picture of which species, what dose 15 DR. SEIDMAN: I think that's on the

16 levels, how many animals, is what she would like 16 table. And it's on the table for different

17 to see. So she went ahead and made tables for 17 endpoints. You might not want verbiage for acute,

18 but you might want it for subacute or chronic.

18 both Smith in '53 and Mingrone in whatever year. 19 DR. BELSITO: Right.

19 And those tables 5A and B or whatever -- 20 DR. SEIDMAN: Because I think that's

20 DR. SEIDMAN: 5D and C. 21 going to be a case-by-case depending on the

21 DR. BELSITO: 5C. 22 endpoint.

22 DR. ANDERSEN: D and C show how we would ANDERSON COURT REPORTING

ANDERSON COURT REPORTING 706 Duke Street, Suite 100

706 Duke Street, Suite 100 Alexandria, VA 22314

Alexandria, VA 22314 Phone (703) 519-7180 Fax (703) 519-7190

Phone (703) 519-7180 Fax (703) 519-7190

CIR Panel Book Page 31 209 210

1 SPEAKER: But it's up for discussion. 1 SPEAKER: (off mike)

2 DR. BELSITO: No, I mean, I think 2 DR. SEIDMAN: And (off mike) the summary

3 certainly for the acute. I mean, we all know what 3 statement and address that in the text.

4 an acute study is. And, you know, basically we're 4 DR. SNYDER: Yeah, up front.

5 looking at, you know, the -- 5 DR. SEIDMAN: And then up front in the

6 DR. SNYDER: The most important thing

7 for an acute study is the LD50. 6 table.

8 DR. SEIDMAN: If it's reported. We 7 MS. ROBINSON: Like the kojic acid.

9 don't always get an LD50. 8 DR. SNYDER: Yeah, like kojic acid,

10 DR. SNYDER: Right, but that kind of 9 right up front.

11 goes to (off mike). 10 MS. ROBINSON: Okay.

12 DR. ANDERSEN: It's two animals. We're 11 DR. SNYDER: There's three or four (off

13 not very interested anymore. 12 mike) around that.

14 DR. SNYDER: But under the Acute section 13 DR. BELSITO: Then no verbiage. Just

15 we have basically, at least I can count, four or 14 your summary statement, no verbiage, and a

16 five different summary statements where we could 15 referral to the tables.

17 have just put that all in one saying there was low 16 DR. SEIDMAN: That's what I would prefer

18 toxicity based upon LD50s and an oral (off mike) 17 (off mike).

19 and -- 18 DR. ANDERSEN: Yeah, definitely.

20 DR. ANDERSEN: As shown in tables. 19 SPEAKER: Really shortened this stuff.

21 DR. SNYDER: Exactly. 20 And that's certainly a case for a lot of the

22 SPEAKER: Right. 21 animal tox studies.

ANDERSON COURT REPORTING 22 DR. BELSITO: Okay.

706 Duke Street, Suite 100 ANDERSON COURT REPORTING

Alexandria, VA 22314 706 Duke Street, Suite 100

Phone (703) 519-7180 Fax (703) 519-7190 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190

211 212

1 DR. SEIDMAN: Well, there's a third 1 that when they start drawing conclusions from the

2 wrinkle on this, if I can just raise the issue, 2 data, that deserved to be in the discussion

3 because it came up in the other group (off mike) 3 section. They really didn't go to whether the

4 has to do with editorializing a bit. So like 4 staff should be doing conclusions or not but

5 under Azelaic Acid, I put -- this was how I 5 rather where they should be included, particularly

6 summarized this particular study, given the low 6 because if we start parsing it too finely we'll

7 toxicity for azelaic acid by the oral and (off 7 end up drawing conclusions about energenicity and

8 mike) routes of exposure and as (off mike) dermal 8 outside of the entire scope of evidence that might

9 route of exposure in concentration used in 9 be relevant to a conclusion of carcinogenicity.

10 cosmetics, because azelaic acid presents a very 10 So we thought within the data section it

11 low risk of causing acute toxicity (off mike). 11 was appropriate to have summaries of the data, but

12 Now, in the -- when Jay brought up the 12 that when you went the further step and said so

13 -- he mentioned that he'd prefer no conclusion on 13 I'm concluding that this presents a low risk, that

14 the data, but we always are considering dose. So 14 that should not be in the data section; that

15 here we -- well, we are considering dose of, you 15 should be in the discussion section.

16 know, use in cosmetics, and that's what we always 16 DR. LIEBLER: I agree with that.

17 are comparing our data to in the literature. So 17 DR. BELSITO: Yeah.

18 we're just editorializing what we put in the 18 DR. SNYDER: I think we should just

19 beginning and end, you know, summarizing the data 19 stick to summarizing the data, not interpreting

20 and then editorializing. 20 the data.

21 Where does one begin and the other end? 21 DR. BELSITO: Mm-hmm.

22 MR. ANSELL: I think it was our position 22 DR. SEIDMAN: In the individual

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

CIR Panel Book Page 32 213 214

1 sections. 1 uncomfortable in preloading the discussion section

2 DR. BELSITO: In the individual 2 with that information for your consideration.

3 sections. 3 It's a toxicologist's review of the information.

4 DR. SEIDMAN: Let's discuss the 4 It sure looks that way and then you guys get to

5 discussion section then at the end of the 5 look at it.

6 document. Do we consider -- in the CIR, is it 6 DR. SNYDER: Well, when we make a

7 appropriate to consider, which I think it is, 7 comment on kojic acid it makes writing the summary

8 concentration of use relative to the data? 8 very easy because you just basically merge all

9 DR. BELSITO: We always do. 9 those summaries (off mike) flow in a summary

10 SPEAKER: Absolutely. 10 statement.

11 DR. SEIDMAN: Okay, so that's fair 11 DR. ANDERSEN: You bet. So I think we

12 enough. 12 have in the past not touched anything relating to

13 DR. BELSITO: Yeah. 13 the discussion section until we have a couple of

14 DR. SEIDMAN: Just let me get a feel for 14 panel discussions under our belt. But I'm

15 that. Okay. 15 wondering whether we may not be able to just be

16 DR. ANDERSEN: Yeah. And I think if I 16 able to prime it a little bit. If you disagree,

17 can take that one step further. Even at this 17 you're not going to be reluctant to say so. If

18 early stage of the document -- let's say that this 18 you agree and say attaboy, then you will have

19 is now December and we got this and we had this 19 accomplished something. But I think, you know,

20 discussion back in September -- that language that 20 Jay's message -- the reinforcement to the other

21 Brenda developed in the front end relating to 21 group that to the extent that we possibly can,

22 azelaic acid, I'm not sure I would be 22 those are discussion elements.

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

215 216

1 SPEAKER: Message received. 1 products were driven by a forward mutation (off

2 DR. BELSITO: On page 11, the Leong 2 mike). That's not in the right place.

3 Study, I don't think it belongs there, but I don't 3 That's got to go.

4 know really where to place it other than perhaps 4 MS. ROBINSON: That's part of the Leong

5 to eliminate it completely. It says the 5 Study.

6 biocompatibility of bioerodible polyanhydrides and 6 DR. BELSITO: That's part of what we're

7 the toxicology of the polymer breakdown products. 7 deleting. We're deleting that whole thing.

8 That certainly doesn't belong under chronic 8 Leong. We're deleting the whole bottom two-thirds

9 toxicity, I don't think. 9 of page 11.

10 SPEAKER: We'd be comfortable removing 10 DR. SNYDER: Okay. Right.

11 it. 11 DR. BELSITO: And the top part of page

12 DR. BELSITO: Yeah. I don't think it 12 12. And Table 6, it says summary of mammalian

13 adds anything to the document at all. 13 effects.

14 DR. SNYDER: I agree. There's lots of 14 And then under Genotoxicity it has

15 stuff related to designating time versus 15 negative aims, so that's not mammalian. So do we

16 short-term studies in that section. And also on 16 want to get rid of mammalian and just say summary

17 that same page right above it, that looks like 17 of effects?

18 aminotricity acids (off mike) that should go back 18 DR. ANDERSEN: Very advanced salmonella.

19 to the genotox section. 19 (Laughter)

20 MS. ROBINSON: I'm sorry. Which page? 20 DR. BELSITO: Point well taken. Page

21 DR. SNYDER: On page 11. The (off mike) 21 20, that's just typos.

22 toxicity and mutagenicity of the degradation 22 SPEAKER: (off mike)

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

CIR Panel Book Page 33 217 218

1 DR. BELSITO: On page 26, the third 1 DR. BELSITO: Right. It just did not

2 paragraph up from the bottom, it says basic acid 2 show fetal toxicity, teratogenicity, and neonatal

3 did not show any teratogenic effect on fetal 3 toxicity in rabbit studies.

4 toxicity. Is that redundant, did not show any 4 SPEAKER: Right.

5 fetal toxicity? 5 DR. ANDERSEN: (off mike) did not cause

6 DR. LIEBLER: So just remove any 6 fetal toxicity (off mike).

7 teratogenic effect. 7 SPEAKER: Yeah, "did not cause" would be

8 DR. BELSITO: Right. Just any fetal 8 better to show --

9 toxicity. Did not show fetal toxicity. 9 DR. BELSITO: And "separate?"

10 DR. ANDERSEN: You know, I think 10 SPEAKER: Yes.

11 actually, Curt, checking on this I'm not sure what 11 DR. ANDERSEN: And that should be "or"

12 the truth is, but those are two separate 12 instead of "and."

13 endpoints. I mean, I could have either yes or no 13 DR. BELSITO: A question that I had

14 for fetal toxicity and still have some increased 14 throughout this report is that we've already

15 incidence in birth defects, so. 15 looked at some other aliphatic diesters, for

16 DR. BELSITO: So did not show 16 instance, maleic acid. And did we want to bring

17 teratogenicity? 17 in at least a summary of any of that data into

18 SPEAKER: (off mike) in developmental 18 this report to help us out in terms of the safety

19 studies that separate out the two. 19 assessment or --

20 DR. ANDERSEN: So what should be done 20 DR. ANDERSEN: I forget. What's the

21 there is take out the word "teratogenic effects 21 (off mike)?

22 in." 22 DR. LIEBLER: Maleic acid is two carbons

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

219 220

1 with two carboxins with a double bond between (off 1 concentration.

2 mike) three carbons. 2 SPEAKER: (off mike) I thought it was a

3 DR. ANDERSEN: So it's about as short as 3 2.2 negative (off mike).

4 you can get. 4 DR. BELSITO: It may be already in the

5 DR. LIEBLER: Yeah. 5 document. I didn't see it.

6 DR. ANDERSEN: And these are a bit 6 DR. ANDERSEN: No, the new thing that

7 longer. It's got eight carbons including the two. 7 was sent out was September 18th. I had it dated

8 I'm not sure -- 8 when the council provided it. It isn't in the

9 DR. LIEBLER: You've got adipic acid 9 book.

10 (off mike) I don't know if that's used. 10 DR. BELSITO: Right. Okay.

11 DR. ANDERSEN: -- how instructive it is. 11 DR. SNYDER: August 18th and September

12 DR. BELSITO: Okay. So I'll strike that 12 24th. There's a memo from John Bailey on August

13 question. Okay. And I guess the only new data we 13 18th that we got prior to coming to the meeting.

14 got were 2 additional sensitization studies at 1.2 14 For this meeting we got one on 9-16 to 9-18.

15 percent. Is that -- I'm trying to look at what we 15 DR. BELSITO: Yeah. And then we got

16 sent as new. That's the only thing I picked up. 16 another one September 16th that I guess was handed

17 It was sent by e-mail or some fashion, 17 out today. Is that right?

18 some additional studies, and I just made a note 18 MS. ROBINSON: Yes.

19 that there were -- in addition to what we have 19 DR. BELSITO: And 18th. So the 16th is

20 here in the document, there were two additional 20 7.8 percent --

21 sensitization studies done at 1.2 percent, which 21 SPEAKER: Two.

22 doesn't really help us since it's used at a higher 22 DR. BELSITO: 7.2 (off mike). And then

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

CIR Panel Book Page 34 221 222

1 it was a human repeat in self patch testing. 1 glance you can see that there are positive

2 SPEAKER: Yes. 2 reactions. You don't have to dig for it.

3 DR. BELSITO: Fifty-one subjects 3 DR. BELSITO: Yeah. Well --

4 completed. And then -- 4 DR. SEIDMAN: I'm going to pass out this

5 SPEAKER: And they all take (off mike). 5 table. (off mike) I don't think you're going to

6 DR. BELSITO: Another study, Primary 6 find it of value, but I'm going to pass it out.

7 Skin Irritation, and this is 100 percent diethyl 7 This is case reports.

8 and 30 percent diethyl, and 8 male Japanese White 8 DR. LIEBLER: I think they've got them.

9 Strain rabbits. At that high concentration there 9 DR. BELSITO: We've got them, yeah.

10 were some allergic reactions. 10 DR. SEIDMAN: Okay, I'm sorry. It's

11 SPEAKER: (off mike) 100 percent. 11 really not -- it's not for induction --

12 DR. BELSITO: A hundred percent, yeah. 12 DR. BELSITO: Right.

13 And then case reports. What is (off mike)? Oh, 13 DR. SEIDMAN: -- so I think this has a

14 (off mike) motion, okay. 14 policy and the (off mike) do not.

15 DR. ANDERSEN: That chart is also 15 DR. BELSITO: Well, looking at what we

16 something we put together that basically 16 have for these and the new data that we got, we

17 summarizes what's in the report. Again, it takes 17 don't have impurities.

18 a long time to read through the case literature. 18 We don't have UV absorption. But

19 Again, we're wondering whether a table that shows 19 looking at the structure, there are no rings.

20 it -- in this case shows that there were some 20 They're not likely to absorb. We have a negative

21 positive reactions in the case literature. You 21 Ames but no mammalian genotoxicity. And then do

22 can do what you will with it, but it's -- at a 22 we have sufficient carcinogenicity? No.

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

223 224

1 If we have sufficient carcinogenicity 1 genotoxicity.

2 and you don't feel we need mammalian genotoxicity 2 SPEAKER: (off mike)

3 and we have negative repro tox, then do we really 3 SPEAKER: All we have so far are (off

4 need impurities? Because presumably whatever 4 mike), but the American Chemistry Council has

5 impurities were there were tested and weren't 5 provided us with data before. So if you wanted

6 giving problems, so. 6 these expanded, it's page 15 in the (off mike) HPB

7 DR. SNYDER: We only really have one 7 test plan that was provided by the council in one

8 carcinogenicity study. 8 of their submissions. It's kind of like that

9 DR. BELSITO: Right. 9 page, a quarter of an inch.

10 DR. SNYDER: The other one is actually 10 SPEAKER: (off mike) looks like a lot

11 renal processing (off mike). 11 that they have. There was a 13-week -- or the 90

12 DR. LIEBLER: Is the one you're talking 12 days thing.

13 about called the BIBRA 96 reference? 13 SPEAKER: Well, those should probably be

14 DR. SNYDER: Yes. 14 summarized.

15 DR. LIEBLER: Which has very little 15 SPEAKER: Yeah. Yeah, we need to bring

16 information provided, right? 16 those in.

17 DR. SNYDER: 10 milligrams per kilogram 17 DR. BELSITO: Right.

18 for (off mike). 18 SPEAKER: Yeah, I think the question the

19 DR. LIEBLER: And the only other one is 19 other group had related to is this enough or do

20 around liver phocyte tests. 20 you want to see the real studies? They were

21 SPEAKER: In the (off mike) in the test 21 inclined to see the real studies, but I'll leave

22 plan, someplace in here a whole bunch of data on 22 it up to you as to what your comfort level is.

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

CIR Panel Book Page 35 225 226

1 DR. BELSITO: Well, I think we'd 1 see in more detail exactly what was done in the

2 probably want to see the real studies, but the 2 studies. I'm assuming, but let me let Curt, Paul,

3 question is I'm assuming since this is being put 3 and Dan discuss that.

4 together for HPV and REACH, that these are 4 DR. SNYDER: It's always a case-by-case

5 accurate summaries or statements of what was 5 basis. So if we have 10 chronic studies, even

6 found. And assuming that's the case, is this safe 6 with low numbers of animals, you get a lot more

7 as used? And do we have enough information? 7 confidence than if you have one study. So if you

8 Assuming that, in fact, when we get the real 8 only have one study, you'd like to have pretty

9 studies they confirm the summary. 9 good information to make sure the methodology and

10 DR. SEIDMAN: Are you proposing that we 10 the valuations and things like that were

11 get the real studies? 11 appropriate. So I think on a case-by-case basis

12 DR. BELSITO: I think it's always been, 12 it's when there is minimal data, I want to see

13 I mean, you know, they're not studies that are my 13 more. When there's minimal data I want to see

14 area of expertise. 14 more data. And when there's lots of data, I don't

15 So, you know, from my standpoint, I 15 need to see lots of data, so to speak, of how it's

16 would read them, but I'd be relying on my 16 done.

17 colleagues to guide me. I think it's always been 17 DR. SEIDMAN: And so lots of data --

18 the policy of the panel, particularly maybe in 18 lots of data coming from one source, is that

19 this case where we don't have impurities and we're 19 right? I haven't reviewed this document.

20 going to be assuming that these are -- we don't 20 MR. ANSELL: No, no, it's a consortium

21 need the impurities because whatever the 21 of producers that come together.

22 impurities are, they were studied. We'd want to 22 DR. SEIDMAN: It's a summary from that

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

227 228

1 consortium and there are no original data which 1 didn't result in mutagenic or genotoxic effects or

2 were published or which were available to CIR? 2 carcinogenic effects if we have that data. So

3 MR. ANSELL: You'd have to look at the 3 that the data that we're missing right now are the

4 data area because they tried to minimize animal 4 mammalian, genotoxin, carcinogenicity. And then

5 testing, so they uses confrontational methods, 5 the question becomes are these summaries adequate

6 they used all sorts of things. But I think if you 6 for my colleagues or do my colleagues want to see

7 were to identify what particular effect you're 7 the actual studies. And I leave that up to them

8 concerned about, we can see if we could get 8 to comment.

9 additional data. 9 MR. RE: Just to note the actual study

10 The package itself goes into, you know, 10 reports that went to ZIAM and OECD programs (off

11 an entire analysis of toxicity and carcinogenicity 11 mike) letting your (off mike). I'm not certain

12 and chronic toxicity. So, I think getting the 12 that we could obtain them (off mike).

13 full package would be -- 13 DR. KLAASSEN: Apparently there's not a

14 DR. BELSITO: I don't think we need the 14 whole lot of carcinogenicity studies that have

15 full package. In my assessment, what we're 15 been done according to their document either. So,

16 lacking right now are impurities and 16 I guess it would be nice to know is that one the

17 genotoxicity/carcinogenicity. And therefore, we 17 only one that's been done? And that one that has

18 would be relying on, you know, if we don't get 18 been done should have more data. Definitely look

19 impurities that's fine because we, you know, again 19 at that data. Did they use two mice?

20 we can say in the discussion, you know, whatever 20 MR. ANSELL: You know, if you look in

21 the impurities are they didn't result to any 21 the report, you know, page 14, 15, the results

22 reproductive or teratogenic effects and they 22 from (off mike) genotoxicity material, mammalian,

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

CIR Panel Book Page 36 229 230

1 in vitro chromosome, in vivo chromosome, so I 1 important questions.

2 think the question is highly relevant. 2 DR. LIEBLER: And if that's, you know --

3 And let us see what data responsive to 3 SPEAKER: And at this stage in the

4 that question we can get. But it's unlikely to be 4 review, (off mike).

5 the entire package (off mike). 5 DR. LIEBLER: It may be, you know, many

6 DR. LIEBLER: We may not need the whole 6 yards of data or whatever you said, but, on the

7 package. In fact, the compound space -- chemical 7 other hand, it may be something that we can get

8 space that this is describing is larger than what 8 our hands on and look at and we should at least

9 we're considering in these ingredients any way. 9 try to do that.

10 There are a lot of shorter chain things here. 10 DR. SNYDER: And when there's a NOEL or

11 There's some unsaturated things that might have 11 a NOAEL, it's always nice to know what it was

12 different properties. 12 based on rather than just -- because these all

13 I just feel it sounds like there are 13 just say there was a NOAEL, but they don't say

14 data out there that we're not getting. I don't 14 what was the basis. Was it decreased weight gain

15 think it's appropriate for us to just decide that 15 or was it some, you know, toxicity endpoint that

16 we shouldn't look at any of it. So, I think we 16 might pop up that we can look at someplace else.

17 need to see if we can get some of this. It sounds 17 So, again, we just need a little bit more

18 like from looking at this Table 3, at the end of 18 information than what's in this summary.

19 the HPV report, I don't see any references for 19 DR. BELSITO: So, where are we going?

20 anything. This looks like it's stuff that's been 20 Are we saying insufficient for impurities,

21 done and not published. 21 genotox, carcinogenicity?

22 MR. ANSELL: Yeah, I mean, those are all 22 Are we tabling to get more information

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

231 232

1 from the summary reports that we see here in the 1 but whatever impurities were there didn't result

2 diester HPV test plan? Where do we want to go? 2 in problems in the genotox, carcinogenicity, and

3 This is the first time we're seeing this document. 3 reproductive studies. We've done that before.

4 DR. SNYDER: I prefer tabling it and 4 SPEAKER: That's a wonderfully empirical

5 asking for the additional data that we know 5 approach --

6 exists. We're going to get it. 6 DR. BELSITO: Right.

7 DR. BELSITO: Okay. 7 SPEAKER: -- resolving (off mike).

8 DR. SNYDER: I think that's a more 8 DR. BRESLAWEC: May I ask your opinion

9 logical (off mike). 9 on one format matter here? If you look at page 21

10 DR. BELSITO: So we're going to table it 10 and 22 and 23 and the top of section 24, it

11 and ask for additional genotoxin carcinogenicity 11 summarizes what are called "skin treatments."

12 data that appears to be from the diester HPV test

13 plan available to us. And if it's linear feet or 12 These are essentially efficacy studies involving

14 data, send it electronically and let us shift 13 one or more of these components. In the past

15 through it. 14 we've summarized them. An alternate way of

16 MS. ROBINSON: Do we need impurities? 15 dealing with these studies is presented in italics

17 No? Impurities? 16 in the middle of page 24 and we'd like your

18 DR. BELSITO: I mean, if they have 17 opinion on that.

19 impurities, that would be great, but, again, I 18 DR. ANDERSEN: It takes much less space.

20 think we can handle -- I mean, we handle 19 (Laughter)

21 reproductive toxicity. You know, we can handle 20 DR. BELSITO: I think when you're

22 impurities by saying we didn't have impurities, 21 dealing with something like azelaic acid that also

ANDERSON COURT REPORTING 22 has a drug use, you know, you're, I think,

706 Duke Street, Suite 100 ANDERSON COURT REPORTING

Alexandria, VA 22314 706 Duke Street, Suite 100

Phone (703) 519-7180 Fax (703) 519-7190 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190

CIR Panel Book Page 37 233 234

1 summarizing it as you did in the alternate skin 1 MR. RE: -- (off mike) part of the (off

2 study section -- is fine. I don't think we need 2 mike).

3 all of the details as to whether they wanted to 3 MR. ANSELL: You know, that's all true.

4 treat acne with minocycline plus azelaic acid and 4 Let us look first and then we will come back and

5 whether there was a difference in those two 5 say if there's a problem or not. You know, I

6 treatment groups. I mean, that's not -- 6 expect that there are robust summaries available

7 DR. BRESLAWEC: We have the same issue 7 (off mike). Let us look first.

8 in things like botanicals where there are 8 SPEAKER: You know, we pay for ASTM

9 thousands of studies done on a variety of 9 standards, IOC standards (off mike), so we'll need

10 conditions from growing hair to losing hair. And 10 to do that.

11 we prefer not to deal with the effectiveness 11 DR. BELSITO: Okay. So --

12 question. 12 DR. SNYDER: I have one more question.

13 DR. BELSITO: Yeah. Right. 13 On page 25, there's a chronic toxicity study in 40

14 DR. BRESLAWEC: Great. Thanks. 14 rats and 40 rabbits. Do you see it there?

15 MR. RE: Just getting back to the 15 MS. ROBINSON: Oh, yeah.

16 availability for a moment, would the CIR entertain 16 DR. SNYDER: They present the results of

17 the idea of joining the aliphatic esters panel? 17 the rabbits, but there are no rats. Is it in the

18 Because that is the usual mechanism on which you 18 table there?

19 gain access to this data (off mike). So if you'd 19 SPEAKER: (off mike)? I'm sorry.

20 like to go that way, that would be the traditional 20 DR. SNYDER: The chronic toxicity was

21 way of gaining access to these data (off mike) -- 21 investigated in 40 rats and 40 rabbits. And then

22 SPEAKER: Never done it. 22 we go on to say that chronic toxicity (off mike)

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

235 236

1 no significant difference in (off mike) and 1 DR. SEIDMAN: If we put it in a table,

2 rabbits, but we don't get any data results for 2 we'd have to say, of course, reference the table.

3 rats. 3 DR. KLAASSEN: Yeah, right.

4 DR. SEIDMAN: Oh. 4 DR. BELSITO: Any other comments? Okay.

5 SPEAKERS: Oops. 5 Why don't we take a 10-minute break before we move

6 DR. SEIDMAN: Yeah, oops. 6 to PEGs, get some coffee, relieve our bladders.

7 DR. SNYDER: I didn't look in the table, 7 (Recess)

8 (off mike) to the table then.  9 DR. SEIDMAN: The two tables I prepared

10 (off mike).

11 DR. BELSITO: Okay. That's what I

12 thought. Okay.

13 DR. SEIDMAN: So I didn't touch this.

14 DR. BELSITO: Okay.

15 DR. SEIDMAN: But that's kind of why I

16 think we do need tables because sometimes the

17 reviewers have a lot to do and they might not get

18 all the information. If we have a table, then

19 we're sure to get it.

20 DR. KLAASSEN: That needs to say Table 3

21 right there if it's going to be in Table 3, so you

22 don't have to guess.

ANDERSON COURT REPORTING

706 Duke Street, Suite 100

Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190

CIR Panel Book Page 38 TEAM MEETING MINUTES ON SEBACATES – DR. MARKS’S TEAM 17 Tables 5-A, 5-B and 5-C. There's nothing much in

18 those tables, so I would just convert that 19 information to text and delete the tables. 14 DR. MARKS: It will be quick, Alan. Is 20 DR. MARKS: Ron, did you feel the 15 everybody fine with that, Ron, Ron and Tom? 21 impurities were okay in that section? Did we have 16 Table. Next we're to the sebacates or sebacates? 22 any needs in the impurities? 17 Which is it? ANDERSON COURT REPORTING 18 DR. ANDERSEN: Is there a linguist in 706 Duke Street, Suite 100 19 the house? Alexandria, VA 22314 20 DR. BERGFELD: It's in the Green Book. Phone (703) 519-7180 Fax (703) 519-7190 21 DR. MARKS: This is the first time that

22 the panel has seen this report, and obviously with

127

1 that, one of the things is do we want to group all

2 these ingredients together, should we delete some

3 of them, and then obviously move on to data needs?

4 And Ron Hill and Ron Shank also, how do you like

5 Acute Toxicity Tables 5-B, 5-C? Was that helpful?

6 We'll open it up for discussion.

7 DR. SHANK: I guess I'll start. I would

8 go as insufficient data. We need genotoxicity

9 data for microbial and mammalian. However, Table

10 3 from the American Chemistry Council report

11 indicates that there may be some data, but it's

12 not in our report other than the table. Maybe

13 it's already there. My second comment is that

14 there are five other dicarboxyl acids listed in

15 the dictionary. Why are they not included? And

16 the rest are just editorial. You did ask about

128 129

1 DR. SHANK: I don't have that marked. 1 do that and we'll go over it? So far we have the

2 DR. MARKS: There are no impurities, so 2 needs, although the table indicates mutagenicity

3 I guess would that be a need we have to move as 3 data and genotox data, that doesn't appear in the

4 insufficient? 4 report itself. Shall we assume that that's going

5 DR. BERGFELD: Ron? 5 to appear in the report and not put that out as an

6 DR. MARKS: Ron? 6 insufficient at this point?

7 DR. SHANK: I don't have any concerns, 7 DR. SLAGA: It's in the data in the

8 but maybe others do. 8 back.

9 DR. SLAGA: I don't have any concerns 9 DR. MARKS: So it's in the data in the

10 here, but mutagenicity in the new data in the back 10 back.

11 there is some mammalian genotoxicity, so there is 11 DR. ANDERSEN: I'm not finding it.

12 genotoxicity, both bacterial as well as mammalian. 12 DR. BERGFELD: It's in the additions

13 DR. SHANK: But it's not in the report, 13 behind everything there.

14 so do we actually have the studies or is this just 14 DR. SHANK: That is a page 15, diesters.

15 a quotation from some report? 15 There's the high production volume. There is a

16 DR. HILL: Which specific compounds are 16 report called "Diesters: High Production Volume

17 in that report? 17 Test Plan" in the back of the book and on pages 14

18 DR. MARKS: We'll want to go over that 18 and 15. It gives mutagenicity data which

19 and make sure because Ron Shank raised the issue 19 apparently the American Chemistry Council has

20 of should there be five more ingredients listed 20 those data. So if those data are there, can they

21 and are the ingredients that are already in the 21 be made available to us?

22 report ones that we want to include. Why don't we 22 DR. ANDERSEN: I got you now. So

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

CIR Panel Book Page 39 130 131

1 there's a whole series of studies that are 1 anyway. But if the panel decides that it's

2 described on pages 14 and 15, the details of 2 irrelevant, then we'll take it out, but there were

3 which, were they captured and put in the document 3 very limited details described by the authors.

4 could resolve the concerns. 4 MS. WEINTRAUB: I was wondering if there

5 DR. MARKS: Rachel? 5 was some reason to include it just to make clear

6 MS. WEINTRAUB: In terms of a lack of 6 that we've seen them but find it inclusive.

7 data, I just wanted to make sure that we consider 7 DR. ANSELL: We support that comment.

8 that there's no phototoxocity data and no 8 The study goes to the drug efficacy and doesn't

9 subchronic toxicity data. So the question is do 9 really report on safety.

10 you think we need that kind of data? Another 10 DR. MARKS: Correct. Actually starting

11 comment that I had is that on page 24 under the 11 with page 21. Is that where you started, Rachel?

12 Alternate Skin Study section, there are a few 12 Eliminate everything from 21 to 24, so Skin

13 studies mentioned, but then it says, "The safety 13 Treatments all the way to the Alternative Skin

14 information from the studies was not provided, or 14 Study section because it all relates to the

15 provided in a manner that was not conducive to 15 efficacy of these drugs and acne. Valerie, I'll

16 evaluation." Why include it if it provides 16 give you two printouts here, once on Azelex, the

17 absolutely no information? Page 24. 17 package insert which isn't very helpful, but

18 MS. ROBINSON: The Alternate Skin 18 there's a much better one on Finacea.

19 Studies? 19 Interestingly, I had the same concerns about

20 MS. WEINTRAUB: Yes. 20 phototox data, and they have a number of

21 MS. ROBINSON: With these studies there 21 irritations, sensitization, photosensitization and

22 were limited details, so we just included it 22 phototox studies on Finacea which is the 15

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

132 133

1 percent azelaic acid preparation, so that I'm 1 be?

2 going to give these to you. The references are in 2 DR. MARKS: The 1.5 percent. That's

3 there even though I think that's helpful data in 3 what was found in the cream which they did an RIPT

4 terms of the safety for this group. The rest of 4 on, so we have that data that we know at that

5 these I had concern about no photo absorption, but 5 limit that it was safe in that cream.

6 then again that would speak to the safety. What's 6 MS. SEIDMAN: The 1.5 percent?

7 your feeling, Ron? 7 DR. MARKS: Yes.

8 MS. SEIDMAN: If you look at the table

8 DR. SHANK: I don't think these 9 that I prepared, I prepared a table on the diethyl

9 compounds would absorb in the important areas of 10 sebacate allergic contact case reports. So if you

10 UV for skin toxicity. 11 look down there, there is a 1 percent diethyl

11 DR. MARKS: Under the Clinical 12 sebacate -- positive. I think there is something

12 Assessments, the diisopropyl sebacate is fine. I 13 less than that. But just look down that table and

13 had there were a number of severe case reports of 14 see what you think.

14 allergic contact dermatitis to diethyl sebacate. 15 DR. MARKS: Which page?

15 What was reassuring was they had a RIPT of this 16 MS. SEIDMAN: This is separate. It's a

16 compound with a 1.5 percent concentration of 17 handout.

17 diethyl sebacate, and I would suggest that we put 18 DR. MARKS: The handout?

18 a limit on that particular ingredient to that 19 MS. SEIDMAN: Yes.

19 concentration because of the alerts for these 20 DR. MARKS: Thank you.

20 multiple cases of severe allergic contact 21 MS. SEIDMAN: You're welcome.

21 dermatitis. 22 DR. SLAGA: So these are case report

22 DR. ANDERSEN: What would a good number ANDERSON COURT REPORTING

ANDERSON COURT REPORTING 706 Duke Street, Suite 100

706 Duke Street, Suite 100 Alexandria, VA 22314

Alexandria, VA 22314 Phone (703) 519-7180 Fax (703) 519-7190

Phone (703) 519-7180 Fax (703) 519-7190

CIR Panel Book Page 40 134 135

1 individuals? Is that correct? 1 memorandum from John Bailey dated August 18 and

2 MS. SEIDMAN: Yes. 2 you'll see in the fourth study down, KGL 2003, a

3 DR. MARKS: So your conclusion from 3 cream containing 1.5 percent diethyl sebacate in

4 these? Take me through it. These were mostly 4 human skin by means of the maximization assay was

5 medications as I read the case reports, hence some 5 okay. So that's why I thought that that would be

6 had, yes, very high concentrations which isn't 6 safe in light of having this alert from the case

7 surprising that they would sensitize at those 7 reports.

8 concentrations. 8 DR. ANSELL: You will also see a report

9 MS. SEIDMAN: I think there was a.1. 9 dated September 18 for diethyl.

10 DR. MARKS: Was that for patch testing 10 DR. MARKS: What does that add, Jay?

11 though? It went down. For actually patch testing 11 DR. ANSELL: Just that it was induced at

12 they went down as I recollect to.01, but that to 12 higher percents than that.

13 me just is a little bit different than what 13 DR. MARKS: This was with?

14 concentration could you feel safe going 14 DR. ANSELL: Diethyl.

15 prospectively and not sensitize individuals, not 15 DR. MARKS: Diethyl. Then I guess you

16 if they've already become sensitized. So that's 16 have the contradiction of which limit level are

17 why I thought the 1.5 percent cream from the 17 you going to use.

18 maximization test, and that was probably one of 18 DR. ANSELL: Yes.

19 these added at the end. 19 DR. MARKS: I think I'd like to err to

20 MS. SEIDMAN: You're just talking about 20 that lower concentration. Are there any other

21 induction. 21 comments? Ron Shank, going back to the original,

22 DR. MARKS: Yes. That was from the PCPC 22 do you think if we have all this data included in

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

136 137

1 the body and not just in the tables that we could 1 there are four CH2 groups, or I think we start

2 move forward without insufficient data conclusion? 2 with five. So if you look at the alkyl with four

3 DR. SHANK: Yes. I assume you have 3 and three, two, one and zero, all of those are

4 those data, it's just not a quote from a review or 4 listed in the dictionary as pH adjusters and

5 something, but if the mutagenicity data are 5 fragrance. So if we're including all of the

6 available, then they should be included into the 6 larger ones, is there a reason that we're not

7 report. 7 including those smaller ones? And I couldn't find

8 DR. ANDERSEN: The American Chemistry 8 that they had been reviewed before.

9 Council has provided the data whenever we've asked 9 DR. HILL: Could I respond to that

10 for it in the past, so I don't anticipate that 10 briefly? The major issue in my mind with lumping

11 that's a glitch. In a sense, I'm thinking that 11 these compounds as a class, the salts to me were

12 tabling this to allow those data to be gathered 12 no-brainer additions, but these's a fair amount of

13 makes sense. I'm also concerned about the idea 13 read-across data or by analogy data that I think

14 that there are four other dicarboxyl acids that we 14 makes the implicit assumptions that the starting

15 didn't capture, I think potentially do that 15 point in all of the biological handling of these

16 homework as well if this were tabled. 16 compounds if they're absorbed is ester hydrolysis,

17 DR. SHANK: The dictionary lists five, 17 actually twice, and so at least in terms of the

18 phthalic acid, malonic acid, succinic acid, 18 ones that were already there and these others that

19 glutaric acid and adipic acid. 19 are not added, for example, oxalic acid, there are

20 DR. ANDERSEN: I see what you're saying. 20 no carbons between the carbonyls, and maleic acid

21 DR. SHANK: It's just you take the alkyl 21 ones, I don't think that's on your list but that's

22 group, and we start with butyl dicarbonyl where 22 in the table here, there are just two carbons in

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

CIR Panel Book Page 41 138 139

1 between. So the ester hydrolysis could go very 1 officially table it.

2 differently there, and I haven't had a chance to 2 DR. BERGFELD: Can just hold it for that

3 digest it because even in the HPV report there's 3 information. I'm not sure there's a difference,

4 quite a bit of listed read-across data and I'm not 4 but when we table things they've been a little bit

5 sure how read across that data actually is until I 5 more worrisome or waiting for a piece of industry

6 have a matter since to map, and since we just got 6 rather than in-house sort of clarifications.

7 this a short time ago, I didn't have access to 7 DR. HILL: We have our marching orders,

8 that information because if there's penetration of 8 so I know what it is we're going to do and I guess

9 the diester or even monoesters into dermal layers 9 from that standpoint it doesn't matter what we

10 where there are potentially precancerous cells, 10 call it, just postponing further discussion until

11 then that changes everything in my mind in terms 11 these data and the question of the further

12 of what you can lump together as a class so that 12 expansion can be resolved is what we're going to

13 then you need information about the biological 13 do, whether it's tabled or not. Just in terms of

14 handling of that to know. 14 the expansion, simply that there may be a place

15 DR. BERGFELD: May I get a point of 15 where the line has to be drawn in terms how far

16 clarification? Alan, you're urging tabling for 16 apart the dicarboxylic acids groups are on these,

17 all the needs that we're assessing and these are 17 and there may be a reason why these are there and

18 needs for further staff work as I see it. Is that 18 the others are not.

19 official that we would table or we would just hold 19 DR. ANDERSEN: I think that's the answer

20 this to the next time to clarify those? Do we 20 to the question, but without getting down and

21 have to officially table this? I mean, we can. 21 dirty and having all of the staff talk that

22 DR. HILL: No, you don't have to 22 through, I didn't want to commit to that.

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

140 141

1 DR. HILL: Then furthermore, if that is 1 going to include in this report. Are those the

2 the case and there is significant absorption in 2 two things? And then Valerie, just capture also

3 the toxicology of the resultant alcoholics that 3 in there the concerns I had about the sensitivity

4 come from the ester hydrolysis in some cases come 4 and potentially having a maximum with a diethyl

5 into play and there's data for some of these 5 sebacate concentration of 2.5 percent.

6 compounds that are pretty comfortable for not for 6 DR. HILL: From my perception, this is a

7 all if the class is expanded the way it's given, 7 fairly complex class as expanded even without

8 but then again my digestion of the HPV information 8 adding the others, and just to make sure that

9 is still limited. I haven't had enough time with 9 Valerie gets the support she needs to wade her way

10 it. 10 through all of those complexities.

11 DR. MARKS: So I think if I understand 11 DR. ANDERSEN: In looking at the, if you

12 where we're going with this, it is the idea to 12 will, parent compound and focusing on the fact

13 table it or postpone it or however you want to 13 that there was an 8 carbon chain between the two

14 talk about this first look at these cosmetic 14 carboxylic acid groups, I think part of the logic

15 ingredients, and I guess are we still going to use 15 was that would give us a certain family depending

16 sebacic acid as our lead ingredient on this, then 16 on whether people are comfortable with the esters

17 as it was put in the memo related dicarboxylic 17 that are on the list. It begged the question how

18 acids and their salts and simple esters? We need 18 different is azelaic acid? It's 7, not that

19 a genotox carcinogenicity, the primary references, 19 different from 8. We would have included one with

20 and get that into the document itself, and then 20 9 except that's not a cosmetic ingredient, and

21 the second big issue is to firm up which 21 then we upped it to 10 on the high side. We can

22 ingredients we're really going to include in this 22 certainly examine the question of can we go a

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

CIR Panel Book Page 42 142 143

1 little lower and can we go a little bit higher 1 DR. HILL: I'm talking about

2 with that trepidation of how distance is going to 2 chain-shortening metabolism, yes. So there's the

3 start to matter depending on what you're doing. 3 parent dicaroxylic acid and then assuming the

4 DR. HILL: And I would also remind you 4 esters are hydrolyzed which there's a lot of

5 that the biologic handling of even-chain and 5 implicit assumption that that is the case, where

6 odd-chain carboxylic acids and I think also 6 does that happen and when? Does it happen in skin

7 dicarboxylic acids ends up quite differently, and 7 so that nothing systemic leaves other than

8 actually there are steps that are different, so 8 dicarboxylic acid in each case? Or we have

9 that probably also needs to be recognized and 9 monoesters? Or we have diesters making it into

10 accounted for. 10 other tissues? Then if you liberate the alcohols,

11 DR. ANDERSEN: So 7 may be more 11 and that happens immediately and fast wherever the

12 different from 8. 12 compound is dosed, then the toxicology of the

13 DR. HILL: Than 6 is or 10 is. 13 alcohols comes into play depending on how much

14 MS. SEIDMAN: Excuse me, handling in 14 dermal penetration if it's dermal.

15 what respect? 15 DR. MARKS: I think, Valerie, when the

16 DR. HILL: If you look at the biological 16 nuances start as we decide on what ingredients

17 handling of fatty acids, and I think the analogy 17 that we include in this report, we're going to

18 here of carboxylic acids, even chain and odd chain 18 have to catch those nuances that Dr. Hill

19 are handled very differently by biologically and 19 mentioned. Ron Shank, going back, and one of the

20 double bonds also have some nuances of handling. 20 questions was Table 5 and Table 5-B and 5-C. I

21 MS. SEIDMAN: You're talking about 21 don't think you found them particularly helpful.

22 metabolism? 22 Could you help clarify that so they could be

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

144 145

1 improved in the future? 1 unless there's just really not much data at all.

2 DR. SHANK: There's not enough 2 Even if there are two data lines, still you can

3 information in my opinion in those three tables to 3 compare numbers of animals, doses, route of

4 warrant the space of the tables, so I would just 4 administration, all those things you mentioned.

5 include that information in the text. 5 MS. SEIDMAN: We think alike.

6 MS. SEIDMAN: I tried to do a table here 6 DR. MARKS: Ron Shank, what more would

7 just because I personally am frustrated as a 7 you liked to have seen?

8 toxicologist in going through text because I 8 DR. SHANK: I don't put that much

9 cannot compare doses, I cannot compare species, I 9 importance on acute toxicity tests when we're

10 can't get a quick read on the data. So personally 10 talking about cosmetics products. We never have a

11 I felt it was really useful to put things in 11 problem with acute toxicity. So to put that

12 tables. This table might not have a lot, so you 12 information in a table, yes, it makes it easier to

13 might say for acute maybe we can just do text. 13 read. It's good to have, but I don't use it in my

14 Still, for me I find it useful because I can see 14 safety evaluation for a cosmetic product because

15 the doses readily and the species readily and the 15 we never get to acute toxicity problems.

16 numbers of animals used. It's all right there. 16 MS. SEIDMAN: So the relevance of the

17 But think about it maybe more for the longer- term 17 acute for cosmetic products is not important.

18 studies if you're not happy with it for acute. I 18 That I can accept. But think about other end

19 think most toxicologists find it very useful to 19 points as you go through this. This is just a

20 put things in tables. 20 trial balloon, so for other end points that might

21 DR. HILL: I would echo the benefits of 21 be more relevant to cosmetic products.

22 tables for me for the same exact reasons you said, 22 DR. SHANK: Absolutely, yes.

ANDERSON COURT REPORTING ANDERSON COURT REPORTING

706 Duke Street, Suite 100 706 Duke Street, Suite 100

Alexandria, VA 22314 Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190 Phone (703) 519-7180 Fax (703) 519-7190

CIR Panel Book Page 43 146 147

1 DR. MARKS: I think the conclusion is 1 Correct.

2 that we will recommend tabling this. Actually 2 DR. ANDERSEN: Thank you. Halyna will

3 it's the Belsito team which will report first, but 3 be pleased.

4 we will recommend tabling it to get the genotox 4 DR. MARKS: Shall we take a break for

5 and carcinogenicity data in the text and also to 5 lunch? Do you think we'll be able to do kojic

6 firm up what ingredients are actually going to 6 acid in 5 minutes?

7 appear in this report. Are there any other 7 DR. BERGFELD: No.

8 comments? 8 DR. ANDERSEN: The audience is on their

9 DR. ANDERSEN: I think in terms of some 9 own for lunch. The panel will be going downstairs

10 of the lessons for this, I want to make sure it 10 to the lobby, make a left, go past the desk, down

11 gets captured in terms of the data that had been 11 to the end to the Carleton Room for lunch.

12 in the back of the report on all of the clinical 12 (Recess)

13 testing, the skin treatments. You're reaffirming

14 that those, to the extent that they describe the

15 clinical effectiveness, have no place in a safety

16 assessment and if there were safety data such as

17 may be gleaned from what you've provided, Valerie,

18 those will stand on their own.

19 DR. MARKS: Correct.

20 DR. ANDERSEN: So we can really shorten

21 that section down to almost zero.

22 DR. MARKS: Yes. I have it deleted.

ANDERSON COURT REPORTING

706 Duke Street, Suite 100

Alexandria, VA 22314

Phone (703) 519-7180 Fax (703) 519-7190

CIR Panel Book Page 44 Report

Draft Report

Dicarboxylic Acids and Their Salts as Used in Cosmetics Esters of Dicarboxylic Acids as Used in Cosmetics

August 30, 2010

The 2010 Cosmetic Ingredient Review Expert Panel members are: Chairman, Wilma F. Bergfeld, M.D., F.A.C.P.; Donald V. Belsito, M.D.; Curtis D. Klaassen, Ph.D.; Daniel C. Liebler, Ph.D.; Ronald A Hill, Ph.D. James G. Marks, Jr., M.D.; Ronald C. Shank, Ph.D.; Thomas J. Slaga, Ph.D.; and Paul W. Snyder, D.V.M., Ph.D. The CIR Director is F. Alan Andersen, Ph.D. This report was prepared by Monice M. Fiume, Scientific Analyst/Writer, and Bart A. Heldreth, Ph.D., Chemist.

© Cosmetic Ingredient Review 1101 17th Street, NW, Suite 412 " Washington, DC 20036-4702 " ph 202.331.0651 " fax 202.331.0088 " [email protected]

CIR Panel Book Page 45 TABLE OF CONTENTS

Introduction...... 1 Chemistry ...... 1 Definition, Structure and Manufacture ...... 1 Physical and Chemical Properties ...... 2 Analytical Methods ...... 3 Impurities ...... 3 Ultraviolet Absorption ...... 4 Use ...... 4 Cosmetic ...... 4 Non-Cosmetic ...... 5 DICARBOXYLIC ACIDS AND THEIR SALTS ...... 6 General Biology ...... 6 Absorption, Distribution, Metabolism, and Excretion ...... 6 Percutaneous Absorption ...... 10 Peroxisome Proliferation...... 11 Cellular Effects ...... 11 Animal Toxicology ...... 12 Acute Toxicity ...... 12 Short-Term Oral Toxicity ...... 12 Short-Term Inhalation Toxicity ...... 12 Subchronic Oral Toxicity ...... 13 Subchronic Inhalation Toxicity ...... 13 Chronic Oral Toxicity ...... 14 Ocular Irritation ...... 14 Dermal Irritation/Sensitization ...... 15 Mucosal Irritation...... 16 Reproductive and Developmental Toxicity ...... 16 Genotoxicity...... 18 In Vitro ...... 18 In Vivo ...... 19 Carcinogenicity ...... 19 Tumor Promotion ...... 19 Clinical Assessment of Safety ...... 20 Dermal Irritation ...... 20 Case Reports ...... 21 ESTERS OF DICARBOXYLIC ACIDS ...... 22 General Biology ...... 22 Absorption, Distribution, Metabolism, and Excretion ...... 22 Penetration Enhancement ...... 26 Peroxisome Proliferation...... 26 Mechanism ...... 27 DNA Binding/DNA Synthesis ...... 29 Hepatic Lipid Metabolism ...... 30 Cellular Effects ...... 30 Animal Toxicology ...... 30 Acute Toxicity ...... 30 Short-Term Oral Toxicity ...... 31 Short-Term Dermal Toxicity ...... 32 Subchronic Oral Toxicity ...... 33 Subchronic Dermal Toxicity ...... 34 Subchronic Inhalation Toxicity ...... 34 Chronic Oral Toxicity ...... 34 Inhalation Toxicity ...... 35 Ocular Irritation ...... 35 Dermal Irritation ...... 36 Dermal Sensitization ...... 38

ii

CIR Panel Book Page 46 Phototoxicity ...... 38 Mucous Membrane Irritation ...... 38 Reproductive and Developmental Toxicity ...... 39 Endocrine Disruption ...... 44 Genotoxicity...... 44 Carcinogenicity ...... 46 Tumor Promotion ...... 47 Clinical Assessment of Safety ...... 47 Human Exposure ...... 47 Dermal Irritation and Sensitization ...... 47 Phototoxicity and Photosensitization ...... 49 Ocular Irritation ...... 49 Comedogenicity ...... 50 Case Reports ...... 50 Risk Assessment ...... 50 Summary ...... 51 Discussion ...... 56 Conclusion ...... 56 References ...... 57

iii

CIR Panel Book Page 47

INTRODUCTION This safety assessment includes sebacic acid and other alkyl α,ω-dicarboxylic acids, salts, monoesters and diesters. The dicarboxylic acids are terminally functionalized straight alkyl chains characterized by a separation between the acid functional groups of one to 10 carbons (1 carbon = malonic acid; 2 carbons = succinic acid; 3 carbons = glutaric acid; 4 carbons = adipic acid; 5-6 carbons = no representative cosmetic ingredients; 7 carbons = azelaic acid; 8 carbons = sebacic acid; 9 carbons = no representative cosmetic ingredients; and 10 carbons = dodecanedioic acid). The simple alkyl di-esters are the result of the condensation of alkyl dicarboxylic acids and two equivalents of alkyl alcohols. These ingredients can be metabolized via hydrolysis back to the parent alcohol, the mono-ester, and the parent dicarboxylic acid (Figure 1). The simple alkyl esters (mono- and di-) of these dicarboxylic acids have straight or branched side chains ranging in length from one to 18 carbons. Throughout this report, the data are presented by order of acid chain length (i.e., beginning with malonic acid and ending with dodecanedioic acid; beginning with dimethyl malate and ending with diisocetyl dodecanedioate). Accordingly, this draft report presents available information pertinent to the safety of 56 cosmetic ingredients in two groups, first, the 12 alkyl dicarboxylic acids/salts and, second, the 44 corresponding esters (mono- and di-). The alkyl dicar- boxylic acids and salts include: malonic acid azelaic acid succinic acid dipotassium azelate sodium succinate disodium azelate disodium succinate sebacic acid glutaric acid disodium sebacate adipic acid dodecanedioic acid.

The esters include: diethyl malonate dihexyl adipate diisostearyl adipate decyl succinate dicapryl adipate isostearyl sebacate dimethyl succinate di-C12-15 alkyl adipate diethyl sebacate diethyl succinate ditridecyl adipate dibutyl sebacate dicapryl succinate dicetyl adipate dicaprylyl/capryl sebacate dicetearyl succinate diisopropyl adipate diisopropyl sebacate diisobutyl succinate diisobutyl adipate diethylhexyl sebacate diethylhexyl succinate diethylhexyl adipate dibutyloctyl sebacate dimethyl glutarate diisooctyl adipate diisooctyl sebacate dibutyl glutarate diisononyl adipate dihexyldecyl sebacate diisostearyl glutarate diisodecyl adipate dioctyldodecyl sebacate dimethyl adipate dihexyldecyl adipate isostearyl sebacate diethyl adipate diheptylundecyl adipate dioctyldodecyl dodecanedioate dipropyl adipate dioctyldodecyl adipate diisocetyl dodecanedioate dibutyl adipate diisocetyl adipate

1

CIR Panel Book Page 48 The structures and functions of these ingredients are presented in Table 1. A safety assessment of diethylhexyl adipate (often inaccurately named dioctyl adipate)1 and diisopropyl adipate was published in 1984 with the conclusion that these ingredients are safe as used in cosmetics.2 The safety of these ingredients was reviewed and confirmed in 20053 and 2006.4 Additionally, dibutyl adipate was previously reviewed in 1996 and the available data were found insufficient to support the safety of dibutyl adipate in cosmetic formulations. When re-reviewed in 2006, additional data were made available to address the needs identified by the CIR Expert Panel, and an amended conclusion was issued stating that dibutyl adipate is safe for use in cosmetic formulations.5 The ingredients in this report function in cosmetics as pH-adjusters, fragrance ingredients, plasticizers, skin- conditioning agents and/or solvents and corrosion inhibitors.

CHEMISTRY Definition, Structure and Manufacture The CAS numbers, definitions, structures and functions for the alkyl dicarboxylic acid, salt and ester ingredients included in this report are given in Table 1. Alkyl Dicarboxylic Acids While many of the alkyl dicarboxylic acids are natural products, commercial production of these acids has historical- ly occurred via alkali pyrolysis of lipids.6 For example, when castor oil (a lipid which is comprised of approximately 84% ricinoleic acid-sidechain bearing triglycerides) is pyrolyzed with sodium hydroxide, some of the major products are sebacic acid and 2-octanol (Figure 2).6 Sodium and potassium salts of the alkyl dicarboxylic acids are readily prepared via addition to the appropriate stoichiometric equivalent(s) of sodium hydroxide or potassium hydroxide, respectively. Malonic Acid (C3) Malonic acid, first prepared by malic acid oxidation, is commonly manufactured by more recent methods including the ozonolysis of cyclopentadiene or the air oxidation of 1,3-propanediol.7 Succinic Acid (C4) Succinic acid is an intermediate of the citric acid cycle and is found in almost all plant and animals cells, although at very low concentrations.8 Succinic acid is commonly produced synthetically by catalytic (e.g., nickel or palladium catalyst) hydrogenation of maleic anhydride. Glutaric (C5) and Adipic(C6) Acids Although glutaric acid is often encountered in nature, adipic acid is not commonly encountered in nature. Glutaric and adipic acids were first synthesized by oxidation of castor oil with nitric acid. However, adipic acid is now more common- ly manufactured by oxidation of cyclohexane, cyclohexanol, or cyclohexanone, and glutaric acid may be manufactured by ozonolysis of cyclopentene.9 Azelaic Acid (C9) Azelaic acid, first detected in rancid fats, was originally produced via nitric acid oxidation of oleic acid. 10 Azelaic acid is a naturally-occurring dicarboxylic acid that can be found in dietary sources, such as whole grains.11 Azelaic acid is commonly manufactured by oxidative cleavage of oleic acid (obtained from grease or tallow) with chromic acid, nitric acid or by ozonolysis.10,7 Sebacic Acid (C10) Sebacic acid was originally isolated from distillation products of beef tallow. More recently, however, sebacic acid has been manufactured via alkali pyrolysis of castor oil, as mentioned above and drawn in Figure 2, or by alkali pyrolysis of 1

CIR Panel Book Page 49 ricinoleic acid.12,7 Dodecanedioic Acid (C12) Dodecanedioic acid can be manufactured by fermentation of long-chain alkanes with a specific strain of Candida tropicalis.13 Another method of manufacture involves the nitric acid oxidation of a mixture of cyclododecanone and cyclododecanol.7

Alkyl Dicarboxylic Acid Esters The alkyl dicarboxylic acids are easily esterified with the appropriate alcohol, with our without acid or metal catalyst (Fischer esterification).9 For example, diethylhexyl adipate can be manufactured from adipic acid and ethylhexanol with an acid catalyst (Figure 3). Diethyl Malonate Malonic acid esters can be produced either by cobalt-catalyzed alkoxycarbonylation of chloroacetates with carbon monoxide in the presence of the appropriate alcohol, or by hydrolysis of cyanoacetic acid followed by esterification with the respective alcohol.14 Diethyl malonate is prepared from chloroacetic acid and sodium cyanide followed by esterification with ethanol and sulfuric acid.15 Diisopropyl Adipate Diisopropyl adipate is produced by esterification of adipic acid with an excess of isopropanol. The excess alcohol is removed by vacuum stripping and the ester is then alkali-refined and filtered. From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2

Dibutyl Adipate Adipic acid is esterified with butyl alcohol by a continuous distillation process.16 Diethylhexyl Adipate Diethylhexyl adipate can be prepared by the reaction of adipic acid and 2-ethylhexanol in the presence of an esterifi- cation catalyst such as sulfuric acid or para-toluenesulfonic acid (Figure 3).17 Purification of the reaction product includes removal of the catalyst, alkali refining, and stripping.2 Alkyl Succinates Succinic anhydride reacts readily with alcohols to give monoesters of succinic acid (e.g., decyl succinate from decanol), which are readily further esterified to the diesters by Fischer methods.7 Dimethyl succinate can be produced from methanol and succinic anhydride or succinic acid, or by hydrogenation of dimethyl maleate. Diethyl succinate can be prepared by the same methods (from ethanol or diethyl maleate). Physical and Chemical Properties Tables 2a lists physical and chemical properties of the dicarboxylic acids and salts and Table 2b lists the properties of the esters. Charts 1a, 1b, and 2 demonstrate the relationship between molecular weight and the log octanol – water partioning coefficient. Dicarboxylic Acids - General The alkyl dicarboxylic acids vary considerably in their physical properties. The shorter chain (malonic, succinic, and glutaric) members are crystalline solids, very water-soluble and have limited solubility in organic solvents. As the chain length increases through adipic to dodecanedioic, water solubility decreases sharply (although still soluble in hot water). In other words, the water solubility of these acids is inversely proportional to their chain length. There is a marked alternation in melting point with changes in carbon number from even to odd.7 Odd members (e.g., malonic acid and glutaric acid) exhibit 2

CIR Panel Book Page 50 lower melting points and higher solubility than even carbon number alkyl dicarboxylic acids (e.g., succinic acid and adipic acid). These alternating effects are believed to be the result of the inability of odd carbon number compounds to assume an in-plane orientation of both carboxyl groups with respect to the hydrocarbon chain. Dicarboxylic acids react with Brønsted-Lowry bases (e.g., sodium hydroxide) to form carboxylate salts (e.g., sodium succinate or disodium succinate). Dicarboxylic acids also react with alcohols to give mono- and di-esters, such as those in this report. Esters The diesters, in contrast, are much more lipid soluble and more difficult to dissolve in water. The mono-esters, by definition, are hybrids of the acids and diesters, but their physical properties are much more closely related to the diesters. The short-chain alkyl (i.e., methyl, isopropyl, and butyl) mono- and diesters are more soluble in water, less lipo- philic, and relatively more volatile than the corresponding longer-chain alkyl (i.e., C8-C13 alcohol) esters.18 Most esters with molecular weights greater than 340 have boiling points greater than 300°C and are relatively non-volatile and lipophilic (log

Kow >7). Analytical Methods Succinic Acid Methods used to analyze succinic acid include acidimetric titration for acidity; comparison with Pt-Co standard calibrated solutions for color; oxidation with potassium permanganate for detection of unsaturated compounds; atomic absorption or plasma spectroscopy for metals; and titration with silver nitrate or barium chloride for chloride or sulfate detection, respectively.7 Small concentrations of succinic acid can be detected by common instrumentation such as gas/liquid chromatography and polarography. Adipic Acid Adipic acid can be extracted from a water sample and analyzed by gas chromatography/mass spectrometry.17 Sebacic Acid Gas chromatography can be used to identify sebacic acid in air.19 Diisopropyl Adipate and Diethylhexyl Adipate Diisopropyl adipate and diethylhexyl adipate can be identified through standard infrared (IR) spectroscopy. Gas-liquid chromatography (GLC), liquid-liquid extraction, mass spectrometry, and high-pressure liquid chromatography (HPLC) are also methods of analysis for the adipates. From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2

Impurities Diethyl Malonate Diethyl malonate is a colorless organic liquid with an ester like odor.14 The purity is typically > 99 %. Impurities from the production process include ethanol (ca. 0.1 % w/w), ethyl acetate (ca. 0.05 % w/w), and ethyl methyl malonate (ca. 0.05 % w/w). Dibutyl Adipate Impurities are generally not found due to the manufacturing process, but available data demonstrate that arsenic levels are below a detection limit of 1 ppm, heavy metals (as lead) are below a detection limit of 10 ppm, and sulfated ash is below a detection limit of 0.1%.16 Diisopropyl Adipate and Diethylhexyl Adipate Diisopropyl adipate and diethylhexyl adipate are considered stable; however, hydrolysis of the ester groupings may occur in the presence of aqueous acids or bases. No known impurities occur in either 3

CIR Panel Book Page 51 diisopropyl adipate or diethylhexyl adipate, although the acid values imply the presence of adipic acid or of the monoester in both. From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2

Diethylhexyl adipate is commercially available with the following specifications: purity – 99 to 99.9%; acidity – 0.25 µg/100g max; moisture – 0.05 to 0.10% max.17 Diisopropyl Sebacate A supplier reported that the expected impurities in diisopropyl sebacate are the starting material sebacic acid, <0.3%, and isopropyl alcohol, <0.2%.20 Ultraviolet Absorption Absorption of ultraviolet (UV) radiation was not detected in the 280 - 500 nm range.21 USE Cosmetic The ingredients included in this safety assessment have a variety of functions in cosmetics.22 The majority of the dicarboxylic acids function in cosmetics as pH adjusters and fragrance ingredients. The functions of most of the salts are not reported, but it is stated that sodium succinate functions as a buffering agent and pH adjuster. For the esters, some of the common functions include skin conditioning agents, fragrance ingredients, plasticizers, solvents, and emollients. The functions of all ingredients are listed in Table 1. Six of the 12 dicarboxylic acids and their salts and 24 of the 44 esters included in this safety assessment are reported to be used in cosmetic formulations. The frequency of use of the acids and salts, as supplied to the Food and Drug Admini- stration (FDA) by industry as part of the Voluntary Cosmetic Registration Program (VCRP),23 and the concentration of use, as supplied by industry in response to a Personal Care Products Council (Council) survey, are found in Table 3a. The frequency and concentration of use of the esters, with the exception of dibutyl, diisopropyl, and diethylhexyl adipate, which have previously been reviewed, are found in Table 3b. The current and historical use data for the 3 previously reviewed esters are found in Table 3c. The 6 acids and salts and 20 esters not currently reported to be used are listed in Table 3d. For the dicarboxylic acids and their salts, disodium succinate has the greatest number of reported uses, with a total of 45. The acid with the greatest concentration of use is succinic acid, 26%; use at this concentration is in rinse-off products. The greatest leave-on concentration is 0.4%, disodium succinate, with dermal contact exposure. For the esters, diisopropyl adipate has the greatest number of uses, with 70 reported. The concentration of use is greatest for dimethyl glutarate, 15% in a dermal rinse-off product. The ingredients with the greatest leave-on use concentra- tions, which are all dermal contact exposures, are diethylhexyl adipate, 14%, diisostearyl adipate, 10%, and diisopropyl sebacate, 10%. A few of the ingredients are applied around the eye, can possibly be ingested, or involve mucous membrane expo- sure, and some are used in underarm deodorant. None are reported to be used in baby products. Dicapryl and diethylhexyl succinate, dibutyl, dicapryl, diisopropyl, diisobutyl, and diethylhexyl adipate, diisopropyl, diethylhexyl, and dioctyldodecyl sebacate, and dioctyldodecyl and diisocetyl dodecanedioate are used in hair sprays, and effects on the lungs that may be induced by aerosolized products containing this ingredient, are of concern. The aerosol properties that determine deposition in the respiratory system are particle size and density. The para- meter most closely associated with deposition is the aerodynamic diameter, da, defined as the diameter of a sphere of unit density possessing the same terminal settling velocity as the particle in question. In humans, particles with an aerodynamic diameter of ≤ 10µm are respirable. Particles with a da from 0.1 - 10µm settle in the upper respiratory tract and particles with a 4

CIR Panel Book Page 52 24,25 da < 0.1 µm settle in the lower respiratory tract Particle diameters of 60-80 µm and ≥80 µm have been reported for anhydrous hair sprays and pump hairsprays, respectively.26 In practice, aerosols should have at least 99% of their particle diameters in the 10 – 110 µm range and the mean particle diameter in a typical aerosol spray has been reported as ~38 µm.27 Therefore, most aerosol particles are deposited in the nasopharyngeal region and are not respirable. With the exception of dipotassium azelate, disodium sebacate, and di-C12-15 alkyl adipate, the dicarboxylic acids and their salts and esters are listed for use by the European Union (EU) without restriction.28 Adipic acid is on the EU Dangerous Substances List; it is classified as Xi (irritant) and R36 (irritating to eyes). Non-Cosmetic Many of the dicarboxylic, their salts, and their esters are used in many foods as direct or indirect food additives. The alkyl dicarboxylic acids are unusually versatile because of their two carboxyl groups.9 This enables many additional types of useful reactions, particularly the manufacture of polymers (e.g., nylon). The most common uses include functions as plastici- zers, lubricants and building blocks in the manufacture of polyesters, polyamides and other plastics. The alkyl dicarboxylic acid salts are used to synthesize cyclic ketones, including commercially used macrocyclic musk compounds.29 The diesters have widespread use as lubricants, plasticizers, and solvents.30 Malonic Acid Malonic acid is a useful intermediate in the manufacture of barbiturates.31 Succinic Acid Succinic acid is listed by the FDA as a food additive that is Generally Recognized as Safe (GRAS).32 Succinic acid is also utilized in detergents, pigments, toners, cement additives, soldering fluxes and as an intermediate in the synthesis of a number of pharmaceutical products.7 Adipic Acid Adipic acid is listed as a GRAS food additives by the FDA.33 Adipic acid has several industrial uses in the production of adhesives, plasticizers, gelatinizing agents, hydraulic fluids, lubricants, emollients, polyurethane foams, leather tanning, and urethane.7 However, the bulk of the industrial production of adipic acid is driven by its usefulness in the manufacture of nylon-6,6 (in combination with 1,6-hexanediamine). Azelaic Acid FDA has approved azelaic acid for use in treating acne and rosacea. A skin cream containing 20% (w/w) azelaic acid, is indicated for the topical treatment of mild-to-moderate inflammatory acne vulgaris,34 and a gel containing 15% azelaic acid is approved for treating rosacea.35 These drugs are available by prescription only. Azelaic acid is used in the manufacture of plasticizers, lubricants, and greases. Azelaic acid was identified as a molecule that accumulated at elevated levels in some parts of plants and was shown to be able to enhance the resistance of plants to infections.36 Sebacic Acid Sebacic acid was widely used in the U.S. as an aromatic in food before 1973.37 Sebacic acid is used in resorbable polymer systems that deliver chemotherapeutic agents (e.g. cisplatin, carboplatin) that are implanted at the site of tumors to provide for sustained release of the drugs.38 Sebacic acid and its derivatives have a variety of industrial uses as plasticizers, lubricants, diffusion pump oils, candles and as intermediates in the synthesis of polyamides and various alkyd resins.7

5

CIR Panel Book Page 53 Dodecanedioic Acid Dodecanedioic acid is used in the production of nylon (nylon-6,12), polyamides, coatings, adhesives, greases, polyesters, dyestuffs, detergents, flame retardants, and fragrances. Diethyl Malonate Diethyl malonate finds great utility as the starting material in Malonic Ester Synthesis, a classic organic chemistry reaction wherein a very wide variety of esters can be synthesized.29 Diisobutyl Adipate The FDA has included diisobutyl adipate in Part 181 of Title 21 of the Code of Federal Regulations (CFR) – Prior- Sanctioned Food Ingredients, Subpart B--Specific Prior-Sanctioned Food Ingredients includes Sec. 181.27, Plasticizers. In this section, “substances classified as plasticizers, when migrating from food-packaging material shall include...diisobutyl adipate...” (21 CFR § 181.27). Diethylhexyl Adipate Diethylhexyl adipate is used as a plasticizer for polyvinyl chloride (PVC) plastics.39 Diethyl Sebacate Diethyl sebacate was widely used in the U.S. as an aromatic in food before 1973.37 Dibutyl Sebacate Dibutyl sebacate is a component of PVC.40

DICARBOXYLIC ACIDS AND THEIR SALTS Much of the information on the dicarboxylic acids was obtained from summary documents that mostly contained unpublished data. The summary documents are therefore listed as the citation, and, when there are numerous studies described, will only be cited at the beginning of that section. GENERAL BIOLOGY Absorption, Distribution, Metabolism, and Excretion Dicarboxylic acids are natural metabolic products of the ω-oxidation of monocarboxylic acids when the β-oxidation of free fatty acids is impaired.41 Under normal physiological conditions, dicarboxylic acids are rapidly β-oxidized, resulting in very low cellular concentrations and practically non-detectable concentrations in the plasma.42 Medium-chain dicarboxylic acids (up to 12 carbon atoms) are β-oxidized in mitochondria and peroxisomes. Oxidation of odd- and even-numbered chains proceeds to different end points. Odd-chain dicarboxylic acids are β-oxidized, giving acetyl Co-A and malonic acid (C3). Oxidation can then go no further, and malonic acid is the starter of fatty acid synthesis. Even-chain carboxylic acids are com- pletely oxidized and produce succynil-CoA, a gluconeogenic substrate, as an intermediate metabolite. Dicarboxylic acids are more polar than their esters, therefore they will diffuse less readily through normal cell membranes.43 Malonic Acid Malonic acid can be activated to malonyl-CoA and undergoes decarboxylation to acetyl-CoA by various mammalian tissues.14 Adipic Acid In animals, after oral dosing with adipic acid, 70% of the dose was exhaled as carbon dioxide. Adipic acid and a number of metabolites was recovered in the urine, while very little radioactivity was found in the tissues. After oral dosing in conjunction with i.p. sodium malonate, the presence of radioactive adipic and succinic acid was an indication of β-oxidation. Oral studies have reported recovering 53-67% unchanged adipic acid in the urine, 6

CIR Panel Book Page 54 while 59-71% was recovered in the urine following i.v. dosing and 61% was recovered following s.c. dosing, with an increase in urinary oxalic acid. In humans, 6.76-61% of the dose was recovered unchanged following oral dosing with adipic acid. A summary document on adipic acid contained a number of excretion and metabolism studies that were performed between 1918-1960.44 These studies are described below. Animal Adipic acid metabolism was studied using fasted male albino rats. In one study, in which the rats were dosed orally, by gavage, with 50 mg radioactive adipic acid (labeled on C1 or C2), 70% of the dosed was exhaled as carbon dioxide. Adipic acid and the metabolites urea, glutamic acid, lactic acid, β-ketoadipic acid, and citric acid, were recovered in the urine. Very little radioactivity was found in the tissues. Fasted male rats were also given a solution containing 50 mg radioactive adipic acid (labeled on C1), by gavage, in conjunction with 2 ml of 0.5 M sodium malonate, given by intraperitoneal (i.p.) injection. Both radioactive adipic acid and succinic acid were found in the urine, an indication that adipic acid underwent β-oxidation. By feeding rats 25 mg radioactive adipic acid (labeled on C1) and 100 mg γ-phenyl-α-aminobutyric acid, it was determined that acetate is a metabolite of adipic acid. Finally, rats were given radioactive sodium bicarbonate with non- radioactive adipic acid. Radioactive citric acid was formed, which suggested that carbon dioxide interacted with a metabolite of adipic acid. Two rats were dosed orally by gavage with 2.43 g/kg partially neutralized adipic acid for 28 days. In the urine, 67% of the dose was recovered unchanged. There was no change in excretion pattern over time during the study. Rabbits were dosed orally by gavage (n=4) or by intravenous (i.v.) administration (n=2) with 2.43 g/kg partially neutralized adipic acid for 2 days. Following oral administration, 53-61% of the dose was recovered unchanged in the urine. With i.v. administration, 59-71% was recovered unchanged in the urine. In another study using rabbits, animals were given a subcutaneous (s.c.) dose of 2000 mg adipic acid; 3 rabbits were given a single dose, one was dosed on days 1 and 5, and one was dosed on days 1, 5, 9, 13, and 15. On average, 61% of the dose was recovered unchanged in the urine. There was an increase in urinary oxalic acid concentrations. A female dog was fed either 150 mg/kg adipic acid (in 2 feedings) for 5 days or 750 mg/kg (in 2 feedings) for 7 days. In the urine, 18% and 63.6% of the low and high doses were recovered unchanged. Rabbits (number not stated) were given up to 4 s.c. injections of ≤2000 mg sodium adipate.45 An average of 61% of the dose was recovered unchanged in the urine. Oxalic acid was increased in the urine. Human In a study in which one subject was given 33 mg/kg sodium adipate, orally, for 5 days (10 g total), 6.76% of the dose was recovered in the urine. In another study in which one person was given 100 mg/kg adipic acid for 10 days (70 g total), 61% of the dose was recovered in the urine. Administration of 19.0 g adipic acid over 5 days or 23.4 g over 6 or 9 days (1 subject per dose) resulted in 53% of the administered dose recovered in the urine. C9 to C12 Dicarboxylic Acids For rats dosed orally with azelaic, sebacic, undecanedioic, and dodecanedioic acid, 2.5, 2.1, 1.8, and 1.6% of the respective acid was found in the urine unchanged. In humans the amount recovered decreased with increasing chain length. After oral dosing, 60, 17, 5, and 0.1% of azelaic, sebacic, decanedioic, and undecanedioic acids, respectively, were recovered unchanged in the urine. In the plasma of both animals and humans, dicarboxylic acid catabolites that were 2-, 4-, or 6- carbons shorter than the corresponding dicarboxylic acid were found. Animal Groups of 30 male Wistar rats were dosed orally, by gavage, with azelaic (C9), sebacic (C10), undecanedioic (C11),

7

CIR Panel Book Page 55 or dodecanedioic (C12) acid.46 Ten rats in each group were dosed with 20, 50, or 100 mg of the respective acid. Blood, urine, and feces from the treated rats were analyzed and compared to the blank control obtained from untreated rats. (None of the C9-C12 acids were found in the blank controls.) In urine, approximately 2.5% of azelaic, 2.1% of sebacic, 1.8% of unde- canedioic, and 1.6% of dodecanedioic acid was recovered after 5 days; the amount recovered was not affected by dosage. The dicarboxylic acids were not excreted in conjugated form. None of the C9-C12 dicarboxylic acids were recovered in the feces. In the plasma, dicarboxylic acid catabolites that were 2-, 4-, or 6-carbons shorter than the corresponding dicarboxylic acid were detected. Human Groups of 3 male and 2 female subjects were also dosed with C9-C12 acids orally, in gelatin capsules, once a wk for 5 wks.46 The dose administered increased each week, from 0.5 g at wk 1 to 5.0 g at wk 5. None of the C9-C12 acids were found in the blank control samples of blood, urine, and feces obtained from non-treated humans. In urine, approximately 60% of azelaic, 17% of sebacic, 5% of undecanedioic, and 0.1% of dodecanedioic acid was recovered after 12 h; the amount re- covered was not affected by dosage. At 24 h, the amounts recovered were not much increased. Initially, undecanedioic and dodecanedioic acid administration raised the urinary pH to a value of 7.4-8.5; the pH returned to normal with 3-6 h. The di-carboxylic acids were not excreted in conjugated form. None of the C9-C12 dicarboxylic acids were recovered in the feces. In the plasma, dicarboxylic acid catabolites that were 2-, 4-, or 6-carbons shorter than the corresponding dicarboxylic acid were detected. Plasma levels of azelaic acid peaked at 2 h, while the levels of the other three acids peaked at 3 h. Recovery in the plasma was greatest for azelaic acid, 74.6 µg/ml with the 5 g dose, and the amount detected decreased with increasing chain length. Azelaic Acid In animals, after oral dosing with azelaic acid, 40% of the radioactivity was recovered in the urine over 5 days, and 14.5% was found in expired carbon dioxide at 48 h. Mostly dicarboxylic acid metabolites were found in the blood for up to 72 h after dosing. Radioactivity was found in all tissues, with the greatest levels in the liver, lungs, and kidneys; levels then decreased in all organs, except the adipose tissue for which an increase continued. In humans, the dermal and oral administration of a 20% azelaic acid cream was compared. A total of 2.2% of the dose was recovered in the urine after dermal administration, as compared to 61.2% following oral administration. The calculated percutaneous absorption was determined to be 3.6%. Azelaic acid is a dietary constituent found in whole grain cereals and animal products.47 It can be formed endogenously from longer-chain dicarboxylic acids, metabolism of oleic acid, and ψ-oxidation of monocarboxylic acids.48 Endogenous plasma concentration and daily urinary excretion of azelaic acid are highly dependent on dietary intake. Azelaic acid crosses the blood-brain barrier.49 Animal A group of 25 male Wistar rats were dosed orally, by gavage, with 100 µCi of [1,9-14C]azelaic acid, and the animals were killed at various intervals 1-96 h after dosing.46 After 12 and 48 h, 13 and 14.5% of the radioactivity was found in expired carbon dioxide, respectively. Approximately 40% of the radioactivity was recovered in the urine over 5 days. The C7 and C5 dicarboxylic acid metabolites were found in the urine up to 72 h after dosing. Very little was recovered in the feces. Labeled dicarboxylic acids were present in the blood for up to 72 h, and consisted mainly of dicarboxylic acid metabolites. Radioactivity was found in all tissues, with the highest levels present in the liver, lungs, and kidneys after 12 h. Tissue radioactivity levels then decreased slowly in all organs except adipose tissue, in which case increasing levels were still seen at 96 h. Approximately 90% of the radioactivity found in the tissues was present in the lipids, and it was essentially localized in the fatty acid portion of the triglycerides and of the phospholipids. Traces of C9, C5, and C7 dicarboxylic acids were detected in the first 24 h. 8

CIR Panel Book Page 56 Human The percutaneous absorption of azelaic acid was determined using 6 male subjects. A total of 5 g of a cream containing 20% azelaic acid was applied to the face (1 g), chest (2 g) and upper back (2 g) of each subject, giving an area dose of approx 5 mg cream/cm2 skin. The test areas were covered 1 h after dosing with cotton tissues, and washed 24 h after dosing. After 1 wk, 100 ml of an aq. microcrystalline suspension containing 1 g azelaic acid was given orally to each subject Urinary excretion of unchanged azelaic acid was measured after each dose. Following dermal application, 1.29% of the dose was recovered unchanged in the urine in 24 h, and a total of 2.2% was recovered by day 3. Following oral administration, 61.2% of the dose was recovered within 4 h; excretion was complete at this point. Assuming similar rates and pathways in biotransformation following both routes of exposure, percutaneous absorption of azelaic acid was determined to be 3.6% of the dermally applied dose.50 Sebacic Acid Sebacic acid is oxidized to water and carbon dioxide, passing through acetyl-CoA and succinyl-CoA formation.51 Disodium Sebacate In rat, following i.v. administration of disodium sebacate, 34.6% of the dose of the dose was recovered in the urine as sebacate and 5% was recovered as suberic acid. A total of 25% of the sebacate was recovered in expired carbon dioxide. No appreciable radioactivity was found in the body. Following i.p. administration, sebacate renal clearance was a concentration-independent function. With oral administration, the relative bioavailability was 69%. In humans given a steady infusion of disodium sebacate, less than 15% of the dose was recovered in the urine. The percent oxidation of sebacate was 6.14% Animal Disodium sebacate, 80 and 160 mg with 25 µCi of (1,10)14C sebacic acid tracer, was administered by i.v. injection to 14 male Wistar rats, and blood samples were obtained at various intervals 5-320 min after dosing.51 The plasma half-life of radioactive disodium sebacate was 37.86 and 39.82 min for the 80 and 160 mg dose groups, respectively. The apparent volume of distribution was 2.65 ml/100 g body wt. In a second experiment, a group of 4 male Wistar rats were given 160 mg disodium sebacate with 25 µCi sebacic acid tracer by i.v. injection, and expired carbon dioxide, urine, and feces were collected. The carbon dioxide half-life for radioactive sebacate was 93.64 min; 25% of the administered dose was expired in carbon dioxide. A total of 34.6% of sebacate was recovered in the urine in 24 h, while 5.08% suberic acid (C8) was recovered in the same time frame. Most of the excretion occurred in the first 24 h. Radioactivity was not found in the feces. In the third experiment, groups of 10 male Wistar rats were also given 160 mg disodium sebacate with 25 µCi sebacic acid tracer by i.v. injection, and the animals were sacrificed at various intervals from 30-360 min after dosing and the amount of radioactivity in various organs was analyzed. No appreciable radioactivity was found in the body. Sebacate appeared to be in an absorption phase in fat 1 h after dosing, but no radioactivity was found in the body after 24 h. The pharmacokinetics of disodium sebacate was studied in male and female Wistar rats.52 Sebacate was admini- stered either i.p., 6 doses o f 10-320 mg, or orally, 2 doses of 80 or 60 mg. Plasma concentrations of sebacate and urinary concentrations of sebacate and its products of β-oxidation (suberic and adipic acids) were measured using GLC/mass spectrometry. Both renal and non-renal elimination parameters were obtained. The sebacate half-life was 31.5 min. The tissue elimination rate was 0.0122 min-1, and the overall volume of distribution was 26.817 ml/100 g. The renal clearance was 0.291 ml/min/100 g, which was much less than the value of the glomerular filtration rate (GFR) of approximately 1 ml/min/100g reported elsewhere, suggesting the presence of sebacate reabsorption from the ultrafiltrate. Sebacate renal clearance was found to be a concentration-independent function, suggesting the presence of a passive back-diffusion. The relative bioavailability of the oral route compared to the i.p. route was 69.09%, showing an extensive absorption of the 9

CIR Panel Book Page 57 compound. Human The metabolism and excretion of disodium sebacate was studied in 7 fasting male subjects that were given a continuous steady infusion of 20 g unlabeled disodium sebacate over 480 min.53 At 240 min into the infusion, (1,10)[C14]-sebacic acid was infused simultaneously as a tracer (sp. act. 0.416 µCi/min). The was a gradual increase in the amount of sebacate expired in carbon dioxide for the first 300 min; the value remained elevated for an additional 120 min before declining. At 24 h, 11.38 mmol sebacate was recovered in the urine, as well as 2.04 mmol suberic acid and 1.11 mmol adipic acid, which was less than 15% of the dose administered. The serum concentration of unlabeled sebacate reached a plateau after 270 min of infusion. Ten to 15% of serum radioactivity was found in the aq. fraction of serum extracts. The renal clearance rate was 5.67 ml/min. The overall tissue uptake of unlabeled sebacate was 180 µmol/min, and the apparent distribution volume was 12.46 l. The percent oxidation of sebacate was 6.14%. The pharmacokinetic profile of disodium sebacate during a short-time infusion (5 h at 10 g/h) was also studied in 7 male subjects .54 Sebacate in serum and urine was measured by HPLC. The apparent volume of distribution of sebacate was 8.39 l, and the plasma fractional removal rate constant was 0.0086 min-1. Six male subjects were given a single i.v. bolus of 1 g disodium sebacate, while another 6 received 10 g of sebacate in 500 ml of distilled water, i.v., at a rate of 3.33 g/h over 3 h.55 For the group given a bolus dose, the distribution phase had a short half-life, 0.34 h, and a rapid elimination, 2.045 h-1. For the group given the 3 h infusion, 12% of the dose was excreted as sebacic acid in 24 h; suberic acid (C8) and adipic acid were also present in the urine. Dodecanedioic Acid Approximately 50% of an oral dose of dodecanedioic acid was recovered in the urine of rats, and the C10, C8, and C6 metabolites were found up to 72 h after dosing. Labeled dicarboxylic acid was found in the blood for up to 72 h after dosing, mainly as the dicarboxylic acid metabolites. Radioactivity was found in all tissues, with the highest levels in the liver, lungs, and kidneys; after 24 h, the levels declined in all tissues except adipose. A group of 25 male Wistar rats were dosed orally, by gavage, with 100 µCi of [10,11-3H]dodecanedioic acid, and the animals were killed at various intervals 1-96 h after dosing.46 Approximately 50% of the radioactivity was recovered in the urine over 5 days. The C10, C8, and C6 dicarboxylic acid metabolites were found up to 72 h after dosing. Only 2% of the radioactivity was recovered in the feces. Labeled dicarboxylic acids were present in the blood for up to 72 h, and consisted mainly of dicarboxylic acid metabolites. Radioactivity was found in all tissues, with the highest levels present in the liver, lungs, and kidneys after 24 h. Tissue radioactivity levels then decreased slowly in all organs except adipose tissue, in which case an increase in radioactivity was still seen at 96 h. Radioactivity levels were 20-40% lower in the lipid extracts of the tissues than in the residual matter. 3H was distributed in the whole molecule, not only the fatty acid portion, of the phospholipid and triglyceride fractions. Traces of C12, C10, C8,and C6 dicarboxylic acids were detected in the first 24 h. Male Wistar rats were given an i.v. bolus of 800 µmol/kg disodium dodecanedioic acid.56 The apparent volume of distribution was 0.248 l/kg, and the plasma half-life was 12.47 min. The renal clearance was 0.00051 l/kg/min, while systemic clearance was 0.0138 l/kg/min. Only 3-5% of the dose was recovered in the urine. Percutaneous Absorption Azelaic Acid Vehicle affects the absorption of azelaic acid.43 After a 12 h period, absorption from a 15% azelaic gel was 8%, while absorption from a water-soluble polyethylene glycol ointment base was only 3%. (Species and details not given.) The in vitro percutaneous absorption of a 15% azelaic acid gel, prior to or after the application of three different moisturizer formulations, was determined.57 All doses were applied as 5 µl/cm2. The second dose was applied 15 min after 10

CIR Panel Book Page 58 the first. [14C]Azelaic acid had a finite dose absorption profile, with a rise to peak penetration followed by a slow but steady decline. In vitro, 70% of the azelaic acid diffused into the reservoir solution. The application of a moisturizer, and whether it was applied prior to or following azelaic acid administration, did not have a statistically significant effect on the penetration of azelaic acid. However, there was a trend toward greater percutaneous penetration and mass distribution with the application of a moisturizer lotion prior to the azelaic acid gel. Peroxisome Proliferation Adipic Acid The effect of adipic acid on hepatic peroxisome proliferation was evaluated in an in vivo study in which 4 male F344 rats were fed chow containing 2% adipic acid dissolved in alcohol.58 After 3 wks of dosing, the animals were killed. Adipic acid did not induce peroxisome proliferation and did not affect relative liver weight. Cellular Effects Dicarboxylic acids have a cytotoxic effect on the abnormally hyperactive and malignant epidermal melanocyte. Dicarboxylic acids, C8to C13, have been shown to inhibit mitochondrial oxidoreductases,59 and they have been show to reversibly inhibit microsomal NADPH and cytochrome P450 reductase.60 Medium chain length dicarboxylic acids are also competitive inhibitors of tyrosinase in vitro. Adipic Acid The effect of adipic acid on primary keratinocyte cultures was evaluated using epidermal cells from neonatal NMRI mice.61 Concentrations of ≤30 mM did not inhibit 3H-thymidine incorporation or affect DNA synthesis, while 40 and 50 mM inhibited both of these parameters. No effect on labeling indices was observed with 1-30 mM adipic acid. Azelaic Acid Azelaic acid, a naturally occurring competitive inhibitor of tyrosinase, has a cytotoxic effect on malignant melanocytes .62 Azelaic acid is also a competitive inhibitor of a number of oxidoreductive enzymes, enzymes involved in DNA synthesis, and of oxidoreductases of the respiratory chain.63 In vitro, azelaic acid is a scavenger of toxic oxygen species, inhibits oxyradical activity in cell cultures, and inhibits generation of reactive oxygen species by neutrophils. It has been reported that, in vitro, azelaic acid has time- and dose-dependent, reversible, and anti-proliferative and cytotoxic effects on a number of tumoral cell lines. Azelaic acid had no effect on normal cell lines. Disodium Azelate Disodium azelate inhibited cell proliferation and affected viability of Cloudman and Harding-Passey murine melanomata at concentrations ≥10-2 M when incubated over a 3 day period.59 The mitochondria were the prime target of action. The effect of disodium azelate on primary keratinocyte cultures was evaluated using epidermal cells from neonatal NMRI mice.61 A dose-dependent inhibition of 3H-thymidine incorporation into DNA, ranging from 50% inhibition with 20mM to 90% inhibition with 50mM disodium azelate, was observed following a 12 h incubation period. Concentrations of 1 and 10 mM did not affect DNA synthesis, but a marked reduction was seen with 20-50 mM. The effects on DNA synthesis were time-dependent, with the maximum inhibitory effect observed at 4 h; this effect was reversible. RNA and protein synthesis were also inhibited during the first 4 h of incubation with 50 mM disodium azelate. Cellular structure was altered upon incubation with disodium azelate, primarily affecting mitochondria, and the rough endoplasmic reticulum. These effects were also reversible. Dodecanedioic Acid The disodium salt of dodecanedioic acid inhibited cell proliferation and affected viability of Cloudman and Harding- 11

CIR Panel Book Page 59 Passey murine melanomata at concentrations ≥10-2 M when incubated over a 3 day period.59 The mitochondria was the prime target of action. ANIMAL TOXICOLOGY Acute Toxicity

The oral LD50 values of the dicarboxylic acids for rats ranged from 0.94 g/kg adipic acid to ≥4 g/kg azelaic acid (although most reported values were >5 g/kg). The reported dermal LD50 values ranged from >6 g/kg dodecanedioic acid to >10 g/kg glutaric acid.

The acute oral, dermal, inhalation, and parenteral toxicity of the dicarboxylic acids and some of the salts are summarized in Table 4.44,64-69 Short-Term Oral Toxicity

In short-term oral toxicity studies, ≤3000 mg/kg/day adipic acid did not produce significant toxicological effects in rats. Signs of toxicity were seen at >3600 mg/kg/day. No toxicity was observed with guinea pigs fed 400-600 mg/day azelaic acid.

Adipic Acid Groups of 6 male Sprague-Dawley rats were dosed orally (method not specified) with 3600-5600 mg/kg adipic acid as an 18.6-24.9% solution in saline for 14 days.44 Three animals of the 3600 mg/kg group, 5 of the 4000 mg/kg group, and all of the 4500-5600 mg/kg groups died prior to study termination. Signs of toxicity included depressed activity, labored respiration, ataxia, and convulsions. No gross findings were noted at necropsy at study termination. Groups of 5 rats were dosed with 0 or 3000 mg/kg of a neutralized 20% adipic acid solution orally, by gavage, for 4 wks. A non-significant decrease in body weight gain was observed. In a 4 wk study in which a group of 3 rats was dosed orally, by gavage, with 2400 mg/kg adipic acid, no significant toxicological effects were noted. In a 4-wk dietary study in which groups of 17-20 female rats were fed 0-40 mg/day (0-435 mg/kg/day) adipic acid, no effects were reported. The no-observable adverse effect level (NOAEL) was >435 mg/kg/day. In a 5-wk dietary study in which groups of 15-18 male rats were fed 0-800 mg/day (0-13,333 mg/kg/day) decreased body weight gains, an unkempt appearance, and diarrhea were observed for the animals fed 800 mg/day the first 3 wks. In another 5-wk dietary study in which groups of 4 rats, gender not specified, were fed 100 or 200 mg/day (310-922 mg/kg/day) of a 20% adipic acid solution in ethanol, 5 days/wk, no signs of toxicity were observed. Ten rats were dosed orally, method not specified, with 199 mg/day (638-1332 mg/kg/day) sodium adipate, 5 days/wk for 9 wks. No toxicological effects were observed. A group of 5 guinea pigs, gender not specified, were dosed orally using capsules with 400 mg/day (682-942 mg/kg/day) adipic acid for 5 days, followed by dosing with 600 mg/day (1032-1739 mg/kg/day), 5 days/wk for 5 wks. No signs of toxicity were observed. No toxicity was observed in a study in which pigs were fed 1% adipic acid in the diet for 7 days. Short-Term Inhalation Toxicity Short-term inhalation exposure to 126 mg/m3 adipic acid to rats did not produce signs of toxicity, but exposure of mice to 460 mg/m3 did.

12

CIR Panel Book Page 60 Adipic Acid Mice, gender and number per group not specified, were exposed to 460 mg/m3 adipic acid for 1.5 mos.44 (Details of exposure were not specified.) Decreased weight gain, altered oxidase activity, and upper respiratory tract, liver, kidney, and central nervous system effects were observed. (Details were not given.) Two male and 2 female rats were exposed to 126 mg/m3 adipic acid for 15 days, 6 h/day. No signs of toxicity were observed, and no gross or microscopic findings were noted at necropsy. Subchronic Oral Toxicity In a subchronic oral study, 10 male and 10 female rats exposed to 10% sodium succinate in the drinking water died, but no compound-related lesions were found. Body weights were decreased in rats given ≥2.5% sodium succinate for 13 wks, but toxicological treatment-related changes were not observed. Glutaric acid ad a low degree of toxicity to rats (at 2%) and dogs (concentration not specified) when given in the drinking water. Dietary administration of ≤3400 mg/kg/day adipic acid for 19 wks produced slight effects in the liver of male rats; the NOAEL was 3333 mg/kg. A mixture of adipic, glutaric, and succinic acids had a low degree of toxicity in rats when tested at 3% for 90-days.

Sodium Succinate The oral toxicity of sodium succinate was evaluated using F344 rats.68 Groups of 10 males and 10 females were given 0, 0.3, 0.6, 1.25, 2.5, 5 or 10% sodium succinate in the drinking water for 13 wks. All animals were killed at the termination of dosing. Body weight gains of animals of the 10% group were significantly decreased, and all animals of this group died by wk 4. These animals were extremely emaciated; however, no compound-related microscopic lesions were found. Body weight gains were decreased in animals given ≥2.5% sodium succinate, as compared to controls. No toxicologi- cal treatment-related effects were observed. Glutaric Acid The oral toxicity of 1-2% glutaric acid was evaluated in a 90-day study using rats.65 Glutaric acid had a low degree of toxicity at ≤2%; decreased weight gains were seen with higher concentrations. No specific target organs were identified. (Additional details, including the method of oral administration, were not given.) A low degree of toxicity was also reported in a 90-day oral toxicity study using dogs. (Details were not provided.) Adipic Acid Groups of 8-10 male rats were given 0, 420, 840, 1700, or 3400 mg/kg/day sodium adipate for 19 wks in a protein deficient diet.67 Animals were killed after either 7 wks or at study termination. For unexplained reasons, only 5-7 animals/group survived until study termination. Rats of the 3400 mg/kg/day group had decreased body weight gains and decreased body weights. (Statistical significance not stated.) Slight effects were seen in the liver, and the NOAEL was 3333 mg/kg. Adipic/Glutaric/Succinic Acid Mixture The oral toxicity of a mixture of adipic, glutaric, and succinic acids, tested at 3%, was evaluated in a 90-day study.65 The mixture had a low degree of toxicity at this dose. Higher concentrations caused decreased weight gain. No specific target organs were identified. (Additional details, including the method of oral administration and percentage of acids in the mixture, were not given.) Subchronic Inhalation Toxicity Signs of toxicity were reported in a subchronic inhalation study in which mice were exposed to 13 or 120 mg/m3 adipic acid.

13

CIR Panel Book Page 61 Adipic Acid Mice, gender and number per group not specified, were exposed to 13 or 120 mg/m3 adipic acid for 4 mos.44 (Details of exposure were not specified.) Decreased weight gain, altered oxidase activity, and upper respiratory tract, liver, kidney, and central nervous system effects were observed. Chronic Oral Toxicity A low degree of toxicity to sodium succinate was observed in a 2 yr oral study using rats. Slight effects were seen in the livers of rats fed ≤3200 mg/kg/day adipic acid for 33 wks, and the NOAEL for rats fed a diet containing adipic acid for 2 yrs was 1%; no significant toxicological effects were seen at concentrations of ≤5%. No significant toxicological effects were observed for mice fed ≤280 mg/kg or rabbits fed ≤400 mg/kg azelaic acid for 180 days. Disodium sebacate was not toxic to rats or rabbits fed up to 1000 mg/kg for 6 mos.

Sodium Succinate The oral toxicity of 1% sodium succinate was evaluated in rats in a 2-year study.65 A low degree of toxicity was observed with this dose. Higher concentrations caused decreased weight gain. No specific target organs were identified. (Additional details, including the method of oral administration, were not given.) Adipic Acid Groups of 13-15 male and female rats were fed a diet containing 0, 1600, or 3200 mg/kg/day adipic acid for 33 wks.44 Rats were killed at various intervals throughout the study. Ten of 14 rats fed 3200 mg/kg/day died during wks 0-4; surviving rats had decreased weight gains during this time. However, at study termination, body weights were for surviving animals of this group were similar to controls. Slight effects were seen in the liver. (Statistical significance not stated.) In a 2-yr study, groups of 20 male rats were fed a diet containing 0, 0.1, 1, 3, and 5% adipic acid (equiv. to 0, 75, 750, 2250, and 3750 mg/kg/day), and a groups of 10 and 19 females were fed 0 and 1% adipic acid, respectively. Weight gains of male rats fed 3 and 5% adipic acid were significantly less than controls. There were no significant toxicological findings upon gross or microscopic observation. The NOAEL was 1% adipic acid for male and female rats. Azelaic Acid Groups of 15 male and 15 female Wistar rats were fed a diet containing 140 or 280 mg/kg azelaic acid for 180 days, and a control group of 10 males and 10 females was given untreated feed.69 No significant toxicological effects were observed. Growth was similar between test and control groups, as were the microscopic examinations and clinical chemistry parameters. The researchers found similar, negative, results when groups of 10 male and 10 female New Zealand rabbits were fed diets containing 0, 200, or 400 mg/kg azelaic acid for 180 days. Disodium Sebacate Groups of 10 male and 10 female Wistar rats were fed a diet containing 0, 500, or 1000 mg/kg disodium sebacate for 6 mos, after which time they were killed and necropsied.64 Growth was similar between test and control groups, as were the microscopic examinations and clinical chemistry parameters. The researchers found similar, negative, results when groups of 10 male and 10 female New Zealand rabbits were fed diets containing 0, 750, or 1000 mg/kg disodium sebacate for 6 mos. Ocular Irritation For the dicarboxylic acids, the severity of ocular irritation seems to decrease with increasing carbon number. Succinic acid was a severe ocular irritant, glutaric acid was moderately irritating, and dodecanedioic acid was a slight irritant. Ocular irritation produced by adipic acid was dose-dependent.

14

CIR Panel Book Page 62 Succinic Acid Succinic acid was severely irritating in an ocular irritation study.65 Details were not provided. Glutaric Acid Glutaric acid was a moderate ocular irritant.65 Details were not provided. Adipic Acid The ocular irritation of adipic acid was evaluated using groups of 2 albino rabbits.65 Ten or 57.1 mg of adipic acid was placed in the eye of each rabbit, and the eye of 1 animal in each group was rinsed. Using 10 mg, no irritation was seen in the rinsed eye, and the test article was minimally irritating to the unrinsed eye. With 57.1 mg adipic acid, mild to moderate irritation was seen in both the rinsed and unrinsed eyes. Severe irritation was seen upon instillation of 0.1 ml, 100 mg, and 50 mg into the eyes of 6, 3, and 2 rabbits, respectively.44 The irritation had not cleared after 8 days. Dodecanedioic Acid In studies using rabbits that evaluated the ocular irritation of dodecanedioic acid, slight irritation was reported in one study, with a primary irritation index (PII) of 11.96/110, and small areas of corneal opacity and mild conjunctival irritation were seen in the other.66 Details were not provided. Ocular irritation studies are summarized in Table 5. Dermal Irritation/Sensitization Slight to mild dermal irritation was observed for succinic, glutaric, and adipic acid. Adipic acid, dodecanedioic acid, and a mixture of succinic, glutaric, and adipic acids are not sensitizers.

Succinic Acid Succinic acid was a slight to mild irritant to rabbit skin.65 Details were not provided. Glutaric Acid Glutaric acid was a slight irritant to rabbit skin.65 Details were not provided. Adipic Acid A dermal irritation study was performed in which 500 mg of 50% aq. adipic acid was applied under an occlusive patch to a 5 cm x 5 cm area of intact and abraded skin of 6 rabbits for 24 h.44 With intact skin, an erythema score of 2-3/4 was reported, with clearing by day 3. With abraded skin, mild to severe erythema and edema were reported, which cleared by day 7. Adipic acid, undiluted or as an 80% aq. paste, was applied occlusively to the backs or ears of rabbits for 24 h. Two rabbits were used per group. No irritation was observed on the backs of animals. Erythema was observed on the ear, with clearing by 72 h. In another study in which adipic acid was applied occlusively for 24 h, irritation was not observed. Details were not provided. A semi-occlusive application of 500 mg of a paste of 50% adipic acid in propylene glycol to 6 rabbits produced slight to mild irritation in 3 of the rabbits. A semi-occlusive application of undiluted adipic acid was not corrosive. Using 10 guinea pigs, 50% adipic acid in propylene glycol was not irritating. The sensitization potential of adipic acid was evaluated using groups of 10 guinea pigs. For induction, 0.1 ml of 1% aq. adipic acid was given as a sacral intradermal injection, once a week for 4 wks. After a 2-wk non-treatment period, the dermal challenge was performed with 0.05 ml of 50 and 25% adipic acid in propylene glycol. Adipic acid produced very mild or no irritation, and it was not a sensitizer.

15

CIR Panel Book Page 63 Succinic/Glutaric/Adipic Acids Mixture A mixture of succinic, glutaric, and adipic acid (percentages not specified) was evaluated for irritation and for sensi- tization using guinea pigs.67 The mixture produced mild or no irritation, and it was not a sensitizer. Details were not provided. Dodecanedioic Acid Dodecanedioic acid was not a sensitizer in a 4-h exposure study or upon application of 0.5 g.66 In a maximization study using female guinea pigs, 0.5% dodecanedioic acid was injected intracutaneously at induction and 25 and 50% was used for the dermal challenge. Dodecanedioic acid was not a sensitizer. Dermal irritation and sensitization studies are summarized in Table 6. Mucosal Irritation Succinic Acid Succinic acid has been considered to be an exacerbating factor in ulcerative colitis, therefore its influence on rat co- lonic mucosa in terms of mucosal blood flow and superoxide generation was investigated.70 The left side of the colon of 5 male and 5 female rats was exposed, and 0.9-5% succinic acid in physiological saline was instilled into the colonic lumen. A segment of the colon was then ligated as to not include the mesenteric blood vessel. Mucosal blood flow decreased with all dose levels. Microscopically, the higher the concentration of succinic acid, the greater was the erosion formation in the colon- ic mucosa. Significant polymorphonuclear cell infiltration superoxide generation from colon tissue was observed with 0.01% succinic acid, as compared to higher or lower concentrations. Succinic acid, at fecal concentrations found in active stage ulcerative colitis, appears to be implicated in mucosal injury, mediated by a decrease in colonic mucosal blood flow and infil- tration of superoxide-generating polymorphonuclear cells into the mucosa.70 REPRODUCTIVE AND DEVELOPMENTAL TOXICITY Reproductive and developmental effects were not seen upon oral dosing with the dicarboxylic acids or disodium sebacate. Malonic acid has a spermicidal effect on human spermatozoa. Glutaric acid was tested at doses of ≤1300 mg/kg in rats and 500 mg/kg in rabbits, adipic acid at doses of ≤263 mg/kg in mice, 288 mg/kg in rats, 205 mg/kg in hamsters, or 250 mg/kg in rabbits, azelaic acid at doses of ≤140 mg/kg in rats and 200 mg/kg in rabbits, disodium sebacate at 500 mg/kg in rats and 1000 mg/kg in rabbits, and dodecanedioic acid was tested at ≤1000 mg/kg using rats. Embryotoxic effects were reported for in a reproductive study of 2500 mg/kg/day azelaic acid using rats and in reproductive studies with ≤500 mg/kg/day azelaic acid using rabbits and monkey. Sodium salts of some dicarboxylic acid had a specific inhibitory effect on the uterine horn, and this effect progressively increased with chain length. Malonic Acid Malonic acid, 0.1%, reduced the pH of sperm suspensions to 4.-5.5 and rendered human spermatozoa immotile within 30 min.71 A concentration of 1.0% reduced the pH to 1.5-3.0 and was almost instantaneously spermicidal. Succinic Acid Subcutaneous injections of 31 mg/kg/day succinic acid for 3 wks did not change the typical diestrous vaginal smears in 2 mos old ovariectomized rats.72 Glutaric Acid The reproductive toxicity of glutaric acid was evaluated. Rats were dosed with 125, 400, or 1300 mg/kg and rabbits with 40, 160, or 500 mg/kg. No reproductive, embryotoxic or teratogenic effects were observed. No other details were provided.73 Adipic Acid Groups of 20-24 gravid albino CD-1 mice were dosed orally, by gavage, with 0, 2.6, 12, 56, or 263 mg/kg adipic

16

CIR Panel Book Page 64 acid on days 6-15 of gestation.44 All animals were killed on day 17 of gestation. No reproductive, developmental, or maternal effects were observed, and the NOAEL for maternal and developmental toxicity was 263 mg/kg. Similar results were obtained in a study in which gravid Wistar rats were dosed orally, by gavage, with 0, 2.9, 13, 62, or 288 mg/kg adipic acid on days 6-15 of gestation. The NOAEL for maternal and developmental toxicity was 288 mg/kg. Groups of 21-24 gravid hamsters were dosed orally, by gavage, with 0, 2.9, 5, 44, or 205 mg/kg adipic acid on days 6-10 of gestation. A significant increase in resorption per implant site was observed with 205 mg/kg adipic acid, resulting in a decreased number of live fetuses. (This decrease was not evaluated statistically.) No other effects were reported. Groups of 10-14 gravid Dutch-belted rabbits were dosed by oral intubation with 0, 2.5, 12, 54, or 250 mg/kg adipic acid on days 6-18 of gestation. No reproductive, developmental, or maternal effects were observed. The NOAEL for mater- nal toxicity was ≥250 mg/kg and for developmental toxicity was 250 mg/kg. Azelaic Acid Reproductive and teratogenic effects of azelaic acid were evaluated using Wistar rats and New Zealand rabbits.69 A group of 20 gravid rats was fed a diet containing 140 mg/kg/day azelaic acid, and a control group of 10 gravid rats was given untreated feed. Half of each group was killed and necropsied on day 19 of gestation, and the remaining animals continued dosing for 3 mos. The day of gestation that dosing started is not clear. No gross or microscopic lesions were observed for the uteri, placentas, or ovaries. There were no differences in reproductive, teratogenic, or developmental effects between treated and control groups, nor were there any differences in fetal weights of the live fetuses. Similar results were seen using groups of 20 gravid rabbits fed 200 mg/kg/day azelaic acid; 10 untreated gravid rabbits were used as a negative control group. Embryotoxic effects were observed in oral studies with rats receiving 2500 mg/kg/day of azelaic acid.48 Similar effects were observed in studies in rabbits given 150 to 500 mg/kg/day and in monkeys given 500 mg/kg/day. The doses at which these effects were noted were all within toxic dose ranges for the dams. No teratogenic effects were observed. (Details were not provided.) Disodium Sebacate Reproductive, teratogenic, and developmental effects of disodium sebacate were evaluated using Wistar rats and New Zealand rabbits.69 Groups of 20 gravid rats were fed a diet containing 0 or 500 mg/kg/day disodium sebacate, and groups of 20 gravid rabbits were fed 0 or 1000 mg/kg. Half of each group was killed and necropsied on day 19 of gestation, and the remaining animals continued dosing for 3 mos. The day of gestation that dosing started is not clear. No gross or microscopic lesions were observed for the uteri, placentas, or ovaries. There were no differences in reproductive or developmental effects between treated and control groups, nor were there any differences in fetal weights of the live fetuses. Dodecanedioic Acid The reproductive toxicity of 0-1000 mg/kg dodecanedioic acid was evaluated using male and female Crl:CD:BR rats.66 The no-observable effect level (NOEL) for reproductive and developmental toxicity was 1000 mg/kg. The NOELs for toxicity of male and female rats were 100 and 500 mg/kg, respectively. The NOAEL for both male and female rats was 1000 mg/kg. (No other details were provided.) Sodium Salt of Adipic, Azelaic, Sebacic, and Dodecanedioic Acids The influence of the sodium salt of some dicarboxylic acids (adipic acid, azelaic acid, sebacic acid, dodecanedioic acid) on both spontaneous and evoked muscle activity of the uterine horns of 35 female Wistar rats (250-300g) has been studied in vitro.74 Spontaneous activity of uterine muscle was inhibited by dicarboxylic salts causing the total abolition of mechanical events at concentrations of 24,, 32, 40, and 64 x 10-3 M. Dicarboxylic salts antagonized the maximal isometric contraction of the uterine horn induced by administration of acetylcholine, oxytocin or prostaglandins (PGF2-α). The amount 17

CIR Panel Book Page 65 of antagonism was dependent upon the concentration of dicarboxylic salt used. Dicarboxylic salts had an specific inhibitory effect on the uterine horn which progressively increased with their chain length. The results suggested that the inhibitory effects of dicarboxylic salts on smooth muscle could be due to a cellular membrane hyperpolarization. GENOTOXICITY The dicarboxylic acids are not genotoxic, and consistently were not mutagenic in Ames tests. Positive results were seen in a transformation assay on glutaric acid using Balb/c-3T3 cells, both with and without metabolic activation. The results of a mouse lymphoma assay, with and without metabolic activation, on glutaric acid was pH-dependent. Equivocal results were obtained in an in vitro chromosomal aberration assay of ≤15 mg/mg disodium succinate using Chinese hamster fibroblast cells. The dicarboxylic acids were not genotoxic in in vivo assays. In Vitro Malonic Acid Malonic acid, 3333 µg/plate, was not mutagenic in a National Toxicology Program (NTP) preincubation assay, with or without metabolic activation.75 Succinic Acid The genotoxic potential of succinic acid was evaluated in an Ames test and in a chromosomal aberration study using a Chinese hamster fibroblast cell line.76 Succinic acid, at a concentration of ≤5.0 mg/plate in phosphate buffer, was not muta- genic in the Ames test. (Whether metabolic activation was used is not stated.) Concentrations of ≤1.0 mg/ml in saline were not genotoxic in the chromosomal aberration assay. Sodium succinate, ≤10 /plate, was negative in an Ames test, with and without metabolic activation.77 Disodium Succinate The genotoxic potential of disodium succinate was evaluated in an Ames test and in a chromosomal aberration study using a Chinese hamster fibroblast cell line.76 In the Ames test, disodium succinate was not mutagenic at concentration up to 5.0 mg/plate in phosphate buffer. (Whether metabolic activation was used is not stated.) Equivocal genotoxic results were obtained in the chromosome aberration assay using concentrations of ≤15.0 mg/ml disodium succinate in saline. Glutaric Acid Glutaric acid was evaluated in vitro in a standard Ames assay, the L5178Y/TK ± mouse lymphoma assay with and without metabolic activation, and the mammalian in vitro Balb/c-3T3 cell transformation assay with and without metabolic activation.73 The Ames tests were negative. However, the cell transformation assay was positive both in the presence and absence of metabolic activation and the results in the mouse lymphoma assay were dependent upon pH of the culture medium. The researchers stated that the variable response in the mouse lymphoma assay and the positive effect in the cell transforma- tion assay may have been an indirect effect of other factors (such as the pH or osmolarity of the media in which the cells were exposed), rather than a direct effect of glutaric acid. Adipic Acid Adipic acid was evaluated in a number of Ames assays using Salmonella typhimurium and Escherichia coli; results were negative, with or without metabolic activation, at concentrations as high as 10,000 mg/plate.44,78,79 Negative results were also obtained in an Ames test with 0-200 mg/l adipic acid using S. typhimurium TA1530 and G-46 without metabolic activa- tion44. Results were negative in a yeast gene mutation assay using Saccharomyces cerevisiae without metabolic activation at concentrations ≤200 mg/l. A mouse lymphoma assay using L5178Y/TK ± cells was negative with and without metabolic acti- vation at concentrations of ≤2000 µg/plate,79 as was a cytogenetic assay using human embryonic lung fibroblast cells with ≤200 mg/l adipic acid.44 In a viral enhanced cell transformation assay using Syrian hamster embryo cells at dosed of 62-1000

18

CIR Panel Book Page 66 µg/ml adipic acid, results were negative. Azelaic Acid Azelaic acid, 20%, was not mutagenic or genotoxic in an Ames assay, HGPRT test in Chinese hamster ovary cells, or human lymphocyte test.48 Details were not provided. Dodecanedioic Acid Dodecanedioic acid was not mutagenic in an Ames assay at concentrations of ≤5000 µg/plate, with and without metabolic activation.66 Toxicity occurred at ≥500 µg/plate. In Vivo Glutaric Acid Glutaric acid was evaluated in a mammalian micronucleus cytogenetic assay in mice.73 Glutaric acid was not genotoxic in this assay. (Details not specified.) Adipic Acid Adipic acid was not genotoxic in in vivo cytogenetic assays using chromosomes from rats dosed orally, by gavage, with a single dose of 5000 mg/kg or daily for 5 days with 2500 mg/kg.44 Adipic acid was also not genotoxic in dominant lethal studies with doses up to 5000 mg/kg.. Azelaic Acid Azelaic acid was not genotoxic in a dominant lethal assay in mice.48 (Details not specified.) Dodecanedioic Acid Dodecanedioic acid, at concentrations of ≤5000 mg/kg, was not mutagenic in a micronucleus assay using mice.66 Available details for the genotoxicity studies are summarized in Table 7. CARCINOGENICITY Carcinogenic results were not seen in rats given up to 2% sodium succinate in the drinking water or 5% adipic acid in feed for 2 yrs. An increase in the incidence of C-cell adenoma/carcinoma of the thyroid in females given 2% sodium succinate, and a positive trend in the occurrence of this tumor, was considered a function of experimental variability and not related to dosing. Sodium Succinate Groups of 50 male and 50 female F344 rats were given drinking water containing 0, 1, or 2% sodium succinate for 2 yrs, and the carcinogenic potential was determined.68 Dosing was discontinued after 104 wks, and, after a 9-wk recovery period, the rats were killed at wk 113. Body weights of the high dose animals were decreased by 10% as compared to con- trols. There were no statistically significant differences in overall tumor incidence or mean survival time between treated and control animals. An increase in the incidence of C-cell adenoma/carcinoma of the thyroid in females of the 2% group, and a positive trend in the occurrence of this tumor, was considered a function of experimental variability and not related to dosing. Sodium succinate was not toxic or carcinogenic to male or female F344 rats when given in the drinking water for 2 yrs. Adipic Acid Adipic acid was not carcinogenic in the 2-yr chronic oral toxicity study (described previously) in which groups of 20 male rats were fed diets containing 0, 0.1, 1, 3, and 5% adipic acid, and groups of 10 and 19 females were fed 0 and 1% adipic acid, respectively.67 Tumor Promotion Succinic Acid, Sodium Succinate, Disodium Succinate The promotion of urinary bladder carcinogenesis by sodium succinate was evaluated using male F344 rats.80 Groups

19

CIR Panel Book Page 67 of 16 male F344 rats were given 5% succinic acid, sodium succinate, or disodium succinate with 0.05% N-butyl-N- (4- hydroxybutyl)nitrosamine (BBN) in the drinking water for 4 wks, followed by dietary administration of 5% of the respective test article without BBN for 32 wks. Negative controls were given water with BBN only and untreated feed. Groups of 8 male F344 rats followed the same protocol without the addition of BBN to the drinking water, as did a group of non-BBN- treated negative controls. The animals were killed at wk 37. In the BBN-pretreated groups, many rats given sodium or disodium succinate developed hematuria towards the end of the study. There were no statistically significant differences in body or organ weights between the control and test groups. (Information on organ and body weights was not provided for the non-BBN groups.) Large tumors were found on the urinary bladders of the BBN-pretreated animals given sodium and disodium succinate; tiny lesions were found in the control or succinic acid BBN-pretreated animals. The incidence and number of urinary bladder carcinomas and papillomas and of papil- lary or nodular hyperplasia (preneoplastic lesions) were statistically significantly increased in the sodium and disodium succinate BBN-pretreated groups as compared to the succinic acid and control BBN-pretreated groups. The incidence and numbers observed in the sodium and disodium succinate groups were not statistically significantly different from each other. An association between tumor area and sodium intake was noted. Urinary bladder lesions were not observed in any of the animals that were not pretreated with BBN. Urinary pH and electrolyte concentrations were affected by dosing with sodium or disodium succinate with BBN, as compared to the control and succinic acid groups, and statistically significant differences between these two groups were observed as well. The researchers also evaluated cell proliferation and DNA synthesis in the urinary bladder epithelium. Groups of 20 male F344 rats were given 5% succinic acid, sodium succinate, or disodium succinate in the feed, without BBN pretreatment for 8 wks. Negative controls were given basal diet. Five rats per group were given an i.p. injection of 50 mg/kg 5-bromo-2’- deoxyuridine (BrdU) 1 h prior to being killed. Compared to control values, BrdU uptake was statistically significantly increased by increased disodium succinate and was increased, but not in a statistically significant manner, by sodium succinate. Succinic acid did not have any effect on DNA synthesis. Microscopically, simple hyperplasia was observed in the urinary bladders of animals given sodium and disodium succinate. The appearance of the urinary bladder epithelial surface was altered by sodium and disodium succinate. Spermidine/spermine N1-acetyltransferase activity in the urinary bladder epi- thelium was increased for disodium succinate, but not sodium succinate, when compared to controls. Urinary pH and electrolyte concentrations were affected as described previously. CLINICAL ASSESSMENT OF SAFETY In a cumulative irritancy test, the cumulative irritation of a 15% azelaic acid gel increased with successive patching. It is not known if the vehicle played a role in the irritation scores. Daily application of a 20% azelaic cream causes erythema and irritation. Dermal Irritation Azelaic Acid The cumulative irritation potential of a 15% azelaic acid gel (prescription formulation; vehicle not identified) was determined in a study using 31 female and 2 male subjects.81 (During the study, 1 subject withdrew for personal reasons.) White petrolatum was used as a negative control. Azelaic acid and petrolatum, 0.2 g of each, were applied under occlusion to 2 cm x 2 cm sites on the back of each subject 3 times per week for 3 wks. Weekday patches were removed after 24 h, while the patches applied on Fridays were removed after 72 h. The test sites were evaluated 15-30 min after removal of the patch, and then a new patch was applied. Application was discontinued if severe irritation, which was designated by a maximum erythema score of 3, was observed. A 15% azelaic acid gel was statistically significantly more irritating than the negative

20

CIR Panel Book Page 68 control, with a mean cumulative irritancy index of 1.05/3. Individual reaction scores for the test article ranged from 0 to 3, and 5 subjects discontinued patching with azelaic acid due to an irritation score ≥3. Cumulative irritancy increased with suc- cessive patching. The researchers noted that since the vehicle used for azelaic acid was not tested, there was uncertainty as to whether the vehicle components affected the irritation scores. Twice daily application of a cream containing 20% azelaic acid has been reported to cause erythema, irritation, pruritus, dryness, scaling, and burning.82 Case Reports Adipic Acid In two case reports with industrial exposure to adipic acid, positive sensitization reactions were reported with follow- up testing.44

21

CIR Panel Book Page 69 ESTERS OF DICARBOXYLIC ACIDS Much of the information on the esters of dicarboxylic acids was obtained from summary documents that mostly contained unpublished data. The summary documents are therefore listed as the citation, and, when there are numerous studies described, will only be cited at the beginning of that section. Data on esterase metabolites other than the parent dicarboxylic acid (i.e. parent alcohol and monoester) are summarized in Appendix I, immediately following the reference section. It is important to note that this is merely for support and not for review as ingredients themselves. GENERAL BIOLOGY Absorption, Distribution, Metabolism, and Excretion Metabolism of diesters in animals is expected to occur, initially, via enzymatic hydrolysis, leading to the correspond- ing dicarboxylic acids and the corresponding linear or branched alcohol.83 These dicarboxylic acids and alcohols can be fur- ther metabolized or conjugated to polar products that are excreted in urine. However, other studies have shown that enzy- matic hydrolysis of at least some diesters may be incomplete and result, instead, in the production of monoesters.84 Diethyl Malonate In in vitro absorption studies using pig skin, 8.8 and 3% of undiluted diethyl malonate was found in the skin and receptor fluid, respectively. Absorption was enhances when diethyl malonate was diluted with ethanol and reduced when diluted in acetone Using human skin, 16% of the applied diethyl malonate penetrated. In vivo, absorption of diethyl malonate, estimated from urinary and fecal recovery, was 15% in nude mice, 4 % in human skin grafted to nude mice, 6% in pig skin grafted to nude mice, 2.5% in pigs, and 4% in dogs. Diethyl malonate is hydrolyzed via a two-step reaction to malonic acid and the corresponding alcohol, ethanol.14 Dimethyl malonate, which is not listed in the International Cosmetic Ingredient Dictionary, has similar physico- chemical properties and hydrolyzes in the same manner to malonic acid and methanol. Because of this similarity, data on dimethyl malonate will be included in this safety assessment to provide read-across data. Distribution of diethyl malonate (and dimethyl malonate) is likely to occur in the water compartments, and accumulation in fat is unlikely based on physical and chemical properties. Both esters are likely to be metabolized by unspecific (serine-) esterases of different tissues, in particular, in the liver to the mono- esters and then to malonic acid and ethanol (or methanol). The hydrolysis product is likely to be metabolized via physiological pathways, such as the tricarboxylic acid cycle, as they are part of the normal intermediate metabolism. Both are assumed to readily absorb via mucous membranes. In Vitro - Animal The percutaneous absorption of radiolabeled diethyl malonate was determined in vitro using skin from Yorkshire pigs. [2-14C]Diethyl malonate was applied either undiluted (100 g/cm2) or diluted in ethanol at 12.5 mg/ml with an applied dose of 100 g/cm2 or as 0.5 mg/ml with an applied dose of 4 g/cm2. At 50 h, with undiluted diethyl malonate, 8.8% of the radioactivity was found in the skin and 3% was in the receptor fluid. With 100 µg in ethanol, 13% of the radioactivity was found in the skin and 6% in the receptor fluid and with 4 µg in ethanol, 30% was found in the skin and 10% in the receptor fluid. Absorption appeared to be enhanced with ethanol. The percutaneous absorption of 1 mg/cm2 [2-14C]diethyl malonate in 10 µl acetone was determined in vitro also using skin from Yorkshire pigs. At 24 h, 0.2-1.6% of the diethyl malonate was found in the receptor fluid, 0.2-0.9% was found in the skin, and 0.6-0.7% was found on the skin surface. Skin mediated hydrolysis amounted to 15-35% of the applied dose. In the receptor fluid, 20-21% of the applied dose was present as hydrolysis products. In the skin and on the skin surface, 3-5% and 2-4%, respectively, of the applied dose was present as hydrolysis products.

22

CIR Panel Book Page 70 In Vivo - Animal The percutaneous penetration of radiolabeled diethyl malonate was studied in vivo in the following animal models: athymic nude mouse, human, and pig skin grafted to athymic nude mice, in weanling pigs, and in hairless dogs. [2-14C]Diethyl malonate was applied at a dose of 0.1 mg/cm2 for 24 h to a 1.27 cm2 area of mouse skin or for 48 h to a 25 cm2 area of pigs and hairless dogs using non-occluded applications. Absorption, estimated using urinary and fecal recovery after s.c. administration, was 15% in nude mice, 4 % in human skin grafted to nude mice, 6% in pig skin grafted to nude mice, 2.5% in pigs, and 4% in dogs. In Vitro - Human An in vitro skin absorption study was performed using diethyl malonate, no vehicle given. Human cadaver split thickness skin was used in flow through cells. Diethyl malonate (4 l) was applied to the skin samples. After 24 h, 16% of the applied dose had penetrated through the skin. The maximum flux rate was reached after 5 h and amounted to 280 g/h (350 g/cm2/h); the mean penetration rate was 99 g/h (120 g/cm2/h). Much of the test substance, 45 to 50%, evaporated from the skin, and 34 to 39% remained on the skin. Ditridecyl Adipate Approximately 11% of ditridecyl adipate was absorbed through the skin of rats; 5.5-7.4% of the applied dose was found in the tissues, 3.5-4.7%was found in the urine, and 0.4-0.7% was found in the feces. Prior dosing with ditridecyl adipate did not significantly affect absorption. The percutaneous absorption of [14C]ditridecyl adipate was determined using groups of 10 male and 10 female Sprague-Dawley rats that were untreated or that had previously been exposed to unoccluded dermal applications of 0 or 2000 mg/kg ditridecyl adipate, 5 days/wk for 13 wks.85 (This study is described in the section on ‘Subchronic Dermal Toxicity’.) A single 58 µl dose of 2000 mg/kg [14C]ditridecyl adipate was applied topically (size of test site not specified), and urine and feces were collected for 4 days. In the previously untreated rats, a total of 11.6 and 10.6% of the [14C] solution was absorbed by male and female rats, respectively, over 4 days. Approximately 63 and 52% of the absorbed dose (7.4 and 5.5% of the applied dose, respectively) was found in the tissues of males and females, respectively. A total of 3.5-4.7% of the applied dose was recovered in the urine and 0.4-0.7% in the feces of previously untreated rats. The values for the animals previously dosed with 2000 mg/kg ditridecyl adipate were not statistically significantly different from the controls. In the previously dosed animals, a total of 10.8 and 9.1% of the dose was absorbed by males and females, respectively, over the 4 days, with approximately 87 and 81% of the absorbed dose (9.4 and 7.4% of the applied dose, respectively) found in the tissues of the male and female rats, respectively. A total of 0.7-1.3% of the [14C] was recovered in the urine and 0.4-0.6% in the feces. Based on the radioactivity recovered in the urine, the bioavailability of ditridecyl adipate was 2-6%, and previous dosing did not significantly affect absorption. Diethylhexyl Adipate In vitro, diethylhexyl adipate was readily hydrolyzed to MEHA or adipic acid in rat liver, pancreas, and small intestine tissue preparations. In animals, diethylhexyl adipate is hydrolyzed to adipic acid and 2-ethylhexanol or MEHA. 2-Ethylhexanol is converted to 2-ethylhexanoic acid, which may form a glucuronide conjugate or may be subjected to ω- and (ω-1)-oxidation and further metabolism. In rats, carbonyl-labeled diethylhexyl adipate rapidly hydrolyzed to adipic acid and MEHA. More than 98% of orally administered diethylhexyl adipate was excreted in 48 h; 21-45% of the radioactivity was expired in carbon dioxide and 34-52% was excreted in the urine. Metabolism studies have shown that excretion in the urine is not as unchanged diethylhexyl adipate; mostly adipic acid is found. In body tissues, the highest levels of radioactivity were found in the liver, kidneys, blood, muscle, and adipose tissue; elimination from the tissues was rapid, with no affinity for a specific organ. In the tissues, diethylhexyl adipate, adipic acid, and/or MEHA were found. When comparing distribution with labeling on the acid versus the alcohol, radioactivity was observed in the ovaries of gravid mice and some fetal tissues following dosing with [carbonyl-14C]diethylhexyl adipate, but none was detected in the ovaries of gravid mice after dosing with [2-ethylhexyl-1-14C]diethylhexyl adipate, and very little 23

CIR Panel Book Page 71 radioactivity was seen in fetal tissues. Vehicle affects absorption, but not urinary excretion. In humans dosed orally with diethylhexyl adipate labeled on the ethyl side chains, unconjugated 2-ethylhexanoic acid was the only measurable compound in plasma, and the rate of elimination was rapid. In urine, 2-ethylhexanoic acid was again the principal metabolite, and was probably eliminated in the conjugated form. Peak urinary elimination of all metabolites occurred within 8 h of dosing. In Vitro The in vitro hydrolysis of diethylhexyl adipate (and mono-(2-ethylheyxl) adipate [MEHA]) using tissue preparations from the liver, pancreas, and small intestine of 2 rats was examined, as were the effects of diethylhexyl adipate on serum and hepatic enzymatic activities in vitro.86 Diethylhexyl adipate was readily hydrolyzed to MEHA or adipic acid by each tissue preparation. The formation of adipic acid was rapid and approximately the same for all three tissues, while the formation of MEHA was rapid only in pancreatic tissue and was negligible in the intestine. The rate of hydrolysis from MEHA to adipic acid was greater that than from diethylhexyl adipate and the highest activity was found in intestinal tissue. In examining the effects on serum and hepatic enzymes, only N-demethylase activity was considerably inhibited by diethylhexyl adipate. In Vivo - Animal The elimination, distribution, and metabolism of diethylhexyl adipate was investigated using male Wistar rats.86 In these studies, diethylhexyl adipate was labeled at the carbonyl carbon. In elimination studies, 2 rats were dosed by gavage with 500 mg/kg [14C]diethylhexyl adipate (1.26 µCi/rat) as a saturated solution in dimethyl sulfoxide (DMSO), and respired carbon dioxide, urine, and feces were collected for 2 days. At 24 h after dosing, 86% of the administered dose was excreted, and at 48 h, more than 98% of the dose was excreted. In one animal, 44.8% of the dose was excreted in expired carbon dioxide and 33.9% in the urine at the 48 h measurement, while in the other rat, 21.1% and of the dose was excreted in expired carbon dioxide and 52.2% in the urine. Little (1.4 or 5% of the dose) was excreted in the feces. In the distribution study, 3 rats per group were given a single dose as described above. The animals were killed at various intervals, and blood, organ, and tissue samples were collected. Not taking into account the stomach and intestines, the greatest levels of radioactivity, as a percent of dose administered, were found in the liver, kidney, blood, muscle, and adipose tissue. These values ranged from 0.34-8.21% at 6 h, with the greatest percentage found in the adipose tissue, and from 0.54- 3.44% at 12 h, with the greatest percentage found in the muscle. In most tissues, the amount of residual radioactivity reached a peak by 6 h, except for the liver, kidneys, testicles, and muscle, which reached a peak at 12 h. The researchers stated that the elimination of radioactivity from the tissues and organs was very rapid, and there was no specific organ affinity. The metabolism of diethylhexyl adipate was examined in rats dosed orally, by gavage, with 100 mg of non-labeled diethylhexyl adipate as a 5% solution in DMSO. A control group was dosed with vehicle only. The rats were killed 1, 3, or 6 h after dosing. The metabolites were determined using GLC. Diethylhexyl adipate was rapidly hydrolyzed to adipic acid, the main intermediate metabolite, and MEHA. In the urine, adipic acid was detected at 1 h, and excretion as adipic acid in the urine reached 20-30% at 6 h. Diethylhexyl adipate and MEHA were not detected in the urine. Adipic acid only also was de- tected in the blood and the liver, with constant excretion of 0.5-0.7% of the dose in the blood and excretion in the liver in- creasing with time, with 2-3.3% excreted in the liver at 6 h. In the stomach, diethylhexyl adipate, adipic acid, and MEHA were found. The concentrations of diethylhexyl adipate declined rapidly, while the levels of adipic acid (9-10%) and MEHA (6-11.5%) peaked at 3 h. Adipic acid, but not MEHA, was found in the intestine and increased with time, reaching 19% at 6 h. The absorption, distribution, and elimination of diethylhexyl adipate was examined using radioactive labeling on the acid [carbonyl-14C] (specific activity 39.5 mCi/mmol) or the alcohol [2-ethylhexyl-1-14C] (44.1 mCi/mmol).39 The research- ers used both DMSO and commercial corn oil as vehicles for all tests. (DMSO is an active penetrant and carrier of other

24

CIR Panel Book Page 72 substances through tissue membranes; a fat-soluble substance, such as diethylhexyl adipate, is more realistically studied dis- solved in corn oil.) The following groups of animals were dosed with 84.3 µg (9 µCi) [carbonyl 14C]diethylhexyl adipate or 84.3 µg (10 µCi) [2-ethylhexyl-1-14C]diethylhexyl adipate in both vehicles: 12 male NMRI mice were dosed i.v. and killed at intervals from 5 min to 4 days after dosing; 10 male NMRI mice were dosed intragastrically (i.g.) and killed at intervals from 20 min to 4 days after dosing; 12 gravid NMRI mice were dosed i.v. or i.g. on day 17 of gestation and killed at intervals from 20 min to 24 h. Six male rats were dosed i.g. with 843 µg (90 µCi) [carbonyl 14C]diethylhexyl adipate or 843 µg (100 µCi) [2-ethylhexyl-1-14C]diethylhexyl adipate and killed at intervals from 20 min to 4 h. Whole body autoradiography was used to determine tissue distribution. Following dosing with [carbonyl-14C]diethylhexyl adipate, distribution was similar in male mice, male rats, and gravid mice. The amount of radioactivity in the tissues peaked at a later time following i.g. dosing as compared to i.v. dosing. The presence of radioactivity in the gastrointestinal tract following i.v dosing indicated biliary excretion. Four h following both i.v. and i.g. dosing, the greatest uptake of radioactivity was found in the liver, bone marrow, brown fat, adrenal cortex, kidneys, and a few other tissues. At 24 h after i.g. dosing, significant levels of radioactivity remained in several tissues, including the liver, of both rats and mice. In gravid mice, a “remarkable strong uptake” of radioactivity in the corpora lutea of the ovary was observed at all time intervals with both i.v. and i.g. dosing, and some radioactivity was found in the fetal intestine, liver, and bone marrow. Similar distribution patterns were seen following dosing with [2-ethylhexyl-1-14C]diethylhexyl adipate as were seen with [carbonyl-14C]diethylhexyl adipate. Following i.g. dosing, the appearance of radioactivity was lessened and not as great as it was with i.v. dosing. Very high radioactivity levels were seen in the liver and kidney at 5 min-1 h after i.v. dosing and at 20 min-4 h after i.g. dosing. The radioactivity in the liver was still high at 24 h after i.g. dosing in mice and rats. Radioactivity was also seen in the intestinal contents at 1-4 h after i.v. dosing, again indicating biliary excretion. At longer intervals after i.v. injection, 4 h-4 days, radioactivity was detected in the bronchi of mice. While radioactivity was observed in the ovaries of gravid mice and some fetal tissues following dosing with [carbonyl-14C]diethylhexyl adipate, none was detected in the ovaries of gravid mice after dosing with [2-ethylhexyl-1-14C]diethylhexyl adipate, and very little radioactivity was seen in some fetal tissues. The effect of vehicle on the absorption and biliary and urinary excretion of diethylhexyl adipate was also examined using rats in a gavage study with [14C]diethylhexyl adipate. Radioactivity was measured every 30 min for 7.5 h. The times and extent of absorption were different for all four preparations of [14C]diethylhexyl adipate. Radioactivity levels in the blood increased faster and were greater with DMSO as the vehicle, as compared to corn oil. The highest blood radioactivity levels were found with [carbonyl-14C]diethylhexyl adipate in DMSO. Biliary excretion of [14C]diethylhexyl adipate was greatly affected by vehicle; with DMSO, 41% of the dose was detected in the bile, while only 10% of the dose was found with the corn oil vehicle. This difference was not seen with [carbonyl-14C]diethylhexyl adipate. Finally, vehicle did not have much influence on urinary excretion. However, unlike the results reported by the previous researchers, little radioactivity was excreted in the urine. The researchers hypothesized that since the study duration was only 7.5 h, urinary excretion may not have been complete. The metabolism of diethylhexyl adipate was examined in vivo using male Wistar rats and compared to in vitro metabolism using hepatocytes.87 In vivo, rats were dosed with 0.665 or 1.5 g/kg diethylhexyl adipate in corn oil by gavage for 5 days, and the controls were given vehicle only. Urine was collected daily. Diethylhexyl adipate was not recovered in the urine after 24 h. Adipic acid was the main metabolite of diethylhexyl adipate. In vitro, the first hydrolysis of diethylhexyl

25

CIR Panel Book Page 73 adipate appears to be a rate-limiting step. In vivo, it was thought that this hydrolysis probably occurs in the gastrointestinal tract. Metabolic pathways (ω and ω-1oxidations, glucuronidation) seemed to prove that transformations of diethylhexyl adipate are localized mainly in the liver. Oral administration of diethylhexyl adipate to cynomolgus monkeys results in rapid elimination, with 47-57% of the dose excreted in the urine.17 Unchanged diethylhexyl adipate is absorbed from the gastrointestinal tract, and the glucuronide of MEHA and traces of unchanged diethylhexyl adipate were found in the urine. (Details were not provided.) In Vivo - Human 2 The pharmacokinetics of [ H10]diethylhexyl adipate, labeled on the ethyl side-chains, were examined using 6 male 88 2 3 subjects. A dose of 46 mg [ H10]diethylhexyl adipate in corn oil, for a total volume of 0.5 cm , was administered orally in a gelatin capsule. Blood samples were taken for up to 31 h after dosing, and urine samples were taken at intervals for up to 96 2 h after dosing. In the plasma, unconjugated [ H5]2-ethylhexanoic acid was the only measurable diethylhexyl adipate-related compound. This compound appeared rapidly in the plasma, and the peak concentrations (1.6 ±0.5 µg/cm3) occurred between 2 1 and 2 h. [ H5]2-Ethylhexanol was detected, but it was below the limit of quantification. The rate of formation was calcu- lated, since there was no evidence of diethylhexyl adipate absorption, as 1.63 ± 1.19 hr-1. The rate of elimination from the plasma was also rapid and estimated to be 0.42 ± 0.15 h-1., which corresponded to an elimination half-life of 1.65 h. 2 Although there were inter-individual differences in the rate and extent of [ H5]2-ethylhexanoic acid formation, it was below the limit of detection in all subjects by 31 h. 2 In the urine, [ H5]2-ethylhexanoic acid was again the principal metabolite, and it was probably eliminated as a 2 conjugated product. This conjugated form, most likely the glucuronide, accounted for up to 99% of the total [ H5]2-ethyl- hexanoic acid measured. Conjugation of the other urinary metabolites was minimal. Peak urinary elimination of the measured metabolites occurred within 8 h of dosing, and no metabolites were detected in the urine after 36 h. The rates of elimination were similar for all metabolites, with a mean elimination half-life of 1.5 h. The measured urinary metabolites accounted for 12.1% of the dose, with the majority being eliminated in 24 h. Fecal analysis determined that a minor portion 2 of the dose was present as diethylhexyl adipate (0.43%) and [ H5]MEHA (0.27%). The researchers noted that recovery of the administered dose was incomplete and hypothesized that it was most probably due to further systemic metabolism. Diethylhexyl Sebacate Diethylhexyl sebacate is not readily absorbed through the skin of guinea pigs (no further details were provided).1 It was noted that the metabolism of diethylhexyl sebacate in rodents and humans may follow partially common pathways,. Penetration Enhancement Diethyl Sebacate The addition of diethyl sebacate to an antifungal agent (ME1401) increased its in vivo antifungal activity and its penetration.89 Peroxisome Proliferation Peroxisome proliferation causes an increase in liver weights and can induce hepatocarcinogenicity in rats and mice. Diethylhexyl adipate is a peroxisome proliferator requiring extensive phase I metabolism to produce the proximate peroxisome proliferator, which in both mice and rats appears to be 2-ethylhexanoic acid. Studies conducted to explain the species difference in liver tumors seen in mice, but not rats, in the NTP carcinogenicity study on diethylhexyl adipate, suggested that diethylhexyl adipate-induced cell replication, rather than hepatic peroxisome proliferation, provided a better correlation with tumor formation. Diethylhexyl adipate is not as potent a proliferator as diethylhexyl phthalate. Peroxisome proliferation is not believed to pose the risk of inducing hepatocarcinogenesis in humans.

26

CIR Panel Book Page 74 Mechanism Peroxisomes, single membrane-limited cytoplasmic organelles present in cells, have several important functions in intermediary metabolism.90 Peroxisome proliferators can induce hepatocarcinogenicity in mice and rats, and may also pro- duce tumors in other organs, such as the testes and pancreas, of these species. Some compounds require extensive phase I metabolism to produce the proximate peroxisome proliferator. Rat liver peroxisomes contain both catalase, which destroys hydrogen peroxide, and a number of hydrogen peroxide- generating oxidase enzymes. Liver peroxisomes contain a complete fatty acid β-oxidation cycle. The stimulation of micro- somal fatty acid-oxidizing enzymes is caused by the induction of cytochrome P450 isoenzymes in the CYP4A subfamily. Good correlations have been reported for rat livers between the induction of peroxisomal fatty acid β-oxidation and organelle proliferation, as well as between the induction of peroxisomal and microsomal fatty acid-oxidizing activities. Peroxisome proliferators can stimulate DNA synthesis in rat hepatocytes, but do not bind covalently to DNA and are not considered to be genotoxic carcinogens. Proposed mechanisms of liver tumor formation include induction of sustained oxidative stress, a role for enhanced cell replication, and the promotion of spontaneous preneoplastic lesions. Sustained oxi- dative stress does not appear to be solely responsible for peroxisome proliferator-induced hepatocarcinogenesis. In general, there is a poor quantitative correlation between markers of oxidative stress and compound potency to produce tumors. The administration of peroxisome proliferators in rodents results in marked increases in liver weights, which is associated with both morphological and biochemical changes. Liver enlargement is due to both hepatocyte hyperplasia and hypertrophy. Morphologically, the size and the number of peroxisomes increase. Although potent peroxisome proliferators are more likely to produce tumors, the magnitude of proliferation does not always correlate with tumor formation. Peroxi- some proliferators that produce a sustained stimulation of cell replication produce tumors more rapidly than other agents that produce a similar magnitude of proliferation but do not produce sustained stimulation of cell replication. Transformation of hepatocytes, by either oxidative stress or alternative mechanisms, may cause initiated cells to be promoted and progress to tumors by enhanced cell replication. Both peroxisome proliferation and cell replication are impor- tant biomarkers of peroxisome proliferator-induced tumor formation in rodent livers. A species difference in response to peroxisome proliferators exists. Humans do not react to peroxisome proliferators in the manner that rodents do, and peroxisome proliferators do not appear to pose any serious hazard for humans. There seems to be a lack of effect on organelle proliferation and induction of peroxisomal and microsomal fatty acid-oxidizing enzymes in species other than rats and mice, and it is suggested that other species are not susceptible to peroxisome prolifera- tor-induced liver tumor formations. Diethylhexyl Adipate In male rats fed 2% diethylhexyl adipate in the feed for 3 wks, hepatic peroxisome proliferation, an increase in liver size, and an increase in two hepatic activities of peroxisome-associated enzymes, catalase and carnitine acetyl transferase, were observed. It was postulated that a metabolite, 2-ethylhexyl alcohol, may be responsible for the induction of hepatic peroxisome proliferation. From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2

Cultured hepatocytes from male Swiss mice and Wistar rats were used to identify the proximate peroxisome prolif- erator derived from diethylhexyl adipate.91 Diethylhexyl adipate did not induce peroxisome proliferation in mouse or rat hepatocyte cultures. However, the primary metabolites of diethylhexyl adipate, MEHA and 2-ethylhexanol, produced similar results in both mouse and rat hepatocyte cultures, with a 5-fold induction of peroxisomal β-oxidation (measured as cyanide- insensitive palmitoyl CoA oxidase [PCO] activity) in mouse hepatocytes and a 4-5-fold increase in rat hepatocytes at concen- trations of 0.5 mM. The most potent peroxisome proliferation occurred in cultured mouse hepatocytes with the secondary 27

CIR Panel Book Page 75 metabolite of diethylhexyl adipate, 2-ethylhexanoic acid, which resulted in a 25-fold induction at a concentration of 1 mM. In cultured rat hepatocytes, 2-ethylhexanoic acid again induced the greatest stimulation of PCO, but only a 9-fold increase was observed with a concentration of 1 mM. 2-Ethyl-5-hydroxyhexan-1-oic acid, at a concentration of 2mM, stimulated PCO approximately 5-fold. (Higher concentrations of these agents were cytotoxic.) In both mice and rats, 2-ethylhexanoic acid was the proximate peroxisome proliferator, but mouse hepatocytes were approximately twice as sensitive as rat hepatocytes. The effects of diethylhexyl adipate and its metabolites on cultured guinea pig and marmoset hepatocytes were also examined. Unlike mouse and rat hepatocytes cultures, neither diethylhexyl adipate nor it metabolites, at concentrations up to 2 mM, stimulated PCO. In another study investigating the effect of diethylhexyl adipate on PCO induction, 2-ethylhexanoic acid was again found to be the proximate peroxisome proliferator.92 In this in vivo study, male and female Wistar rats and male and female Swiss mice, 5 per gender per group, were dosed orally by gavage with 0-2.5 g/kg (0-6.74 mmol/kg) diethylhexyl adipate, 0- 1.75 g/kg (0-13.49 mmol/kg) 2-ethylhexanol, or 0-1.0 g/kg (0-13.49 mmol/kg) 2-ethylhexanoic acid in corn oil for 14 days. The animals were killed 24 h after the final dose and the livers excised. Relative liver weights were increased in a dose- dependent manner for rats and mice. On a molar basis, diethylhexyl adipate was approximately twice as potent as 2-ethylhex- anol or 2-ethylhexanoic acid. Diethylhexyl adipate, 2-ethylhexanol, and 2-ethylhexanoic acid induced peroxisomal β-oxida- tion in a linear dose-response manner in F344 rats and Swiss mice. PCO was stimulated to the greatest extent in male mice. In an attempt to explain the species difference in an NTP study93 that used these strains of rat and mouse, the peroxi- some proliferation induced by diethylhexyl adipate was examined, with the same dosing scheme, using male and female F344 92 rats and male and female B6C3F1 mice (The NTP study found that diethylhexyl adipate induced hepatocellular tumors, and was carcinogenic, in mice but not rats.) In contrast to the expected results, it was found that PCO activity was increased to the greatest extent, up to 15-fold, in male F344 rats. However, catalase activity was statistically significantly increased in male and female mice at all doses, but this was not seen in rats at any dose. Using electron microscopy, a dose-related peroxisome proliferation was observed for rats and mice. This increase was statistically significant at all dose levels for male rats, and all but the lowest dose level for female rats and male and female mice. Relative liver weights were increased in a dose-dependent manner for rats and mice. Another in vivo study was performed in which the species difference on the hepatic effects of diethylhexyl adipate 94 was evaluated using female F344 rats and female B6C3F1 mice. Groups of 5-8 rats were fed diets containing 0-4.0% di- ethylhexyl adipate for 1, 4, or 13 wks, and 5-8 mice were fed diets containing 0-2.5% for the same time periods. Marked dose-dependent increases in relative liver weights were reported for rats given 1.2-4.0% and mice given 0.6-2.5% diethyl- hexyl adipate for 1 and 4 wks and for rats given 0.6-4.0% and mice given 1.2 and 2.5% diethylhexyl adipate for 13 wks. Di- ethylhexyl adipate markedly increased PCO induction in rats and mice at all 3 time periods, and microsomal lauric acid 11- and 12-hydroxylase activities were also increased. Diethylhexyl adipate had little effect on microsomal cytochrome activity in the rat, but significant increases were observed in mice given 1.2 and 2.5% diethylhexyl adipate for 1 and 4 wks and those given 0.6-2.5% for 13 wks. Replicative DNA synthesis was determined using 5-bromo-2’-deoxyuridine. Diethylhexyl adipate produced a sustained stimulation of replicative DNA synthesis in mice given 1.2 and 2.5%, but this effect was not seen in rats. The researchers suggested that diethylhexyl adipate-induced cell replication, rather than hepatic peroxisome proliferation, provided a better correlation with tumor formation (in the NTP study). Toxic effects are described in “Short- Term Oral Toxicity” for the 1 and 4 wk studies and “Subchronic Oral Toxicity” for the 13 wk study. The effect of diethylhexyl adipate on in vitro steady-state levels of hydrogen peroxide was also evaluated as an

28

CIR Panel Book Page 76 95 attempt to explain the species diference. Groups of 5 male F344 rats and female B6C3F1 mice were dosed with 5 ml/kg of 0 or 2 g/kg diethylhexyl adipate daily for 14 days. (Route of dosing not specified.) Statistically significant increases in PCO and catalase activity, but not glutathione peroxidase activity, were observed for both rats and mice. As compared to controls, a 5-fold increase was seen in PCO activity with rats, and a 4-fold increase with mice. Catalase activity, as compared to controls, was 2-times greater in both rats and mice. Steady-state hydrogen peroxide concentrations, determined by measuring rates of hydrogen peroxide production from palmitoyl CoA oxidation, was also increased 2-fold for male rats and female mice. However, absolute values for steady-state hydrogen peroxide concentrations were greater in the livers of treated mice, compared to rats. The researchers stated the difference in the absolute concentration may be important and possibly a contributing factor to the species difference. The extent of peroxisome proliferation induced by diethylhexyl adipate was compared to the proliferative effects of diethylhexyl phthalate and ciprofibrate, a very effective peroxisome proliferator.96 Groups of 3-4 male F344 rats were fed a diet containing 0.25-2% diethylhexyl adipate, 0.25-2% diethylhexyl phthalate, or 0.001-0.02% ciprofibrate for 30 days. With diethylhexyl adipate, a statistically significant increase in liver weight was seen at the 2% dose level, but not at lower dose levels. Dose-dependent increases in liver weight were seen at all dose levels with the other two test compounds, and the hepatomegalic potencies of diethylhexyl phthalate and ciprofibrate were approximately 200 and 1000-fold greater than di- ethylhexyl adipate, respectively. The researchers found a close relationship between hepatomegalic and peroxisome prolifera- tive effects. Diethylhexyl adipate induced a moderate degree of peroxisome proliferation at the 2% concentration, but not at lower dose levels, where as a greater induction in peroxisome proliferation was observed with diethylhexyl phthalate, and a much greater induction with ciprofibrate. The researchers also stated that the degree of peroxisome proliferation also corresponded with hepatocarcinogenic potential in rodents. Peroxisome induction by diethylhexyl adipate was again determined and compared to a number of phthalate esters, including diethylhexyl phthalate.97 When groups of 2 male and female rats were fed a diet containing ≤2.5% diethylhexyl adipate for 21 days, a marked increase in peroxisome proliferation was observed for males and a moderate increase for females of the 2.5% group. Overall, however, diethylhexyl adipate consistently showed weak activity. The peroxisome proliferation was analyzed using multivariate analysis.98 Diethylhexyl adipate ranked lower than diethylhexyl phthalate, di(isodecyl)phthalate, di(isononyl)- phthalate, and di(n-butyl)phthalate, but greater than di(undecyl)phthalate, butyl benzyl phthalate, di(heptyl, nonyl, undecyl) phthalate, and di(n-hexyl, n-octyl, n-decyl) phthalate. However, in terms of 99.9% statistically predicted dose, diethylhexyl adipate was the least potent, presumably due to a threshold effect. Histological findings of reduced basophilia or increased eosinophilia were highly correlated with peroxisome proliferation. Diethylhexyl Sebacate The effect of diethylhexyl sebacate on hepatic peroxisome proliferation was evaluated in an in vivo study in which 4 male F344 rats were fed chow containing 2% diethylhexyl adipate.58 After 3 wks of dosing, the animals were killed. Hepatic peroxisome proliferation was found, as evidenced by an increase in liver size, hepatic activities of peroxisome-associated enzymes, and hypolipidemia. Relative liver weights were statistically significantly increased in test animals compared to con- trols. (The conclusion regarding the involvement of 2-ethylhexyl alcohol is described earlier under ‘Diethylhexyl Adipate’.) DNA Binding/DNA Synthesis Diethylhexyl adipate did not bind covalently to hepatic DNA in mice. It did stimulate DNA synthesis in livers of rats. In another study, a significant increase in 8-OH-dG was seen in rat liver, but not kidney DNA. The IARC remarked that the weight of evidence for diethylhexyl adipate demonstrated that rodent, peroxisome proliferators do not act as direct DNA-damaging agents.

29

CIR Panel Book Page 77 Diethylhexyl Adipate The potential of diethylhexyl adipate to bind to liver DNA of female NMRI mice was evaluated by administering a solution of 119 mg diethylhexyl adipate/ml with 3.85 mCi/ml of [14C]diethylhexyl adipate (labeled at C1 of the alcohol moiety) and 27.7 mCi/ml of [3H]diethylhexyl adipate (tritiated at positions 2 and 3 of the alcohol moiety) in olive oil.99 The animals were dosed by gavage, and the livers were excised 16 h after dosing. Some animals were pretreated with 10 g/kg of unlabeled dietary diethylhexyl adipate for 4 wks. Diethylhexyl adipate did not covalently bind to hepatic DNA in mice. Pre- treatment with diethylhexyl adipate caused an increase in liver weight, but no increase in DNA binding. The researchers stated that tumorigenicity of diethylhexyl adipate must be due to an activity other than DNA binding. The ability of diethylhexyl adipate to stimulate liver DNA synthesis in male F344 rats was investigated using radio- labeled thymidine.100 Contrary to expected results, diethylhexyl adipate did stimulate DNA synthesis. The stimulation factor, which is indicated by the ratio of the thymidine incorporation in treated animals compared to controls, was 10.5 and the doubling dose, which is the dose that produced a doubling of the control level DNA synthesis, was 0.7 mmol/kg. The effect of dosing with diethylhexyl adipate on 8-hydroxydeoxyguanosine (8-OH-dG) in liver and kidney DNA of rats was examined.101 Groups of 10 male F344 rats were fed a diet containing 0 or 2.5 diethylhexyl adipate. Five animals per group were killed after 1 wk, and the other 5 after 2 wks of dosing. Relative liver weights were statistically significantly increased after 1 and 2 weeks of dosing, and the relative kidney weights were statistically significantly increased only after 2 wks. A statistically significant increase in 8-OH-dG was increased in the liver DNA, but not the kidney DNA, at wk 1 and 2. The IARC remarked that the weight of evidence for diethylhexyl adipate, and other rodent peroxisome proliferators in general, demonstrated that rodent peroxisome proliferators do not act as direct DNA-damaging agents.17 Hepatic Lipid Metabolism Diethylhexyl Adipate Dietary administration of diethylhexyl adipate affects hepatic lipid metabolism.17 Hepatic fatty acid-binding protein and microsomal stearoyl-CoA desaturation were increased in Wistar rats fed 2% diethylhexyl adipate for 7 days.102,103 When fed to rats for 14 days, an increase in hepatic phospholipid levels and a decrease in phosphatidylcholine:phosphatidylethanol- amine ratio was reported.104 In male NZB mice fed 2% diethylhexyl adipate for 5 days, an induction of fatty acid translocase, fatty acid transporter protein, and fatty acid binding protein in the liver was reported.105 Cellular Effects Dibutyl Adipate Dibutyl adipate was tested for cytotoxicity in the metabolic inhibition test. A dilution series of dibutyl adipate was suspended in HeLa cells. Dibutyl adipate had no acute toxicity to the cells, which was attributed to its insolubility in water. From the Amended Final Report on Dibutyl Adipate5

Dibutyl Sebacate The toxic effects of dibutyl sebacate on cultures of human KB cells, monkey Vero cells, and dog MDKC cells was examined.40 Dibutyl sebacate, in ethyl alcohol and diluted in Eagle’s minimum essential medium, caused a dose-dependent inhibition of growth in all three cell types. ANIMAL TOXICOLOGY Acute Toxicity

The oral and dermal LD50 values are greater than 2 g/kg. Mostly, acute exposure via inhalation to diethyl malonate, dibutyl adipate, and diethylhexyl sebacate did not result in death of rats. Acute toxicity data on esters of dicarboxylic acids are presented in Table 8.1,14,40,85,106-117 Data from the original 30

CIR Panel Book Page 78 safety assessments on dibutyl, diisopropyl and diethylhexyl adipate are included in italics.2,5 The acute toxicity of esterase metabolites are also summarized in this table.118,119 Short-Term Oral Toxicity Oral administration of ≤1000 mg/kg dibutyl adipate for 28 days was not toxic effects in rats. In short-term oral dosing with diethylhexyl adipate, decreased weight gain was reported for rats and mice. The NOELs for rats and mice were 2 and 0.63%, respectively, in feed; administration of 10% in feed killed 5/5 female mice. In 2- and 4-wk studies of diethylhexyl adipate, the oral NOAEL for ovarian toxicity was 200 mg/kg in rats; an increase in atresia of the large follicle and a decrease in currently formed corpora lutea were seen in females dosed with 1000 and 2000 mg/kg diethylhexyl adipate. Dibutyl Adipate Male and female Crj:CD(SD) rats, number per group not specified, were dosed orally, by gavage, with 0, 20, 140, or 1000 mg/kg dibutyl adipate in olive oil daily for 28 days.106 No clinical, hematological, or microscopic test-article related changes were observed. Diethylhexyl Adipate In a 14-day dietary study, groups of 5 male rats and mice were given ≤50,000 ppm and groups of 5 female rats and mice were given ≤100,000 ppm diethylhexyl adipate. Male rats and mice fed 50,000 ppm and female rats and mice fed ≥25,000 ppm had decreased weight gains or weight loss. (It is not specified whether the results were statistically significant.) One female rat and all female mice of the 100,000 ppm group died. From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2 Groups of 5 male and 5 female F344 rats were fed a diet containing 0-5.0% (males) or 0-10% (females) diethylhexyl adipate for 14 days, after which time they were killed for necropsy.117 One female of the 10% group died. Weight gain was decreased for male rats of the 5% group and female rats of the 2.5 and 5% groups. Feed consumption was decreased for the animals of the 5 and 10% groups. Females of the 10% group were lethargic, and piloerection, wetness of hair, and ataxia were observed. Microscopically, gray-white casts were observed in the livers one female of both the 2.5 and 5% groups and for 2 females of the 10% group, and atrophy of the spleen in one female of the 10% group. These findings were not considered test-article related. The NOEL was 2.5% and the lowest observable effect level (LOEL) was 5% diethylhexyl adipate.

Groups of 5 male and 5 female B6C3F1 mice were dosed following the same protocol, with the same dose concentrations, as described above. All females of the 10% dose group died, and all animals were lethargic and emaciated prior to death. Feed consumption was decreased in this group. Weight loss was observed for males of the 5% group and female of the 2.5% group. No compound-related microscopic effects were found. The NOEL was 0.63% and the LOEL was 2.5% diethylhexyl adipate. In the 14-day study described in the section on “Peroxisome Proliferation” in which 5 male and 5 female Wistar and

F344 rats and Swiss and B6C3F1 mice were dosed with 0-2.5 g/kg diethylhexyl adipate in corn oil for 14 days, diethylhexyl 92 adipate was toxic to female B6C3F1 mice, causing mortality, at a dose level of 2.5 g/kg. The toxicity of two metabolites of diethylhexyl adipate, 2-ethylhexanol and 2-ethylhexanoic acid, was also examined using Wistar rats and Swiss mice. 2-Ethyl- hexanol was toxic to male and female rats, with mortality reported at doses >1.05 g/kg in male and female rats. 2-Ethylhexa- noic acid was toxic to female rats, with mortality reported at doses ≥1.9 g/kg; mortality was not reported for male rats.. These effects were not reported in mice. In a 1 and 4-wk dietary study described in the section on “Peroxisome Proliferation” in which groups of 5-8 rats and mice were fed diets containing 0-4.0% and 0-2.5% diethylhexyl adipate, respectively, feed consumption by rats was decreased in the 4.0% group at 1 wk and in the 2.5 and 4.0% dose groups at 4 wks.94 Body weights were significantly reduced in these 31

CIR Panel Book Page 79 groups. Feed consumption by mice was not affected, but a significant decrease in body weights was seen in the 1.2 and 2.5% dose groups at 4 wks. Toxicity was evaluated in a study in which groups of 10 female Crl:CD(SD) rats were dosed, by gavage, with 5 ml/kg of 0, 200, 1000, or 2000 mg/kg diethylhexyl adipate in corn oil for 2 or 4 wks.120 All animals survived until study termination. In the 2-wk study, no statistically significant findings were observed for the animals dosed with 200 mg/kg, and the only statistically significant finding in the 1000 mg/kg dose group was an increase in relative liver weight. In the 2000 mg/kg dose group, there was staining around the perineum, statistically significant increases in relative liver and kidney weights, and a statistically significant decrease in the relative weight of the left ovary. Microscopically, abnormal findings were reported for both the ovary and kidney. In the ovary, an increase in atresia of the large follicle and a decrease in current- ly formed corpora lutea were seen in animals dosed with 1000 and 2000 mg/kg, and in the 2000 mg/kg group, an increase in follicular cysts was observed. In the kidney, an eosinophilic change of the proximal tubule was observed for the 2000 mg/kg dose group. The NOAEL was 200 mg/kg. In the rats dosed for 4 wks, similar observations were made. There was staining around the perineum of animals dosed with 1000 and 2000 mg/kg diethylhexyl adipate, and final body weights of animals dosed with 2000 mg/kg were statistically significantly decreased. The relative kidney weights were statistically significantly increased in animals at all dose levels, and liver weights were statistically significantly increased in animals of the 1000 and 2000 mg/kg dose groups. The mean estrous cycle length was statistically significantly decreased in the 200 mg/kg dose group, but this was not considered treatment-related since a dose-response was not seen. The same microscopic abnormalities reported in the 2 wk study were seen in the ovaries and kidneys of the animals dosed with 1000 and 2000 mg/kg in the 4-wk study. As in the 2-wk study, the NOAEL for ovarian toxicity was 200 mg/kg. Short-Term Dermal Toxicity In a short-term dermal study in which 10 rabbits were dosed dermally with 0.5 or 1.0 ml/kg of a 20% dispersion of dibutyl adipate for 6 wks, there was a significant reduction in body weights in the high dose group, and renal lesions in one animal of each group. There were no signs of toxicity in guinea pigs in an immersion study with 20.75% diisopropyl adipate, diluted to an actual concentration of 0.10% adipate. Dermal administration of diethylhexyl adipate to rabbits for 2 wks resulted in slight to moderate erythema at the test site, but toxic effects were not reported for most of the animals. Dibutyl Adipate Groups of 10 rabbits were dosed dermally with 0.5 or 1.0 ml/kg/day of a 20% dispersion of dibutyl adipate, 5x/wk for 6 wks. A significant reduction in body weight gain was seen for animals of the 1.0 ml/kg/day group, and renal lesions were seen in one animal of each group. From the Amended Final Report on Dibutyl Adipate5

Diisopropyl Adipate An immersion study was performed using guinea pigs in which a product containing 20.75% diisopropyl adipate was diluted, giving an actual adipate concentration of 0.10%. The animals were immersed 4 h/day for 3 days. There were no signs of systemic toxicity, and the degree of dermal irritation was considered minimal. From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2

Diethylhexyl Adipate Diethylhexyl adipate, 410 or 2060 mg/kg, was applied to the shaved abdomen of male rabbits, 4 per group, 5 days per wk for 2 wks.117 Mineral oil was applied in the same manner to a group of 4 rabbits as a negative control. A collar was used to restrict ingestion. One animal in the 410 mg/kg group died during wk 2 of the study. All other animals in this group appeared normal. Slight to moderate erythema was observed at the test site. No animals of the 2060 mg/kg group died, but 3 32

CIR Panel Book Page 80 of the 4 did not gain weight, and they had labored breathing and were lethargic during wk 2. Moderate erythema was observed in this group. Microscopically, one animal of the 2060 mg/kg group had altered cytology of the liver parenchymal cells. No other microscopic lesions were noted. Diethylhexyl Sebacate No deaths occurred when 4 rats, 2 guinea pigs, 2 rabbits and 1 cat were exposed to 400 mg diethylhexyl sebacate/m3, 7 hrs/day, for 10 days.1 Details were not provided. Subchronic Oral Toxicity In a subchronic oral toxicity study using rats, the NOEL for diethyl malonate was 1000 mg/kg/day. Dietary administration of ≤2.5% di-C7-9 branched and linear alkyl esters of adipic acid (approx. 1500 and 1900 mg/kg/day for males and females, respectively) for 90 days did not result in systemic toxicity. The NOAELS for male and female rats were 1500 and 1950 mg/kg/day, respectively. Subchronic oral administration of diethyl- hexyl adipate to rats caused significant decreases in body weight gains and increases in liver and kidney weights. The dietary NOEL for rats in a 90-day study was 610 mg/kg. In mice, no lesions were induced by dietary administration of ≤25,000 ppm diethylhexyl adipate for 13 wks. A decrease in body weights was seen in mice fed a diet with 1.2 and 2.5% diethylhexyl adipate. For diisononyl adipate, dietary administration of up to 500 mg/kg for 13 wks, a statistically significant increase in relative kidney weights was reported, but there were not toxicological findings. With dogs, 3.0% dietary diisononyl adipate resulted in a decrease in body weights, testes weight, and feed consumption, increased liver weight, elevated enzyme levels, liver and kidney discolora- tion, and microscopic changes in the liver, testes, spleen, and kidneys. Diethyl Malonate Groups of 10-16 male and female CD rats were fed diets containing either 0, 36 (males) or 41 mg/kg/day (females) diethyl malonate.14 No treatment related effects were observed, and the NOEL was 1000 mg/kg/day. Di-C7-9 Branched and Linear Alkyl Esters of Adipic Acid Groups of 15 male and 15 female Sprague-Dawley rats were fed a diet containing 0, 0.1, 0.5, or 2.5% di-C7-9 branched and linear alkyl esters of adipic acid for 90 days, corresponding to approximately 1500 mg/kg/day for high dose males and 1900 mg/kg/day for high dose females.85 All rats were killed for necropsy at study termination. No systemic toxicity was reported. Small, but significant, increases in absolute and relative kidney weights reported for females of the 2.5% dose group were not considered treatment-related. The NOAELs for male and female rats were 1500 and 1950 mg/kg/day, respectively. Diethylhexyl Adipate In a 13-wk dietary study, groups of 10 rats and 10 mice were fed ≤25,000 ppm diethylhexyl adipate. With the exception of decreased weight gain for some of the groups, no compound-related toxicologic effects were observed. From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2 In a 90-day dietary study, groups of 10 rats per group were fed 0-4740 mg/kg diethylhexyl adipate for 90 days.117 Mortality occurred in the 4740 mg/kg group, but the number of deaths was not specified. Decreased growth and feed con- sumption was reported for animals fed 2920 mg/kg. Changes in kidney and liver weights were noted, but no details were given. The NOEL was 610 mg/kg, and the LOEL was 2920 mg/kg diethylhexyl adipate. Groups of 10 male and 10 female F344 rats and B6C3F1 mice were fed a diet containing 0-25,000 ppm for 13 wks, after which time the surviving animals were necropsied.121 No compound-related deaths occurred. No lesions were induced by administration of ≤25,000 ppm diethylhexyl adipate. Groups of 15 male and 15 female Sprague Dawley rats were fed 0 or 2.5% diethylhexyl adipate for 90 days.85 At study termination, all animals were killed for necropsy. Body weight gains were statistically significantly decreased for treated males and females, and relative kidney and liver weights were statistically significantly increased for treated females,

33

CIR Panel Book Page 81 when compared to controls. In a 13-wk dietary study described in the section on “Peroxisome Proliferation” in which groups of 5-8 rats and mice were fed diets containing 0-4.0% and 0-2.5% diethylhexyl adipate, feed consumption by rats was decreased in the 2.5 and 4.0% dose groups, and body weights were significantly reduced in these groups.94 Feed consumption by mice was not affected, but a significant decrease in body weights was seen in the 1.2 and 2.5% dose groups Diisononyl Adipate Groups of 10 male and 10 female rats were fed 0, 50, 150, or 500 mg/kg diisononyl adipate for 13 wks.85 A statistically significant increase in relative kidney weights was reported for males and females given 500 mg/kg, but absolute kidney weights were not affected and no significant microscopic effects were seen. Microscopic changes in any of the organs, including the testes and epididymis of males and ovaries of females, were not observed. There were not significant toxicological findings, and the NOAEL was 500 mg/kg/day. In another 13-wk study, groups of 4 male and 4 female beagle dogs were fed 0, 0.3, 1.0, or 3.0% diisononyl adipate; the high dose was increased to 6% during wks 9-13. No significant findings were reported for the 0.3 or 1.0% groups. In the high dose group, decreased body weight, testes weight, and feed consumption, increased liver weight, elevated enzyme levels, liver and kidney discoloration, and microscopic changes in the liver, testes, spleen, and kidneys were reported. The dietary NOAEL for diisononyl adipate was 1.0%. Subchronic Dermal Toxicity No adverse effects were reported with whole-body application of an 6.25% emulsion of dibutyl adipate to dogs for 3 mos. Unoccluded dermal application of up to 2000 mg/kg ditridecyl adipate for 13 wks to rats produced slight erythema, but no systemic toxicity. Dibutyl Adipate No adverse effects were reported in a study using 4 dogs in which entire-body applications of an emulsion containing 6.25% dibutyl adipate were made 2x/wk for 3 mos. From the Amended Final Report on Dibutyl Adipate5

Ditridecyl Adipate Ditridecyl adipate, 0, 800, or 2000 mg/kg, was applied to the backs of groups of 10 male and 10 female Sprague- Dawley rats, 5 days/wk for 13 wks.85 The test sites were not occluded, but the animals wore Elizabethan collars. Slight erythema and flaking of the skin was observed in the treated groups, with hyperplasia of the sebaceous glands in the dermis, but otherwise no significant differences were observed between test and control animals. Differences in relative organ weights were not statistically significant, and ditridecyl adipate did not appear to cause systemic toxicity. Subchronic Inhalation Toxicity Diethylhexyl Sebacate Groups of 12 F344 rats, gender not specified, were exposed 4 h/day, 5 days/wk, to 25 or 250 mg/m3 diethylhexyl sebacate for ≤13 wks.85 No adverse systemic or lung effects were observed. Chronic Oral Toxicity In a 6-mos study in which rats were dosed intragastrically with ≤2.0 g/kg diethylhexyl adipate, hepatic detoxification appeared depressed at the beginning of the study, while in a 10-mos study, a decrease in central nervous system excitability was noted. Dietary administration of ≤1.25% dibutyl sebacate for 1 yr or ≤6.25% for 2 yrs did not have an effect on growth Diethylhexyl Adipate Intragastric doses of ≤2.0 g/kg diethylhexyl adipate to rats (number not stated) for 6 mos produced no enzy- matic changes, but levels of sulphydryl compounds in the blood were increased. Hepatic detoxification 34

CIR Panel Book Page 82 appeared depressed at the onset of the study, but it was accelerated after 6 mos. Administration of 0.1 g/kg for 10 mos decreased central nervous system excitability. From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2

Dibutyl Sebacate Groups of 5 male and 5 female Sprague-Dawley rats were fed a diet containing 0, 0.01, 0.05, 0.25, or 1.25% dibutyl sebacate for 1 yr.116 Necropsies were performed whenever rats exhibited significant weight losses or other evidences of severe concurrent infection. Dibutyl sebacate had no effect on growth or well-being. The researches then fed groups of 16 male Sprague Dawley rats a diet containing 0.01, 0.05, 0.25, 1.25, or 6.25% dibutyl sebacate for 2 yrs.116 Two control groups were given untreated feed. Necropsies were performed on 3 rats from each group after 1yr, and the experiment was terminated at the end of the 2-year feeding period. Interim, animals were killed whenever they became moribund. In such instances the rats usually had incapacitating tumors or severe intercurrent infec- tions. Dibutyl sebacate did not adversely affect growth or survival, and it did not produce significant hematological changes in peripheral blood. As the rats increased in age, slight changes in distribution of leukocytes were found, but these trends occurred in both the control and treatment groups. Inhalation Toxicity In a 4-hr inhalation toxicity study of 5.9 mg/l of a mixture of 66.0% dimethyl glutarate, 16.5% dimethyl succinate, and 17.0% dimethyl adipate, the anterior and posterior nasal passageways were affected. Regeneration of damaged olfactory epithelium was related to the severity of the initial tissue damage. Dimethyl Glutarate/Dimethyl Succinate/Dimethyl Adipate Mixture Male Crl:Cd/BR rats were used to study the development of nasal lesions upon exposure to a solvent mixture of 66.0% dimethyl glutarate, 16.5% dimethyl succinate, and 17.0% dimethyl adipate, termed dibasic esters.122 A group of 42 rats were exposed, nose-only, to dibasic esters at an aerosol concentration of 5.9 mg/l for 4 hr, and a negative control group of 24 rats was exposed to air only. The mean dibasic esters aerosol concentration was 5900 mg/m3, and the mass median aero- dynamic diameter was 3.9 µm. Seven test animals and 4 controls were killed at 1, 4, 7, 14, 21, or 42 days post-exposure. Three test and 3 control animals were injected i.p. with BrdU 2 h prior to being killed. Both the anterior and posterior nasal passages were affected. Nasal lesions were distributed along major inspiratory airflow routes, and the lesions were markedly less severe in the posterior nasal cavity. Regeneration of damaged olfactory epithelium was related to the severity of the initial tissue damage, with extensively damaged epithelium failing to regain a normal structure at 6 wks. Numerous mitotic figures and BrdU labeling were found in the regenerating basal cells, stem cells, and sustentacular cells at 4 and 7 days. Ocular Irritation Ocular irritation appeared to lessen in severity as chain length of the dicarboxylic acid esters increased. Diethyl malonate was slightly to moderately irritating to rabbit eyes. Dibutyl, diisopropyl, and diethylhexyl adipate were non- or minimal ocular irritants. Diisopropyl sebacate was minimally irritating, while diethylhexyl sebacate was non-irritating to rabbit eyes. Dioctyldodecyl and diisocetyl dodecanedioate were not irritating to rabbit eyes. Ocular irritation data on esters of dicarboxylic acids are presented in Table 9. Data from the original safety assess- ments on dibutyl, diisopropyl and diethylhexyl adipate are included in italics. The available ocular irritation data on esterase metabolites are also summarized in this table. Diethyl Malonate The ocular irritation potential of diethyl malonate was evaluated using rabbits, number and gender not specified.14 A volume of 0.1 ml was instilled into the conjunctival sac of one eye, which was not rinsed, and the contralateral eye was untreated and served as the negative control. Diethyl malonate produced slight to moderate irritation. 35

CIR Panel Book Page 83 Dimethyl Malonate In a similar study as described above, undiluted dimethyl malonate produced slight to moderate irritation in rabbit eyes.14 All signs of irritation were cleared by day 8. Dibutyl Adipate Undiluted dibutyl adipate was minimally irritating to they eyes of rabbits, and 0.1% in olive oil was non- irritating. From the Amended Final Report on Dibutyl Adipate5

Diisopropyl Adipate The ocular irritation potential of 2 lots of undiluted diisopropyl adipate was evaluated using rabbits. One caused negligible irritation, while the other was non-irritating. A formulation containing 0.7% diisopropyl adipate produced some corneal stippling in rabbit eyes, while a formulation containing 5.0% and one containing 20.75% were non-irritating to rabbit eyes. From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2

The ocular irritation of undiluted diisopropyl adipate was evaluated using 3 albino rabbits.114 A volume of 0.1 ml was instilled into the conjunctival sac of one eye, which was not rinsed. The contralateral eye was untreated and served as the negative control. Diisopropyl adipate was not irritating. Diethylhexyl Adipate Undiluted diethylhexyl adipate was non-irritating to rabbit eyes and a formulation containing 0.0175 was, at most, a mild transient irritant. From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2

Diisopropyl Sebacate A primary ocular irritation study was performed using 6 New Zealand white to determine the ocular irritation potential of diisopropyl sebacate.123 A volume o f 0.1 ml was applied to one eye of each animal, which was not rinsed, and the contralateral eye of each animal served as the control. The average Draize scores were 2.0 at 24 and 48 h, 0.3 at 72 h, and 0.0 at 4 days. Diisopropyl sebacate was a minimal ocular irritant. Diethylhexyl Sebacate The ocular irritation o f a cream containing 1.2% diethylhexyl sebacate was evaluated using the in vitro EpiOcular MTT viability assay .124 The tissue samples were exposed to undiluted test material for 64 min, 256 min, or 1200 min.

Following treatment, the viability of those tissues were calculated. The ET50 (time for tissue viability to be reduced by 50%) was 484.9 min, and diethylhexyl sebacate was considered to be non-irritating. Dioctyldodecyl Dodecanedioate The primary eye irritation of dioctyldodecyl dodecanedioate was evaluated using 6 albino rabbits.125 A volume of 0.1 ml was applied to one eye of each animal, which was not rinsed. and the contralateral eye served as a negative control. They eyes were evaluated at 24, 48, and 72 h. At 24 h, the maximum mean total score (MMTS) was 0.00, and dioctyldodecyl dodecanedioate was considered not irritating. Diisocetyl Dodecanedioate The primary eye irritation of diisocetyl dodecanedioate was evaluated using the procedure described above.126 The MMTS was 0.00, and diisocetyl dodecanedioate was considered not irritating to the eyes of rabbits. Dermal Irritation The esters of dicarboxylic acids were mostly non or mildly irritating to rabbits. Some minimal irritation was seen with diisopropyl adipate, undiluted or at 5-20.75% in formulation, and moderate erythema was reported with undiluted dibutyl adipate.

36

CIR Panel Book Page 84 Diethyl Malonate The dermal irritation potential of diethyl malonate was evaluated using a 24 h occlusive application.14 Diethyl malonate was slightly irritating to rabbit skin. Dimethyl Malonate Dimethyl malonate was applied undiluted to rabbit skin for 4 h under a semi-occlusive patch.14 Slight erythema was observed only at 30-60 min after patch removal, and dimethyl malonate was considered non-irritating to rabbit skin. Dibutyl Adipate Application of undiluted butyl adipate to rabbit skin resulted in a primary irritation score of 2/8. Undiluted dibutyl adipate caused moderate erythema in rabbits following repeated dermal exposure. However, material impregnated with dibutyl adipate was not irritating to the skin of rabbits. Application of dibutyl adipate at 10% in acetone produced no observable adverse effect when applied to rabbit ears, and no dermal reaction was observed following twice daily application for 14 days to the backs of hairless mice Two perfume formulations containing 1.1% diisopropyl adipate were not primary dermal irritants using rabbits. From the Amended Final Report on Dibutyl Adipate5

Diisopropyl Adipate Draize tests of undiluted diisopropyl adipate resulted in, at most, mild irritation of rabbit skin. In Draize tests with formulations containing 5.0% or 20.75% diisopropyl adipate, minimal irritation was reported with both formulations. From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2

Diethylhexyl Adipate Undiluted diethylhexyl adipate was a very mild irritant when applied under occlusion to intact and abraded rabbit skin). A formulation containing 0.175% diethylhexyl adipate had an irritation index of 1.6/4. From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2

Diisodecyl Adipate The dermal irritation potential of diisodecyl adipate was determined using 3 albino rabbits.112 Undiluted test material was applied to the skin for 4 h under a semi-occlusive patch. The erythema scores were 0-1 during 1-72 h, and the edema scores were 0. Diisodecyl adipate was considered non-irritating to rabbit skin. Dioctyldodecyl Adipate The dermal irritation of dioctyldodecyl adipate was evaluated using the same procedure.113 The erythema scores were 0-2 during 1-72 h, and the edema scores were 0-1. Dioctyldodecyl adipate was considered non-irritating to rabbit skin. Diisocetyl Adipate The dermal irritation of diisocetyl adipate was evaluated using the same procedure.111 The erythema scores were 0-2 during 1-72 h, and the edema scores were 0-1. Diisocetyl adipate was considered non-irritating to rabbit skin. Diethyl Sebacate Undiluted diethyl sebacate and 30% diethyl sebacate in ethanol were tested on 8 male Japanese White strain rabbits (gender not specified).127 The flank of the animals was clipped free of hair 1 day prior to application of test substance. The skin of 4 animals abraded. The test substance, 0.3 ml, was applied occlusively to the back of all animals for 24 h. The skin reactions were evaluated at 24 h and 72 h. The primary irritation score was 0.0 (none to weak irritant) in undiluted diethyl sebacate and 0.3 (none to weak irritant) in 30% diethyl sebacate. These results suggest that 100% diethyl sebacate has no primary skin irritation under these test conditions. Diisopropyl Sebacate A primary dermal irritation study on diisopropyl sebacate was performed using 6 New Zealand white rabbits.123 A dermal application of 0.5 ml of undiluted test material was applied to an abraded and an intact site on each animal. The test

37

CIR Panel Book Page 85 sites were occluded for 24 h and observed individually for erythema, edema, and other effects 24 and 72 h after application. Mean scores from the 24 and 72 h reading were averaged to give a primary irritation index (PII) of 2.88. Diisopropyl sebacate was not considered a primary dermal irritant. The dermal irritation potential of diisopropyl sebacate was determined using 3 albino rabbits.128 Undiluted test material was applied to the skin for 4 h under a semi-occlusive patch. The erythema scores were 1 during 1-72 h, and the edema scores were 0-1. Diisopropyl sebacate was considered non-irritating to rabbit skin. Diethylhexyl Sebacate The dermal irritation potential of diethylhexyl sebacate was evaluated using the same procedure.110 The erythema scores were 1 during 1-72 h, and the edema scores were 0. Diethylhexyl sebacate was considered non-irritating to rabbit skin. Patch tests with diethylhexyl sebacate (neat; 48-hr occluded) did not irritate the skin of 2-4 rabbits.1 It was also reported that diethylhexyl sebacate was non-irritating to the skin of guinea pigs. No further study details were provided Dermal Sensitization Dimethyl malonate, dibutyl and diethylhexyl adipate, diethylhexyl sebacate, and dioctyldodecyl dodecanedioate were not sensitizers in guinea pigs or rabbits. Dimethyl Malonate Dimethyl malonate was not a sensitizer in a Buehler guinea pig sensitization test.14 Details were not provided. Dibutyl Adipate Dibutyl adipate was not a dermal sensitizer in guinea pigs when tested at 25% in a maximization test. From the Amended Final Report on Dibutyl Adipate5

Diethylhexyl Adipate Diethylhexyl adipate, 0.1%, was not a sensitizer in a maximization study using guinea pigs. From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2

Diethylhexyl Sebacate A limited attempt was made to sensitize a group of 2-4 rabbits by applying diethylhexyl sebacate using occlusive patches.1 No reactions were seen in an occlusive challenge with the undiluted test article 2 weeks later. Details were not provided. Dioctyldodecyl Dodecanedioate A maximization test was performed to evaluate the sensitization potential of dioctyldodecyl dodecanedioate.129 Ten female guinea pigs were used. The dose used at intradermal injection was 0.1 ml, and 0.5 ml was used for the topical challenge. Slight erythema was observed at induction, but a sensitization reaction was not observed. Dermal irritation and sensitization data on esters of dicarboxylic acids are presented in Table 10. Data from the original safety assessments on dibutyl, diisopropyl and diethylhexyl adipate are included in italics. The available dermal irritation and sensitization data on esterase metabolites are also summarized in this table. Phototoxicity Diisopropyl Adipate Two perfume formulations containing 1.1% diisopropyl adipate were not phototoxic using rabbits. From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2

Mucous Membrane Irritation Diethylhexyl Adipate A product containing 0.175% diethylhexyl adipate did not produce mucous membrane irritation in rabbits.

38

CIR Panel Book Page 86 From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2

REPRODUCTIVE AND DEVELOPMENTAL TOXICITY Oral administration of up to 1000 mg/kg dimethyl malonate did not have an effect on fertility, and no development toxicity was reported. The NOAEL was 300 mg/kg for repeated dose and maternal toxicity and 1000 mg/kg for fertility and developmental toxicity. Oral administration of up to 100 mg/kg dibutyl adipate did not cause any reproductive effects, and the NOEL for parental and offspring toxicity was 300 mg/kg/day and for reproductive toxicity was 100 mg/kg/day. Oral administration of ≤7000 mg/kg di-C7-9 branched and linear alkyl esters of adipic acid branched and linear alkyl esters of adipic acid did not result in developmental toxicity. Dietary administration of up to 1.2% diethylhexyl adipate did not affect fertility when fed to rats prior to mating. Fetal weight, total litter weight, and litter size were reduced with 1.2% diethylhexyl adipate. In a study in which gravid rats were fed the same doses during gestation, no significant effects on fetal weight or litter size were reported. An increased incidence of minor skeletal abnormalities was attributed to fetotoxicity. . In a study in which diethylhexyl adipate was given orally to rats from day 7 of gestation until post-natal day 17, antiandrogenic effects were not observed, although some increase in post-natal death were observed. Administration of up to 2000 mg/kg diethylhexyl adipate prior to dosing and through day 7 of gestation did have an effect on the mean estrus cycle length at a dose of 1000 and 2000 mg/kg, and did appear to disturb ovulation. Significant decreases were also seen in implantation rate and number of live embryos, as well as an increase in pre-implantation loss. Diethylhexyl adipate did not produce testicular toxic effects when fed at up to 25,000 ppm in the diet for 4 wks. Dietary administration of 6.25% dibutyl sebacate to male and female for 10 wks prior to mating had no adverse effects on fertility, litter size, or survival of offspring. Diethylhexyl sebacate, 200 ppm in the diet, did not produce reproductive or developmental effects. Dermal applications of up to 2000 mg/kg ditridecyl adipate affected mean fetal body weights and crown-rump lengths. Ditridecyl adipate, 2000 mg/kg, did not have an effect on sperm morphology. Some visceral anomalies were reported. The NOAELs for maternal toxicity and developmental and reproductive effects were 2000 and 800 mg/kg/day, respectively. Dimethyl, diethyl, dipropyl dibutyl, diisobutyl, and diethylhexyl adipate were evaluated for fetotoxic and terato- genic effects in rats when administered i.p. at 1/3 – 1/30 of the i.p. LD50 values. Some effect on resorptions and abnormalities were seen with all but diethyl adipate. Inhalation by rats of up to 1.0 mg/l of a mixture of dimethyl glutarate, dimethyl succinate, and dimethyl adipate on days 7-16 of gestation or for 14 days prior to mating, during mating and gestation, and lactation, produced no observed adverse developmental or reproductive effects. The only exception was a statistically significant decrease in pup weight at birth and day 21. Dimethyl Malonate The reproductive and developmental toxicity of dimethyl malonate was evaluated using groups of 10 male and 10 female Wistar rats.14 The animals were dosed with 0, 100, 300, or 1000 mg/kg dimethyl malonate orally, by gavage. Males were dosed for 2 wks prior to mating, during mating, and 2 wks after mating, for a total of 39 doses. Females were dosed 2 weeks prior to mating, during mating, and through day 4 of lactation. A recovery group of 5 male and 5 female high dose animals were observed for 14 days after the termination of dosing. Microscopically, the incidence of treatment-related hepatocellular hypertrophy of the liver was observed for males and females given 1000 mg/kg dimethyl malonate. This effect was not observed in the recovery animals or in the other test groups. No other significant toxicological effects were observed. Performance in a functional observation battery was similar for test and control animals. There was no effect on fertility. An increase in post-implantation loss was increased in the 100 mg/kg group, resulting in a statistically significant decrease in the number of live pups. This effect was not considered treatment related, and no developmental toxicity was reported. The NOAEL was 300 mg/kg for repeated dose and maternal toxicity and 1000 mg/kg for fertility and developmental toxicity. Dimethyl Adipate The fetotoxic and teratogenic effects of dimethyl adipate were evaluated in a study in which groups of 5 gravid

Sprague Dawley rats were dosed i.p. with 0.0603-0.6028 ml/kg (1/30, 1/10, 1/5, and 1/3 of the i.p. LD50 value) on days 5, 10, and 15 of gestation.115 A pooled volume control consisted of animals dosed with 10 ml/kg distilled water, saline, or cotton- 39

CIR Panel Book Page 87 seed oil. A positive control group was not used. All animals were killed and examined on day 20 of gestation. The mean fetal weights and the numbers of live fetuses were not statistically significantly different between treated and blunt-needle control groups. Resorptions in animals dosed with 0.1809 ml/kg were statistically significantly increased when compared to the pooled controls, but not the blunt-needle controls. Gross and skeletal abnormalities, but not visceral, were statistically significantly increased in fetuses of the 0.3617 and 0.6028 ml/kg groups. Diethyl Adipate The fetotoxic and teratogenic effects of diethyl adipate were evaluated following the same procedure described above.115 These rats were dosed i.p. with 0.0837-0.8373 ml/kg diethyl adipate. The mean fetal weight and the number of live fetuses were not statistically significantly different between treated and blunt-needle control groups, and the number of resorp- tions was similar between treated animals and both the blunt needle and pooled controls. There were no differences in the incidences of gross, skeletal, or visceral abnormalities in fetuses of the treated groups compared to pooled controls. Dipropyl Adipate The fetotoxic and teratogenic effects of dipropyl adipate were evaluated following the same procedure described earlier.115 These rats were dosed i.p. with 0.1262-1.2619 ml/kg dipropyl adipate. The numbers of live and dead fetuses were not statistically significantly different between treated and blunt-needle control groups, but there was a statistically significant decrease in the mean fetal weight of the 0.7572 ml/kg group. Resorptions in animals dosed with 1.2619 ml/kg were statisti- cally significantly increased when compared to the pooled controls, but not the blunt-needle controls. Gross abnormalities, but not skeletal or visceral, were statistically significantly increased in fetuses of the 1.2619 ml/kg group. Dibutyl Adipate A reproductive toxicity study was performed in which groups of 5 gravid Sprague Dawley rats were dosed i.p. with 0.1748-1.7480 ml/kg dibutyl adipate on days 5, 10, and 15 of gestation. The incidence of gross abnormalities was only statistically significantly increased in the high dose group when compared to pooled controls. From the Amended Final Report on Dibutyl Adipate5

The reproductive toxicity of dibutyl adipate was evaluated in a study Sprague-Dawley rats.106 Groups of 13 males and 13 female’s rats were dosed with 0, 100, 300, or 1000 mg/kg dibutyl adipate orally, by gavage, for 14 days prior to mating through parturition; males were dosed for a total of 42 days and female dams were dosed until day 3 of lactation. The test article had no effect on fertility. Body weight gains of males of the 1000 mg/kg group were slightly decreased. Kidney weights of the high dose males and females sere increased compared to controls. No gross or microscopic effects were noted at necropsy, and the internal genitalia were normal. Dosing with dibutyl adipate did not produce any reproductive effects. The only effect on the offspring was a decrease in pup weight on post-natal days 0 and 4 and in viability on post-natal day 4. The NOEL for parental and offspring toxicity was 300 mg/kg/day. The reproductive NOEL was 1000 mg/kg/day. Di-C7-9 Branched and Linear Alkyl Esters of Adipic Acid Groups of 24 gravid Sprague Dawley rats were dosed orally by gavage with 0, 1000, 4000, or 7000 mg/kg/day di- C7-9 branched and linear alkyl esters of adipic acid on days 6-19 of gestation, and all animals were killed and examined on day 20.85 All dams survived until study termination. Body weights were significantly decreased for dams of the 7000 mg/kg group. Weights of male and female fetuses of the 7000 mg/kg group were slightly, but not statistically significantly, decreased compared to the other groups. A greater incidence of rudimentary structures was observed for high dose fetuses as compared to the other groups in this study, but the incidence was within the range of historical controls. There was no evidence of developmental toxicity at any dose tested.

40

CIR Panel Book Page 88 Ditridecyl Adipate The reproductive and developmental toxicity of ditridecyl adipate was evaluated using groups of 15 mated female Sprague-Dawley rats.85 Doses of 0, 800, and 2000 mg/kg were applied dermally without occlusion on days 0-19 of gestation, and the dams were killed on day 20. Mild skin irritation consisting of erythema and flaking were observed at the test sites of the treated animals. No maternal mortality was reported. Weight gains were statistically significantly decreased for the 2000 mg/kg group during days 0-3 and 16-20 of gestation. Weight gain was statistically significantly decreased in the 800 mg/kg group during days 0-3 of gestation. Mean fetal body weights and crown-rump lengths were affected exposure to the test article. No differences in skeletal anomalies were observed, but there were some differences in visceral anomalies, including increased incidence of levocardia at 2000 mg/kg. These anomalies were not considered treatment-related. The NOAEL for maternal toxicity was 2000 mg/kg/day, and for developmental and reproductive effects it was 800 mg/kg/day. Groups of 25 mated female rats were dosed dermally with 0 and 2000 mg/kg ditridecyl adipate following the same study protocol as above. Again, there were no signs of maternal toxicity. No developmental toxicity was reported, and there were no visceral anomalies. Tridecyl adipate, 2000 mg/kg, was applied, unoccluded, to groups of 10 male Sprague-Dawley rats, 5 days/wk for 13 wks, and the effect on sperm morphology was evaluated.85 (The 'Subchronic Dermal Toxicity’ study was described earlier.) No differences in sperm morphology were observed between control and test animals. Diisobutyl Adipate The fetotoxic and teratogenic effects of diisobutyl adipate were evaluated following the procedure described in the earlier i.p. study.115 These rats were dosed i.p. with 0.1983-1.9833 ml/kg diisobutyl adipate. The numbers of live and dead fetuses were not statistically significantly different between treated and blunt-needle control groups, but there was a statistical- ly significant decrease in the mean fetal weight of the 1.1900 and 0.9833ml/kg dose groups. The number of resorptions was similar between treated animals and both the blunt needle and pooled controls. Gross abnormalities, but not skeletal or vis- ceral, were statistically significantly increased in fetuses of the 0.5950 and 1.9833 ml/kg groups. Diethylhexyl Adipate The reproductive effects of diethylhexyl adipate were studied in Swiss mice. Groups of 10 male mice were dosed i.p. with ≤9.3 g/kg and then mated with undosed females. A reduction in the number of gravid females was considered an anti-fertility effect, and the dominant lethal mutation was determined directly from the dose-dependent increase in the number of early fetal deaths and indirectly from the dose- and time- dependent decrease in implantations. There were no test article-related changes in the incidence of late fetal deaths. It was noted that the experimental design and interpretation have been questioned by some. Diethylhexyl adipate, ≤9.3 g/kg, was administered by i.p. injection to groups of 5 gravid Sprague Dawley rats on day 5, 10, and 15 of gestation. Resorption rates were similar to controls. A decrease in the mean fetal body weight and a significant increase in gross fetal abnormalities at the high dose were observed when compared to pooled control values. However teratogenic effects were not observed when compared to concurrent controls. It was stated that the lack of historical and positive controls affected the validity of the results. From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2

Groups of 15 male and 30 female Wistar rats were fed a diet containing 0, 0.03, 0.18, or 1.2% diethylhexyl adipate (calculated as 28, 170, or 1080 mg/kg/day) for 10 weeks prior to mating.85 Dosing was terminated, and the animals were mated. (A different source indicated that dosing continued throughout the study.130) A reduction in body weight gain was reported during gestation for the dams of the 1.2% group. No test article-related effects on fertility were observed. Fetal weight, total litter weight, and litter size were reduced in the 1.2% group, but the number of pups born live, or their survival, was not affected. The NOAEL was 170 mg/kg/day and the LOAEL was 1080 mg/kg/day.

41

CIR Panel Book Page 89 In another study in which gravid females were fed the same doses as above on days 1-22 of gestation, maternal body weight and feed consumption were statistically decreased in the 1.2% group. No significant effects on fetal weight or litter size were reported. Animals of the 0.18 and 1.2% groups had slightly increased incidences of minor skeletal abnormalities; this increase was attributed to fetotoxicity. The NOEL for maternal toxicity was 170 mg/kg/day. The NOAELs for develop- mental toxicity and fetotoxicity were 170 and 28 mg/kg/day, respectively. The LOAEL was 1080 mg/kg day. A dose-range finding study was performed using groups of 8 gravid Wistar rats that were dosed by gavage with 2 ml/kg of 0, 800, or 1200 mg/day diethylhexyl adipate, in peanut oil, from day 7 of gestation until day 17 after parturition.131 No signs of toxicity were reported in any of the groups. In the 800 mg/kg group, the only statistically significant observation made was decreased body weights of male and female pups on day 3. In the 1200 mg/kg group, statistically significant effects were observed for a number of parameters, including decreased maternal weight gain during days 7-21 of gestation, increased length of gestation (by 1 day), decreased pup body weights at birth and day 3, and an increase in perinatal loss per litter. (Perinatal loss was 42% in the 1200 mg/kg groups, as compared to 4.6% in controls.) Based on the results of the dose-range finding study, groups of 20 gravid Wistar rats were dosed with 2 ml/kg of 0, 200, 400, or 800 mg/kg diethylhexyl adipate, in peanut oil, from day 7 of gestation until post-natal day 17. At postnatal day 21, all dams and pups were killed, with the exception that one male and one female pup per litter was kept for further evalua- tion. No signs of toxicity were reported in any of the groups. No significant effects were observed in the 200 mg/kg group. In the 400 mg/kg dose groups, the number of postnatal deaths per number of pups was statistically significant increased. In the 800 mg/kg group, statistically significant effects were observed for a number of parameters, including increased length of gestation (by 1 day), decreased pup body weights at birth and days 3 and 13, increased mean number of postnatal deaths, and an increase in postnatal death per number of pups. The percentage of perinatal loss per litter was twice as high in the 400 and 800 mg/kg groups (23%) as compared to controls (11%), but the change was not statistically significant. Testicular testoster- one levels were unaffected in any of the pups that were killed on postnatal day 21 or the adult male offspring, and all other hormones that were measured were similar to controls. None of the sperm parameters that were evaluated were affected by dosing. The only statistically significant effects, noted in the 800 mg/kg group, were increased relative liver weights in male pups on day 21 and increased body weights and decreased adrenal weights in adult male offspring. Diethylhexyl adipate did not produce any antiandrogenic effects in the study. Fetal steroidogenesis was not evaluated. NOAEL was 200 mg/kg. Groups of 10 female Crl:CD(SD) rats were dosed with 5 ml/kg, by gavage, of 0, 200, 1000, or 2000 mg/kg diethyl- hexyl adipate in corn oil for 2 wks prior to mating with undosed males, throughout mating, and until day 7 of gestation day.120 The dams were killed on day 14 of gestation. All animals survived until study termination. Body weights and body weight gains were significantly decreased in the 2000 mg/kg dose group prior to mating. Staining around the perineum was observed in the 1000 and 2000 mg/kg dose groups. No statistically significant differences were observed for the 200 mg/kg group compared to controls. The mean estrus cycle length was statistically significantly increased in the 1000 and 2000 mg/kg groups, and the post-implantation loss rate was also statistically significantly increased in these groups. Additionally, in the 2000 mg/kg group, there was a significant decrease in implantation rate, and the number of live embryos was statistically significantly decreased and the pre-implantation loss rate statistically significantly increased. The researchers stated that the effects observed in this fertility study, in conjunction with the ovarian effects described earlier in the repeated dose study, suggest that diethylhexyl adipate disturbed ovulation. This correlated with the effect on estrus cycle length. The testicular toxicity of diethylhexyl adipate was examined using male F344 rats.132 Groups of six rats were fed a diet containing 6000 or 25,000 ppm diethylhexyl adipate for 4 wks, and the controls were given untreated feed. Some groups

42

CIR Panel Book Page 90 were dosed i.p. with 200 mg/kg thioacetamide, 3x/wk for 4 wks, and prior to dosing with diethylhexyl adipate to evaluate whether liver disease enhanced testicular effects. (There was a 1-wk rest period prior to dosing with diethylhexyl adipate.) The final body weights of animals given 25,000 ppm diethylhexyl adipate, with and without prior administration of thioceta- mide, were statistically significantly decreased compared to their respective controls. The relative liver weights of these animals were statistically significantly increased. No significant effect on the relative weights of the testes or epididymis were seen for any of the test groups. Diethylhexyl adipate did not have any testicular toxic effects, with or without the induction of hepatic damage. Dibutyl Sebacate A test group of 20 male and 20 female Sprague-Dawley rats was fed a diet containing 6.25% dibutyl sebacate for 10 wks, while a control group of 12 male and 12 female rats were fed the basal diet, and then animals of each group were then mated.116 The dams were allowed to deliver their litters, and at weaning, 24 male and 24 female offspring were randomly chosen, fed the test diet for 21 days, and then killed for necropsy. The study results indicated that ingestion of a diet contain- ing 6.25% dibutyl sebacate had no adverse effect on fertility, litter size, or survival of offspring. Growth was decreased dur- ing the pre-weaning and post-weaning periods. However, no gross pathological changes were found among young rats killed at the end of the 21-day post-weaning period. Diethylhexyl Sebacate Reproduction, suckling and growth were normal in a four-generation study of rats fed a diet containing 200 ppm diethylhexyl sebacate (~10 mg/kg/day).1 No reproductive or developmental toxicity was observed. Dimethyl Glutarate/Dimethyl Succinate/Dimethyl Adipate Mixture The developmental toxicity produced by the inhalation of dibasic esters (mixture of 65.1% dimethyl glutarate, 17.8% dimethyl succinate, and 16.8% dimethyl adipate) was evaluated in rats.133 Groups of 24 gravid Crl:CD rats were exposed for 6 h/day to 0, 0.16, 0.4, or 1.0 mg/l dibasic esters, by whole body inhalation, on days 7-16 of gestation. The aerosol particle size in the 1.0 mg/l chamber was 5.3-5.4 µm, with 72-74% of the aerosol <10 µm. The animals were killed on day 21 of gestation. All animals survived until study termination. Body weight gains were statistically significantly decreased in the 0.4 and 1.0 mg/l groups. Feed consumption by these groups was reduced during the first 6 exposures; statistical significance was not given. Statistically significant differences in absolute and relative liver weights were not observed, but there was a significant trend of decreased absolute, but not relative, liver weights. The only significant clinical signs observed were perinasal staining and wet fur of rats in the 1.0 mg/l group. Reproductive and developmental effects were not observed, and dibasic esters was not a developmental toxicant in rats following inhalation of ≤1.0 mg/l. Groups of 20 Crl:CD(SD)BR rats/gender were exposed for 6 h/day, 5 days/wk, to 0, 0.16, 0.40, or 1.0 mg/l dibasic esters by whole body inhalation for 14 wks prior to mating, and then 7 days/wk for 8 wks of mating, gestation, and lactation. 134 The mean aerosol particle size in the 1.0 mg/l chamber was 6.2 µm, with 69% of the aerosol <10 µm. Exposure was discontinued from day 109 of gestation through day 3 post-partum. All parental rats and 10 pups/gender were killed and necropsied on day 21 post-partum. The remaining pups were not necropsied. Maternal body weights in the 0.40 mg/l group were decreased during the last week of the study, while body weights of male and female rats of the 1.0 mg/l group were slightly decreased from wk 7 on. Relative liver weights were slightly, but not significantly, decreased in the 0.4 and 1.0 mg/l groups. Other differences in organ weights were not considered dose-related. With the exception of a statistically significant decrease in pup body weights at birth and day 21, no reproductive or developmental effects were observed. The only microscopic findings were squamous metaplasia in the olfactory epithelium of all treated parental rats. This effect was

43

CIR Panel Book Page 91 minimal in the 0.16 mg/l group and mild to moderate in the 0.4 and 1.0 mg/ml groups. The NOEL for reproductive parameters was 1.0 mg/l. Endocrine Disruption Diethylhexyl adipate appeared to have endocrine-mediated effects in a 28-day oral study; however, it was stated that the findings may be attributable to the disturbance ovarian function according to the hypothalamic- pituitary-gonad axis. Diethylhexyl Adipate A 28-day repeated-dose toxicity study was performed to determine whether diethylhexyl adipate has endocrine- mediated activities.135 Groups of 10 male and 10 female Crj:CD (SD) rats were dosed orally by gavage with 0, 40, 200, or 1000 mg/kg diethylhexyl adipate in corn oil, at a volume of 10 ml/kg, for a minimum of 28 days. In addition to clinical observations, a functional observation battery was performed during wk 4, estrous cycling was assessed from day 22, hormone analysis was measured at the end of the test period, and sperm morphology and sperm count were examined. Male animals were killed and necropsied on day 29, while females were killed and necropsied on days 30-34 when in diestrous. Signs of toxicity were not observed, and no clinical chemistry or hematological findings were recorded. Hormonal and spermatological analyses were normal. Statistically significant increased were seen in relative kidney weights in males of the 200 and 1000 mg/kg groups, relative liver weights of males in the 1000 mg/kg group, and in relative liver, kidney, and adrenal weights in females of the 1000 mg/kg group. Microscopically, increased eosinophilic bodies and hyaline droplets were seen in the kidneys of male rats of the 1000 mg/kg group. Ovarian follicle atresia was observed in 4 females of the 1000 mg/kg group, accompanied by a prolonged estrous cycle in 2 of these rats. A change in the estrous cycle is an important endpoint for determination of endocrine-mediated effects in the enhanced TG 407 assay. The researchers stated that this effect, in conjunction with the microscopic findings, appears to be related to endocrine-mediated effects of diethylhexyl adipate. However, it was also stated that these finding may be attributable to the disturbance of ovarian function according to the hypothalamic-pituitary-gonad axis. The changes in relative organ weights were considered toxic effects, and the NOEL was 40 mg/kg/day. The effect of diethylhexyl adipate on estrogen receptor and thyroid hormone (TH) functions was also examined.136 The TH-like activity was assessed using the rat pituitary tumor cell line Gh3 expressing intracellular TH and estrogen receptors and responding to physiological concentration of TH by proliferation. At low potency, diethylhexyl adipate stimulated the TH- dependent rat pituitary GH3 cell proliferation in a concentration-dependent manner. Cotreatment of GH3 cells with diethylhexyl adipate potentiated the L-3,5,3’-triiodothyronine (T3)-EC50 potentiated the T3-induced GH3 cell proliferation. Diisononyl Adipate In a subchronic dietary study described earlier, groups of male and female beagle dogs were fed 0, 0.3, 1.0, or 3.0% (wks 1-8) and 6.0% (wks 9-13) diisononyl adipate for 13 wks.85 Reproductive tissues were evaluated. No significant findings were reported for the 0.3 and 1.0% groups. In the high dose group, testes weight was decreased. At microscopic examina- tion, it was found that the epididymal ducts were devoid of spermatozoa, the seminiferous tubules were composed of sertoli cells and spermatogonia, spermatocytes and spermatids were not evident, and there was almost total aspermatogenesis. Ovaries were not weighed at necropsy. There were no gross or microscopic changes in any of the test groups upon compari- son to controls. GENOTOXICITY The esters of dicarboxylic acids are not mutagenic or genotoxic a battery of in vitro and in vivo tests. The only

44

CIR Panel Book Page 92 non-negative results reported were equivocal results in a sister-chromatid exchange assay with ≤400 µg/ml diethylhexyl adipate in the presence of metabolic activation and a dose-dependent inhibition of 3H-thymidine into replicating DNA, with a dose-dependent increase in the ratio of acid-incorporated 3H-thymidine with ≤0.01 M diethylhexyl adipate.. (The same effect was seen in the 3H-thymidine assay with 2-ethylhexanol.) Diethyl Malonate Diethyl malonate was not mutagenic in an Ames test or a cytogenetic assay using human peripheral lymphocytes at concentrations ≤5000 g/plate.14 Dimethyl Malonate Dimethyl malonate was not mutagenic in an Ames test at concentrations ≤5000 g/plate.14 Dimethyl Succinate Dimethyl succinate was not mutagenic in an Ames tests with concentrations of ≤ 20,000 µg/plate137 or in a preincubation assay with concentrations of ≤10,000 g/plate.138 Dimethyl Glutarate Dimethyl glutarate was not mutagenic in a preincubation assay with concentrations of ≤10,000 g/plate.139 Dimethyl Adipate Dimethyl adipate was not mutagenic in a preincubation assay with concentrations of ≤10,000 g/plate.140 Dibutyl Adipate Dibutyl adipate was mutagenic in an Ames test at concentrations of ≤5000 µg/plate. It was not genotoxic in an in vivo mouse micronucleus assay in which the animals were dosed with ≤2000 mg/kg. From the Amended Final Report on Dibutyl Adipate5

Di-C7-9 Branched and Linear Alkyl Esters of Adipic Acid Di-C7-9 branched and linear alkyl esters of adipic acid were not mutagenic in an Ames test at concentrations of ≤10.0 µl/plate.85 Ditridecyl Adipate Ditridecyl adipate was not mutagenic in an Ames test at concentrations of 0-10 µl/plate, and it was not clastogenic in an in vivo micronucleus assay using rats dosed dermally with 0, 800, or 2000 mg/kg ditridecyl adipate.85 Diethylhexyl Adipate Diethylhexyl adipate was not mutagenic in an Ames. (The specific concentrations tested were not provided.) From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2

Diethylhexyl adipate was not mutagenic in a number of genotoxicity studies. In vitro, negative results were reported in Ames tests at concentrations ranging from ≤150 -10,000 µg/plate,85,141-143 in an NTP preincubation assay,144 in a liquid suspension assay,145 and in a forward mutation assay using L5178Y cells at concentrations ≤1000 µg/ml .146 In an assay for sister chromatid exchanges and chromosomal aberrations using concentrations of ≤200 g/plate, results were negative,147 while in another assay with ≤400 l/plate, results were negative without, but equivocal with, metabolic activation in the sister chromatid exchange assay and there was some evidence of genotoxicity without, but none with, metabolic activation in the chromosomal aberration assay.148 In a 3H-thymidine assay, there was a dose-dependent inhibition of 3H-thymidine into repli- cating DNA, with a dose-dependent increase in the ratio of acid-soluble DNA-incorporated 3H-thymidine.142 In vivo, results were negative in micronucleus tests85,149 and chromosomal aberration assays.150,151 An Ames test was performed on urine of rats dosed with diethylhexyl adipate to assess whether mutagenic substances occur in the urine following diethylhexyl adipate adminstration.152 Groups of ≥6 male Sprague-Dawley rats were dosed orally by gavage with 0 or 2000 mg/kg diethylhexyl adipate in corn oil for 15 days. Urine was collected daily. The urine was not

45

CIR Panel Book Page 93 mutagenic in the Ames test, indicating that it is not converted to metabolic urinary metabolites. The urine of rats dosed with 1000 mg/kg 2-ethylhexanol by gavage for 15 days was also tested in an Ames assay. The urine of these rats also was not mutagenic. Urine from rats that were dosed with a known mutagen gave a positive response in an Ames test. Diisononyl Adipate Diisononyl adipate was not mutagenic in an Ames assay at ≤1000 g/plate, and it was not genotoxic in a mouse lymphoma assay, a transformation assay, or a BALB/3t3 assay at concentrations of ≤100, 1000, or 1.3 g/ml, respectively.153 Diethyl Sebacate Diethyl sebacate was non-mutagenic in an Escherichia coli Sd-4-73 reversion (streptomycin dependence to independence) assay.154 Dibutyl Sebacate Dibutyl sebacate, ≤10,000 g/plate, was not mutagenic in the Ames assay.155,156 Diethylhexyl Sebacate Diethylhexyl sebacate was not mutagenic in an Ames assay at concentrations of ≤10,000 µg/plate.141,157 In the rat liver foci test, diethylhexyl sebacate demonstrated no evidence of activity when administered orally at 500 mg/kg 3x/wk for 11 wks, following a single oral treatment with a known carcinogen.158 Details of the genotoxicity studies on esters of dicarboxylic acids are described in Table 11. Data from the original safety assessments on dibutyl and diethylhexyl adipate are included in italics. Details of the available genotoxicity data on esterase metabolites are also summarized in this table. CARCINOGENICITY In a 2-yr NTP study, ≤2.5% diethylhexyl adipate did not produce tumors in male or female rats, but it did increase the incidence of hepatocellular adenoma and carcinoma in male and female mice. Diethylhexyl adipate did not cause skin tumors with weekly application of 10 mg to the back of mice in a lifetime study. Other compounds with a 2-ethylhexyl group that have been evaluated for carcinogenicity had some evidence of hepatocarcinogenicity, ranging from very strong to equivocal, in rodents. The IARC has stated that diethylhexyl adipate is not classifiable as to its carcinogenicity in humans. However, feeding of diethylhexyl sebacate to rats for 19mos did not result in carcinogenic effects. Diethylhexyl Adipate In an NTP carcinogenicity study, administration of ≤25,000 ppm diethylhexyl adipate to rats for 103 wks did not produce carcinogenic effects. However, mice fed the same amount for 103 wks had dose-related body weight reductions and a higher incidence of hepatocellular adenoma and carcinoma than the controls. In another study in which rats were fed ≤2.5% diethylhexyl adipate for 2 yrs, tumor incidence for the test animals was similar to that of controls. The same researchers found no tumors in dogs fed up to 0.2% diethylhexyl adipate for 1 yr. A single 10 mg dose of diethylhexyl adipate given by s.c.. injection was not carcinogenic in mice. In a lifetime study, diethylhexyl adipate caused no skin tumors when 10 mg was applied weekly to the back skin of mice. From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2

Research has shown that other compounds with a 2-ethylhexyl group that have been evaluated for carcinogenicity had some evidence of hepatocarcinogenicity, ranging from very strong to equivocal, in rodents. 121 In an evaluation of the carcinogenic risk of diethylhexyl adipate, the IARC stated that there was limited evidence in experimental animals for the carcinogenicity of diethylhexyl adipate.17 Therefore, the overall evaluation of diethylhexyl adipate was not classifiable as to its carcinogenicity to humans (Group 3). Diethylhexyl Sebacate No evidence of carcinogenicity was observed in an unspecified number of rats fed a diet providing about 10 mg diethylhexyl sebacate/kg/day for up to 19 months.1 No further study details were provided. 46

CIR Panel Book Page 94 Tumor Promotion Diethylhexyl Adipate A group of 14 male F344 rats were used to assess the carcinogenic potential of diethylhexyl adipate in a medium- term liver bioassay.159 The rats were given a single i.p. dose of diethylnitrosamine, and 2 wks later they were given 20,000 ppm diethylhexyl adipate in the diet. At wk 3, a partial hepatectomy was performed. Positive results for carcinogenic potential were indicated by a significant increase in GST-P positive foci. Diethylhexyl adipate did not have an enhancing effect on the development of GST-P-positive foci. CLINICAL ASSESSMENT OF SAFETY Human Exposure Diethylhexyl Adipate Diethylhexyl adipate can migrate into food, and it is most marked when plasticized PVC film comes in direct contact with fatty foods.88 Using the analyses of a range of typical food, a maximum intake of 16 mg/person/day for diethylhexyl adipate was estimated. The amount of diethylhexyl adipate used in PVC films was reduced, and the estimate was revised to 8.2 mg/day. Dibutyl Sebacate Dibutyl sebacate, a component of PVC, can pass from the packing films to the enclosed food.40 Dermal Irritation and Sensitization In a number of irritation and sensitization studies, the diesters of dicarboxylic acids are not irritants or sensitizers. The only exception noted was that undiluted diisopropyl adipate was moderately irritating in one cumulative irritancy test, and some slight irritation was seen with formulations containing diethylhexyl adipate. Dimethyl Malonate The sensitization potential of 8% dimethyl malonate in petrolatum was evaluated in a maximization test using 25 subjects.14 Dimethyl malonate was not a sensitizer. Dibutyl Adipate Undiluted dibutyl adipate was not irritating in a 24-hr clinical patch test with 10 subjects. Slight reactions (not defined) were reported for 4 of 18 subjects in a 24-h patch test with dibutyl adipate, 20% in alcohol. From the Amended Final Report on Dibutyl Adipate5

Diisopropyl Adipate The dermal irritation and sensitization o f diisopropyl adipate was evaluated in a number of studies. Undiluted diisopropyl adipate produced no irritation in 4 h patch tests, but was moderately irritating in a 21- day cumulative irritancy test. Formulations containing 0.26-20.75% diisopropyl adipate caused minimal to mild irritation, but no sensitization. From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2

Diethylhexyl Adipate The dermal irritation and sensitization o f diethylhexyl adipate was evaluated in a number of studies. Clini- cal assessment of diethylhexyl adipate at concentrations of 0.01-9.0% in formulation showed, at most, erythema and papules when applied under occlusion for extended periods of time. From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2

Diisostearyl Adipate A human repeat insult patch test (HRIPT) using 50 subjects was used to evaluate the irritation and sensitization potential of diisostearyl adipate.160 Two-tenths ml was applied neat to the back of each subject under an occlusive patch for 24 h, after which time the subject removed the patch. This procedure was performed 3 times per wk for 3 wks, for a total of 9

47

CIR Panel Book Page 95 induction patches. Following a 10-14 day non-treatment period, a 24 h challenge patch was applied to a previously untreated site, and reactions were scored at 24 and 48 h. No adverse reactions were observed, and diisostearyl adipate was not a primary irritant or a sensitizer. Diethyl Sebacate A single insult occlusive patch test (SIOPT) was performed using 20 subjects to determine the irritation potential of a body cream containing 1.5% diethyl sebacate.161 The test patch was applied for 24 h. The PII was 0.00, and the body cream containing 1.5% diethyl sebacate was non-irritating. The sensitization potential of a body cream containing 1.5% diethyl sebacate was evaluated in a maximization study.162 During induction, 0.05 ml of 0.25% aq. sodium lauryl sulfate (SLS) was applied under an occlusive patch for 24 h. At that time, the patch was removed and 0.05 ml of the test material was applied to the same site under an occlusive patch for 48-72 h. If no irritation was present at the test site upon patch removal, an occlusive patch with 0.25% aq. SLS was applied for 24 h, followed by a patch of the test material. This sequence was used for 5 induction patches. If irritation developed during induction, the SLS patch was eliminated. After a 10-day non-treatment period, a challenge was performed at a pre- viously untreated site. The challenge site was pretreated with 0.05 ml of 5.0% aq. SLS under an occlusive patch for 1 h, followed by an occlusive patch of the test material for 48 h. Twenty-five subjects completed the study. No reactions were seen at challenge, and a body cream containing 1.5% diethyl sebacate did not have contact-sensitizing potential. Diisopropyl Sebacate An SIOPT was performed using 20 subjects to determine the irritation potential of a foundation containing 1.8% diisopropyl sebacate.163 The patch was applied for 24 h. The foundation containing 1.8% diisopropyl sebacate was not irritating. The irritation and sensitization potential of diisopropyl sebacate was evaluated in a patch test that consisted of four 24-h applications of diisopropyl sebacate as supplied (approximately 100%) during weeks 1, 2, 3, and 6 on a 2 cm x2 cm area of skin on the right upper arm of each subject.164 Examinations were performed immediately after patch removal. The induction phase was performed during wks 1-4 using 107 subjects. No clinically significant effects were detected on any of the subjects during this phase. During wk 6, the challenge phase was conducted on 105 subjects. No clinically significant effects were noted in any of the subjects during this phase. Diisopropyl sebacate was not observed to have any significant skin-irritating or sensitizing activity under the conditions of this study. A maximization assay was performed, using a modified protocol of the maximization assay procedure described earlier, to determine the contact-sensitization potential of a foundation containing 2.2% diisopropyl sebacate.165 In this study, the test material was allowed to volatilize for 30 min before the occlusive patch was applied. Twenty-five subjects completed the study. No reactions were seen at challenge, and a foundation containing 2.2% diisopropyl sebacate did not have contact- sensitizing potential. Two heat protection hair spray products containing 1% diisopropyl sebacate were tested using a modified Draize HRIPT procedure to determine the potential of those products to induce irritation and contact sensitization.166 The products were tested neat and allowed to volatilize prior to patch application. Samples were patched under semi-occlusive conditions. Approximately 0.2ml was used in each patch. One hundred ten subjects completed the study. Generally transient, barely perceptible (0.5-level) to mild (1-level) patch test responses on 22 test subjects for one formulation and only barely per- ceptible (0.5-level) patch test response on 15 test subjects with the other formulation during the induction and/or challenge phases of the study were reported. Both products were considered to be non-irritating and non-sensitizing.

48

CIR Panel Book Page 96 A heat protection hair spray product containing 7.2% diisopropyl sebacate was tested using an HRIPT to determine the potential of this product to induce irritation and contact sensitization.167 The product was tested neat under semi-occlusive conditions. Approximately 0.2 ml sample was used in each patch. Fifty-one subjects completed the study. No skin reactivity was observed in any of the test subjects during the course of the study. Diethylhexyl Sebacate Diethylhexyl sebacate was applied neat using occlusive patches to the skin of 15-30 subjects (sex not specified) for 48-h.1 No local reactions were observed in the challenge phase (48-h covered contact with neat liquid) that was carried out 2 weeks later, presumably due to limited induction. Dioctyldodecyl Dodecanedioate An HRIPT with 50 subjects was performed to evaluate the irritation and sensitization potential of dioctyldodecyl dodecanedioate.160 Two-tenths ml was applied to the back of each subject under an occlusive patch for 24 h, after which time the subject removed the patch. This procedure was performed 3 times per wk for 3 wks, for a total of 9 induction patches. Following a 10-14 day non-treatment period, a 24 h challenge patch was applied to a previously untreated site, and reactions were scored at 24 and 48 h. No adverse reactions were observed, and dioctyldodecyl dodecanedioate was not a primary irritant or a sensitizer. Diisocetyl Dodecanedioate An HRIPT with 50 subjects was performed as described above to evaluate the irritation and sensitization potential of diisocetyl dodecanedioate.160 No adverse reactions were observed, and diisocetyl dodecanedioate was not a primary irritant or a sensitizer. Clinical dermal irritation and sensitization data on esters of dicarboxylic acids are presented in Table 12. Data from the original safety assessments on dibutyl, diisopropyl and diethylhexyl adipate are included in italics. The available dermal irritation and sensitization data on esterase metabolites are also summarized in this table. Phototoxicity and Photosensitization A 10% dilution of dibutyl adipate and formulations containing 0.7-17% diisopropyl adipate and 9% diethylhexyl adipate were not phototoxic. Dibutyl Adipate Dibutyl adipate, as a 10% dilution in liquid paraffin, was not phototoxic in a clinical study.5

Diisopropyl Adipate In photopatch test studies, formulations containing 0.7-17.0% diisopropyl adipate were not phototoxic, primary irritants, or sensitizers. From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2

Diethylhexyl Adipate In a photopatch test with 9.0% diethylhexyl adipate, no phototoxic or photoallergic reactions were observed. From the Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate2

Ocular Irritation Dibutyl Adipate Dibutyl adipate, 0.1% in paraffin oil, was not an ocular irritant in two subjects. From the Amended Final Report on Dibutyl Adipate5

49

CIR Panel Book Page 97 Comedogenicity Dibutyl Adipate Dibutyl adipate, 10-100% (vehicle not stated), was not comedogenic in clinical testing. From the Amended Final Report on Dibutyl Adipate5

Case Reports A number of investigators have reported cases of allergic contact dermatitis in response to diethyl sebacate-contain- ing products, and have demonstrated diethyl sebacate to be the substance, or one of several substances in the products, elicit- ing the dermatitis.37,168-172 Two case studies were reported of allergic reactions to lotion containing diisopropyl sebacate.173,174 In a case study where one patient was sensitized to other sebacate esters, diethylhexyl sebacate was not irritating.173 For stearyl alcohol, a metabolite of distearyl succinate, contact sensitization was reported in 3 individuals.175 These case reports are described in Table 13. Risk Assessment Diethylhexyl Adipate According to the Integrated Risk Information System of the EPA, the weight-of-evidence classification for diethyl- hexyl adipate was “possible human carcinogen”.130 The classification was based on an absence of human data and increased liver tumors in female mice. The only genotoxic effect was a positive dominant lethal assay. It was noted that diethylhexyl adipate exhibits structural relationships to other non-genotoxic compounds that are classified as probable and possible carcinogens.

50

CIR Panel Book Page 98

SUMMARY This safety assessment includes sebacic acid and other alkyl α,ω-dicarboxylic acids, salts, monoesters and diesters, for a total of 56 ingredients. The dicarboxylic acids are terminally functionalized straight alkyl chains characterized by a separation between the acid functional groups of one to 10 carbons. The simple alkyl di-esters are the result of the condensa- tion of alkyl dicarboxylic acids and two equivalents of alkyl alcohols. These ingredients can be metabolized via hydrolysis back to the parent alcohol, the mono-ester, and the parent dicarboxylic acid. The simple alkyl esters (mono- and di-) of these dicarboxylic acids have straight or branched side chains ranging in length from one to 18 carbons. This safety assessment is divided into two parts – (1) the dicarboxylic acids and their salts and (2) esters of dicarboxylic acids A safety assessment of diethylhexyl adipate and diisopropyl adipate was published in 1984 with the conclusion that these ingredients are safe as used in cosmetics. Additionally, dibutyl adipate was previously reviewed in 1996 and the avail- able data were found insufficient to support the safety of dibutyl adipate in cosmetic formulations. When re-reviewed in 2006, additional data were made available to address the needs identified by the CIR Expert Panel, and an amended conclu- sion was issued stating that dibutyl adipate is safe for use in cosmetic formulations. While many of the alkyl dicarboxylic acids are natural products, commercial production of these acids has historical- ly occurred via alkali pyrolysis of lipids. A relationship exists between the molecular weight and the log octanol – water partioning coefficient. Physical properties change as chain length increases, and the water solubility of these acids is inversely proportional to their chain length. Odd versus even chain length also plays a role. The alternating effects are believed to be the result of the inability of odd carbon number compounds to assume an in-plane orientation of both carboxyl groups with respect to the hydrocarbon chain. The diesters, in contrast, are much more lipid soluble and more difficult to dissolve in water. The short-chain alkyl mono- and diesters are more soluble in water, less lipophilic, and relatively more volatile than the corresponding longer-chain alkyl esters. The ingredients in this report function in cosmetics as pH-adjusters, fragrance ingredients, plasticizers, skin-condi- tioning agents and/or solvents and corrosion inhibitors. The majority of the dicarboxylic acids function in cosmetics as pH adjusters and fragrance ingredients. Six of the 12 dicarboxylic acids and their salts and 24 of the 44 esters included in this safety assessment are reported to be used in cosmetic formulations. For the dicarboxylic acids and their salts, disodium succi- nate has the greatest number of reported uses, with a total of 45. The acid with the greatest concentration of use is succinic acid, 26%; use at this concentration is in rinse-off products. The greatest leave-on concentration is 0.4%, disodium succinate, with dermal contact exposure. For the esters, diisopropyl adipate has the greatest number of uses, with 70 reported. The con- centration of use is greatest for dimethyl glutarate, 15% in a dermal rinse-off product. The ingredients with the greatest leave- on use concentrations, which are all dermal contact exposures, are diethylhexyl adipate, 14%, diisostearyl adipate, 10%, and diisopropyl sebacate, 10%. With the exception of dipotassium azelate, disodium sebacate, and di-C12-15 alkyl adipate, the dicarboxylic acids and their salts and esters are listed for use by the European Union (EU) without restriction. Dicarboxylic Acids and Their Salts Dicarboxylic acids are natural metabolic products of the ω-oxidation of monocarboxylic acids when the β-oxidation of free fatty acids is impaired. Under normal physiological conditions, dicarboxylic acids are rapidly β-oxidized, resulting in very low cellular concentrations and practically non-detectable concentrations in the plasma. Oxidation of odd- and even- numbered chains proceeds to different end points; even chains are completely, while odd-number chains are not completely oxidized. 51

CIR Panel Book Page 99 Unchanged dicarboxylic acid was found in the urine of rats. With oral dosing, approximately 53-67% adipic acid, 40% azelaic acid, and 50% dodecanedioate was recovered with the respective acid. With i.v. dosing, 59-71% adipic acid and 35% sebacate was recovered. In humans, 6.76-61 adipic acid, and 61% azelaic acid were found in the urine after dosing with the respective acid. With azelaic acid and dodecanedioic acid, radioactivity was found in all tissues, and it decreased after 24 h in all tissues except adipose tissue. Radioactivity was found in expired carbon dioxide following dosing adipic acid (70%), azelaic acid (14.5%), and disodium sebacate (25%). For rats dosed orally with azelaic, sebacic, undecanedioic, and dodecanedioic acid, 2.5, 2.1, 1.8, and 1.6% of the respective acid was found in the urine unchanged. The amount recovered decreased with increasing chain length. After oral dosing, 60, 17, 5, and 0.1% of azelaic, sebacic, decanedioic, and undecane- dioic acids, respectively, were recovered unchanged in the urine. In the plasma of both animals and humans, dicarboxylic acid catabolites that were 2-, 4-, or 6- carbons shorter than the corresponding dicarboxylic acid were found. Adipic acid did not induce peroxisome proliferation. Dicarboxylic acids did have some cellular effects and inhibited mitochondrial oxidoreductases, reversibly inhibited microsomal NADPH and cytochrome P450 reductase ,and competitively inhibited tyrosinase in vitro.

The oral LD50 values of the dicarboxylic acids for rats ranged from 0.94 g/kg adipic acid (although most reported values were >5 g/kg) to ≥4 g/kg azelaic acid. The reported dermal LD50 values ranged from >6 g/kg dodecanedioic acid to >10 g/kg glutaric acid. In short-term oral toxicity studies, ≤3000 mg/kg/day adipic acid did not produce significant toxicological effects in rats. Signs of toxicity were seen at >3600 mg/kg/day. No toxicity was observed with guinea pigs fed 400-600 mg/day azelaic acid. Short-term inhalation exposure to 126 mg/m3 adipic acid to rats did not produce signs of toxicity, but exposure of mice to 460 mg/m3 did. In a subchronic oral study, 10 male and 10 female rats exposed to 10% sodium succinate in the drinking water died, but no compound-related lesions were found. Body weights were decreased in rats given ≥2.5% sodium succinate for 13 wks, but toxicological treatment-related changes were not observed. Glutaric acid ad a low degree of toxicity to rats (at 2%) and dogs (concentration not specified) when given in the drinking water. Dietary administration of ≤3400 mg/kg/day adipic acid for 19 wks produced slight effects in the liver of male rats; the NOAEL was 3333 mg/kg. A mixture of adipic, glutaric, and succinic acids had a low degree of toxicity in rats when tested at 3% for 90-days. Signs of toxicity were reported in a sub- chronic inhalation study in which mice were exposed to 13 or 120 mg/m3 adipic acid. A low degree of toxicity to sodium succinate was observed in a 2 yr oral study using rats. Slight effects were seen in the livers of rats fed ≤3200 mg/kg/day adipic acid for 33 wks, and the NOAEL for rats fed a diet containing adipic acid for 2 yrs was 1%; no significant toxicological effects were seen at concentrations of ≤5%. No significant toxicological effects were observed for mice fed ≤280 mg/kg or rabbits fed ≤400 mg/kg azelaic acid for 180 days. Disodium sebacate was not not toxic to rats or rabbits fed up to 1000 mg/kg for 6 mos. For the dicarboxylic acids, the severity of ocular irritation seems to decrease with increasing carbon number. Suc- cinic acid was a severe ocular irritant, glutaric acid was moderately irritating, and dodecanedioic acid was a slight irritant. Ocular irritation produced by adipic acid was dose-dependent. Slight to mild dermal irritation was observed for the succinic, glutaric, and adipic acid. Adipic acid, dodecanedioic acid, and a mixture of succinic, glutaric, and adipic acids are not sensitizers. Reproductive and developmental effects were not seen upon oral dosing with the dicarboxylic acids or disodium sebacate. Malonic acid has a spermicidal effect on human spermatozoa. Glutaric acid was tested at doses of ≤1300 mg/kg in

52

CIR Panel Book Page 100 rats and 500 mg/kg in rabbits, adipic acid at doses of ≤263 mg/kg in mice, 288 mg/kg in rats, 205 mg/kg in hamsters, or 250 mg/kg in rabbits, azelaic acid at doses of ≤140 mg/kg in rats and 200 mg/kg in rabbits, disodium sebacate at 500 mg/kg in rats and 1000 mg/kg in rabbits, and dodecanedioic acid was tested at ≤1000 mg/kg using rats. Embryotoxic effects were reported for in a reproductive study of 2500 mg/kg/day azelaic acid using rats and in reproductive studies with ≤500 mg/kg/day azelaic acid using rabbits and monkey. Sodium salts of some dicarboxylic acid had a specific inhibitory effect on the uterine horn, and this effect progressively increased with chain length. The dicarboxylic acids are not genotoxic, and consistently were not mutagenic in Ames tests. Positive result were seen in a transformation assay on glutaric acid using Balb/c-3T3 cells, both with and without metabolic activation. The results of a mouse lymphoma assay, with and without metabolic activation, on glutaric acid was pH-dependent. Equivocal results were obtained in an in vitro chromosomal aberration assay of ≤15 mg/mg disodium succinate using Chinese hamster fibroblast cells. The dicarboxylic acids were not genotoxic in in vivo assays. Carcinogenic results were not seen in rats given up to 2% sodium succinate in the drinking water or 5% adipic acid in feed for 2 yrs. An increase in the incidence of C-cell adenoma/carcinoma of the thyroid in females given 2% sodium succinate, and a positive trend in the occurrence of this tumor, was considered a function of experimental variability and not related to dosing. In a cumulative irritancy test, the cumulative irritation of a 15% azelaic acid gel increased with successive patching. It is not known if the vehicle played a role in the irritation scores. Daily application of a 20% azelaic cream causes erythema and irritation. Esters of Dicarboxylic Acids The metabolism of diesters in animals is expected to occur, initially, via enzymatic hydrolysis, leading to the corre- sponding dicarboxylic acids and the corresponding linear or branched alcohol. These dicarboxylic acids and alcohols can be further metabolized or conjugated to polar products that are excreted in urine, or, the enzymatic hydrolysis may be incomplete and result, at least for some diesters, in the production of monoesters. In vitro using pig skin, 8.8 and 3% of undiluted diethyl malonate was found in the skin and receptor fluid, respective- ly, 30 and 10% of diethyl malonate in ethanol was found in the skin and receptor fluid, respectively, and 0.2-0.9 and 0.2-1.6% diethyl malonate in acetone was found in the skin are receptor fluid, respectively. Using human skin, 16% of the applied di- ethyl malonate penetrated. In vivo, absorption of diethyl malonate, estimated from urinary and fecal recovery, was 15% in nude mice, 4 % in human skin grafted to nude mice, 6% in pig skin grafted to nude mice, 2.5% in pigs, and 4% in dogs. Approximately 11% of ditridecyl adipate was absorbed through the skin of rats; 5.5-7.4% of the applied dose was found in the tissues, 3.5-4.7%was found in the urine, and 0.4-0.7% was found in the feces. Prior dosing with ditridecyl adipate did not significantly affect absorption. In vitro, diethylhexyl adipate was readily hydrolyzed to MEHA or adipic acid in rat liver, pancreas, and small intestine tissue preparations. In animals, diethylhexyl adipate is hydrolyzed to adipic acid and 2-ethylhexanol or MEHA. 2- Ethylhexanol is converted to 2-ethylhexanoic acid, which may form a glucuronide conjugate or may be subjected to ω- and (ω-1)-oxidation and further metabolism. More than 98% of diethylhexyl adipate administered orally to rats was excreted in 48 h; 21-45% of the radioactivity was expired in carbon dioxide and 34-52% was excreted in the urine. Diethylhexyl adipate and MEHA are not found in the blood or urine; diethylhexyl adipate or the metabolites are recovered in the tissues. Metabo- lism studies have shown that excretion in the urine is not as unchanged diethylhexyl adipate; mostly adipic acid is found. In humans, peak urinary elimination of all metabolites occurred within 8 h of dosing.

53

CIR Panel Book Page 101 Diethylhexyl sebacate is not readily absorbed through the skin of guinea pigs. Metabolism in rodents and humans may follow partially common pathways, producing 2-ethylhexanol as an intermediary metabolite. Peroxisome proliferation causes an increase in liver weights and can induce hepatocarcinogenicity in rats and mice. Diethylhexyl adipate is a peroxisome proliferator requiring extensive phase I metabolism to produce the proximate peroxi- some proliferator, which in both mice and rats appears to be 2-ethylhexanoic acid. Studies conducted to explain the species difference in liver tumors seen in mice, but not rats, in the NTP carcinogenicity study on diethylhexyl adipate suggested that diethylhexyl adipate-induce cell replication, rather than hepatic peroxisome proliferation, provided a better correlation with tumor formation. Diethylhexyl adipate is not as potent a proliferator as diethylhexyl phthalate. Peroxisome proliferation is not believed to pose the risk of inducing hepatocarcinogenesis in humans. Diethylhexyl adipate did not bind covalently to hepatic DNA in mice. It did stimulate DNA synthesis in livers of rats. In another study, a significant increase in 8-OH-dG was seen in rat liver, but not kidney DNA. The IARC remarked that the weight of evidence for diethylhexyl adipate demonstrated that rodent, peroxisome proliferators do not act as direct DNA- damaging agents.

The oral and dermal LD50 values of esters of dicarboxylic acids are greater than 2 g/kg. Mostly, acute exposure via inhalation to diethyl malonate, dibutyl adipate, and diethylhexyl sebacate did not result in death of rats Oral administration of ≤1000 mg/kg dibutyl adipate for 28 days was not toxic effects in rats. In short-term oral dos- ing with diethylhexyl adipate, decreased weight gain was reported for rats and mice. The NOELs for rats and mice were 2 and 0.63%, respectively, in feed; administration of 10% in feed killed 5/5 female mice. In 2- and 4-wk studies of diethylhexyl adipate, the oral NOAEL for ovarian toxicity was 200 mg/kg in rats; an increase in atresia of the large follicle and a decrease in currently formed corpora lutea were seen in females dosed with 1000 and 2000 mg/kg diethylhexyl adipate. In a short-term dermal study in which 10 rabbits were dosed dermally with 0.5 or 1.0 ml/kg of a 20% dispersion of dibutyl adipate for 6 wks, there was a significant reduction in body weights in the high dose group, and renal lesions in one animal of each group. There were no signs of toxicity in guinea pigs in an immersion study with 20.75% diisopropyl adipate, diluted to an actual concentration of 0.10% adipate. Dermal administration of diethylhexyl adipate to rabbits for 2 wks resulted in slight to moderate erythema at the test site, but toxic effects were not reported for most of the animals. In a subchronic oral toxicity study using rats, the NOEL for diethyl malonate was 1000 mg/kg/day. Dietary admini- stration of ≤2.5% di-C7-9 branched and linear alkyl esters of adipic (approx. 1500 and 1900 mg/kg/day for males and females, respectively) for 90 days did not result in systemic toxicity. The NOAELS for male and female rats were 1500 and 1950 mg/kg/day, respectively. Subchronic oral administration of diethylhexyl adipate to rats caused significant decreases in body weight gains and increases in liver and kidney weights. The dietary NOEL for rats in a 90-day study was 610 mg/kg. In mice, no lesions were induced by dietary administration of ≤25,000 ppm diethylhexyl adipate for 13 wks. A decrease in body weights was seen in mice fed a diet with 1.2 and 2.5% diethylhexyl adipate. For diisononyl adipate, dietary administration of up to 500 mg/kg for 13 wks, a statistically significant increase in relative kidney weights was reported, but there were not toxicological findings. With dogs, 3.0% dietary diisononyl adipate resulted in a decrease in body weights, testes weight, and feed consumption, increased liver weight, elevated enzyme levels, liver and kidney discoloration, and microscopic changes in the liver, testes, spleen, and kidneys. No adverse effects were reported with whole-body application of an 6.25% emulsion of dibutyl adipate to dogs for 3 mos. Unoccluded dermal application of up to 2000 mg/kg ditridecyl adipate for 13 wks to rats produced slight erythema, but no systemic toxicity.

54

CIR Panel Book Page 102 In a 6-mos study in which rats were dosed intragastrically with diethylhexyl adipate, hepatic detoxification appeared depressed at the beginning of the study, while in a 10-mos study, a decrease in central nervous system excitability was noted. Dietary administration of ≤1.25% dibutyl sebacate for 1 yr or ≤6.25% for 2 yrs did not have an effect on growth In a 4-hr inhalation toxicity study of 5.9 mg/l of a mixture of 66.0% dimethyl glutarate, 16.5% dimethyl succinate, and 17.0% dimethyl adipate, the anterior and posterior nasal passageways were affected. Regeneration of damaged olfactory epithelium was related to the severity of the initial tissue damage. In a 13-wk study with ≤250 mg/m3 diethylhexyl sebacate, no systemic or lung effects were observed. Ocular irritation appeared to lessen in severity as chain length of the dicarboxylic acid esters increased. Diethyl malonate was slightly to moderately irritating to rabbit eyes. Dibutyl, diisopropyl, and diethylhexyl adipate were non- or min- imal ocular irritants. Diisopropyl sebacate was minimally irritating, while diethylhexyl sebacate was non-irritating to rabbit eyes. Dioctyldodecyl and diisocetyl dodecanedioate were not irritating to rabbit eyes. The esters of dicarboxylic acids were mostly non or mildly irritating to rabbits. Some minimal irritation was seen with diisopropyl adipate, undiluted or at 5-20.75% in formulation, and moderate erythema was reported with undiluted dibutyl adipate. Dimethyl malonate, dibutyl and diethylhexyl adipate, diethylhexyl sebacate, and dioctyldodecyl dodecanedioate were not sensitizers in guinea pigs or rabbits. Perfume formulations containing 1.1% diisopropyl adipate were not phototoxic. Oral administration of up to 1000 mg/kg dimethyl malonate did not have an effect on fertility, and no development toxicity was reported. The NOAEL was 300 mg/kg for repeated dose and maternal toxicity and 1000 mg/kg for fertility and developmental toxicity. Oral administration of up to 100 mg/kg dibutyl adipate did not cause any reproductive effects, and the NOEL for parental and offspring toxicity was 300 mg/kg/day and for reproductive toxicity was 100 mg/kg/day. Oral administration of ≤7000 mg/kg di-C7-9 branched and linear alkyl esters of adipic acid branched and linear alkyl esters of adipic acid did not result in developmental toxicity. Dietary administration of up to 1.2% diethylhexyl adipate did not affect fertility when fed to rats prior to mating. Fetal weight, total litter weight, and litter size were reduced with 1.2% diethylhexyl adipate. In a study in which gravid rats were fed the same doses during gestation, no significant effects on fetal weight or litter size were reported. An increased incidence of minor skeletal abnormalities was attributed to fetotoxicity. . In a study in which diethylhexyl adipate was given orally to rats from day 7 of gestation until post-natal day 17, antiandrogenic effects were not observed, although some increase in post-natal death were observed. Administration of up to 2000 mg/kg diethyl- hexyl adipate prior to dosing and through day 7 of gestation did have an effect on the mean estrus cycle length at a dose of 1000 and 2000 mg/kg, and did appear to disturb ovulation. Significant decreases were also seen in implantation rate and number of live embryos, as well as an increase in pre-implantation loss. Diethylhexyl adipate did not produce testicular toxic effects when fed at up to 25,000 ppm in the diet for 4 wks. Dietary administration of 6.25% dibutyl sebacate to male and fe- male for 10 wks prior to mating had no adverse effects on fertility, litter size, or survival of offspring. Diethylhexyl sebacate, 200 ppm in the diet, did not produce reproductive or developmental effects. Dermal applications of up to 2000 mg/kg ditridecyl adipate affected mean fetal body weights and crown-rump lengths. Ditridecyl adipate, 2000 mg/kg, did not have an effect on sperm morphology. Some visceral anomalies were reported. The NOAELs for maternal toxicity and developmental and reproductive effects were 2000 and 800 mg/kg/day, respectively. Dimethyl, diethyl, dipropyl dibutyl, diisobutyl, and diethylhexyl adipate were evaluated for fetotoxic and teratogenic effects in rats when administered i.p. at 1/3 – 1/30 of the i.p. LD50 values. Some effect on resorptions and abnormalities were seen with all but diethyl adipate.

55

CIR Panel Book Page 103 Inhalation by rats of ≤1.0 mg/l of a mixture of dimethyl glutarate, dimethyl succinate, and dimethyl adipate on days 7-16 of gestation or for 14 days prior to mating, during mating and gestation, and lactation, no adverse developmental or re- productive effects were observed. The only exception was a statistically significant decrease in pup weight at birth and day 21. Diethylhexyl adipate appeared to have endocrine-mediated effects in a 28-day oral study; however, it was stated that the findings may be attributable to the disturbance ovarian function according to the hypothalamic-pituitary-gonad axis. The esters of dicarboxylic acids are not mutagenic or genotoxic a battery of in vitro and in vivo tests. The only non- negative results reported were equivocal results in a sister-chromatid exchange assay with ≤400 µg/ml diethylhexyl adipate in the presence of metabolic activation and a dose-dependent inhibition of 3H-thymidine into replicating DNA, with a dose- dependent increase in the ratio of acid-incorporated 3H-thymidine with ≤0.01 M diethylhexyl adipate.. (The same effect was seen in the 3H-thymidine assay with 2-ethylhexanol.) In a 2-yr NTP study, ≤2.5% diethylhexyl adipate did not produce tumors in male or female rats, but it did increase the incidence of hepatocellular adenoma and carcinoma in male and female mice. Diethylhexyl adipate did not cause skin tumors with weekly application of 10 mg to the back of mice in a lifetime study. Other compounds with a 2-ethylhexyl group that have been evaluated for carcinogenicity had some evidence of hepatocarcinogenicity, ranging from very strong to equivocal, in rodents. The IARC has sated that diethylhexyl adipate is not classifiable as to its carcinogenicity in humans. However, feeding of diethylhexyl sebacate to rats for 19mos did not result in carcinogenic effects. In a number of irritation and sensitization studies, the diesters of dicarboxylic acids are not irritants or sensitizers. The only exception noted was that undiluted diisopropyl adipate was moderately irritating in one cumulative irritancy test, and some slight irritation was seen with formulations containing diethylhexyl adipate. A 10% dilution of dibutyl adipate and formulations containing 0.7-17% diisopropyl adipate and 9% diethylhexyl adipate were not phototoxic. Cases of allergic contact dermatitis in response to diethyl sebacate-containing products have been reported, and it has been demonstrated diethyl sebacate was the substance, or one of several substances in the products, eliciting the derma- titis. Two case studies were reported of allergic reactions to lotion containing diisopropyl sebacate. According to the Integrated Risk Information System of the EPA, the weight-of-evidence classification for diethyl- hexyl adipate was “possible human carcinogen”. The classification was based on an absence of human data and increased liver tumors in female mice.

DISCUSSION To be determined at the meeting. CONCLUSION To be determined at the meeting.

56

CIR Panel Book Page 104 REFERENCES 1. BIBRA Working Group. 1996. Diethylhexyl Sebacate Toxicity Profile. 6 pages.

2. Elder RL,ed. Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate. J Am Coll Toxicol. 1984;3:(3):101-129.

3. Andersen FA (ed). Annual review of Cosmetic Ingredient Safey Assessments - 2002/2003. Int J Toxicol. 2005;24:(Suppl 1):1-102.

4. Andersen FA (ed). Annual review of Cosmetic Ingredient Safety Assessments - 2004/2005. Int J Toxicol. 2006;25:(Suppl 1):1-89.

5. Andersen FA,ed. Amended final report of the safety assessment of dibutyl adipate as used in cosmetics. Int J Toxicol. 2006;25:(Suppl 1):129-134.

6. Vasihtha, A. K, Trivedi, R. K., and Das, G. Sebacic Acid and 2-Octanol from Castor Oil. Journal of the American Oil Chemists' Society. 1990;67:(5):333-337.

7. Editor: Kroschwitz, J. Concise Encyclopedia od Chemical Technology. 4th ed. 2001.

8. Nghiem NP, Davison BH, and Thomspon JE. Effect of biotin on the production of succinic acid by anaerobiospirillum succinicproducents. Gov't Reports Announcements & Index. Appl.Biochem.Biotechnol. 1995;633:57-58.

9. Kadesch, Richard G. Dibasic Acids. Journal of the American Oil Chemists' Society. 1954;31:568-573.

10. Breathnach, A. S. Azelaic Acid - Biological Activities and Therapeutic Applications. Drugs of Today. 1989;25:(7):463-472.

11. Aditya K.Gupta, Melissa D. Gover. Azelaic acid (15% gel) in the treatment of acne rosacea. International Journal of Dermatology. 2007;46:(5):533-538.

12. Kadesch, Richard G. Dibasic Acids. Journal of the American Oil Chemists' Society. 1954;31:(11):568-573.

13. Ayorinde et al. Synthesis of Dodecanedioic Acid from Vernonia galamensis Oil. Jounal of the American Oil Chemists' Society. 1989;66:(5):690.

14. Organisation for Economic Co-Operation and Development.SIDS Initial Assessment for SIAM 22 -Malonic Acid Diesters, Dimethylmalonate, 108-59-8, and Diethylmalonate, 105-53-3. 2005. http://www.chem.unep.ch/irptc/sids/OECDSIDS/Malonates.pdf. Accessed 7-29-2010.

15. Gattermann, T. Wieland. Praxis des Organischen Chemikers. 40th ed. 1961.

16. Cognis Deutchsland GmbH & Co. 2002. Data Profile: Cetiol® B. 4 pages.

17. International Agency for Research on Cancer (IARC). Di(2-ethylhexyl) adipate. IARC Monographs on the evaluation of the carcinogenic risk of chemicals to humans. 2000;77:149-175.

18. American Chemistry Council Aliphatic Esters Diesters Task Group.High Production Volume (HPV) Chemical Challenge Program test plan for th diesters category of the aliphatic esters chemicals. Dated June 2, 2010. 6-28-2010. http://www.epa.gov/oppt/chemrtk/pubs/summaries/alipestr/c13466rt8.pdf. Accessed 7-8-0010.

19. Kvatadze MK. The gas-chromatographic detection of Sebacic Acid in the air. Gig.Tr.Prof.Zabol. 1989;9:45-46.

20. Hokoku Corporation. Impurity data on diisopropyl sebacate. Unpublished data submitted by the Personal Care Products Council (1 p). 2010.

21. Cognis Deutchsland GmbH & Co. 2002. Cetiol® B Toxicological Evaluation. 8 pages. 57

CIR Panel Book Page 105 22. Gottschalck TE and Bailey J, eds. International Cosmetic Ingredient Dictionary and Handbook. 2010. 13:(1- 3):Washington DC: Personal Care Products Council.2977.

23. Food and Drug Administration.Frequency of use of cosmetic ingredients. 2010.

24. James, A. C., Stahlhofen, W, Rudolf, G, Kobrich, R, Briant, J. K., Egan, M. J., Nixon, W, and Birchall, A. Annexe D. Deposition of inhaled particles. Annals of the ICRP. 1994;24:(1-3):231-2.

25. Oberdorster, G, Oberdorster, E, and Oberdorster, J. Nanotoxicology: An Emerging Discipline Evolving from Studies of Ultrafine Particles. Environmental Health Perspectives. 2005;113:(7):823-839.

26. Bower, D. 1999. Unpublished information on hair spray particle sizes provided at the September 9, 1999 CIR Expert Panel meeting.

27. Johnson, M. A. The Influence of Particle Size. Spray Technology and Marketing. 2004;November:24-27.

28. European Commission.Cosing database on cosmetic ingredient and substances. 2010. http://ec.europa.eu/consumers/cosmetics/cosing/. Accessed 7-27-2010.

29. Dekker. Ingredients. The Chemistry and Manufacture of Cosmetics. 1999. III:(2):

30. American Chemistry Council Aliphatic Esters Diesters Task Group.High Production Volume (HPV) Chemical Challenge Program test plan for th diesters category of the aliphatic esters chemicals. Dated June 2, 2010. 6-28-2010. http://www.epa.gov/oppt/chemrtk/pubs/summaries/alipestr/c13466rt8.pdf. Accessed 7-8-0010.

31. The Merck Index. 2009. http://themerckindex.cambridgesoft.com/TheMerckIndex/index.asp. Accessed 10-20-2009.

32. Food and Drug Administration.GRAS status sof succinic acid. 10-31-2006. http://www.accessdata.fda.gov/scripts/fcn/fcnDetailNavigation.cfm?rpt=scogsListing&id=339. Accessed 7- 28-2010.

33. Food and Drug Administration.GRAS status of adipic acid. 10-31-2006. http://www.accessdata.fda.gov/scripts/fcn/fcnDetailNavigation.cfm?rpt=scogsListing&id=9.

34. Food and Drug Administration.Azelaic Acid 20% Cream – Drug Approval Label. 1999. http://www.accessdata.fda.gov/drugsatfda_docs/label/2003/20428slr016_azelex_lbl.pdf.

35. Food and Drug Administration.Azelaic Acid 15% Gel – Drug Approval Label. 2003. http://www.accessdata.fda.gov/drugsatfda_docs/label/2005/021470s003lbl.pdf.

36. Jung, H. W., Tschaplinski, T. J., Wang, L., Glazebrook, J., and Greenberg, J. T. Priming in Systemic Plant Immunity. Science. 2009;324:(5923):89-91.

37. Schneider KW. Contact dermatitis due to diethyl sebacate. Cont.Derm. 1980;6:506-507.

38. Shikani AH, Eisele DW, and Domb AJ. Polymer delivery of chemotherapy for squamous cell carcinoma of the head and neck. Arch.Otolaryngol.Head and Neck Surg. 1994;120:1242-1247.

39. Bergman K and Albanus L. Di-(2-ethylhexyl) adipate: absorption, autoradiographic distribution and elimination in mice and rats. Fd Chem Toxicol. 1987;25:(4):309-316.

40. Mochida K, Gomyoda M, and Fujita T. Acetyl tributyl citrate and Dibutyl Sebacate inhibit the growth of cultured mammalian cells. Bulletin of Environ.Contamin.and Toxicol. 1996;56:635-637.

41. Greco AV and Mingrone G. Dicarboxylic acids, an alternate fuel substrate in parental nutrition: an update. Clin.Nutr. 1995;14:143-148.

42. Bertuzzi A, Gandolfi A, Salinari S, Mingrone G, and Greco AV. Pharmacokinetics of dicarboxylic acids in man. Engng Med Biol. 1994;13:472-478. 58

CIR Panel Book Page 106 43. Nguyen, Q. H. and Bui, T. P. Azelaic acid: pharmacokinetic and pharmacodynamic properties and its therapeutic role in hyperpigmentary disorders and acne. Int.J.Dermatol. 1995;34:(2):75-84.

44. Organisation for Economic Co-Operation and Development.IUCLID Data Set for Adipic Acid. 2-15-2006. http://www.chem.unep.ch/irptc/sids/OECDSIDS/124049.pdf. Accessed 7-29-2010.

45. Kennedy GL. Toxicity of adipic acid. Drug.Chem.Toxicol. 2002;25:191-202.

46. Passi S, Nazzaro-Porro M, Picardo M, Mingrone G, and Fasella P. Metabolism of straight saturated medium chain length (C9 to C12) dicarboxylic acids. J Lipid Res. 1983;24:1140-1147.

47. Akhavan A and Bershad S. Topical acne drugs: review of clinical properties, systemic exposure, and safety. Am.J.Clin.Dermatol. 2003;4:473-492.

48. Allergan Inc. 2004. Package Insert: AZELEX – Azelaic Acid cream (20%).

49. Passi S. Pharmacology and pharmacokinetics of azelaic acid. (Secondary reference in Breathnach 1999). Rev Comtemp Pharmacother. 1993;4:441-447.

50. Täuber U, Weiss C, and Matthes H. Percutaneous absorption of azelaic acid in humans. Exp Dermatol. 1992;1:176- 179.

51. Tataranni PA, Mingrone G, DeGaetano A, Raguso C, and Greco AV. Tracer study of metabolism and tissue distributon of sebacic acid in rats. Ann Mutr Metab. 1992;36:296-303.

52. Favuzzi AM, Mingrone G, Bertuzzi A, and et al. Pharmacokinetics of Sebacic Acid in rats. Eur Rev Med Pharmacol Sci. 1999;3:119-125.

53. Mingrone G, Greco AV, Castegneto M, De Gaetano A, Raguso C, and et al. Tissue uptake and oxdation of Disodium Sebacate in man. J.Parenter.Enteral.Nutr. 1991;15:(4):454-459.

54. Greco AV, Mingrone G, Raguso C, and et al. Metabolic effects and disposition of sebacate, an laternate dicarboxylic fuel substrate. Ann.Nutr.Metab. 1992;36:1-11.

55. Mingrone, G., Tacchino, R. M., Castagneto, M., Finotti, E., and Greco, A. V. Use of even-numbered carbon atom dicarboxylic salts in parenteral nutrition as fuel substrate. JPEN J.Parenter.Enteral Nutr. 1992;16:32-38.

56. Mingrone G, Greco AV, Tataranni PA, DeGaetano A, and Castagneto M. Pharmacokinetic profile of dodecanedioic acid, a proposed alternative fuel substrate. JPEN. 1994;18:225-230.

57. Del Rosso, J. Q., Lehman, P. A., and Raney, S. G. Impact of order of application of moisturizers on percutaneous absorption kinetics: evaluation of sequential application of moisturizer lotions and azelaic acid gel 15% using a human skin model. Cutis. 2009;83:119-124.

58. Moody DE and Reddy, J. K. Hepatic peroxisome (microbody) proliferation in rats fed plasticizers and related compounds. Toxicol Appl Pharmacol. 1978;45:497-504.

59. Robins, E. J., Breathnach, A. S., Ward, B. J., Bhasin, Y. P., Ethridge, L., Nazzaro-Porro, M., Passi, S., and Picardo, M. Effect of dicarboxylic acids on Harding-Passey and Cloudman S91 melanoma cells in tissue culture. J.Invest Dermatol. 1985;85:(3):216-221.

60. Passi S, Picardo M, Mingrone G, Breathnach AS, and Navarro-Porro M.Azelaic Acid - biochemistry and metabolism. 1989.

61. Detmar, M., Mayer-da-Silva, A., Stadler, R., and Orfanos, C. E. Effects of azelaic acid on proliferation and ultrastructure of mouse keratinocytes in vitro. J.Invest Dermatol. 1989;93:(1):70-74.

62. Nazzaro-Porro, M., Passi, S., Zina, G., Bernengo, A., and Breathnach, A. Effect of azelaic acid on human malignant melanoma. Lancet. 1980;1:1109-1111. 59

CIR Panel Book Page 107 63. Breathnach, A. S. Azelaic acid: potential as a general antitumoural agent. Med.Hypotheses. 1999;52:(3):221-226.

64. Greco AV, Mingrone G, Arcieri Mastromattei E, Finotti E, and Castegneto M. Toxicity of Disodium Sebacate. Drug Exp.Clin.Res. 1990;16:531-536.

65. Environmental Protection Agency.Robust summary for dicarboxylic acid category. 7-10-2001. http://www.epa.gov/HPV/pubs/summaries/dicarbx/c13108.pdf. Accessed 7-19-2010.

66. Organisation for Economic Co-Operation and Development.SIDS report on Dodecanedioic Acid, CAS No. 693-23- 2. 1994. http://www.chem.unep.ch/irptc/sids/OECDSIDS/693232.pdf. Accessed 7-29-2010.

67. Organisation for Economic Co-Operation and Development.SIDS Initial Report for SIAM 18 - Adipic Acid, CAS No. 124-04-9. 2004. http://www.chem.unep.ch/irptc/sids/OECDSIDS/124049.pdf. Accessed 7-29-2010.

68. Maekawa, A., Todate, A., Onodera, H., Matsushima, Y., Nagaoka, T., Shibutani, M., Ogasawara, H., Kodama, Y., and Hayashi, Y. Lack of toxicity/carcinogenicity of monosodium succinate in F344 rats. Food Chem.Toxicol. 1990;28:(4):235-241.

69. Mingrone G, Greco AV, Navarro-Porro M, and Passi S. Toxicity of Azelaic Acid. Drugs Exp.Clin.Res. 1983;9:447- 455.

70. Fukui S, Shimoyama T, Tamura K, Yamamura M, and Satomi M. Mucosal blood flow and generation of superoxide in rat experimental colitis induced by Succinic Acid. J.Gastroenterol. 1997;32:464-471.

71. Brown-Woodman PD, Post EJ, Chow PY, and White IG. Effects of malonic, citric and caffeic acids on the motility of human sperm and penetration of cervical mucous. Int.J.Fertil. 1985;30:38-44.

72. Organisation for Economic Co-Operation and Development.SIDS Initial Assessment for SIAM 16 - Disodium Succinate, CAS No. 150-90-3. 2003. http://www.chem.unep.ch/irptc/sids/OECDSIDS/150903.pdf. Accessed 7-29-2010.

73. Bradford JC, Brown GL, Caldwell JA, and Drobeck HP. Teratology and mutagenicity studies with glutaric acid. Teratol. 1984;29:19A.

74. Mingrone G, Manicelli R, and Metro D. Influence of sodium salts of saturated medium chain length C6, C9, C10 and C12 dicarboxylic acids on the uterine horn of rat in vitro. Q.J.Exp.Physiol. 1988;73:153-162.

75. National Toxicology Program.Salmonella assayon malonic acid. Study ID: 228078. 1989. http://ntp- apps.niehs.nih.gov/ntp_tox/index.cfm?fuseaction=salmonella.salmonellaData&endpointlist=SA&study%5F no=228078&cas%5Fno=141%2D82%2D2&activetab=detail. Accessed 7-26-2010.

76. Ishidate M Jr, Sofuni T, Yoshikawa K, Hayaski M, Nohmi T, Sawada M, and MAtsouka A. Primary mutagenicity screening of food additives currently used in Japan. Food Chem Toxicol. 1984;22:(8):623-636.

77. TOXNET.CCRIS database - Sodium Succinate. CAS No. 2292-54-5. 2-14-1997. Accessed 7-26-2010.

78. National Toxicology Program.Salmonella assay on adipic acid. Study ID: A86767. 1997. http://ntp- apps.niehs.nih.gov/ntp_tox/index.cfm?fuseaction=salmonella.salmonellaData&endpointlist=SA&study%5F no=A86767&cas%5Fno=124%2D04%2D9&activetab=detail. Accessed 7-26-2010.

79. TOXNET.CCRIS database - Adipic acid. CAS No. 124-04-9. 10-1-2009. http://toxnet.nlm.nih.gov/cgi- bin/sis/search/f?./temp/~vDEUtx:1.

80. Otoshi T, Iwata H, Yamamoto S, Murai T, Yamaguchi S, Matsui-Yuasa I, Otani S, and Fukushima S. Severity of promotion by sodium salts of succinic acid in rat urinary bladder carcinogenesis correlates with sodium ion concentration under conditions of equal urinary pH. Carcinogenesis. 1993;14:(11):2277-2281.

81. Ziel, K., Yelverton, C. B., Balkrishnan, R., and Feldman, S. R. Cumulative irritation potential of metronidazole gel compared to azelaic acid gel after repeated applications to healthy skin. J.Drugs Dermatol. 2005;4:(6):727- 60

CIR Panel Book Page 108 731.

82. Feldman, S., Careccia, R. E., Barham, K. L., and Hancox, J. Diagnosis and treatment of acne. Am.Fam.Physician. 2004;69:(9):2123-2130.

83. American Chemistry Council Aliphatic Esters Diesters Task Group.High Production Volume (HPV) Chemical Challenge Program test plan for th diesters category of the aliphatic esters chemicals. Dated June 2, 2010. 6-28-2010. http://www.epa.gov/oppt/chemrtk/pubs/summaries/alipestr/c13466rt8.pdf. Accessed 7-8-0010.

84. Dirven, H. A., Theuws, J. L., Jongeneelen, F. J., and Bos, R. P. Non-mutagenicity of 4 metabolites of di(2- ethylhexyl)phthalate (DEHP) and 3 structurally related derivatives of di(2-ethylhexyl)adipate (DEHA) in the Salmonella mutagenicity assay. Mutat.Res. 1991;260:(1):121-130.

85. Environmental Protection Agency.Robust summaries for substances in the HPV test plan for the diesters category of the aliphatic esters chemicals. 6-2-2010. http://www.epa.gov/oppt/chemrtk/pubs/summaries/alipestr/c13466rr8.pdf. Accessed 7-8-2010.

86. Takahashi T, Tanaka A, and Yamaha T. Elimination, distribution, and metabolsim of di-(2-ethylhexyl)adipate (DEHA) in rats. Toxicology. 1981;22:223-233.

87. Cornu, M. C., Keith, Y., Elcombe, C. R., and Lhuguenot, J. C. In vivo and in vitro metabolism of di-(2-ethylhexyl) adipate a peroxisome proliferator, in the rat. Arch.Toxicol. 1988;Suppl 12:265-268.

88. Loftus, N. J., Laird, W. J., Steel, G. T., Wilks, M. F., and Woollen, B. H. Metabolism and pharmacokinetics of deuterium-labelled di-2-(ethylhexyl) adipate (DEHA) in humans. Food Chem.Toxicol. 1993;31:(9):609- 614.

89. Fukuyasu H, Orikasa Y, Kai F, Inouye S, Yamaguchi H, and et al. In vivo activity and metabolic fate of 2-(2,3,3- triiodoallyl)tetrazole (ME1401), a novel antifungal agent. Drugs Exp.Clin.Res. 1989;15:(8):335-347.

90. Lake BG. Mechanisms of hepatocarcinogenicity of peroxisome-proliferating drugs and chemicals. Annu Rev Pharmacol Toxicol. 1995;35:483-507.

91. Cornu, M. C., Lhuguenot, J. C., Brady, A. M., Moore, R., and Elcombe, C. R. Identification of the proximate proliferator(s) derived from di (2-ethylhexyl) adipate and species differences in response. Biochem Pharmacol. 1992;43:(10):2129-2134.

92. Keith, Y., Cornu, M. C., Canning, P. M., Foster, J., Lhuguenot, J. C., and Elcombe, C. R. Peroxisome proliferation due to di (2-ethylhexyl) adipate, 2-ethylhexanol and 2-ethylhexanoic acid. Arch.Toxicol. 1992;66:(5):321- 326.

93. National Toxicology Program.Carcinogenesis bioassay of di(2-ethylhexyl)adipate (CAS No. 103-23-1). F344 and B6C3F1 mice. (Feed study). 1982. http://ntp.niehs.nih.gov/ntp/htdocs/LT_rpts/tr212.pdf. Accessed 6-20- 2010.

94. Lake, B. G., Price, R. J., Cunninghame, M. E., and Walters, D. G. Comparison of the effects of di-(2- ethylhexyl)adipate on hepatic peroxisome proliferation and cell replication in the rat and mouse. Toxicology. 1997;123:(3):217-226.

95. Tomaszewski, K. E., Agarwal, D. K., and Melnick, R. L. Invitro steady-state levels of hydrogen eroxide after exposure of male F344 rats and female B6C3F1 mice to hepatic peroxisome proliferators. Carcinogenesis. 1986;7:(11):1871-1876.

96. Reddy, J. K., Reddy, M. K., Usman, M. I., Lalwani, N. D., and Rao, M. S. Comparison of hepatic peroxisome proliferative effect and its implication for hepatocarcinogenicity of phthalate esters, di(2-ethylhexyl) phthalate, and di(2-ethylhexyl) adipate with a hypolipidemic drug. Environ.Health Perspect. 1986;65:317- 327.

97. Barber, E. D., Astill, B. D., Moran, E. J., Schneider, B. F., Gray, T. J., Lake, B. G., and Evans, J. G. Peroxisome 61

CIR Panel Book Page 109 induction studies on seven phthalate esters. Toxicol.Ind.Health. 1987;3:7-22.

98. Lin, L. I. The use of multivariate analysis to compare peroxisome induction data on phthalate esters in rats. Toxicol.Ind.Health. 1987;3:(2):25-48.

99. von Däniken A., Lutz, W. K., Jâckh, R., and Schlatter, C. Investigation of the potential for binding of di(2- ethylhexyl) phthalate and di(2-ethylhexyl) adipate to liver DNA in vivo. Toxicol Appl Pharmacol. 1984;73:(3):373-387.

100. Büsser, M.-T. and Lutz, W. K. Stimulation of DNA Synthesis in Rat and Mouse Liver by Various Tumor Promoters. Carcinogenesis. 1987;8:(10):1433-1437.

101. Takagi, A., Sai, K., Umemura, T., Hasegawa, R., and Kurokawa, Y. Significant increase of 8- hydroxydeoxyguanosine in liver DNA of rats following short-term exposure to the peroxisome proliferators di(2-ethylhexyl)phthalate and di(2-ethylhexyl)adipate. Jpn.J.Cancer Res. 1990;81:213-215.

102. Kawashima, Y., Hanioka, N., Matsumura, M., and Kozuka, H. Induction of microsomal stearoyl-CoA desaturation by the administration of various peroxisome proliferators. (Secondary reference in IARC 2000). Biochim.Biophys.Acta. 1983;752:259-264.

103. Kawashima Y, Nakagawa S, Tachibana Y, and Kozuka H. Effects of peroxisome proliferators on fatty acid-binding protein in rat liver. (Secondary reference in IARC 2000). Biochim Biophys Acta. 1983;754:21-27.

104. Yanagita T., Satoh M., Nomura H., Enomoto N., and Sugano M. Alteration of hepatic phospholipids in rats and mice by feeding di-(2-ethylhexyl)adipate and di-(2-ethylhexyl)phthalate. (Secondary reference in IARC 2000.). Lipids. 1987;22:572-577.

105. Motojima, K., Passilly, P., Peters, J. M., Gonzalez, F. J., and Latruffe, N. Expression of putative fatty acid transporter genes are regulated by peroxisome proliferator-activated receptor alpha and gamma activators in a tissue- and inducer-specific manner. (Secondary reference in IARC 2000). J.Biol.Chem. 1998;273:16710-16714.

106. Organisation for Economic Co-Operation and Development.SIDS Initial Report for Siam 4 - Dibutyl Adipate CAS No. 105-99-7. 1996. http://www.chem.unep.ch/irptc/sids/OECDSIDS/105997.pdf. Accessed 7-29-2010.

107. AMA Laboratories. Acute oral toxicity of Liquiwax DIADD (Dioctyldodecyl Dodecanedioate). AMA Ref No. WPAT90-59/OT5982.II. Unpublished data submitted by the Council (3 pp). 12-13-1990.

108. AMA Laboratories. Acute oral toxicity of Liquiwax DICDD (Diisocetyl Dodecanedioate). AMA Ref No: WPAT90- 59/OT6098.II Unpublished data submitted by the Council (3 pp). 1-3-1991.

109. AMA Laboratories. Acute dermal toxicity - limit test - Liquiiwax DIADD (Dioctyldedecyl Dodecanediaote). AMA Ref No. WPAT93-155/AD1076.BII. Unpublished data submitted by Council (4 pp). 6-14-1993.

110. EviC-CEBA. Attestation of biological significance (acute oral toxicity and dermal irritation) on diethylhexyl sebacate. Study ref. T 207/4072. Unpublished data submitted by the Council (2 pp). 4-7-1994.

111. EviC-CEBA. Attestation of biological significance (acute oral toxicity and dermal irritation) on diisocetyl adipate. Study ref. T 207/4073. Unpublished data submitted by the Council (2 pp). 4-7-1994.

112. EviC-CEBA. Attestation of biological test (acute oral toxicity and dermal irritation) on diisodecyl adipate. Study ref. T 207/4076. Unpublished data submitted by the Council (2 pp). 5-9-1994.

113. EviC-CEBA. Attestation of biological significance (acute oral toxicity and dermal irritation) on dioctyldodecyl adipate. Study ref. T 207/4071. Unpublished data submitted by the Council (2 pp). 5-9-1994.

114. EviC-CEBA. Study of acute toxicity and local tolerance (acute oral, acute dermal, and ocular irritation) on diisopropyl adipate. Study report: Te 297/98-1589 and Te 298/98-1589. Unpublished data submitted by the Council (4 pp). 5-27-1998. 62

CIR Panel Book Page 110 115. Singh AR, Lawrence WH, and Autian J. Embryonic-fetal toxicity and teratogenic effects of adipic acid esters in rats. J Pharm Sci. 1973;62:(10):1596-1600.

116. Smith CC. Toxicity of butyl stearate, dibutyl sebacate, dibutyl phthalate, and methoxyethyl oleate. AMA Arch.Ind.Hyg.Occup.Med. 1953;7:310-318.

117. Organisation for Economic Co-Operation and Development.SIDS Initial Report for SIAM 10 - Bis(2-ethylhexyl) Adipate (DEHA), CAS No. 103-23-1. 2000. http://www.chem.unep.ch/irptc/sids/OECDSIDS/103231.pdf. Accessed 7-29-2010.

118. ExxonMobil Chemical Company.High Production Volume (HPV) Chemical Challenge Program. Test plan for the alkyl alcohols C6-C13 Category. 2-5-2002. http://www.epa.gov/HPV/pubs/summaries/alkal613/c13590rs.pdf. Accessed 2010.

119. Smyth HF, Carpenter CP, and Weil CS. Range-finding toxicity data: List IV. AMA Arch.Ind.Hyg.Occup.Med. 1949;4:119-122.

120. Wato, E., Asahiyama, M., Suzuki, A., Funyu, S., and Amano, Y. Collaborative work on evaluation of ovarian toxicity. 9) Effects of 2- or 4-week repeated dose studies and fertility study of di(2-ethylhexyl)adipate (DEHA) in female rats. J.Toxicol.Sci. 2009;34:(Special Issue I):SP101-SP109.

121. Kluwe, W. M. Carcinogenic potential of phthalic acid esters and related compounds: structure-activity relationships. Environ.Health Perspect. 1986;65:271-278.

122. Lee, K. P., Valentine, R., and Bogdanffy, M. S. Nasal lesion development and reversibility in rats exposed to aerosols of dibasic esters. Toxicol.Pathol. 1992;20:(3 Part 1):376-393.

123. Consumer Product Testing. 1991. Primary dermal irritation in rabbits, primary ocular irritation in rabbits of Schermcemol DIS (Diisopropyl Sebacate). Experiment Reference Number: 91055. (Material tested as supplied - approx. 100%). 3 pages.

124. MB Research Laboratories. 2003. EpiOcular MTT viability assay of a cream containing 1.2% Diethylhexyl Sebacate. 10 pages.

125. AMA Laboratories. Primary eye irritation of liquiwax DIADD (Dioctyldodecyl Dodecanedioate). AMA Lab No. WPAT90-59/PE5982.II. Unpublished data submitted by theCouncil (5 pp). 12-3-1990.

126. AMA Laboratories. Primary eye irritation of liquiwax DICDD (Diisocetyl Dodecanedioate). AMA Ref No: WPAT90-59/PE6098.II. Unpublished data submitted by the Council (3 pp). 12-22-1990.

127. Personal Care Products Council. 1989. Summary of safety data on Diethyl Sebacate (primary skin irritation, cumulative skin irritation, contact allergenicity). 8 pages.

128. EviC-CEBA. Attestation of biological significance (acute oral toxicity and dermal irritation) on diisopropyl sebacate. Study ref. T 207/4078. Unpublished data submitted by the Council (2 pp). 4-7-1998.

129. AMA Laboratories. Guinea pig maximization test (Kligman) on liquiwax DIADD (Dioctyldodecyl Dodecanedioate). AMA Ref No: WPAT93-GPMB1076.BII. Unpublished data submitted by the Council (6 pp). 6-14-1993.

130. Environmental Protection Agency.Integrated Risk Information System. Di(2-ethylhexyl)adipate. (CASRN 103-23- 1). 3-6-2010. N:\CIR\DicarboxylicAcids\DatabaseSearchResults\EPA\IRIS-searched7-8-10_updatred3-16- 10.mht. Accessed 7-8-2010.

131. Dalgaard, M., Hass, U., Vinggaard, A. M., Jarfelt, K., Lam, H. R., rensen, I. K., Sommer, H. M., and Ladefoged, O. Di(2-ethylhexyl) adipate (DEHA) induced developmental toxicity but not antiandrogenic effects in pre- and postnatally exposed Wistar rats. Reprod.Toxicol. 2003;17:(2):163-170.

132. Kang, J. S., Morimura, K., Toda, C., Wanibuchi, H., Wei, M., Kojima, N., and Fukushima, S. Testicular toxicity of DEHP, but not DEHA, is elevated under conditions of thioacetamide-induced liver damage. 63

CIR Panel Book Page 111 Reprod.Toxicol. 2006;21:253-259.

133. Alvarez, L., Driscoll, C., Kelly, D. P., Staples, R. E., Chromey, N. C., and Kennedy, G. L., Jr. Developmental toxicity of dibasic esters by inhalation in the rat. Drug Chem.Toxicol. 1995;189:(4):295-314.

134. Kelly, D. P., Kennedy, G. L., Jr., and KEENAN, C. M. Reproduction study with dibasic esters following inhalation in the rat. Drug Chem.Toxicol. 1998;21:(3):253-267.

135. Miyata, K., Shiraishi, K., Houshuyama, S., Imatanaka, N., Umano, T., Minobe, Y., and Yamasaki, K. Subacute oral toxicity study of di(2-ethylhexyl)adipate based on the draft protocol for the "Enhanced OECD Test Guideline no. 407". Arch.Toxicol. 2006;80:181-186.

136. Ghisari, M. and Bonefeld-Jorgensen, E. C. Effects of plasticizers and their mixtures on estrogen receptor and thyroid hormone functions. Toxicol.Lett. 2009;189:(1):67-77.

137. Andersen, P. H. and Jensen, N. J. Mutagenic investigation of flavourings: dimethyl succinate, ethyl pyruvate and aconitic acid are negative in the Salmonella/mammalian-microsome test. Food Addit.Contam. 1984;1:(3):283-288.

138. National Toxicology Program.Salmonella assay on dimethyl succinate. Study ID: 947738. 1988. http://ntp- apps.niehs.nih.gov/ntp_tox/index.cfm?fuseaction=salmonella.salmonellaData&endpointlist=SA&study%5F no=947738&cas%5Fno=106%2D65%2D0&activetab=detail. Accessed 7-26-2010.

139. National Toxicology Program.Salmonella assay on dimethyl glutarate. Study ID A20348. 1995. http://ntp- apps.niehs.nih.gov/ntp_tox/index.cfm?fuseaction=salmonella.salmonellaData&endpointlist=SA&study%5F no=A20348&cas%5Fno=1119%2D40%2D0&activetab=detail. Accessed 7-26-2010.

140. National Toxicology Program.Salmonella assay on dimethyl adipate. Study ID: A45330. 1994. http://ntp- apps.niehs.nih.gov/ntp_tox/index.cfm?fuseaction=salmonella.salmonellaData&endpointlist=SA&study%5F no=A45330&cas%5Fno=627%2D93%2D0&activetab=detail. Accessed 7-26-2010.

141. Zeiger E, Haworth E, Mortelmans S, and et al. Mutagenicity testing of di(2-ethylhexyl) phthalate and related chemicals in Salmonella. Environ Mutagen. 1985;7:213-232.

142. Warren JR, Lalwani, N. D., and Reddy, J. K. Phthalate esters as peroxisome proliferator carcinogens. Environ.Health Perspect. 1982;45:35-40.

143. Simmon VF, Kauhanen K, and Tardiff R.G. Mutagenic activity of chemicals identified in drinking water. Dev Toxicol Environ Sci. 1977;2:249-258.

144. National Toxicology Program.Salmonella assay eith diethylhexyl adipate. Study ID: 963935. 1988. http://ntp- apps.niehs.nih.gov/ntp_tox/index.cfm?fuseaction=salmonella.salmonellaData&endpointlist=SA&study%5F no=963935&cas%5Fno=103%2D23%2D1&activetab=detail. Accessed 7-26-2010.

145. Seed JL. Mutagenic activity of phthalate estes in bacterial seed liquid suspension assays. Environ.Health Perspect. 1982;45:111-114.

146. McGregor, D. B., Brown, A., Cattanach, P., Edwards, I., McBride, D., Riach, C., and Caspary, W. J. Responses of the L5178Y tk+/tk- mouse lymphoma cell forward mutation assay: III. 72 coded chemicals. (Secondary reference in IARC 2000). Environ.Mol.Mutagen. 1988;12:(1):85-154.

147. Reisenbichler, H. and Eckl, P. M. Genotoxic effects of selected peroxisome proliferators. (Secondary reference in IARC 2000). Mutat.Res. 1993;286:135-144.

148. Galloway SM, Armstrong MJ, Rueben C, Colman S, Brown B, CAnnon C, Bloom AD, Nakamura F, Ahmed M, Duk S, Rimpo J, Margolin BH, REsnick MA, Anderson B, and Zeiger E. Chromosome aberrations and sister chromatid exchanges in Chinese hamster ovary cells: Evaluation of 108 chemicals. Environ Mol Mutagen. 1987;10:(Suppl 10):1-175.

64

CIR Panel Book Page 112 149. Shelby, MD, Erexson GL, Hook GH, and Tice RR. Evaluation of a three-exposure mouse bone marrow micronucleus protocol: Results with 49 chemicals. Environ Mol Mutagen. 1993;21:160-179.

150. Shelby, MD and Witt, KL. Comparison of results from mouse bone marrow chromosome aberration and micronucleus tests. Environ Mol Mutagen. 1995;25:302-313.

151. National Toxicology Program.Chromosome aberration assay of diethylhexyl adipate. Study ID: 959525. 1988. http://ntp- apps.niehs.nih.gov/ntp_tox/index.cfm?fuseaction=invivoca.casummary&study_no=959525&cas_no=103% 2D23%2D1&endpointlist=CA. Accessed 7-26-2010.

152. DiVincenzo, G. D., Hamilton, M. L., Mueller, K. R., Donish, W. H., and Barber, E. D. Bacterial mutagenicity testing of urine from rats dosed with 2-ethylhexanol derived plasticizers. Toxicology. 1985;34:(3):247-259.

153. McKee, R. H., Lington, A. W., and Traul, K. A. An evaluation of the genotoxic potential of di-isononyl adipate. Environ.Mutagen. 1986;8:(6):817-827.

154. Szybalski W. Special microbiological systems - Observations on chemical mutagenesis in microorganisms. Ann.NY Acad.Sci. 1958;76:475-489.

155. Wild, D., King, M. T., Gocke, E., and Eckhardt, K. Study of artificial flavoring substances for mutagenicity in the salmonella/microsome, basc and micronucleus tests. Fd Chem Toxicol. 1983;21:707-719.

156. TOXNET.CCRIS database - Dibutyl Sebacate. CAS No. 109-43-3. 2-12-2001. http://toxnet.nlm.nih.gov/cgi- bin/sis/search/f?./temp/~LfAk2H:2. Accessed 7-26-2010.

157. TOXNET.CCRIS database - Diethylhexyl Sebacate. CAS NO. 122-62-3. 6-2-2010. http://toxnet.nlm.nih.gov/cgi- bin/sis/search/f?./temp/~J98xpD:1. Accessed 7-26-2010.

158. Osterle D and Deml E. Promoting activity of di(2-ethylhexyl)phthalate in rat liver foci bioassay. J.Cancer Res.Clin.Oncol. 1988;114:(2):133-136.

159. Hasegawa R and Ito N. Liver medium-term bioassay in rats for screening of carcinogenesis and modifying factors in hepatocarcinogenesis. Fd Chem Toxicol. 1992;30:(11):979-992.

160. AMA Laboratories. 50 Human subject repeat insult patch test skin irritation/sensitization evaluation on Liquiwax DISA (Diisostearyl Adipate), DICDD (Diisocetyl Dodecanedioate), and DIADD (Dioctyldodecyl Dodecanedioate). AMA Ref No: WP96-BERN1-18/RIPT4618.BII. Unpublished data submitted by the Council. 4-11-1996.

161. Anonymous. Clinical evaluation report: human patch test of a body cream containing 1.5% diethyl sebacate. Unpublished data submittd by the Personal Care Products Council. 2003.

162. KGL Inc. 2003. An evaluation of the contact-sensitization potential of a topical coded product (body cream containing 1.5% diethyl sebacate) in human skin by means of the maximization assay. 10 pages.

163. Anonymous. Clinical evaluation report: human patch test of a foundation containing 1.8% diisopropyl sebacate. Unpublished data submitted by the Council.. 2000.

164. Product Investigations Inc. 2005. Determination of the irritating and sensitizing propensities of Schercemol DIS (Diisopropyl Sebacate) on human skin. Report: PII No. 19745. (Material tested as supplied - approx. 100%). 12 pages.

165. KGL Inc. 2006. An evaluation of the contact-sensitization potential of a topical coded product (foundation containing 2.2% diisopropyl sebacate) in human skin by means of the maximization assay. 10 pages.

166. Reliance Clinical Testing Services. 2007. Summary of HRIPT of two heat protection hair sprays containing 1% Diisopropyl Sebacate. 1 pages.

65

CIR Panel Book Page 113 167. Essex Testing Clinic Inc. 2007. Summary of HRIPT of a heat protection hair spray containing 7.2% Diisopropyl Sebacate.

168. Kimura M and Kawada A. Contact dermatitis due to Diethyl Sebacate. Contact Dermatitis. 1999;40:48-49.

169. Narita T, Oiso N, Ota T, Kawara S, and Kawada A. Allergic contact dermatitis due to Diethyl Sebacate in a hand cream. Contact Dermatitis. 2006;55:117.

170. Sasaki E, Hata M, Aramaki J, and Honda M. Allergic contact dermatitis due to Diethyl Sebacate. Contact Dermatitis. 1997;36:172.

171. Soga F, Katoh N, and Kishimoto S. Contact dermatitis due to lanoconazole, cetyl alcohol and Diethyl Sebacate in lanoconazole cream. Contact Dermatitis. 2004;50:49-50.

172. Tanaka M, Kobayashi S, Murata T, Tanikawa A, and Nishikawa T. Allergic contact dermatitis from Diethyl Sebacate in lanoconazole cream. Contact Dermatitis. 2000;43:233-234.

173. De Groot AC, Conemans JM, and Schutte T. Contact allergy to Diisopropyl Sebacate in Zineryt lotion. Contact Dermatitis. 1991;25:260-261.

174. Dooms-Goossens A, De Veyhlder H, De Boulle K, and Maertens M. Allergic contact dermatitis due to Diisopropyl Sebacate. Contact Dermatitis. 1986;15:192.

175. Elder RL (ed). Final Report on the Safety Assessment of Stearyl Alcohol, Oleyl Alcohol, and Octyl Dodecanol. Journal of the American College of Toxicology. 1985;4:(5):1-29.

176. Opdyke, D. L. J. Fragrance Raw Materials Monographs - Supplement to Earlier Monographs on Fragrance Materials. 1973. 1079

177. Andersen, F. A. Final Report on the Safety Assessment of Formic Acid. International Journal of Toxicology. 1997;16:(3).

178. International Programme on Chemical Safety.Methanol: Poisons Information Monograph 335. 2009. http://www.inchem.org/documents/pims/chemical/pim335.htm. Accessed 10-20-2009.

179. D'Alessandro, A., Osterloh, J. D., Chuwers, P., Quinlan, P. J., Kelly, T. J., and Becker, C. E. Formate in serum and urine after controlled methanol exposure at the threshold limit value. Environ.Health Perspect. 1994;102:(2):178-181.

180. Heldreth BA. Final report on Methyl Acetate, Simple Alkyl Acetate Estes and Related Alcohols (Draft). Report in progress. Available from the Cosmetic Ingredient Review. 2010.

181. Andersen, F. A. Final Report on the Safety Assessment of Methyl Alcohol. International Journal of Toxicology. 2001;20:((Suppl. 1)):57-85.

182. American Chemistry Council and Oxo Process Panel. Screening Information Data Set (SIDS) Initial Assessment Report for SIDS Initial Assessment Meeting (SIAM) 27: n-Propyl Acetate. OECD. 2008.

183. Nelson BK, Brightwell WS, Taylor BJ, Khan A, Burg JR, Kreig EF Jr, and Massari VJ. Behavioral teratology investigatoi of 1-propanol administered by inhalation to rats. Neurotoxicol Teratol. 1989;11:(2):153-159.

184. Stillman MA, Maiback HI, and Shalita AR. Relative irritancy of free fatty acids of different chain length. Contact Derm. 1975;1:65-69.

185. Wahlberg JE and MAibach HI. Nonanoic acid irrigation: A positive control at routine patch testing? Contact Derm. 1980;6:(2):128-130.

186. Agner, T. and Serup J. Skin reactions to irritants assessed by polysulfide rubber replica. Contact Dermatitis. 1987. 17:(4):205-211. 66

CIR Panel Book Page 114 187. Agner T and Serup J. Nonanoic acid irrigation: A positive contorl at routine patch testing? Contact Derm. 1987;17:(4):20-211.

188. Willis CM, Stephens JM, and Wilkinson JD. Experimentally-induced irritant contact dermatitis. Determination of optimum irritant concentrations. Contact Derm. 2010;1:20-24.

189. Agner T and Serup J. Seasonal variation ofskin resistance to irritants. Br J Dermatol. 2010;121:(3):323-328.

190. Elder RL (ed). Final Report on the Safety Assessment of Cetearyl Alcohol, Cetyl Alcohol, lsostearyl Alcohol, Myristyl Alcohol, and Behenyl Alcohol. Journal of the American College of Toxicology. 1988;7:(3):359- 413.

191. Martinez, T. T., Jaeger, R. W., deCastro, F. J., Thompson, M. W., and Hamilton, M. F. A comparison of the absorption and metabolism of isopropyl alcohol by oral, dermal and inhalation routes. Vet Hum Toxicol. 1986;28:(3):233-236.

192. Burleight-Flayer H, Garman R, Neptun D, Bevan C, Gardiner T, Kapp R, Tyler T, and Wright G. Isopropanol vapor inhalaiton oncogenicity study in Fischer 344 rats and CD-1 mice. Fund Appl Toxicol. 1997;36:(2):95-111.

193. International Programme on Chemical Safety.Isopropyl Alcohol: Poisons Information Monograph 290. 2009. http://www.inchem.org/documents/pims/chemical/pim290.htm. Accessed 10-12-2009.

194. Tyl, R. W., Masten, L. W., Marr, M. C., Myers, C. B., Slauter, R. W., Gardiner, T. H., Strother, D. E., McKee, R. H., and Tyler, T. R. Developmental toxicity evaluation of isopropanol by gavage in rats and rabbits. Fundam.Appl Toxicol. 1994;22:(1):139-151.

195. Damato JM, Martin DM, and Fehn PA. Allergic contact sensitization test of a spray concentrate containining 80.74% isopropyl alcohol. 1979.

196. Anonymous. Final Report Repeated Insult Patch Test of a Hair Dye Base (3373) Containing 2.85% Isopropyl Alcohol and 1.95% Isopropyl Acetate. Unpublished data submitted by teh Council. 2010.

197. Suihko C and Serup J. Fluorescence confocal laser scanning microscopy for in vivo imaging of epidermal reactions to two experimental irritants. Skin Res Technol. 2008;14:(4):498-503.

198. High Production Volume (HPV) Chemical Challenge Program. Test Plan for the Alkyl Alcohols C6 - C13 Category. 2002.

199. International Program on Chemical Safety.Butyl acetates. 2005. http://www.inchem.org/documents/cicads/cicad64.htm. Accessed 11-20-2009.

200. Kawaguchi, M., Yamazaki, T., and Nakazawa, H. Biological effects of di(2-ethylhexyl)adipate. Environmental.Sciences.: an.International.Journal of Environmental.Physiology.and Toxicology. 2002;9:(2-3):198.

67

CIR Panel Book Page 115 APPENDIX I – ESTEARSE METABOLITE SUMMARY DATA

Decyl Alcohol – metabolite of Decyl Succinate

Clinical Irritation and Sensitization Tested in at a concentration of 3% in petrolatum, decyl alcohol produced no irritation in a 48 h closed-patch test in 25 human subjects.176

Methanol – metabolite of Dimethyl Succinate, Dimethyl Glutarate, Dimethyl Adipate

Absorption, Distribution, Metabolism, and Excretion Methanol is further metabolized to formaldehyde and then to formic acid. The CIR Expert Panel concluded that formic acid is safe where used in cosmetic formulations as a pH adjustor with a 64 ppm limit for the free acid.177 The main toxicological risks in humans are severe metabolic acidosis with increased anion gap, typically following oral exposure resulting in > 100 mg/L of formate in the urine.178 The acidosis and the formic acid metabolite are believed to play a central role in both the central nervous system toxicity and the ocular toxicity.

A study to determine the formate levels that resulted from exposure of human volunteers to 200 ppm of methanol for 4 h was conducted. Human volunteers (n=27; age 20-55 y) were exposed to 200 ppm methanol (the Occupational Safety and Health Administration [OSHA] Permissible Exposure Limit) for 4 h and to water vapor for 4 h in a double-blind, random study.179 Urine samples were collected at 0, 4 and 8 h and blood samples were collected from the subjects before they entered the chamber, every 15 min for the first hour, every 30 min from the first to the third hour and at 4 h. Urine and serum samples were analyzed for formate (LOD 0.5 mg/L). Twenty-six of 27 enrolled subjects completed the study (11 females and 15 males). The researchers did not find any statistically significant differences in serum or urine formate levels between the two exposure conditions at any time point. At the end of the 4 h methanol exposure, formate concentrations of 14.28 ± 8.90 and 7.14 ± 5.17 mg/L were measured in serum and urine, respectively. Under control conditions, formate concentrations of 12.68 ± 6.43 (p=0.38; n=26) and 6.64 ± 4.26 (p=0.59; n=25) mg/L were measured in serum and urine respectively. After 8 h (4 h of no exposure) the serum concentrations were not statistically different with 12.38 ± 6.53 mg/L under methanol exposure conditions and 12.95 ± 8.01 (P=0.6; n=26) under control conditions. Urine formate concentrations after 8 h were 6.08 ± 3.49 and 5.64 ±3.70 mg/L (p=0.6; n=25) in exposed and control conditions, respectively, and were not statistically significantly different. From CIR Final Report on Methyl Acetate180

Clinical Irritation and Sensitization Methyl Alcohol caused primary irritation to the skin; prolonged and repeated contact with Methyl Alcohol resulted in de- fatting and dermatitis. In one occupational study, 3.2% of 274 metalworkers with dermatitis had positive results to a patch test of 30% Methyl Alcohol. Typical allergic responses observed after contact with alcohols were eczematous eruption and wheal and flare at the exposure sites. Eczema and erythema were reported after the consumption of alcoholic beverages by persons sensitized to ethyl alcohol. Five percent Methyl Alcohol caused a slight positive (+) reaction in a closed patch test for allergic contact dermatitis, and concentrations of 7% and 70% caused (+++) reactions. From the CIR final report on methyl alcohol181

Clinical Assessment of Safety Clinical data show that Methyl Alcohol can cause severe metabolic acidosis, blindness, and death: toxicity was manifested earlier and at a lower dose compared to ethyl alcohol, but the comparative fatal dose was the same for both alcohols. All routes of exposure were toxicologically equivalent, as the alcohol distributed readily and uniformly throughout all tissues and organs. Individual susceptibilities to Methyl Alcohol varied, but typically, the ingestion of 80 to 150 ml of 80% Methyl Alcohol was fatal. Symptoms of Methyl Alcohol intoxication after ingestion were delayed for 12 to 18 hours; afterwards, the symptoms included headache, anorexia, weakness, fatigue, leg cramps, and/or pain and vertigo. Severe gastrointestinal pain, nausea, vomiting, diarrhea, mania, failed vision, and convulsions could occur. Chronic exposure to Methyl Alcohol could cause edema, granular degeneration, and necrosis of heart muscle fibers, as well as fatty degeneration of the heart muscle; sudden cardiac failure was associated with Methyl Alcohol intoxication. The liver and kidneys often had parenchymatous degeneration, and the liver had focal necrosis and fatty infiltration. Severe acidosis was necessary for the development of blindness. Similar symptoms were observed after percutaneous or inhalation exposure to Methyl Alcohol. From the CIR final report on methyl alcohol181 68

CIR Panel Book Page 116

Propyl Alcohol – metabolite of Dipropyl Adipate

Absorption, Distribution, Metabolism, and Excretion Rats (strain/sex/number not specified) were exposed via inhalation to 2,000 ppm (8360 mg/m3) for 90 min.182 Propyl acetate was rapidly hydrolyzed to propyl alcohol. During the 90 min exposure period, blood levels of propyl alcohol were between 2.6 and 7.7 fold greater than propyl acetate. From the CIR final report on Methyl Acetate180

Reproductive and Developmental Toxicity The effects of propyl alcohol on fertility were investigated by exposing male Sprague-Dawley rats (18/group) to 0, 3500 or 7000 ppm (0, 8.61 or 17.2 mg/L) propyl alcohol vapor via inhalation 7 h/day, 7days/week for 62 days, prior to mating with unexposed virgin females.183 Female Sprague-Dawley rats (15/group) were similarly exposed and mated with unexposed males. Following parturition, litters were culled to 4/sex and the pups fostered by unexposed dams. The pups were weaned on post natal day (PND) 25 and weighed on PND’s 7, 14, 21, 28 and 35. Male rats exposed to 7000 ppm exhibited a decrease in mating success with 2/16 producing a litter (1 male died as a result of a cage fight and 1 male did not mate). Mating success was not affected in 3500 ppm exposed males or in females. Six males from the 7000 ppm group were retained to determine if this effect was reversible. All 6 males successfully mated 15 weeks after exposure. The authors reported that weight gain was not affected in 7000 ppm exposed females (data not shown), but feed intake was decreased in this treatment group. Crooked tails were observed in 2-3 offspring in 2 of 15 litters from the 7000 ppm maternally exposed group. No other effects on female fertility were reported. No significant differences resulted between offspring of the 7000 ppm group and controls on several behavioral toxicology measures including the Ascent test, Rotorod test, Open Field test, activity test, running wheel activity, avoidance conditioning, and operant conditioning. Activity measures were significantly different between offspring of the 3500 ppm exposure group and controls. From the CIR final report on Methyl Acetate180

Clinical Irritation and Sensitization A cumulative irritation study was conducted involving 20 male subjects, where the relative irritancy of free fatty acids of different chain lengths was evaluated.184 Equimolar concentrations (0.5 M and 1.0 M) of even- and odd- numbered - straight chain saturated fatty acids were dissolved in propanol. Each Al-test® patch containing a fatty acid (0.5 M) was applied to the interscapular area of 10 subjects, and, similarly, each fatty acid was applied at a higher concentration (1.0 M) to the remaining 10 subjects. A control patch containing propanol was also applied to each subject. Patches remained in place for 24 h and reactions were scored 30 minutes after patch removal. This procedure was repeated daily for a total of 10 applications. In both groups of 10 subjects, there were no reactions to propanol.

In an irritation study, wherein 116 healthy male subjects (21 to 55 years old) were patch tested with pelargonic acid at concentrations of 5%, 10%, 20%, and 39.9% in propanol, a propanol-treated control patch was used.185 Dose response curves were developed. Patches (Al-test® discs) were saturated with 0.04 ml of a test solution and applied to the upper back for 48 h. Reactions were scored at 48 h and 96 h post-application. There were no reactions to propanol.

In an another irritation study, wherein 16 volunteers (10 females, 6 males; median age of 29.5 years) were patch tested (closed patches, Finn chambers) with 20% pelargonic acid in propanol (pH of 4.3), propanol was one of the controls used.186 Patches were applied to the anterolateral surface of both upper arms for 24 h. Reactions were scored at 24, 48, and 96 h post-application according to the following scale: 0 (no reaction) to 3 (strong positive reaction: marked erythema, infiltration, possibly vesicles, bullae, pustules and/or pronounced crusting). There were no reactions to propanol.

A skin irritation study was conducted using 42 healthy, non-atopic male volunteers (mean age = 34 years; skin types: II [20 subjects], III [17 subjects], and IV [5 subjects]).187 Pelargonic acid was patch-tested (Finn chambers, volar forearm) at the following concentrations (in propanol): 40% (12 subjects), 60% (32 subjects), 70% (32 subjects), and 80% (28 subjects), and propanol was used as a control. Each subject received between 3 and 10 patch tests. The patches remained in place for 48 h, and reactions were scored 1 h later according to the following scale: - (no visible reaction) to 4+ (intense erythema with bullous formation). There were no reactions to propanol.

In an irritation study, wherein 16 healthy subjects (ages not stated) were patch tested with pelargonic acid (20% in 69

CIR Panel Book Page 117 propanol), propanol was used as a control.188 Closed patches (Finn chambers) containing the test substance were applied to the anterolateral surface of both upper arms. The patches were removed at 24 h post-application and reactions were scored at 24 h and 96 h post-application. There were no reactions to propanol.

In study conducted to investigate a possible seasonal variation in the skin response to pelargonic acid during the winter and summer, propanol was used as a control.189 The study was conducted using 17 healthy volunteers (10 males, 7 females; mean age = 27 years). The test substance was applied (closed patch, Finn chamber) to each arm for 24 h. Reactions were scored at 30 min post-removal. Reactions were not observed at sites treated with propanol, water, or to which an empty chamber was applied. From CIR final report on Methyl Acetate180

Cetyl Alcohol – metabolite of Dicetyl Succinate and Dicetyl Adipate

Clinical Irritation and Sensitization A topical tolerance study involving an 11.5% Cetyl Alcohol cream base was conducted with 80 male subjects, ranging in age from 21 to 52 years and in weight from 120 to 220 pounds. The preparations were applied five times daily (every 3 hours) for 10 days. One subject had erythema, folliculitis, and pustule formation (forearm site).

A formulation containing 6.0% Cetyl Alcohol was tested for its skin irritation potential in 20 subjects according to the protocol stated above. The product did not induce skin irritation. In another study, the skin irritation potential of a cream containing 6.0% Cetyl Alcohol was evaluated in 12 female subjects (18-60 years old). The total irritation score (all panelists) for the 21 applications was 418, indicating mild cumulative irritation.

The skin irritation and sensitization potential of a product containing 8.4% Cetyl Alcohol was evaluated in 110 female subjects. Fourteen days after scoring of the tenth application site, a challenge patch was applied to each subject and removed after 48 h; sites were scored after patch removal. The product did not induce primary irritation or sensitization.

The sensitization potential of a cream containing 3.0% Cetearyl Alcohol was evaluated in 25 subjects (18-25 years old). Following a 10-day non-treatment period, occlusive challenge patches were applied to new sites and removed after 48 h. Sensitization reactions were not observed in any of the subjects. From the CIR final report on cetyl alcohol190

Photosensitization The photosensitization potential of a lipstick product containing 4.0% Cetyl Alcohol was evaluated in 52 subjects. The experimental procedure was not stated. Photosensitization reactions were not noted in any of the subjects. In another study, a skin care preparation containing 1.0% Cetyl Alcohol did not induce photosensitization in the 407 subjects tested. The experimental procedure was not stated. From CIR final report on cetyl alcohol190

Isopropyl Alcohol – metabolite of Diisopropyl Adipate and Diisopropyl Sebacate

Absorption, Distribution, Metabolism, and Excretion Male rabbits (3/group; strain not specified) were treated by different routes of exposure to compare the absorption and metabolism of isopropyl alcohol.191 Groups 1 and 2 were treated via gavage with the equivalent of 2 and 4 ml/kg absolute isopropyl alcohol, respectively, as a 35% isopropanol/water solution. Groups 3 and 4 were treated via whole-body inhalation for 4 h (towels soaked with isopropyl alcohol were place in the inhalation chamber and replenished at ½ hour intervals to maintained a saturated environment; no exact concentration given), with Group 3 animals receiving an additional dermal exposure in the form of a towel soaked with 70% isopropyl alcohol applied to the animals’ chests and Group 4 animals having plastic barriers on their chests and towels prepared the same way as in Group 3 applied on top of the plastic barriers. The alcohol on the towels was replenished at half hour intervals throughout the duration of the experiment. Blood samples were taken at 0, 1, 2, 3 and 4 h. Samples were analyzed for isopropyl alcohol and the metabolite acetone.

Following gavage exposure to 2 or 4 ml/kg, maximum blood levels of 147 and 282 mg/dl, respectively, of isopropyl alcohol were measured. Concentrations of acetone rose steadily over the 4 h period and were 74 and 73 mg/dl 70

CIR Panel Book Page 118 following exposure to 2 or 4 ml/kg, respectively. The authors stated that the maximum levels of isopropyl alcohol observed in this experiment, correlated with inebriation and near coma in the animals. Following inhalation and dermal exposure, the concentration of isopropyl alcohol in the blood continued to rise and was 112 mg/dl at 4 h while the concentration of acetone was 19 at 4 h. Inhalation exposure with a plastic barrier between the soaked towel and the chest resulted in isopropyl alcohol and acetone blood levels of <10 mg/dl.

The researchers concluded that isopropyl alcohol is absorbed by the dermal route but that prolonged dermal exposure (i.e. repeated sponging or soaking for several hours) would be required to produce significant toxicity. From CIR final report on Methyl Acetate180

Subchronic Inhalation Toxicity Fischer 344 rats and CD-1 mice (10/sex/group) were exposed via inhalation to 0, 100, 500, 1500, or 5000 ppm (0, 246, 1230, 3690, or 12,300 mg/m3) isopropyl alcohol for 6 h/day, 5 days/wk for 13 weeks.192 Ataxia, narcosis, hypoactivity and the lack of a startle reflex were observed during exposure at 5000 ppm. Hypoactivity was observed in animals exposed to 1500 ppm isopropyl alcohol. At 13 weeks, no gross lesions were observed. Microscopic examination of control and 5000 ppm exposed animal tissues showed hyaline droplets within the kidneys of male rats only. The size and frequency of the droplets was increased in the treated group. The authors concluded that the NOAEL for this study was 500 ppm and the lowest-observable adverse effect level (LOAEL) was 1500 ppm based upon clinical signs and changes in hematology at 6 weeks. To evaluate the neurobehavioral effects of isopropanol exposure, an additional 15 rats/sex were exposed (via inhalation) to 0, 500, 1500, or 5000 ppm (0, 1230, 3690, or 12,300 mg/m3) for 6 h/day, 5 days/wk for 13 weeks. Isopropyl alcohol did not produce any changes to the parameters of the functional observations battery which was conducted at 1, 2, 4, 9 and 13 weeks. Clinical signs observed in mice, during the exposure, included ataxia, narcosis, hypoactivity and lack of a startle reflex at 5000 ppm. Narcosis, ataxia and hypoactivity were observed in animals exposed to 1500 ppm isopropyl alcohol. At 5000 ppm, increased body weight and increased rate of weight gain were observed in female mice. At 13 weeks, no gross lesions were observed and no treatment-related microscopic changes were observed. A 10% and 21% increase in relative liver weight was observed in female mice at 1500 and 5000 ppm, respectively. The authors concluded that the NOAEL for this study was 500 ppm and the LOAEL was 1500 ppm based on clinical signs and increased liver weights.

Ocular Irritation Isopropyl alcohol has been labeled a severe ocular irritant based on rabbit ocular irritation tests.193 From CIR final report on Methyl Acetate180

Reproductive and Developmental Toxicity Female Sprague-Dawley rats (25/group) were exposed to 0, 400, 800, or 1200 mg/kg/day isopropyl alcohol via gavage on gestational days (GD) 6 through 15.194 Female New Zealand white rabbits (15/group) were exposed to 0, 120, 240, or 480 mg/kg/day isopropyl alcohol via gavage on GD 6 through 18. Animals were observed for body weight, clinical effects and feed consumption and the fetuses examined for body weight, sex and visceral and skeletal alterations at GD 20 for rats and GD 30 for rabbits. In rats, 2 dams died at the 1200 mg/kg dose and 1 dam died at the 800 mg/kg dose. Maternal gestational weight gain was reduced at the highest dose tested. No other effects were observed on maternal reproductive health. Fetal body weights at the two highest doses were decreased statistically. No evidence of teratogenicity was observed at any dose. In rabbits, four does died at the 480 mg/kg dose. Treatment related clinical signs of toxicity were observed at the 480 mg/kg dose and included, cyanosis, lethargy, labored respiration and diarrhea. No treatment related findings were observed at GD 30. Decreased feed consumption and maternal body weights, at 480 mg/kg, were statistically significant. No other effects were observed on maternal reproductive health. No evidence of teratogenicity was observed in the rabbits at any dose. The authors determined NOAEL’s for both maternal and developmental toxicity of 400 mg/kg, each, in rats and 240 and 480 mg/kg, respectively, in rabbits. From CIR final report on Methyl Acetate180

Carcinogenicity Fischer 344 rats and CD-1 mice (65/rats/sex/group; 55/mice/sex/group) were treated via inhalation with 0, 500, 2500, or 5000 ppm (0, 1230, 6150, or 12,300 mg/m3) isopropyl alcohol for 6 h/day, 5 days/wk for 104 weeks in rats and 78 weeks in mice.192 An additional 10/animals/sex/species were treated with these same concentrations of isopropyl alcohol for 6 h/day, 5 days/wk for 72 weeks in rats and 54 weeks in mice and underwent an interim evaluation. Another 10 mice/sex/group were treated according to the paradigm described above for 54 weeks and then allowed to recover before being killed at 78 weeks. Animals were observed and evaluated for body and organ weights, ophthalmology, and clinical and anatomic pathology.

In rats, increased mortality due to chronic renal disease was observed at 5000 ppm (both sexes) and at 2500 ppm (males only). Hypoactivity and lack of startle reflex were observed in 2500 ppm treated rats and hypoactivity, lack of startle 71

CIR Panel Book Page 119 reflex and narcosis were observed in 5000 ppm treated rats. With the exception of the ataxia, the clinical signs were transient and ceased when the exposure ended. Increases in body weight, body weight gain, and liver weights were observed in 2500 and 5000 ppm treated rats. Chronic renal disease was exacerbated in rats treated with isopropyl alcohol. Male rats had a concentration related increase in absolute and relative testes weights. At the interim euthanasia (after 72 weeks) male rats treated with 5000 ppm had an increased frequency of testicular seminiferous tubule atrophy upon microscopic evaluation. At the terminal euthanasia (104 weeks), male rats had a concentration dependent increase in the incidence of interstitial (Leydig) cell adenomas of the testes at all administered doses. No other tumor types were increased in rats under these treatment conditions as compared to controls.

In mice, no differences in mortality were observed between control and treated animals. Hypoactivity, lack of a startle reflex, narcosis, ataxia, and prostration were observed in 5000 ppm treated mice. Hypoactivity, lack of startle reflex and narcosis were observed in 2500 ppm treated mice. Increases in body weight, body weight gain, and liver weights were observed in 2500 and 5000 ppm treated mice. Male mice in all treatment groups had a decrease in relative testes weights, and female mice exposed to 5000 ppm isopropyl alcohol exhibited decreases in absolute and relative brain weights. At the terminal euthanasia (78 weeks) an increased incidence of minimal to mild renal tubular proteinosis was observed in males and females in all treatment groups. Male mice exposed to 2500 and 5000 ppm exhibited an increased incidence of dilation of the seminal vesicles. No neoplastic lesions were observed in male or female mice. The authors reported a NOAEL for toxic effects of 500 ppm for both rats and mice based on kidney and testicular effects.

IARC (International Agency for Research on Cancer) has determined that isopropyl alcohol is not classifiable as to its carcinogenicity to humans (Group 3). From the CIR final report on Methyl Acetate180

Clinical Irritation and Sensitization According to unpublished data, a 80.74% spray concentrate caused did not exhibit any potential for dermal sensitization in 9 human subjects.195

According to unpublished HRIPT study on 109 test subjects, a 2.85% hair dye base formulation of isopropyl alcohol and a 1.95% isopropyl acetate caused no dermal sensitization in humans.196

The applicability of fluorescence confocal laser scanning microscopy for in situ imaging of irritant contact dermatitis caused by pelargonic acid using 12 healthy individuals (8 males, 4 males; 18 to 64 years old) was studied.197 Using Finn chambers (occlusive patches), the flexor side of the right and left forearm was exposed to 60 µl of 10% (w/v) pelargonic acid in isopropanol solution and isopropanol vehicle. Isopropanol was used as a control. The Finn chambers were removed at 24 h post-application and reactions were scored according to the following scale: 0 (no visible reaction) to 4+ (intense erythema with bullous formation). Reactions were not observed at sites treated with isopropanol. From the CIR final report on Methyl Acetate180

Hexyl Alcohol – metabolite of Dihexyl Adipate

Ocular Irritation Undiluted hexyl alcohol has been labeled as highly irritating on rabbit ocular irritation tests.198

Dermal Sensitization In a maximization test using guinea pigs, hexyl alcohol was not a sensitizer at 1% in petrolatum.198

Caprylic Alcohol – metabolite of Dicapryl Succinate, Dicapryl Adipate and Dicaprylyl/Capryl Sebacate

Dermal Irritation – Animals Caprylic alcohol applied full strength to intact or abraded rabbit skin produced a mild irritation.176

Clinical Irritation and Sensitization Tested in at a concentration of 2% in petrolatum, caprylic alcohol produced no irritation in a 48 h closed-patch test in 25 human subjects.176

72

CIR Panel Book Page 120 Isobutyl Alcohol – metabolite of Diisopropyl Adipate and Diisopropyl Sebacate

Subchronic Inhalation Toxicity Rats (10/sex/group) were exposed via inhalation to isobutyl alcohol vapor concentrations of approximately 0, 770, 3100, or 7700 mg/m3, for 6 h/day, 5 days/week, for 14 weeks.199 The functional observational battery was conducted along with endpoints of motor activity, neuropathology and scheduled-controlled operant behavior. A slight reduction in responsiveness to external stimuli was observed in all treated groups during exposure. This effect resolved upon cessation of exposure to isobutyl alcohol. From the CIR final report on Methyl Acetate180

Ethylhexyl Alcohol – metabolite of Diethylhexyl Succinate, Diethylhexyl Adipate and Diethylhexyl Sebacate Ocular Irritation Instillation of 20 µg of ethylhexyl alcohol into the conjunctival sac of rabbits caused moderately severe irritation of the cornea.198

Dermal Irritation – Animals Ethylhexyl alcohol was applied under occlusion to the skin of 3 male rabbits for 4 hours and found to be irritating.198 In another study with rabbits, 0.5 ml of ethylhexyl alcohol was applied under occlusion on intact skin for 1, 2, 4, and 24 hours. Irritation was considered high, and effects seen after 7 days were not reversible.

Reproductive and Developmental Toxicity A group of female rats was exposed for 7 h per day to 850 mg/m3 of ethylhexyl alcohol on gestation days 1-19.198 Dams were sacrificed at day twenty. Ethylhexyl alcohol reduced maternal feed intake, but did not produce any malformations.

The estrogenic activity of 2-ethylhexanoic acid was examined using an E-SCREEN assay using T47D human breast cancer cells.200 Weak estrogenic activity was observed. (Additional details were not provided.)

Genotoxicity In vitro, ethylhexyl alcohol was negative in a number of Ames assays, a liquid suspension assay, mouse lymphoma assay, and unscheduled DNA synthesis assay.118,141,142,145,153 In a 3H-thymidine assay, there was a dose-dependent inhibition of 3H-thymi- dine into replicating DNA, with a dose-dependent increase in the ratio of acid-soluble DNA incorporated into the thymidine.142 The urine of rats dosed orally with 1000 mg/kg ethylhexyl alcohol was not mutagenic.152 In vivo, ethylhexyl alcohol was not genotoxic in a mouse micronucleus test or a transformation assay.153

MEHA, Mono-(2-Ethyl-5-Hydroxyhexyl)Adipate,Mono-(2-Ethyl-5-Oxohexyl)Adipate - metabolites of Diethylhexyl Adipate

Genotoxicity MEHA was not mutagenic in an Ames assay at concentrations of ≤100084 or 10,000 µg/plate.141 Mono-2(ethyl-5- hydroxy- hexyl)adipate and mono-(2-ethyl-5-oxohexyl)adipate, were not mutagenic in an Ames assay at concentrations of ≤1000 g/plate.84

Isooctyl Alcohol – metabolite of Diisooctyl Adipate and Diisooctyl Sebacate

Subchronic Oral Toxicity In a subchronic gavage toxicity study of a mixture of C7-9, branched alkyl alcohols in rats, a NOEL of 125 mg/kg/day and a lowest-observed effect level of 250 mg/kg/day were determined.118

Reproductive and Developmental Toxicity In an oral gavage developmental toxicity study of a mixture of C7-9, branched alkyl alcohols in rats, a maternal NOAEL of 500 mg/kg and a fetal NOAEL of 1000 mg/kg were reported.198

Genotoxicity A mixture of C7-9, branched alkyl alcohols were not mutagenic in in vitro bacterial and mammalian cell assays.198

Carcinogenicity 73

CIR Panel Book Page 121 Ethylhexyl alcohol was not oncogenic in rats dosed, via gavage, with 0, 50, 150, or 500 mg/kg, in an aqueous vehicle with 0.005% Cremophor EL.198

Nonyl Alcohol – metabolite of Diisononyl Adipate

Reproductive and Developmental Toxicity In an oral gavage developmental toxicity study of a mixture of C8-10, branched alkyl alcohols in rats, maternal and fetal NOAEL values were each reported to be 144 mg/kg. 198

Isodecyl Alcohol – metabolite of Diisodecyl Adipate

Reproductive and Developmental Toxicity In an oral gavage developmental toxicity study of a mixture of C9-11, branched alkyl alcohols in rats, a maternal NOAEL of 158 mg/kg and a fetal NOAEL of 790 mg/kg were reported. 198

Isostearyl Alcohol – metabolite of Diisostearyl Glutarate, Diisostearyl Adipate and Isostearyl Sebacate

Clinical Irritation and Sensitization The skin irritation potential of lsostearyl Alcohol was evaluated in 19 male and female subjects (18-65 years old) at a concentration of 25.0% in petrolatum. The test substance did not induce skin irritation in any of the subjects (Primary Irritation Index = 0.05). In three similar studies, three different lipstick products containing 25.0, 27.0, and 28.0% Isostearyl Alcohol, respectively, were tested according to the same protocol. The three products did not induce skin irritation.

The irritation and sensitization potential of Isostearyl Alcohol (25% v/v in 95.0% isopropyl alcohol) was evaluated in 12 male subjects (21-60 years old). Challenge applications were made to original and adjacent sites 2 weeks after removal of the last induction patch. Three of 12 subjects had slight erythema during induction, and there was no evidence of sensitization.

The sensitization potential of a pump spray antiperspirant containing 5.0% Isostearyl Alcohol was evaluated using 148 male and female subjects. The product was applied via an occlusive patch to the upper arm for a total of nine induction applications (3 times/week for 3 weeks). Each patch remained for 24 h, and sites were scored immediately before subsequent applications. During the challenge phase, a patch was applied to the induction site and to a new site on the opposite arm of each subject. Reactions were scored 48 and 96 h after application. Ten of the twelve subjects with reactions suggestive of sensitization were re-challenged with the product 2 months later. Patches remained for 24 h, and sites were scored at 48 and 96 h post-application. Six subjects had reactions during the re-challenge. Four of the six subjects were then tested with 5.0% Isostearyl Alcohol in solution with ethanol 6 weeks after scoring of the first rechallenge; all had positive responses. Negative responses were reported when the product (without lsostearyl Alcohol) and 100.0% ethanol each were tested. In a second study, the same product was applied to 60 male and female subjects (same protocol). Five of the subjects had positive responses after the first challenge. One of the five was re-challenged with 5.0% Isostearyl Alcohol in ethanol solution, and a positive reaction was observed. From the CIR final report on Isostearyl Alcohol190

Isopropanol

Comedogencity An LDLo of 2-4 ml/kg of isopropyl alcohol has been reported in adults and 6 ml/kg (9 ml/kg 70% isopropyl alcohol) was reported to induce coma in children.

74

CIR Panel Book Page 122 APPENDIX I – ESTEARSE METABOLITE SUMMARY DATA

Decyl Alcohol – metabolite of Decyl Succinate

Clinical Irritation and Sensitization Tested in at a concentration of 3% in petrolatum, decyl alcohol produced no irritation in a 48 h closed-patch test in 25 human subjects.177

Methanol – metabolite of Dimethyl Succinate, Dimethyl Glutarate, Dimethyl Adipate

Absorption, Distribution, Metabolism, and Excretion Methanol is further metabolized to formaldehyde and then to formic acid. The CIR Expert Panel concluded that formic acid is safe where used in cosmetic formulations as a pH adjustor with a 64 ppm limit for the free acid.178 The main toxicological risks in humans are severe metabolic acidosis with increased anion gap, typically following oral exposure resulting in > 100 mg/L of formate in the urine.179 The acidosis and the formic acid metabolite are believed to play a central role in both the central nervous system toxicity and the ocular toxicity.

A study to determine the formate levels that resulted from exposure of human volunteers to 200 ppm of methanol for 4 h was conducted. Human volunteers (n=27; age 20-55 y) were exposed to 200 ppm methanol (the Occupational Safety and Health Administration [OSHA] Permissible Exposure Limit) for 4 h and to water vapor for 4 h in a double-blind, random study.180 Urine samples were collected at 0, 4 and 8 h and blood samples were collected from the subjects before they entered the chamber, every 15 min for the first hour, every 30 min from the first to the third hour and at 4 h. Urine and serum samples were analyzed for formate (LOD 0.5 mg/L). Twenty-six of 27 enrolled subjects completed the study (11 females and 15 males). The researchers did not find any statistically significant differences in serum or urine formate levels between the two exposure conditions at any time point. At the end of the 4 h methanol exposure, formate concentrations of 14.28 ± 8.90 and 7.14 ± 5.17 mg/L were measured in serum and urine, respectively. Under control conditions, formate concentrations of 12.68 ± 6.43 (p=0.38; n=26) and 6.64 ± 4.26 (p=0.59; n=25) mg/L were measured in serum and urine respectively. After 8 h (4 h of no exposure) the serum concentrations were not statistically different with 12.38 ± 6.53 mg/L under methanol exposure conditions and 12.95 ± 8.01 (P=0.6; n=26) under control conditions. Urine formate concentrations after 8 h were 6.08 ± 3.49 and 5.64 ±3.70 mg/L (p=0.6; n=25) in exposed and control conditions, respectively, and were not statistically significantly different. From CIR Final Report on Methyl Acetate181

Clinical Irritation and Sensitization Methyl Alcohol caused primary irritation to the skin; prolonged and repeated contact with Methyl Alcohol resulted in de- fatting and dermatitis. In one occupational study, 3.2% of 274 metalworkers with dermatitis had positive results to a patch test of 30% Methyl Alcohol. Typical allergic responses observed after contact with alcohols were eczematous eruption and wheal and flare at the exposure sites. Eczema and erythema were reported after the consumption of alcoholic beverages by persons sensitized to ethyl alcohol. Five percent Methyl Alcohol caused a slight positive (+) reaction in a closed patch test for allergic contact dermatitis, and concentrations of 7% and 70% caused (+++) reactions. From the CIR final report on methyl alcohol182

Clinical Assessment of Safety Clinical data show that Methyl Alcohol can cause severe metabolic acidosis, blindness, and death: toxicity was manifested earlier and at a lower dose compared to ethyl alcohol, but the comparative fatal dose was the same for both alcohols. All routes of exposure were toxicologically equivalent, as the alcohol distributed readily and uniformly throughout all tissues and organs. Individual susceptibilities to Methyl Alcohol varied, but typically, the ingestion of 80 to 150 ml of 80% Methyl Alcohol was fatal. Symptoms of Methyl Alcohol intoxication after ingestion were delayed for 12 to 18 hours; afterwards, the symptoms included headache, anorexia, weakness, fatigue, leg cramps, and/or pain and vertigo. Severe gastrointestinal pain, nausea, vomiting, diarrhea, mania, failed vision, and convulsions could occur. Chronic exposure to Methyl Alcohol could cause edema, granular degeneration, and necrosis of heart muscle fibers, as well as fatty degeneration of the heart muscle; sudden cardiac failure was associated with Methyl Alcohol intoxication. The liver and kidneys often had parenchymatous degeneration, and the liver had focal necrosis and fatty infiltration. Severe acidosis was necessary for the development of blindness. Similar symptoms were observed after percutaneous or inhalation exposure to Methyl Alcohol. From the CIR final report on methyl alcohol182 68

CIR Panel Book Page 123

Propyl Alcohol – metabolite of Dipropyl Adipate

Absorption, Distribution, Metabolism, and Excretion Rats (strain/sex/number not specified) were exposed via inhalation to 2,000 ppm (8360 mg/m3) for 90 min.183 Propyl acetate was rapidly hydrolyzed to propyl alcohol. During the 90 min exposure period, blood levels of propyl alcohol were between 2.6 and 7.7 fold greater than propyl acetate. From the CIR final report on Methyl Acetate181

Reproductive and Developmental Toxicity The effects of propyl alcohol on fertility were investigated by exposing male Sprague-Dawley rats (18/group) to 0, 3500 or 7000 ppm (0, 8.61 or 17.2 mg/L) propyl alcohol vapor via inhalation 7 h/day, 7days/week for 62 days, prior to mating with unexposed virgin females.184 Female Sprague-Dawley rats (15/group) were similarly exposed and mated with unexposed males. Following parturition, litters were culled to 4/sex and the pups fostered by unexposed dams. The pups were weaned on post natal day (PND) 25 and weighed on PND’s 7, 14, 21, 28 and 35. Male rats exposed to 7000 ppm exhibited a decrease in mating success with 2/16 producing a litter (1 male died as a result of a cage fight and 1 male did not mate). Mating success was not affected in 3500 ppm exposed males or in females. Six males from the 7000 ppm group were retained to determine if this effect was reversible. All 6 males successfully mated 15 weeks after exposure. The authors reported that weight gain was not affected in 7000 ppm exposed females (data not shown), but feed intake was decreased in this treatment group. Crooked tails were observed in 2-3 offspring in 2 of 15 litters from the 7000 ppm maternally exposed group. No other effects on female fertility were reported. No significant differences resulted between offspring of the 7000 ppm group and controls on several behavioral toxicology measures including the Ascent test, Rotorod test, Open Field test, activity test, running wheel activity, avoidance conditioning, and operant conditioning. Activity measures were significantly different between offspring of the 3500 ppm exposure group and controls. From the CIR final report on Methyl Acetate181

Clinical Irritation and Sensitization A cumulative irritation study was conducted involving 20 male subjects, where the relative irritancy of free fatty acids of different chain lengths was evaluated.185 Equimolar concentrations (0.5 M and 1.0 M) of even- and odd- numbered - straight chain saturated fatty acids were dissolved in propanol. Each Al-test® patch containing a fatty acid (0.5 M) was applied to the interscapular area of 10 subjects, and, similarly, each fatty acid was applied at a higher concentration (1.0 M) to the remaining 10 subjects. A control patch containing propanol was also applied to each subject. Patches remained in place for 24 h and reactions were scored 30 minutes after patch removal. This procedure was repeated daily for a total of 10 applications. In both groups of 10 subjects, there were no reactions to propanol.

In an irritation study, wherein 116 healthy male subjects (21 to 55 years old) were patch tested with pelargonic acid at concentrations of 5%, 10%, 20%, and 39.9% in propanol, a propanol-treated control patch was used.186 Dose response curves were developed. Patches (Al-test® discs) were saturated with 0.04 ml of a test solution and applied to the upper back for 48 h. Reactions were scored at 48 h and 96 h post-application. There were no reactions to propanol.

In an another irritation study, wherein 16 volunteers (10 females, 6 males; median age of 29.5 years) were patch tested (closed patches, Finn chambers) with 20% pelargonic acid in propanol (pH of 4.3), propanol was one of the controls used.187 Patches were applied to the anterolateral surface of both upper arms for 24 h. Reactions were scored at 24, 48, and 96 h post-application according to the following scale: 0 (no reaction) to 3 (strong positive reaction: marked erythema, infiltration, possibly vesicles, bullae, pustules and/or pronounced crusting). There were no reactions to propanol.

A skin irritation study was conducted using 42 healthy, non-atopic male volunteers (mean age = 34 years; skin types: II [20 subjects], III [17 subjects], and IV [5 subjects]).188 Pelargonic acid was patch-tested (Finn chambers, volar forearm) at the following concentrations (in propanol): 40% (12 subjects), 60% (32 subjects), 70% (32 subjects), and 80% (28 subjects), and propanol was used as a control. Each subject received between 3 and 10 patch tests. The patches remained in place for 48 h, and reactions were scored 1 h later according to the following scale: - (no visible reaction) to 4+ (intense erythema with bullous formation). There were no reactions to propanol.

In an irritation study, wherein 16 healthy subjects (ages not stated) were patch tested with pelargonic acid (20% in 69

CIR Panel Book Page 124 propanol), propanol was used as a control.189 Closed patches (Finn chambers) containing the test substance were applied to the anterolateral surface of both upper arms. The patches were removed at 24 h post-application and reactions were scored at 24 h and 96 h post-application. There were no reactions to propanol.

In study conducted to investigate a possible seasonal variation in the skin response to pelargonic acid during the winter and summer, propanol was used as a control.190 The study was conducted using 17 healthy volunteers (10 males, 7 females; mean age = 27 years). The test substance was applied (closed patch, Finn chamber) to each arm for 24 h. Reactions were scored at 30 min post-removal. Reactions were not observed at sites treated with propanol, water, or to which an empty chamber was applied. From CIR final report on Methyl Acetate181

Cetyl Alcohol – metabolite of Dicetyl Succinate and Dicetyl Adipate

Clinical Irritation and Sensitization A topical tolerance study involving an 11.5% Cetyl Alcohol cream base was conducted with 80 male subjects, ranging in age from 21 to 52 years and in weight from 120 to 220 pounds. The preparations were applied five times daily (every 3 hours) for 10 days. One subject had erythema, folliculitis, and pustule formation (forearm site).

A formulation containing 6.0% Cetyl Alcohol was tested for its skin irritation potential in 20 subjects according to the protocol stated above. The product did not induce skin irritation. In another study, the skin irritation potential of a cream containing 6.0% Cetyl Alcohol was evaluated in 12 female subjects (18-60 years old). The total irritation score (all panelists) for the 21 applications was 418, indicating mild cumulative irritation.

The skin irritation and sensitization potential of a product containing 8.4% Cetyl Alcohol was evaluated in 110 female subjects. Fourteen days after scoring of the tenth application site, a challenge patch was applied to each subject and removed after 48 h; sites were scored after patch removal. The product did not induce primary irritation or sensitization.

The sensitization potential of a cream containing 3.0% Cetearyl Alcohol was evaluated in 25 subjects (18-25 years old). Following a 10-day non-treatment period, occlusive challenge patches were applied to new sites and removed after 48 h. Sensitization reactions were not observed in any of the subjects. From the CIR final report on cetyl alcohol191

Photosensitization The photosensitization potential of a lipstick product containing 4.0% Cetyl Alcohol was evaluated in 52 subjects. The experimental procedure was not stated. Photosensitization reactions were not noted in any of the subjects. In another study, a skin care preparation containing 1.0% Cetyl Alcohol did not induce photosensitization in the 407 subjects tested. The experimental procedure was not stated. From CIR final report on cetyl alcohol191

Isopropyl Alcohol – metabolite of Diisopropyl Adipate and Diisopropyl Sebacate

Absorption, Distribution, Metabolism, and Excretion Male rabbits (3/group; strain not specified) were treated by different routes of exposure to compare the absorption and metabolism of isopropyl alcohol.192 Groups 1 and 2 were treated via gavage with the equivalent of 2 and 4 ml/kg absolute isopropyl alcohol, respectively, as a 35% isopropanol/water solution. Groups 3 and 4 were treated via whole-body inhalation for 4 h (towels soaked with isopropyl alcohol were place in the inhalation chamber and replenished at ½ hour intervals to maintained a saturated environment; no exact concentration given), with Group 3 animals receiving an additional dermal exposure in the form of a towel soaked with 70% isopropyl alcohol applied to the animals’ chests and Group 4 animals having plastic barriers on their chests and towels prepared the same way as in Group 3 applied on top of the plastic barriers. The alcohol on the towels was replenished at half hour intervals throughout the duration of the experiment. Blood samples were taken at 0, 1, 2, 3 and 4 h. Samples were analyzed for isopropyl alcohol and the metabolite acetone.

Following gavage exposure to 2 or 4 ml/kg, maximum blood levels of 147 and 282 mg/dl, respectively, of isopropyl alcohol were measured. Concentrations of acetone rose steadily over the 4 h period and were 74 and 73 mg/dl 70

CIR Panel Book Page 125 following exposure to 2 or 4 ml/kg, respectively. The authors stated that the maximum levels of isopropyl alcohol observed in this experiment, correlated with inebriation and near coma in the animals. Following inhalation and dermal exposure, the concentration of isopropyl alcohol in the blood continued to rise and was 112 mg/dl at 4 h while the concentration of acetone was 19 at 4 h. Inhalation exposure with a plastic barrier between the soaked towel and the chest resulted in isopropyl alcohol and acetone blood levels of <10 mg/dl.

The researchers concluded that isopropyl alcohol is absorbed by the dermal route but that prolonged dermal exposure (i.e. repeated sponging or soaking for several hours) would be required to produce significant toxicity. From CIR final report on Methyl Acetate181

Subchronic Inhalation Toxicity Fischer 344 rats and CD-1 mice (10/sex/group) were exposed via inhalation to 0, 100, 500, 1500, or 5000 ppm (0, 246, 1230, 3690, or 12,300 mg/m3) isopropyl alcohol for 6 h/day, 5 days/wk for 13 weeks.193 Ataxia, narcosis, hypoactivity and the lack of a startle reflex were observed during exposure at 5000 ppm. Hypoactivity was observed in animals exposed to 1500 ppm isopropyl alcohol. At 13 weeks, no gross lesions were observed. Microscopic examination of control and 5000 ppm exposed animal tissues showed hyaline droplets within the kidneys of male rats only. The size and frequency of the droplets was increased in the treated group. The authors concluded that the NOAEL for this study was 500 ppm and the lowest-observable adverse effect level (LOAEL) was 1500 ppm based upon clinical signs and changes in hematology at 6 weeks. To evaluate the neurobehavioral effects of isopropanol exposure, an additional 15 rats/sex were exposed (via inhalation) to 0, 500, 1500, or 5000 ppm (0, 1230, 3690, or 12,300 mg/m3) for 6 h/day, 5 days/wk for 13 weeks. Isopropyl alcohol did not produce any changes to the parameters of the functional observations battery which was conducted at 1, 2, 4, 9 and 13 weeks. Clinical signs observed in mice, during the exposure, included ataxia, narcosis, hypoactivity and lack of a startle reflex at 5000 ppm. Narcosis, ataxia and hypoactivity were observed in animals exposed to 1500 ppm isopropyl alcohol. At 5000 ppm, increased body weight and increased rate of weight gain were observed in female mice. At 13 weeks, no gross lesions were observed and no treatment-related microscopic changes were observed. A 10% and 21% increase in relative liver weight was observed in female mice at 1500 and 5000 ppm, respectively. The authors concluded that the NOAEL for this study was 500 ppm and the LOAEL was 1500 ppm based on clinical signs and increased liver weights.

Ocular Irritation Isopropyl alcohol has been labeled a severe ocular irritant based on rabbit ocular irritation tests.194 From CIR final report on Methyl Acetate181

Reproductive and Developmental Toxicity Female Sprague-Dawley rats (25/group) were exposed to 0, 400, 800, or 1200 mg/kg/day isopropyl alcohol via gavage on gestational days (GD) 6 through 15.195 Female New Zealand white rabbits (15/group) were exposed to 0, 120, 240, or 480 mg/kg/day isopropyl alcohol via gavage on GD 6 through 18. Animals were observed for body weight, clinical effects and feed consumption and the fetuses examined for body weight, sex and visceral and skeletal alterations at GD 20 for rats and GD 30 for rabbits. In rats, 2 dams died at the 1200 mg/kg dose and 1 dam died at the 800 mg/kg dose. Maternal gestational weight gain was reduced at the highest dose tested. No other effects were observed on maternal reproductive health. Fetal body weights at the two highest doses were decreased statistically. No evidence of teratogenicity was observed at any dose. In rabbits, four does died at the 480 mg/kg dose. Treatment related clinical signs of toxicity were observed at the 480 mg/kg dose and included, cyanosis, lethargy, labored respiration and diarrhea. No treatment related findings were observed at GD 30. Decreased feed consumption and maternal body weights, at 480 mg/kg, were statistically significant. No other effects were observed on maternal reproductive health. No evidence of teratogenicity was observed in the rabbits at any dose. The authors determined NOAEL’s for both maternal and developmental toxicity of 400 mg/kg, each, in rats and 240 and 480 mg/kg, respectively, in rabbits. From CIR final report on Methyl Acetate181

Carcinogenicity Fischer 344 rats and CD-1 mice (65/rats/sex/group; 55/mice/sex/group) were treated via inhalation with 0, 500, 2500, or 5000 ppm (0, 1230, 6150, or 12,300 mg/m3) isopropyl alcohol for 6 h/day, 5 days/wk for 104 weeks in rats and 78 weeks in mice.193 An additional 10/animals/sex/species were treated with these same concentrations of isopropyl alcohol for 6 h/day, 5 days/wk for 72 weeks in rats and 54 weeks in mice and underwent an interim evaluation. Another 10 mice/sex/group were treated according to the paradigm described above for 54 weeks and then allowed to recover before being killed at 78 weeks. Animals were observed and evaluated for body and organ weights, ophthalmology, and clinical and anatomic pathology.

In rats, increased mortality due to chronic renal disease was observed at 5000 ppm (both sexes) and at 2500 ppm (males only). Hypoactivity and lack of startle reflex were observed in 2500 ppm treated rats and hypoactivity, lack of startle 71

CIR Panel Book Page 126 reflex and narcosis were observed in 5000 ppm treated rats. With the exception of the ataxia, the clinical signs were transient and ceased when the exposure ended. Increases in body weight, body weight gain, and liver weights were observed in 2500 and 5000 ppm treated rats. Chronic renal disease was exacerbated in rats treated with isopropyl alcohol. Male rats had a concentration related increase in absolute and relative testes weights. At the interim euthanasia (after 72 weeks) male rats treated with 5000 ppm had an increased frequency of testicular seminiferous tubule atrophy upon microscopic evaluation. At the terminal euthanasia (104 weeks), male rats had a concentration dependent increase in the incidence of interstitial (Leydig) cell adenomas of the testes at all administered doses. No other tumor types were increased in rats under these treatment conditions as compared to controls.

In mice, no differences in mortality were observed between control and treated animals. Hypoactivity, lack of a startle reflex, narcosis, ataxia, and prostration were observed in 5000 ppm treated mice. Hypoactivity, lack of startle reflex and narcosis were observed in 2500 ppm treated mice. Increases in body weight, body weight gain, and liver weights were observed in 2500 and 5000 ppm treated mice. Male mice in all treatment groups had a decrease in relative testes weights, and female mice exposed to 5000 ppm isopropyl alcohol exhibited decreases in absolute and relative brain weights. At the terminal euthanasia (78 weeks) an increased incidence of minimal to mild renal tubular proteinosis was observed in males and females in all treatment groups. Male mice exposed to 2500 and 5000 ppm exhibited an increased incidence of dilation of the seminal vesicles. No neoplastic lesions were observed in male or female mice. The authors reported a NOAEL for toxic effects of 500 ppm for both rats and mice based on kidney and testicular effects.

IARC (International Agency for Research on Cancer) has determined that isopropyl alcohol is not classifiable as to its carcinogenicity to humans (Group 3). From the CIR final report on Methyl Acetate181

Clinical Irritation and Sensitization According to unpublished data, a 80.74% spray concentrate caused did not exhibit any potential for dermal sensitization in 9 human subjects.196

According to unpublished HRIPT study on 109 test subjects, a 2.85% hair dye base formulation of isopropyl alcohol and a 1.95% isopropyl acetate caused no dermal sensitization in humans.197

The applicability of fluorescence confocal laser scanning microscopy for in situ imaging of irritant contact dermatitis caused by pelargonic acid using 12 healthy individuals (8 males, 4 males; 18 to 64 years old) was studied.198 Using Finn chambers (occlusive patches), the flexor side of the right and left forearm was exposed to 60 µl of 10% (w/v) pelargonic acid in isopropanol solution and isopropanol vehicle. Isopropanol was used as a control. The Finn chambers were removed at 24 h post-application and reactions were scored according to the following scale: 0 (no visible reaction) to 4+ (intense erythema with bullous formation). Reactions were not observed at sites treated with isopropanol. From the CIR final report on Methyl Acetate181

Hexyl Alcohol – metabolite of Dihexyl Adipate

Ocular Irritation Undiluted hexyl alcohol has been labeled as highly irritating on rabbit ocular irritation tests.199

Dermal Sensitization In a maximization test using guinea pigs, hexyl alcohol was not a sensitizer at 1% in petrolatum.199

Caprylic Alcohol – metabolite of Dicapryl Succinate, Dicapryl Adipate and Dicaprylyl/Capryl Sebacate

Dermal Irritation – Animals Caprylic alcohol applied full strength to intact or abraded rabbit skin produced a mild irritation.177

Clinical Irritation and Sensitization Tested in at a concentration of 2% in petrolatum, caprylic alcohol produced no irritation in a 48 h closed-patch test in 25 human subjects.177

72

CIR Panel Book Page 127 Isobutyl Alcohol – metabolite of Diisopropyl Adipate and Diisopropyl Sebacate

Subchronic Inhalation Toxicity Rats (10/sex/group) were exposed via inhalation to isobutyl alcohol vapor concentrations of approximately 0, 770, 3100, or 7700 mg/m3, for 6 h/day, 5 days/week, for 14 weeks.200 The functional observational battery was conducted along with endpoints of motor activity, neuropathology and scheduled-controlled operant behavior. A slight reduction in responsiveness to external stimuli was observed in all treated groups during exposure. This effect resolved upon cessation of exposure to isobutyl alcohol. From the CIR final report on Methyl Acetate181

Ethylhexyl Alcohol – metabolite of Diethylhexyl Succinate, Diethylhexyl Adipate and Diethylhexyl Sebacate Ocular Irritation Instillation of 20 µg of ethylhexyl alcohol into the conjunctival sac of rabbits caused moderately severe irritation of the cornea.199

Dermal Irritation – Animals Ethylhexyl alcohol was applied under occlusion to the skin of 3 male rabbits for 4 hours and found to be irritating.199 In another study with rabbits, 0.5 ml of ethylhexyl alcohol was applied under occlusion on intact skin for 1, 2, 4, and 24 hours. Irritation was considered high, and effects seen after 7 days were not reversible.

Reproductive and Developmental Toxicity A group of female rats was exposed for 7 h per day to 850 mg/m3 of ethylhexyl alcohol on gestation days 1-19.199 Dams were sacrificed at day twenty. Ethylhexyl alcohol reduced maternal feed intake, but did not produce any malformations.

The estrogenic activity of 2-ethylhexanoic acid was examined using an E-SCREEN assay using T47D human breast cancer cells.137 Weak estrogenic activity was observed. (Additional details were not provided.)

Genotoxicity In vitro, ethylhexyl alcohol was negative in a number of Ames assays, a liquid suspension assay, mouse lymphoma assay, and unscheduled DNA synthesis assay.118,142,143,146,154 In a 3H-thymidine assay, there was a dose-dependent inhibition of 3H-thymi- dine into replicating DNA, with a dose-dependent increase in the ratio of acid-soluble DNA incorporated into the thymidine.143 The urine of rats dosed orally with 1000 mg/kg ethylhexyl alcohol was not mutagenic.153 In vivo, ethylhexyl alcohol was not genotoxic in a mouse micronucleus test or a transformation assay.154

MEHA, Mono-(2-Ethyl-5-Hydroxyhexyl)Adipate,Mono-(2-Ethyl-5-Oxohexyl)Adipate - metabolites of Diethylhexyl Adipate

Genotoxicity MEHA was not mutagenic in an Ames assay at concentrations of ≤100084 or 10,000 µg/plate.142 Mono-2(ethyl-5- hydroxy- hexyl)adipate and mono-(2-ethyl-5-oxohexyl)adipate, were not mutagenic in an Ames assay at concentrations of ≤1000 g/plate.84

Isooctyl Alcohol – metabolite of Diisooctyl Adipate and Diisooctyl Sebacate

Subchronic Oral Toxicity In a subchronic gavage toxicity study of a mixture of C7-9, branched alkyl alcohols in rats, a NOEL of 125 mg/kg/day and a lowest-observed effect level of 250 mg/kg/day were determined.118

Reproductive and Developmental Toxicity In an oral gavage developmental toxicity study of a mixture of C7-9, branched alkyl alcohols in rats, a maternal NOAEL of 500 mg/kg and a fetal NOAEL of 1000 mg/kg were reported.199

Genotoxicity A mixture of C7-9, branched alkyl alcohols were not mutagenic in in vitro bacterial and mammalian cell assays.199

Carcinogenicity 73

CIR Panel Book Page 128 Ethylhexyl alcohol was not oncogenic in rats dosed, via gavage, with 0, 50, 150, or 500 mg/kg, in an aqueous vehicle with 0.005% Cremophor EL.199

Nonyl Alcohol – metabolite of Diisononyl Adipate

Reproductive and Developmental Toxicity In an oral gavage developmental toxicity study of a mixture of C8-10, branched alkyl alcohols in rats, maternal and fetal NOAEL values were each reported to be 144 mg/kg. 199

Isodecyl Alcohol – metabolite of Diisodecyl Adipate

Reproductive and Developmental Toxicity In an oral gavage developmental toxicity study of a mixture of C9-11, branched alkyl alcohols in rats, a maternal NOAEL of 158 mg/kg and a fetal NOAEL of 790 mg/kg were reported. 199

Isostearyl Alcohol – metabolite of Diisostearyl Glutarate, Diisostearyl Adipate and Isostearyl Sebacate

Clinical Irritation and Sensitization The skin irritation potential of lsostearyl Alcohol was evaluated in 19 male and female subjects (18-65 years old) at a concentration of 25.0% in petrolatum. The test substance did not induce skin irritation in any of the subjects (Primary Irritation Index = 0.05). In three similar studies, three different lipstick products containing 25.0, 27.0, and 28.0% Isostearyl Alcohol, respectively, were tested according to the same protocol. The three products did not induce skin irritation.

The irritation and sensitization potential of Isostearyl Alcohol (25% v/v in 95.0% isopropyl alcohol) was evaluated in 12 male subjects (21-60 years old). Challenge applications were made to original and adjacent sites 2 weeks after removal of the last induction patch. Three of 12 subjects had slight erythema during induction, and there was no evidence of sensitization.

The sensitization potential of a pump spray antiperspirant containing 5.0% Isostearyl Alcohol was evaluated using 148 male and female subjects. The product was applied via an occlusive patch to the upper arm for a total of nine induction applications (3 times/week for 3 weeks). Each patch remained for 24 h, and sites were scored immediately before subsequent applications. During the challenge phase, a patch was applied to the induction site and to a new site on the opposite arm of each subject. Reactions were scored 48 and 96 h after application. Ten of the twelve subjects with reactions suggestive of sensitization were re-challenged with the product 2 months later. Patches remained for 24 h, and sites were scored at 48 and 96 h post-application. Six subjects had reactions during the re-challenge. Four of the six subjects were then tested with 5.0% Isostearyl Alcohol in solution with ethanol 6 weeks after scoring of the first rechallenge; all had positive responses. Negative responses were reported when the product (without lsostearyl Alcohol) and 100.0% ethanol each were tested. In a second study, the same product was applied to 60 male and female subjects (same protocol). Five of the subjects had positive responses after the first challenge. One of the five was re-challenged with 5.0% Isostearyl Alcohol in ethanol solution, and a positive reaction was observed. From the CIR final report on Isostearyl Alcohol191

Isopropanol

Comedogencity An LDLo of 2-4 ml/kg of isopropyl alcohol has been reported in adults and 6 ml/kg (9 ml/kg 70% isopropyl alcohol) was reported to induce coma in children.

74

CIR Panel Book Page 129 Figure 1. Map of the ester ingredients in this assessment, and associated esterase metabolites.

CIR Panel Book Page 130 Figure 1. Map of the ester ingredients in this assessment, and associated esterase metabolites.

Adipic Acid O HO OH O Dimethyl Adipate Methyl Adipate* O O Methyl alcohol O O CH3 H3C O CH3 H3C OH HO O O Diethyl Adipate Ethyl alcohol Ethyl Adipate* O O HO CH H3C O H3C O 3 OH O CH3 O O Dipropyl Adipate Propyl alcohol Propyl Adipate* O O CH O O CH3 HO 3 H3C OH H3C O O O Dibutyl Adipate Butyl alcohol Butyl Adipate* O O HO CH H3C O H3C O 3 OH O CH3 O O Dihexyl Adipate Hexyl alcohol Hexyl Adipate* O O O (CH ) CH HO CH3 O 2 5 3 CH3(CH2)5 OH CH3(CH2)5 O O O Dicapryl Adipate O Caprylic Adipate* O Caprylic alcohol O O (CH2)7CH3 CH (CH ) OH CH3(CH2)7 O HO CH 3 2 7 3 O O Lauryl alcohol

HO CH3 Tridecyl alcohol CH Di-C12-15 Alkyl Adipate HO 3 O Myristyl alcohol O (CH2)nCH3 CH3(CH2)n O HO CH3 O n = 11-14 Heptadecyl alcohol* CH HO 3 C12-15 Alkyl Adipate* O O CH (CH ) OH Ditridecyl Adipate 3 2 n n = 11-14 O O O (CH2)12CH3 CH3(CH2)12 O Tridecyl Adipate O O O * CH3(CH2)12 OH O

Dicetyl Adipate O Cetyl alcohol O (CH ) CH H CH (CH ) O 2 15 3 O CH 3 2 15 3 O Cetyl Adipate O O * CH3(CH2)15 OH O

CIR Panel Book Page 131 Figure 1. Map of the ester ingredients in this assessment, and associated esterase metabolites.

Adipic Acid O HO (Branched esters) OH O

Diisopropyl Adipate Isopropyl alcohol Isopropyl Adipate* O O CH3 CH3 H3C O H3C O OH O CH3 H3C OH CH3 O CH3 O

Isobutyl alcohol * Isobutyl Adipate* Diisobutyl Adipate CH3 O CH3 O CH3 O O CH3 OH H3C OH H3C O H3C O O CH3

* Ethylhexyl Adipate* Diethylhexyl Adipate (DEHA) Ethylhexyl alcohol H C H3C 3 O O H3C O (CH2)3CH3 O CH3(CH2)3 O H C OH CH3(CH2)3 OH 3 O O monoester CH3 Diisooctyl Adipate Isooctanol O CH3 Isooctyl Adipate O H3C H C * O (CH2)5 H C 3 O (CH2)5 O 3 OH (CH ) OH CH3 2 5 H3C O H C CH3 3 O Diisononyl Adipate O Isononyl alcohol * CH3 Isononyl Adipate O H3C O (CH ) CH3 H C * 2 6 3 O (CH2)6 O CH3 (CH2)6 OH H3C O H3C OH H3C O Diisodecyl Adipate O Isodecyl alcohol * CH3 Isodecyl Adipate O H3C * O (CH2)7 H3C (CH2)7 O H3C O CH3 OH (CH2)7 OH H3C O H C CH3 3 O

Hexyldecyl Adipate CH3(CH2)4 * CH (CH ) Dihexyldecyl Adipate Hexyldecanol O 3 2 4 O H C OH O O (CH ) CH 3 CH3(CH2)7 OH CH (CH ) O 2 7 3 H C 3 2 7 3 O O (CH2)4CH3

Heptylundecanol CH (CH ) Heptylundecyl Adipate* 3 2 5 O CH (CH ) Diheptylundecyl Adipate 3 2 5 O H3C O CH (CH ) OH O (CH ) CH OH 3 2 8 CH (CH ) O 2 8 3 H3C O 3 2 8 O (CH2)5CH3

Octyldodecanol CH (CH ) Octyldodecyl Adipate* CH (CH ) Dioctyldodecyl Adipate 3 2 6 O 3 2 6 O H C OH 3 O O (CH ) CH CH (CH ) OH CH (CH ) O 2 9 3 H C 3 2 9 3 2 9 3 O O (CH2)6CH3 Isocetyl Adipate* O H3C O (CH2)13 OH Diisocetyl Adipate H3C O O CH3 H3C Isocetyl alcohol O (CH2)13 H CH3 (CH2)13 O O CH3 H3C O CH3 Isostearyl Adipate O H3C O * Diisostearyl Adipate (CH ) OH 2 15 O CH3 H C H3C 3 O O (CH2)15 (CH2)15 O CH3 Isostearyl alcohol H3C O H CH O 3 CH3

CIR Panel Book Page 132 Figure 1. Map of the ester ingredients in this assessment, and associated esterase metabolites. Sebacic Acid O HO OH O

Diethyl Sebacate O O Ethyl alcohol Ethyl Sebacate* H C O H C O 3 O CH HO CH 3 OH 3 3 O O Dibutyl Sebacate O O Butyl alcohol Butyl Sebacate* O (CH ) CH O CH (CH ) O 2 3 3 HO CH CH (CH ) OH 3 2 3 3 3 2 3 O O Caprylic alcohol Dicaprylyl/Capryl Sebacate O Caprylyl/Capryl Sebacate* O HO CH3 O O (CH2)nCH3 CH (CH ) OH Decyl alcohol CH3(CH2)n O 3 2 n n=7or9 O n=7or9 O HO CH3 CH3 Isostearyl alcohol Diisostearyl Sebacate O O H C H CH3 3 H3C O (CH2)15 (CH2)15 O CH3 Isostearyl Sebacate H3C O O CH3 HO (CH ) O 2 15 CH3 O

Diisopropyl Sebacate O CH O 3 Isopropyl alcohol Isopropyl Sebacate* H C O CH H C O 3 O CH 3 3 OH 3 CH3 O H3C OH CH3 O

H C Diethylhexyl Sebacate 3 O Ethylhexyl alcohol * H C 3 O O (CH ) CH H3C Ethylhexyl Sebacate* CH (CH ) O 2 3 3 O 3 2 3 CH (CH ) OH O H3C OH 3 2 3 CH O 3

Butyloctanol CH (CH ) Dibutyloctyl Sebacate CH3(CH2)2 3 2 2 O Butyloctyl Sebacate* O H C OH + O (CH ) CH 3 O CH (CH ) O 2 5 3 CH3(CH2)5 OH 3 2 5 H3C O O (CH2)2CH3

Diisooctyl Sebacate O Isooctanol CH3 O H3C CH Isooctyl Sebacate* O (CH2)5 3 H3C (CH2)5 O O CH3 OH (CH2)5 OH H3C O H3C H3C O Dihexyldecyl Sebacate CH3(CH2)4 Hexyldecanol CH3(CH2)4 O Hexyldecyl Sebacate* O + O (CH2)7CH3 H3C OH O CH3(CH2)7 O CH3(CH2)7 OH H3C O O (CH2)4CH3

CH (CH ) CH (CH ) Dioctyldodecyl Sebacate Octyldodecanol 3 2 6 3 2 6 O + Octyldodecyl Sebacate* O H3C OH O O (CH2)9CH3 CH3(CH2)9 OH CH3(CH2)9 O H3C O O (CH2)6CH3

CIR Panel Book Page 133 Figure 2. Sebacic acid synthesis from castor oil.

CIR Panel Book Page 134 Figure 3. Diethylhexyl adipate synthesis from adipic acid.

CIR Panel Book Page 135 Chart 1. Log Kow vs Molecular Weight: Alkyl Dicarboxylic Acids

CIR Panel Book Page 136 Chart 2. Log Kow vs Molecular Weight: Alkyl Dicarboxylic Acids and Salts

CIR Panel Book Page 137 Chart 3. Log Kow vs Molecular Weight: Esters of Alkyl Dicarboxylic Acids

CIR Panel Book Page 138 Table 1. Definitions, functions and structures of dicarboxylic acid, salt and ester ingredients in this safety assessment. Ingredient Definition Function(s) Formula/structure CAS No. Dicarboxylic Acids and Metal Salts Malonic Acid Malonic Acid is a three Fragrance 141-82-2 carbon, straight-chain alkyl Ingredients; pH dicarboxylic acid. Adjusters Succinic Acid Succinic Acid is a four Fragrance O 110-15-6 carbon, straight-chain alkyl Ingredients; pH HO OH dicarboxylic acid. Adjusters O Sodium Sodium Succinate is the Buffering Agents; Succinate monosodium salt of pH Adjusters 2922-54-5 succinic acid. Disodium Disodium Succinate is the Fragrance Succinate disodium salt of succinic Ingredients; Not 150-90-3 acid. Reported Glutaric Acid Glutaric Acid is a five Fragrance 110-94-1 carbon, straight-chain alkyl Ingredients; pH dicarboxylic acid. Adjusters Adipic Acid Adipic Acid is a six Fragrance 124-04-9 carbon, straight-chain alkyl Ingredients; pH dicarboxylic acid. Adjusters Azelaic Acid Azelaic Acid is an eight Fragrance 123-99-9 carbon, straight-chain alkyl Ingredients; pH dicarboxylic acid. Adjusters Disodium Disodium Azelate is the Not Reported Azelate disodium salt of azelaic 17265-13-3 acid. Dipotassium Dipotassium Azelate is the Not Reported Azelate dipotassium salt of azelaic 19619-43-3 acid. Sebacic Acid Sebacic Acid is a ten pH Adjusters 111-20-6 carbon, straight-chain alkyl dicarboxylic acid. Disodium Disodium Sebacate is the Not Reported Sebacate disodium salt of sebacic 17265-14-4 acid. Dodecanedioic Dodecanedioic Acid is a Skin- Acid twelve carbon, straight- Conditioning 693-23-2 chain alkyl dicarboxylic Agents - acid. Miscellaneous

Malonic Diester Ingredient Diethyl Diethyl Malonate is the Fragrance Malonate diester of ethanol and Ingredients 105-53-3 malonic acid.

Succinic Ester Ingredients Monoester Decyl Decyl Succinate is the Skin- Succinate monoester of decyl alcohol Conditioning 54482-22-3 and succinic acid. Agents - (wrong CAS Emollient No. 2530-33-8) Diesters Dimethyl Dimethyl Succinate is the Nail Polish and Succinate diester of methyl alcohol Enamel Removers 106-65-0 and succinic acid. Diethyl Diethyl Succinate is the Fragrance Succinate diester of ethyl alcohol and Ingredients; 123-25-1 succinic acid. Plasticizers; Solvents

CIR Panel Book Page 139 Table 1. Definitions, functions and structures of dicarboxylic acid, salt and ester ingredients in this safety assessment. Ingredient Definition Function(s) Formula/structure CAS No. Dicapryl Dicapryl Succinate is the Film Formers; Succinate diester of caprylic alcohol Hair Conditioning 14491-66-8 and succinic acid. Agents; Nail Conditioning Agents; Plasticizers; Skin- Conditioning Agents - Emollient Dicetearyl Dicetearyl Succinate is the Skin- Succinate diester of cetearyl alcohol Conditioning 93280-98-9 and succinic acid. Agents - Miscellaneous wherein n=15 or 17 Branched Diisobutyl Diisobutyl Succinate is the Plasticizers Succinate diester of isobutyl alcohol 925-06-4 and succinic acid. Diethylhexyl Diethylhexyl Succinate is Plasticizers; Skin- Succinate the diester of 2-ethylhexyl Conditioning 2915-57-3 alcohol and succinic acid. Agents - Emollient; Solvents

Glutaric Ester Ingredients Dimethyl Dimethyl Glutarate is the Nail Polish and Glutarate diester of methyl alcohol Enamel Removers 1119-40-0 and glutaric acid. Branched Diisobutyl Diisobutyl Glutarate is the Plasticizers Glutarate diester of isobutyl alcohol 71195-64-7 and glutaric acid. Diisostearyl Diisostearyl Glutarate is Skin- One example of an “iso” Glutarate the diester of isostearyl Conditioning No CAS No. alcohol and glutaric acid. Agents - Emollient

Adipic Ester Ingredients Dimethyl Dimethyl Adipate is the Plasticizers; Skin- Adipate diester of methyl alcohol Conditioning 627-93-0 and adipic acid. Agents - Emollient; Solvents Diethyl Diethyl Adipate is the Fragrance Adipate diester of ethyl alcohol and Ingredients; Skin- 141-28-6 adipic acid. Conditioning Agents - Emollient Dipropyl Dipropyl Adipate is the Skin- Adipate diester of propyl alcohol Conditioning 106-19-4 and adipic acid. Agents - Emollient; Solvents Dibutyl Dibutyl Adipate is the Nail Polish and Adipate diester of butyl alcohol and Enamels; Suntan 105-99-7 adipic acid. Gels, Creams, and Liquids Dihexyl Dihexyl Adipate is the Skin- Adipate diester of hexyl alcohol Conditioning 110-33-8 and adipic acid. Agents - Emollient; Solvents

CIR Panel Book Page 140 Table 1. Definitions, functions and structures of dicarboxylic acid, salt and ester ingredients in this safety assessment. Ingredient Definition Function(s) Formula/structure CAS No. Dicapryl Dicapryl Adipate is the Plasticizers Adipate diester of capryl alcohol 105-97-5 and adipic acid. Di-C12-15 Di-C12-15 Alkyl Adipate Skin- Alkyl Adipate is the diester of C12-15 Conditioning No CAS No. alcohols and adipic acid. Agents - Emollient wherein n=11, 12, 13 or 14 Ditridecyl Ditridecyl Adipate is the Skin- Adipate diester of tridecyl alcohol Conditioning 16958-92-2 and adipic acid. Agents - Emollient; Solvents Dicetyl Dicetyl Adipate is the Skin- Adipate diester of cetyl alcohol and Conditioning 26720-21-8 adipic acid. Agents - Emollient

Branched Diisopropyl Diisopropyl Adipate is the Fragrance Adipate diester of isopropyl alcohol Ingredients; 6938-94-9 and adipic acid. Plasticizers; Skin- Conditioning Agents - Emollient; Solvents Diisobutyl Diisobutyl Adipate is the Fragrance Adipate diester of isobutyl alcohol Ingredients; 141-04-8 and adipic acid. Plasticizers; Skin- Conditioning Agents - Emollient; Solvents Diethylhexyl Diethylhexyl Adipate is Plasticizers; Skin- Adipate the diester of a 2- Conditioning 103-23-1 ethylhexyl alcohol and Agents - adipic acid. Emollient; Solvents Diisooctyl Diisooctyl Adipate is the Skin- One example of an “iso” Adipate diester of isooctyl alcohol Conditioning 108-63-4 and adipic acid. Agents - Emollient; Solvents Diisononyl Diisononyl Adipate is the Plasticizers; Skin- One example of an “iso” Adipate diester of isononyl alcohol Conditioning 33703-08-1 and adipic acid. Agents - Emollient; Solvents Diisodecyl Diisodecyl Adipate is the Plasticizers; Skin- One example of an “iso” Adipate diester of isodecyl alcohol Conditioning 27178-16-1 and adipic acid. Agents - Emollient; Solvents Dihexyldecyl Dihexyldecyl Adipate is Skin- Adipate the diester of hexyldecanol Conditioning 57533-90-1 and adipic acid. Agents - Emollient; Solvents Diheptylundecyl Diheptylundecyl Adipate Skin- Adipate is the diester of Conditioning 155613-91-5 heptylundecanol and Agents - adipic acid. Emollient; Solvents

CIR Panel Book Page 141 Table 1. Definitions, functions and structures of dicarboxylic acid, salt and ester ingredients in this safety assessment. Ingredient Definition Function(s) Formula/structure CAS No. Dioctyldodecyl Dioctyldodecyl Adipate is Plasticizers; Skin- Adipate the diester of Conditioning 85117-94-8 octyldodecanol and adipic Agents - acid. Emollient

Diisocetyl Diisocetyl Adipate is the Plasticizers; Skin- One example of an “iso” Adipate diester of hexadecyl Conditioning 59686-69-0 alcohol and adipic acid. Agents - Emollient; sec: Solvents 58262-41-2 Diisostearyl Diisostearyl Adipate is the Plasticizers; Skin- One example of an “iso” Adipate diester of isostearyl alcohol Conditioning 62479-36-1 and adipic acid. Agents - Emollient

Sebacic Ester Ingredients Diethyl Diethyl Sebacate is the Fragrance Sebacate diester of ethyl alcohol and Ingredients; 110-40-7 sebacic acid. Plasticizers; Skin- Conditioning Agents - Emollient; Solvents Dibutyl Dibutyl Sebacate is the Fragrance Sebacate diester of butyl alcohol and Ingredients; 109-43-3 sebacic acid. Plasticizers; Skin- Conditioning Agents - Emollient; Solvents Dicaprylyl/ Dicaprylyl/Capryl Plasticizers; Skin- Capryl Sebacate the diester of Conditioning Sebacate caprylic and capryl Agents -

No CAS. No. alcohol, and sebacic acid. Emollient; wherein n=7 or 9 Solvents

Branched Monoester Isostearyl Isostearyl Sebacate is the Skin- One example of an “iso” Sebacate monoester of isostearyl Conditioning 478273-24-4 alcohol and sebacic acid. Agents - Miscellaneous Branched Disesters Diisopropyl Diisopropyl Sebacate is the Plasticizers; Skin- Sebacate diester of isopropyl alcohol Conditioning 7491-02-3 and sebacic acid. Agents - Emollient; Solvents Diethylhexyl Diethylhexyl Sebacate is Fragrance Sebacate the diester of 2-ethylhexyl Ingredients; 122-62-3 alcohol and sebacic acid. Plasticizers; Solvents

Dibutyloctyl Dibutyloctyl Sebacate is Skin- Sebacate the diester of butyloctyl Conditioning 184706-97-6 alcohol and sebacic acid. Agents - Emollient; Solvents

CIR Panel Book Page 142 Table 1. Definitions, functions and structures of dicarboxylic acid, salt and ester ingredients in this safety assessment. Ingredient Definition Function(s) Formula/structure CAS No. Diisooctyl Diisooctyl Sebacate is the Antioxidants; One example of an “iso” Sebacate diester of isooctyl alcohol Plasticizers; Skin- 10340-41-7 and sebacic acid. Conditioning Agents - Emollient Dihexyldecyl Dihexyldecyl Sebacate is Skin- Sebacate the diester of hexyldecyl Conditioning 359073-59-9 alcohol and sebacic acid. Agents - Emollient; Solvents

Dioctyldodecyl Dioctyldodecyl Sebacate is Skin- Sebacate the diester of Conditioning 69275-01-0 octyldodecanol and sebacic Agents - acid. Emollient; Solvents

Diisostearyl Diisostearyl Sebacate is the Skin- One example of an “iso” Sebacate diester of isostearyl alcohol Conditioning No CAS No. and sebacic acid. Agents - Emollient

Dodecanoic Ester Ingredients Dioctyldodecyl Dioctyldodecyl Hair Dodecanedioate Dodecanedioate is the Conditioning 129423-55-8 diester of Agents; Skin- octyldodecanol and Conditioning dodecanedioic acid. Agents - Miscellaneous Diisocetyl Diisocetyl Skin- One example of an “iso” Dodecanedioate Dodecanedioate is the Conditioning 131252-83-0 diester of Agents - octyldodecanol and Emollient; dodecanedioic acid. Surfactants - Emulsifying Agents

CIR Panel Book Page 143 Table 2a. Physical and Chemical properties of the alkyl dicarboxylic acid and salt ingredients.

INCI Name Malonic Succinic Sodium Disodium Glutaric Adipic Acid Acid Acid Succinate Succinate Acid

Appearance small colorless crystalline crystalline large white, crystals prisms monoclinic monoclinic prisms prisms

Molecular 104.06 118.09 140.07 162.05 132.11 146.14 Weight (g/mol) Melting/Boiling 135 (dec.)/ 185-187/ 206 (est.)/ 156 (est.)/ 97.5-98/ 152/265 Point (°C) 264 (est.) 235 486 (est.) 426 (est.) 302-304

Density (g/cm3) 1.63 1.56 -- -- 1.429 1.360

Vapor pressure 0.001 (est.) 0.0000002 7.3 E-10 (est.) 8.7 E-8 0.000003 0.07 (mm Hg @ (est.) 25°C) Solubility (g/L 1520 83 1000 (est.) 31 (est.) 639 30 water @ 25°C) Log Kow -0.81 -0.59 -3.98 (est.) -3.98 (est.) -0.29 0.08

INCI Name Azelaic Disodium Dipotassium Sebacic Disodium Dodecanedioic Acid Azelate Azelate Acid Sebacate Acid

Appearance monoclinic crystalline crystalline Monoclinic crystalline -- prismatic prismatic needles tablets

Molecular 188.22 238.18 264.40 202.25 246.21 230.31 Weight (g/mol) Melting/Boiling 106.5/ 186 (est.)/ 186 (est.)/ 134.5/ 194/496 128/383 (est.) Point (°C) 286.5 484 (est.) 484 (est.) 294.5 (est.)

Density (g/cm3) 1.0291 -- -- 1.207 -- 1.16

Vapor pressure 0.00002 1.4 E-9 1.4 E-9 (est.) 0.000007 5.9 E-10 0.000002 (est.) (mm Hg @ (est.) (est.) (est.) (est.) 25°C) Solubility (g/L 2.4 1000 (est.) 1000 (est.) 1.0 1000 (est.) 0.040 water @ 20°C) Log Kow 1.57 -3.56 (est.) -3.56 (est.) 2.19 (est.) -3.01 (est.) 3.17 (est.)

CIR Panel Book Page 144 Table 2b. Physical and Chemical properties of the mono- and di-carboxylic acid esters.

INCI Name Diethyl Decyl Dimethyl Diethyl Dicapryl Dicetearyl Diisobutyl Malonate Succinate Succinate Succinate Succinate Succinate Succinate

Appearance colorless -- -- liquid -- -- liquid liquid Molecular 160.17 258.35 146.14 174.19 342.51 566-623 230.30 Weight (g/mol)

Melting/Boiling -50/ 96/377 19.5/196.1 -21.3/217.7 14 (est.)/ --/-- -48 (est.)/ Point (°C) 198-199 (est.) 375 (est.) 216

Density (g/cm3) 1.055 1.002 1.1 1.04 0.94 (est.) -- 0.967 (est.) Vapor pressure 0.269 0.000001 0.4 (est.) 0.126 0.000008 -- 0.019 (mm Hg @ (est.) (est.) (est.) 25°C) Solubility (g/L 20 20 (est.) 50 (est.) 10 (est.) 0.0015 -- 0.60 (est.) water @ 25°C) (est.)

Log Kow 0.96 4.57 (est.) 0.26 (est.) 1.28 (est.) 7.39 (est.) -- 3.00 (est.)

INCI Name Diethylhexyl Dimethyl Diisobutyl Diisostearyl Dimethyl Diethyl Dipropyl Succinate Glutarate Glutarate Glutarate Adipate Adipate Adipate

Appearance -- liquid ------

Molecular 342.51 160.17 244.33 637.07 174.19 202.25 230.30 Weight (g/mol)

Melting/Boiling -12 (est.)/ -42.5/ -38 (est.)/ 212 (est.)/ 210/229 24-26/ -15.7/ Point (°C) 359 (est.) 214.2 237 600 (est.) (est.) 248-249 274 (est.)

Density (g/cm3) 0.933 1.0876 0.97 (est.) -- 1.062 1.08 0.98

Vapor pressure 0.00002 0.185 0.008 7.8 E-13 0.073 0.027 0.0055 (mm Hg @ (est.) (est.) (est.) (est.) (est.) (est.) 25°C) Solubility (g/L 0.002 (est.) 27 (est.) 0.29 (est.) 1.16 E-16 14 (est.) 2.8 0.62 water @ 25°C) (est.) (est.) (est.)

Log Kow 7.08 (est.) 0.57 (est.) 3.44 (est.) 17.5 (est.) 0.95 1.97 2.99 (est.) (est.) (est.)

CIR Panel Book Page 145 INCI Name Dibutyl Dihexyl Dicapryl Di-C12- Ditridecyl Dicetyl Diisopropyl Adipate Adipate Adipate 15 Alkyl Adipate Adipate Adipate Adipate Appearance -- liquid ------liquid

Molecular 258.35 314.46 426.67 482-567 510.83 594.99 230.30 Weight (g/mol)

Melting/Boiling 37.5/300 -8/351 26.5-27.1/ --/-- 45.9/503 56.5-57/ -1.1/253 Point (°C) (est.) (est.) 442 (est.) (est.) 559 (est.) (est.)

Density (g/cm3) 0.96 0.95 0.92 (est.) -- 0.91 (est.) 0.897 (est.) 0.982 (est.) (est.) Vapor pressure 0.0011 0.00004 0.00000005 -- 3.0 E-10 1.5 E-12 0.0192 (mm Hg @ (est.) (est.) (est.) (est.) (est.) (est.) 25°C) Solubility (g/L 0.14 (est.) 0.0082 0.000041 -- 0.0000011 0.00000005 0.78 (est.) water @ 25°C) (est.) (est.) (est.) (est.)

Log Kow 4.0 (est.) 6.0 (est.) 10.1 (est.) -- 13.8 (est.) 17 (est.) 2.68 (est.)

INCI Name Diisobutyl Diethylhexyl Diisooctyl Diisononyl Diisodecyl Dihexyldecyl Adipate Adipate Adipate Adipate Adipate Adipate

Appearance liquid liquid ------

Molecular 258.35 370.57 370.57 398.62 426.67 594.99 Weight (g/mol)

Melting/Boiling -20/278-280 -67.8/390 9 (est.)/ 56 (est.)/ 51 (est.)/ 181 (est.)/ Point (°C) 382 (est.) 230 426 (est.) 548 (est.)

Density (g/cm3) 0.95 0.925 0.93 (est.) -- -- 0.896 (est.)

Vapor pressure 0.0036 0.0000009 0.000004 3.3 E-6 1.9 E-6 4.6 E-12 (est.) (mm Hg @ (est.) (est.) (est.) (est.) 25°C) Solubility (g/L 0.18 0.00078 0.00067 4.0 E-5 5.2 E-6 0.00000006 water @ 25°C) (est.) (est.) (est.) (est.)

Log Kow 3.70 (est.) 6.11 7.77 (est.) 9.24 (est.) 10.1 (est.) 16.6 (est.)

CIR Panel Book Page 146 INCI Name Diheptylundecyl Dioctyldodecyl Diisocetyl Diisostearyl Diethyl Dibutyl Adipate Adipate Adipate Adipate Sebacate Sebacate

Appearance ------liquid liquid

Molecular 651.10 707.20 594.99 651.10 258.35 314.46 Weight (g/mol)

Melting/Boiling 229 (est.)/ 267 (est.)/ 181 (est.)/ 229 (est.)/ 5/298 -10/ Point (°C) 584 (est.) 619 (est.) 565 (est.) 611 (est.) 344-345

Density (g/cm3) 0.892 (est.) 0.888 (est.) 0.896 -- 0.969 0.94 (est.) (est.) Vapor pressure 1.26 E-13 (est.) 3.17 E-15 (est.) 1.4 E-11 2.4 E-13 0.00054 0.00004 (mm Hg @ (est.) (est.) (est.) (est.) 25°C) Solubility (g/L 9.8 E-9 (est.) 2.1 E-9 (est.) 4.0 E-12 3.6 E-14 0.15 0.0085 water @ 25°C) (est.) (est.) (est.) (est.)

Log Kow 18.7 (est.) 20.9 (est.) 16.0 (est.) 17.9 (est.) 3.92 5.96 (est.) (est.)

INCI Name Dicaprylyl/ Isostearyl Diisopropyl Diethylhexyl Dibutyloctyl Diisooctyl capryl Sebacate Sebacate Sebacate Sebacate Sebacate Sebacate Appearance ------

Molecular 426-482 454.73 286.41 426.67 538.89 426.67 Weight (g/mol)

Melting/Boiling --/-- 215 (est.)/ -7 (est.)/ -48/436 135 (est.)/ 51 (est.)/ Point (°C) 545 (est.) 308 (est.) (est.) 510 (est.) 428 (est.)

Density (g/cm3) -- 0.929 (est.) 0.953 (est.) 0.91 0.901 (est.) 0.916 (est.) Vapor pressure -- 2.5 E-13 0.0007 8.7 E-8 (est.) 1.6 E-10 (est.) 1.6 E-7 (mm Hg @ (est.) (est.) (est.) 25°C) Solubility (g/L -- 0.0013 0.046 0.00006 0.0000006 0.00006 water @ 25°C) (est.) (est.) (est.) (est.)

Log Kow -- 11.2 (est.) 4.63 (est.) 9.72 (est.) 14.1 (est.) 9.72 (est.)

CIR Panel Book Page 147 Table 2b. Physical and Chemical properties of the mono- and di-carboxylic acid esters. (continued)

INCI Name Dihexyldecyl Dioctyldodecyl Diisostearyl Dioctyldodecyl Diisocetyl Sebacate Sebacate Sebacate Dodecanedioate Dodecanedioate

Appearance ------

Molecular 651.10 763.31 707.20 791.36 679.15 Weight (g/mol)

Melting/Boiling 229 (est.)/ 299 (est.)/ 268 (est.)/ 314 (est.)/ 247 (est.)/ Point (°C) 584 (est.) 652 (est.) 568 (est.) 668 (est.) 635 (est.)

Density (g/cm3) 0.892 (est.) 0.885 (est.) -- 0.884 (est.) --

Vapor pressure 1.3 E-13 (est.) 7.4 E-17 (est.) 4.8 E-15 1.1 E-17 (est.) 3.6 E-14 (est.) (mm Hg @ (est.) 25°C) Solubility (g/L 0.0000001 6.8 E-10 (est.) 3.2 E-16 3.6 E-10 (est.) 3.4 E-15 (est.) water @ 25°C) (est.) (est.) Log Kow 18.4 (est.) 22.6 (est.) 19.9 (est.) 23.7 (est.) 18.9 (est.)

“(est.)” = estimated value by EPI Suite

“(dec.)” = some decomposition occured

“--“ = Value not found

“E-13” = divided by 1013

CIR Panel Book Page 148 Table 3a. Frequency and concentration of use by duration and exposure - Dicarboxylic Acids and Their Salts

2010 Uses 2010 Conc. of Use 2010 Uses 2010 Conc. of 2010 Uses 2010 Conc. of Use (VCRP) (%) (Council) (VCRP) Use (%) (Council) (VCRP) (%) (Council) Succinic Acid Sodium Succinate Disodium Succinate Totals 4 0.001-26 7 NR 45 0.0005-0.4 Duration of Use Leave-On 2 0.001-0.2 3 NR 38 0.005-0.4 Rinse Off 2 0.001-26 4 NR 7 0.0005 Exposure Type Eye Area NR NR NR NR 4 NR Possible Ingestion NR NR NR NR NR NR Inhalation NR NR NR NR NR NR Dermal Contact 2 0.01-26 5 NR 40 0.0005-0.4 Deodorant (Underarm) NR NR NR NR NR NR Hair, Non-Coloring 2 0.001-0.2 2 NR 5NR Hair, Coloring NR NR NR NR NR NR Nail NR NR NR NR NR NR Mucous Membrane NR 0.2 1 NR NR NR Bath Products NR 26 1 NR NR NR Baby Products NR NR NR NR NR NR

Adipic Acid Azelaic Acid Sebacic Acid Totals 25 0.000001-18 9 0.007-10 12 0.0009-1 Duration of Use Leave-On 2 0.000001 7 0.007-0.3 9 0.0009-0.03 Rinse Off 23 0.5-18 2 10 3 0.001-1 Exposure Type Eye Area NR 0.000001 NR NR NR NR Possible Ingestion NR 0.000001 NR NR NR NR Inhalation NR NR NR NR NR NR Dermal Contact 1 0.000001-18 25 0.007-10 12 0.0009-1 Deodorant (Underarm) NR NR NR NR NR 0.0009 HiHair, NNon-ColorinCl ig 24 050.5 NNRR NRNR NRNR NRNR Hair, Coloring NR NR NR NR NR NR Nail NR NR NR NR NR NR Mucous Membrane - NR NR NR 1 0.04 Bath Products 1 15-18 NR NR NR NR Baby Products NR NR NR NR NR NR

CIR Panel Book Page 149 Table 3b. Frequency and concentration of use by duration and exposure - Esters of Dicarboxylic Acids

2010 Conc. of 2010 Conc. of 2010 Conc. 2010 Conc. of 2010 Conc. of 2010 Conc. 2010 Uses Use (%) 2010 Uses Use (%) 2010 Uses of Use (%) 2010 Uses Use (%) 2010 Uses Use (%) 2010 Uses of Use (%) (VCRP) (Council) (VCRP) (Council) (VCRP) (Council) (VCRP) (Council) (VCRP) (Council) (VCRP) (Council) Diethyl Malonate Dimethyl Succinate Dicapryl Succinate Diethylhexyl Succinate Dimethyl Glutarate Dimethyl Adipate Totals NR 0.004-0.02 12 0.002-5 12 NR 38 0.02-6 13 0.5-15 12 0.2 Duration of Use Leave-On NR 0.02 NR 0.002 9 NR 34 0.02-6 NR NR NR NR Rinse Off NR 0.004-0.01 12 0.2-5 NR NR 4 3-5 13 0.5-15 12 0.2 Exposure Type Eye Area NR NR NR 0.002 NR NR 1 NR NR NR NR NR Possible Ingestion NR NR NR NR NR NR NR 3 NR NR NR NR Inhalation NR NR NR NR 1 NR NR 1 NR NR NR NR Dermal Contact NR 0.004-0.02 NR 0.002-5 8 NR 34 1-6 NR 15 NR NR Deodorant (Underarm) NR NR NR NR NR NR NR NR NR NR NR NR Hair, Non-Coloring NR NR NR NR 1 NR 4 0.02-5 NR NR NR NR Hair, Coloring NR NR NR NR NR NR NR NR NR NR NR NR Nail NR NR 12 0.2 NR NR NR NR 13 0.5 12 0.2

CIR Mucous Membrane NR NR NR NR 2 NR 1 NR NR NR NR NR Bath Products NR NR NR NR NR NR NR NR NR NR NR NR Panel Baby Products NR NR NR NR NR NR NR NR NR NR NR NR Book

Dihexyl Adipate Dicapryl Adipate Diisobutyl Adipate Diisodecyl Adipate Diheptylundecyl Adipate Dioctyldodecyl Adipate Page Totals 1 3 43 NR 22 0.001-3 1 NR NR 6 3 NR Duration of Use 150 LeaveLeave-OOnn 1 NR 38 NR 22 0.001-3 1 NR NR 6 3 NR Rinse Off NR 3 5 NR NR 0.002-0.5 NR NR NR NR NR NR Exposure Type Eye Area NR 3 NR NR NR NR NR NR NR NR NR NR Possible Ingestion NR NR NR NR NR NR NR NR NR NR 3 NR Inhalation NR NR 1 NR 5 0.05-3 NR NR NR NR NR NR Dermal Contact 1 3 43 NR 8 0.002-3 1 NR NR 6 3 NR Deodorant (Underarm) NR NR 30 NR NR NR NR NR NR NR NR NR Hair, Non-Coloring NR NR NR NR 5 0.05-0.2 NR NR NR NR NR NR Hair, Coloring NR NR NR NR NR NR NR NR NR NR NR NR Nail NR NR NR NR 9 0.001-0.7 NR NR NR NR NR NR Mucous Membrane NR NR NR NR NR 0.009 NR NR NR NR NR NR Bath Products NR NR 5 NR NR 0.5 NR NR NR NR NR NR Baby Products NR NR NR NR NR NR NR NR NR NR NR NR Table 3b. Frequency and concentration of use by duration and exposure - Esters of Dicarboxylic Acids (continued)

2010 Conc. of 2010 Conc. of 2010 Conc. 2010 Conc. of 2010 Conc. of 2010 Conc. 2010 Uses Use (%) 2010 Uses Use (%) 2010 Uses of Use (%) 2010 Uses Use (%) 2010 Uses Use (%) 2010 Uses of Use (%) (VCRP) (Council) (VCRP) (Council) (VCRP) (Council) (VCRP) (Council) (VCRP) (Council) (VCRP) (Council) Diisostearyl Adipate Diethyl Sebacate Isostearyl Sebacate Diisopropyl Sebacate Diethylhexyl Sebacate Diisooctyl Sebacate Totals 6 3-10 NR 1.5 NR 0.005-0.7 30 0.06-10 13 0.5-5 NR 1-3 Duration of Use Leave-On 4 10 NR 1.5 NR 0.005-0.7 29 0.06-10 13 0.5-5 NR 1-3 Rinse Off 2 3 NR NR NR NR 12NR 1 NR NR Exposure Type Eye Area NR NR NR NR NR NR 1NR 4NRNRNR Possible Ingestion 410NRNRNRNRNR NR NR NR NR NR Inhalation NR NR NR NR NR NR 1NRNR 1 NR NR Dermal Contact 6 3-10 NR 1.5 NR 0.005-0.7 23 0.06-10 11 0.5-5 NR 1-3 Deodorant (Underarm) NR NR NR NR NR NR 41NR 0.5 NR NR Hair, Non-Coloring NR NR NR NR NR NR 68 2NRNRNR Hair, Coloring NR NR NR NR NR NR NR NR NR NR NR NR Nail NR NR NR NR NR NR 1 0.08 NR NR NR NR CIR Mucous Membrane NR NR NR NR NR NR NR NR NR NR NR NR

Panel Bath Products NR NR NR NR NR NR NR NR NR NR NR NR Baby Products NR NR NR NR NR NR NR NR NR NR NR NR Book

Page Dioctyldodecyl Sebacate Dioctyldodecyl Diisocetyl Dodecanedioate Dodecanedioate Totals NR 3-8 5 6 2 0.9-7 151 Duration of Use Leave-On NR 3-8 5 6 2 0.9-7 Rinse Off NR NR NR NR NR NR Exposure Type Eye Area NR NR NR NR NR NR Possible Ingestion NR 8 1 6 NR NR Inhalation NR 3-5 2 NR NR 0.9-3 Dermal Contact NR 3-8 5 6 2 0.9-7 Deodorant (Underarm) NR NR NR NR NR NR Hair, Non-Coloring NR NR NR NR NR NR Hair, Coloring NR NR NR NR NR NR Nail NR NR NR NR NR NR Mucous Membrane NR NR NR NR NR NR Bath Products NR NR NR NR NR NR Baby Products NR NR NR NR NR NR

NR - none reported Table 3c. Current and historical frequency and concentration of use according to duration and type of exposure - previously reviewed esters

# of Uses Conc. of Use (%) # of Uses Conc. of Use (%) # of Uses Conc. of Use (%) Dibutyl Adipate Diisopropyl Adipate Diethylhexyl Adipate data year 2002 2010 2002 2010 1981 2010 1981 2010 1981 2010 1981 2010 Totals NR 6 5-8 NR 112 70 ≤0.1-25 0.005-8 27 48 ≤0.1-25 0.6-14 Duration of Use Leave-On NR 6 5-8 NR 92 64 ≤0.1-25 0.005-8 21 39 ≤0.1-10 0.9-14 Rinse Off NR 0 NR NR 20 6 ≤0.1-26 2-7 6 9 1-25 0.6 Exposure Type Eye Area NR 2 NR NR 2 2 1-25 1 NR 3 NR NR Possible Ingestion NR NR NR NR NR 1 NR NR 5 1 1-5 NR Inhalation NR 2 NR NR 47 21 0.1-25 0.005-8 6 5 1-5 NR Dermal Contact NR 3 8 NR 102 50 ≤0.1-25 0.005-8 25 43 ≤0.1-25 0.6-14 Deodorant (underarm) NR NR NR NR NR 6 NR NR 1 NR 0.1-1 0.9 Hair - Non-Coloring NR NR NR NR 10 17 ≤0.1-5 0.5-3 NR 1 NR NR Hair-Coloring NR NR NR NR NR NR NR NR NR NR NR NR Nail NR 1 5 NR NR NR NR NR 2 4 1-5 2-3 Mucous Membrane NR NR NR NR 1 NR 0.1-1 NR NR 1 NR NR

CIR Bath Products NR NR NR NR 8 1 1-25 2 4 NR 10-25 NR Baby Products NR NR NR NR NR NR NR NR NR 1 NR NR Panel

NR - not reported to the VCRP or the Council Book Page 152 Table 3d. Ingredients not reported to be used

Dicarboxylic Acids and Their Salts

Malonic Acid Glutaric Acid Disodium Azelate Dipotassium Azelate Disodium Sebacate Dodecanedioic Acid

Esters of Dicarboxylic Acids Decyl Succinate Diethyl Succinate Dicetearyl Succinate Diisobutyl Succinate Diisobutyl Glutarate Diisostearyl Glutarate Diethyl Glutarate Dipropyl Glutarate Di-C 12-15 Alkyl Adipate Ditridecyl Adipate Dicetyl Adipate Diisooctyl Adipate Diisononyl Adipate Dihexyldecyl Adipate Diisocetyl Adipate Dibutyl Sebacate Dicaprylyl/Capryl Sebacate Dibutyloctyl Sebacate Dihexyldecyl Sebacate Diisostearyl Sebacate

CIR Panel Book Page 153 Table 4. Acute toxicity - Dicarboxylic Acids and Their Salts Animals No./Gender/Group Dose median lethal dose/conc. Reference ORAL Succinic Acid rats not specified not specified 2750 mg/kg EPA 2001 rats not specified not specified 2260 mg/kg OECD 2003 Sodium Succinate rats 4 males, 4 females 0.5-8 g/kg 8000 mg/kg Maekawa et al 1990 Glutaric Acid rats not specified not specified 2260 mg/kg EPA 2001 Adipic Acid mice 13 males 1500-2500 mg/kg of a 6% suspension in 0.5% 1900 mg/kg OECD 2006 methyl cellulose mice not specified not specified 4175 mg/kg OECD 2006 mice not specified not specified 4200 mg/kg OECD 2006 rats not specified not specified 5050 mg/kg EPA 2001 rats 5 or 10 males 100-3000 mg/kg (n=5) or 5000 mg/kg (n=10) adipic 940 mg/kg OECD 2006 acid in 0.85% saline Wistar rats not specified not specified approx. 3600 mg/kg OECD 2006 rats 10 males 5000 mg/kg of a 33.3% suspension in 0.85% saline >5000 mg/kg OECD 2006

rats 5 males, 5 females 14.7-10,000 mg/kg as a 14.7-50% suspension in 5560 mg/kg OECD 2006 carboxymethyl cellulose (CMC) rats not specified 10,000 mg/kg >10,000 mg/kg OECD 2006 rat and rabbit not specified not specified >11,000 mg/kg OECD 2006 rabbits not specified 2430 or 4860 mg/kg of a 20% partially neutralized >2430 and <4860 mg/kg OECD 2006 soln (75% sodium adipate) Adipic/Glutaric/Succinic Mixture (percentages not given) rats not specified not specified 6829 mg/kg EPA 2001 Azelaic Acid Wistar rats 6 males 6 females 500-4000 mg/kg ≥4000 mg/kg Mingrone et al 1983 New Zealand 6 males 6 females 500-4000 mg/kg ≥4000 mg/kg Mingrone et al 1983 rabbits Disodium Sebacate Wistar rats 4 males, 4 females 0-5000 mg/kg >5000 mg/kg Greco et al 1990 New Zealand 4 males, 4 females 0-6000 mg/kg >6000 mg/kg Greco et al 1990 rabbits Dodecanedioic Acid rats m/f; no. not specified not specified >3000 mg/kg OECD 1994 DERMAL Glutaric Acid rabbits not specified not specified >10,000 mg/kg EPA 2001 Adipic Acid rabbits 1- 2/group; male and 5010 (n=1) or 7940 mg/kg (n=2) 40% adipic acid in >7940 mg/kg OECD 2006 female corn oil, with occlusion Adipic/Glutaric/Succinic Mixture (percentages not given) rabbits not specified not specified >7940 mg/kg EPA 2001 Dodecanedioic Acid albino rabbits males; no. not specified not specified >6000 mg/kg OECD 1994 INHALATION Adipic Acid 3 rats 20/group; males and 5.4 or 7.7 mg/l; head/nose-only exposure; MMAD50 >7700 mg/m OECD 2004 females <3.5 µm Adipic/Glutaric/Succinic Mixture (percentages not given) rats not specified not specified >0.03 mg/l EPA 2001 PARENTERAL Disodium Succinate mice not specified i.v. 4500 mg/kg OECD 2003 Adipic Acid mouse not specified i.p., 0.681-50% solution in 0.5% CMC approx. 170 mg/kg OECD 2006 mouse not specified i.p., 600 and 900 mg/kg aq. 600 mg/kg OECD 2006 mouse not specified i.p. admin 4000 mg/kg OECD 2006 rats 7 males i.p., 200-350 mg/kg 275 mg/kg OECD 2006

CIR Panel Book Page 154 Table 4. Acute toxicity - Dicarboxylic Acids and Their Salts Animals No./Gender/Group Dose median lethal dose/conc. Reference mouse not specified i.v., 650-700 mg/kg 2% solution 680 mg/kg OECD 2006 rabbit not specified i.v., 2430 mg/kg 20% soln, partially neutralized 2430 mg/kg OECD 2006 Disodium Azelate rats 6 males, 6 females i.p., 0-1198 mg/kg >1198 mg/kg Mingrone et al 1983 rabbits 6 males, 6 females i.p., 0-1198 mg/kg >1198 mg/kg Mingrone et al 1983 Disodium Sebacate Wistar rats 4 males, 4 females i.p., 0-7000 mg/kg 5500 mg/kg; dehydration and Greco et al. 1990 ascites formation was noted New Zealand 4 males, 4 females i.p., 0-8000 mg/kg 6000 mg/kg; dehydration and Greco et al 1990 rabbits ascites formation was noted Wistar rats 10 i.v. , 0-1000 mg/kg 560 mg/kg; dehydration and Greco et al 1990 ascites formation was noted New Zealand 10 i.v., 0-1800 mg/kg 1400 mg/kg; dehydration and Greco et al 1990 rabbits ascites formation was noted

CIR Panel Book Page 155 Table 5. Ocular Irritation - Dicarboxylic Acid and Their Esters Concentration Animals Procedure Results Reference Succinic Acid not specified not specified ocular irritation study (details not specified) severe ocular irritant EPA 2001 Glutaric Acid not specified not specified ocular irritation study (details not specified) moderate ocular irritant EPA 2001 Adipic Acid undiluted 2 albino rabbits 10 or 57.1 mg placed in eye; eye of 1 animal rinsed 10 mg: no irritation in rinsed eye, minimal in unrinsed eye; EPA 2001 57.1 mg: mild to moderate irritation in rinsed and unrinsed eyes undiluted 6 rabbits; gender not specified 0.1 ml instilled into the eye; eyes were not rinsed severely irritating - primary irritation index of 41.5/110; OECD 2006 irritated conjunctiva and scar formation, increased corneal opacity and iridal inflammation; not cleared by day 8

undiluted 3 rabbits; gender not specified 100m g instilled following GLP; acute eye severe irritation; corneal opacity and iridal irritation; cleared OECD 2006 irritation/corrosion test within 16 days undiluted 2 rabbits; gender not specified 50 mg placed in eye; eyes were not rinsed severely irritating; corneal opacity still present at day 8 OECD 2006 Dodecanedioic Acid

CIR not specified male rabbits, no. not specified ocular irritation study (GLP; details not provided) slight irritant; irritation index 11.96/110 OECD 1994

Panel not specified rabbits; no./gender not specified ocular irritation study (details not provided) small area of corneal opacity and mild conjunctival irritation; OECD 1994 cleared within 7 days Book Page 156 Table 6. Dermal irritation and sensitization - Dicarboxylic Acids and Their Salts Dose/Conc.. Animals Procedure Results Reference IRRITATION Succinic Acid not specified rabbits, no./gender not specified irritation studies (details not provided) slight to mild irritation EPA 2001 Glutaric Acid not specified rabbits, no./gender not specified irritation studies (details not provided) slight irritation EPA 2001 Adipic Acid 500 mg of 50% aq. 6 rabbits occlusive application to a 5 cm x 5 cm area of intact skin: erythema (score 2-3/4), clearedby day 3; abraded skin: OECD 2006 abraded or intact skin for 24 h mild to severe erythema and edema (2/4 at 24 h; 0-2 at 72 h), cleared by day 7 undiluted or 80% aq. paste 2 rabbits/group occlusive application to intact skin on the back and no irritation on the back; erythema on the ear at 24 h (score of 2/4), OECD 2006 the ear for 20 h with clearing by 72 h not specified rabbits, no./gender not specified occlusive application for 24 h not irritating OECD 2006 undiluted or 50% paste in 6 rabbits semi-occlusive application of 500 mg for 24 h slight to mild irritation in 3/6 rabbits with 50%; no corrosion with OECD 2006 propylene glycol (PG) undiluted test material 50% in PG 10 guinea pigs, gender not specified applied to intact skin no irritation OECD 2006 CIR Succinic/Glutaric/Adipic Acids Mixture (percentages not specified) Panel not given guinea pigs, no./gender not specified irritation study (details not provided) no to mild irritation OECD 2004 Dodecanedioic Acid Book not specified male rabbits, no. not specified irritation study; 4 h exposure (GLP; details not not an irritant; irritation index 0/8 OECD 1994

Page provided) 0.5 g male rabbits, no. not specified FHSA procedures not an irritant OECD 1994 157 SENSITIZATIONSENSITIZATION Adipic Acid induction: 0.1 ml of 1.0% aq. 10 guinea pigs/group induction: 4 sacral intradermal injections, 1/wk; very mild to no irritation; no sensitization OECD 2006 soln; challenge: 0.05 ml of 50 challenge: dermal application after a 2 wk rest period and 25% in PG Succinic/Glutaric/Adipic Acids Mixture (percentages not specified) not given guinea pigs, no./gender not specified sensitization study (details not provided) not a sensitizer OECD 2004 Dodecanedioic Acid induction: 0.5%; challenge: 25 female guinea pigs, no. not specified Magnusson-Kligman maximization test not a sensitizer OECD 1994 and 50% (intracutaneous admin at induction; dermal admin at challenge) Table 7. Genotoxicity studies - Dicarboxylic Acids and Their Salts

Concentration Vehicle Procedure Test System Results Reference IN VITRO Malonic Acid ≤3333 µg/plate water NTP preincubation assay, +/- metabolic activation S. typhimurium TA100, TA1535, TA97, TA98 negative NTP 1989 ID: 228078 Succinic Acid ≤5 mg/plate phosphate buffer Ames test S. typhimurium TA92, TA1535, TA100, TA1537, TA94, TA98 negative Ishidate et al. 1984

≤1.0 mg/ml saline chromosomal aberration assay Chinese hamster fibroblasts cells negative Ishidate et al. 1984 Sodium Succinate ≤10 µg/plate distilled water Ames test, +/- metabolic activation S. typhimurium TA97, TA102 negative TOXNET 1997 Disodium Succinate ≤5 mg/plate phosphate buffer Ames test S. typhimurium TA92, TA1535, TA100, TA1537, TA94, TA98 negative Ishidate et al. 1984 ≤10,000 µg/plate distilled water Ames test, +/- metabolic activation S. typhimurium TA97, TA102 negative OECD 2003 ≤15.0 mg/ml saline chromosomal aberration assay Chinese hamster fibroblasts cells equivocal Ishidate et al. 1984 Glutaric Acid not specified not specified Ames test, +/- metabolic activation S. typhimurium , strains not specified negative Bradford 1984 not specified not specified mouse lymphoma assay, +/- metabolic activation L5178Y/TK ± cells dependent on Bradford 1984 CIR culture pH

Panel not specified not specified transformation assay, +/- metabolic activation Balb/c-3T3 cells positive, +/- Bradford 1984 activation ≤10,000 µg/plate water NTP preincubation assay, +/- metabolic activation S. typhimurium TA100, TA1535, TA97, TA98 negative NTP 1989 ID: 994718 Book Adipic Acid

Page ≤10,000 µg/plate DMSO NTP preincubation assay, +/- metabolic activation S. typhimurium TA100, TA1535, TA97, TA98 negative NTP 1997 ID: A86767 ≤10 mg/plate not specified Ames test, +/- metabolic activation S. typhimurium TA1535, TA1537, TA1538, TA98, TA100; E. negative OECD 2006 158 colicoli WP2 ≤5 mg/plate not specified Ames test, +/- metabolic activation S. typhimurium TA1535, TA1537, TA1538, TA98, TA100; E. negative OECD 2006 coli WP2uvrA ≤200 mg/l not specified Ames test, without metabolic activation S. typhimurium TA1530, G-46 negative OECD 2006 ≤200 mg/l not specified yeast gene mutation assay, without metabolic S. cerevisiae D-3 negative OECD 2006 activation ≤2000 µg/plate DMSO mouse lymphoma assay, +/- metabolic activation L5178Y/TK ± cells negative TOXNET 2009 ≤200 mg/l not specified cytogenetic assay, without metabolic activation human embryonic lung fibroblasts negative OECD 2006 ≤1000 µg/ml not specified viral enhanced cell transformation assay Syrian hamster ovary cells negative OECD 2006 Azelaic Acid 20% cream Ames test; no details not specified negative Allegan 2004 20% cream HGRPT test; no details Chinese hamster ovary cells negative Allegan 2004 20% cream human lymphocyte test, no details human lymphocytes negative Allegan 2004 Sebacic Acid ≤5000 µg/plate DMSO Ames test, +/- metabolic activation S. typhimurium TA1535, TA1537, TA1538, TA98, TA100; E. negative TOXNET 1990 coli WP2 Dodecanedioic Acid 10-5000 µg/plate not specified Ames test, +/- metabolic activation S. typhimurium TA1535, TA1537, TA1538, TA98, TA100 negative OECD 1994 Table 7. Genotoxicity studies - Dicarboxylic Acids and Their Salts

Concentration Vehicle Procedure Test System Results Reference IN VIVO Glutaric Acid not specified not specified micronucleus assay mice negative Bradford 1984 Adipic Acid ≤375 mg/kg; 1 or 5 doses not specified cytogenetic assay; animals dosed orally by gavage male rats negative OECD 2006

5000 mg/kg (1 dose); not specified cytogenetic assay; animals dosed orally by gavage male rats negative OECD 2006 2500 mg/kg (5 doses) ≤375 mg/kg; 1 or 5 doses not specified dominant lethal assay; animals dosed orally by gavage male rats negative OECD 2006

5000 mg/kg (1 dose); not specified dominant lethal assay; animals dosed orally by gavage male rats negative OECD 2006 2500 mg/kg (5 doses) Azelaic Acid 20% cream dominant lethal assay mice negative Allergan 2004 Dodecanedioic Acid ≤5000 mg/kg not specified micronucleus assay Crl:CD-1(CR)BR mice negative OECD 1994 CIR Panel Book Page 159 Table 8. Acute toxicity - Esters of Dicarboxylic Acids Animals No./Gender/Group Dose median lethal dose/concentration, or result Reference ORAL Diethyl Malonate rats not specified not specified 15,000 mg/kg OECD 2005 Dimethyl Malonate rats not specified not specified >2000 mg/kg OECD 2005 Diethyl Succinate rats not specified not specified 8530 mg/kg Smyth et al.1949 Dibutyl Adipate rats not specified 20% dispersion 11,260-12,900 mg/kg Andersen 2006 rats not specified undiluted 1520 mg/kg Andersen 2006 rats not specified not specified 1290 mg/kg OECD1996 rats not specified undiluted 12,900 mg/kg EPA 2010 Di-C7-9 Branched and Linear Alkyl Esters of Adipic Acid Sprague-Dawley 5-10; males/females 2000-15,800 mg/kg, undiluted >15,800 mg/kg EPA 2010 Ditridecyl Adipate Sherman Wistar 5/gender 16,000 mg/kg >16,000 mg/kg EPA 2010 Wistar rats 5/gender 15,000 mg/kg >15,000 mg/kg EPA 2010 Diisopropyl Adipate Sprague-Dawley 5 males/5 females formulation containing 1.08% 1 female died Elder 1984 Sprague-Dawley 5 males/5 females formulation containing 1.08% no animals died Elder 1984 Sprague-Dawley 5 males/5 females formulation containing 5% no animals died Elder 1984 rats 5 males/f5 females formulation containing 0.7% >76,800 mg/kg Elder 1984 rats not specified formulation containing 20.75% >15,ooo mg/kg Elder 1984 Diisobutyl Adipate NMRI mice 5 males 2000 mg/kg >2000 mg/kg EvicCEBA 1998 Diethylhexyl Adipate mice 5 males/5 females ≤20,000 mg/kg in corn oil males: 15,000 mg/kg; females: 24,600 mg/kg Elder 1984 rats 5 males/5 females ≤20,000 mg/kg, undiluted 2 males of the 10,000 mg/kg group died; 1 male and 1 Elder 1984 female of the 20,000 mg/kg group died albino rats 5 males/5 femals 7400 mg/kg 1 animal died Elder 1984 rats not specified not specified single oral toxic dose - 9.11 g/kg Elder 1984 rats not specified not specified no-effect dose: 6000 mg/kg; central nervous system Elder 1984 effectsff seen at hihigher h concentrationsi Harlan-Wistar rats 5 males/5 females formulations containing 0.175% >6500 mg/kg Elder 1984 rats not specified not specified 9110 mg/kg OECD 2000 rats 5 males/females 7380 mg/kg, undiluted >7300 mg/kg EPA 2010 rats not specified not specified 9.1 g/kg EPA 2010 Diisooctyl Adipate rats 5/group 2000-64,000 mg/kg, undiluted >64,000 mg/kg EPA 2010 guinea pigs not specified not specified >5 ml/kg EPA 2010 Diisononyl Adipate rats 5/group 0.0346-10 g/kg, undiluted >10,000 mg/kg EPA 2010 Diisodecyl Adipate NMRI mice 5 male 2000 mg/kg >2000 mg/kg EviC-CEBA 1994 rats not specified undiluted 20,500 mg/kg EPA 2010 Dioctyldodecyl Adipate NMRI mice 5 female 2000 mg/kg >2000 mg/kg EvicC-CEBA1994 rats not specified not specified NOAEL <4000 mg/kg Mochida et al 1996 Diisocetyl Adipate NMRI mice 5 males 2000 mg/kg >2000 mg/kg EviC-CEBA 1994 Diisopropyl Sebacate NMRI mice 5 female 2000 mg/kg >2000 mg/kg EvicC-CEBA1994 Diethylhexyl Sebacate NMRI mice 5 female 2000 mg/kg >2000 mg/kg EviC-CEBA 1994 mice not specified undiluted 9.5 g/kg EPA 2010 rats not specified undiluted 5.0 cc/kg EPA 2010 rats not specified undiluted 12.8 g/kg EPA 2010 rats not specified undiluted 17 g/kg EPA 2010 Dioctyldodecyl Dodecanedioate Wistar rats 5 male/5 female 5000 mg/kg >5000 mg/kg AMA 1990

CIR Panel Book Page 160 Table 8. Acute toxicity - Esters of Dicarboxylic Acids Animals No./Gender/Group Dose median lethal dose/concentration, or result Reference Diisocetyl Dodecanedioate Wistar rats 5 male/5 female 5000 mg/kg >5000 mg/kg AMA 1991 Esterase Metabolites Ethylhexyl Alcohol (metabolite of diethylhexyl succinate, diethylhexyl adipate, and diethylhexyl sebacate) rats 1516-7000 mg/kg ExxonMobil 2002 mice 2500-3768 mg/kg ExxonMobil 2002 Hexyl Alcohol (metabolite of diehexyl adipate) rats 3131-4900 mg/kg ExxonMobil 2002 mice 103-1950 mg/kg ExxonMobil 2002 Butyloctanol (metabolite of dibutyloctyl sebacate) rats 12,900 mg/kg Smyth et al 1949 Decyl Alcohol (metabolite of decyl succinate ) rats 9800 mg/kg Smyth et al 1949 Isooctyl Alcohol (metabolite of diisooctyl adipate and diisoctyl sebacate) rats mixture of C7-9 branched alkyl >2000 mg/kg ExxonMobil 2002 Nonyl Alcohol (metabolite of diisononyl adipate) rats mixture of C8-10 branched alkyl 3000 mg/kg ExxonMobil 2002 Isodecyl Alcohol (metabolite of diisodecyl adipate) rats mixture of C9-11 branched alkyl 4600 mg/kg ExxonMobil 2002 DERMAL Diethyl Malonate rabbits not specified not specified 16,700 mg/kg OECD 2005 Dibutyl Adipate rabbits not specified 96% 20 ml/kg Andersen 2006 rats not specified i.m. NOAEL >8000 mg/kg Smith 1953 Ditridecyl Adipate rabbits 3 2000 mg/kg >2000 mg/kg EPA 2010 rabbits 10 5000 m/kg to abraded skin; semi- >5000 mg/kg EPA 2010 occlusive Diethylhexyl Adipate rabbits 8 ≤8700 m/kg to abraded skin; mild irritation; no systemic toxic effects Elder 1984 occlusive rabbitsrabbits 1male/1female1 male/1 female ≤8660 mg/kgmg/kg forfor 24 h, occcludedocccluded, 1 >8670 mg mg/kg/kg OECD 2000 intact and 1 abraded site Diisononyl Adipate rabbits 4/group 50-3160 mg/kg to abraded skin >3160 mg/kg EPA 2010 Diethylhexyl Sebacate guinea pigs not specified not specified <10,000 mg/kg BIBRA 1996 Dioctyldodecyl Dodecanedioate NZW rabbits 5 male/5 female 2000 mg/kg, intact skin, 24 h >2000 mg/kg AMA 1993 Esterase Metabolites Ethylhexyl Alcohol (metabolite of diethylhexyl succinate, diethylhexyl adipate, and diethylhexyl sebacate) rats >3000 mg/kg ExxonMobil 2002 rabbits 1980-2600 mg/kg ExxonMobil 2002 Hexyl Alcohol (metabolite of diehexyl adipate) rats 1500 mg/kg ExxonMobil 2002 rabbits 1500 - >500 mg/kg ExxonMobil 2002 Butyloctanol (metabolite of dibutyloctyl sebacate) rabbits 3.36 ml/kg Smyth et al 1949 Decyl Alcohol (metabolite of decyl succinate ) rabbits 3.5 ml/kg Smyth et al 1949 Isooctyl Alcohol (metabolite of diisooctyl adipate and diisoctyl sebacate) rats mixture of C7-9 branched alkyl >2600 mg/kg ExxonMobil 2002 Nonyl Alcohol (metabolite of diisononyl adipate) rats mixture of C8-10 branched alkyl 3160 mg/kg ExxonMobil 2002 Isodecyl Alcohol (metabolite of diisodecyl adipate) rats mixture of C9-11 branched alkyl >2600 mg/kg ExxonMobil 2002

CIR Panel Book Page 161 Table 8. Acute toxicity - Esters of Dicarboxylic Acids Animals No./Gender/Group Dose median lethal dose/concentration, or result Reference INHALATION Diethyl Malonate rats not specified concentrated vapors for 8 h no deaths OECD 2005 Diethyl Succinate rats not specified concentrated vapors for 8 h no deaths Smyth et al.1949 Dibutyl Adipate albino rats 6 male flowing stream of saturated air, 8 h no mortality Elder 1984 Dethylhexyl Adipate rats not specified concentrated vapors for 8 h no deaths Smyth et al.1949 Diethylhexyl Sebacate rats not specified 250 mg/m3 for 4 h no effect on lung or liver BIBRA 1996 rats 3 saturated vapor, 6 h no lung toxicity BIBRA 1996 rats 4 940 mg/m3, 7 h 3 rats died, may be attributable to thermal decomp BIBRA 1996 guinea pigs 2 940 mg/m3, 7 h no animals died BIBRA 1996 rabbits 4 940 mg/m3, 7 h 2 rabbits died, may be attributable to thermal decomp BIBRA 1996 Esterase Metabolites Ethylhexyl Alcohol (metabolite of diethylhexyl succinate, diethylhexyl adipate, and diethylhexyl sebacate) rats 3 males/3 females vapor conc. of 0.89 mg/l or aerosol/ 0.89 mg/l: all animals survived; 5.3 mg/l: all animals died ExxonMobil 2002 vapor conc of 5.3 mg/l, 4 h mice, rats, and 10 227 ppm, 6 h all animals survived ExxonMobil 2002 guinea pigs Hexyl Alcohol (metabolite of diehexyl adipate) rats 21 mg/l, 1 h >21 mg/l ExxonMobil 2002 Butyloctanol (metabolite of dibutyloctyl sebacate) rats concentrated vapors for 8 h no deaths Smyth et al 1949 Decyl Alcohol (metabolite of decyl succinate ) rats concentrated vapors for 8 h no deaths Smyth et al 1949 PARENTERAL Dimethyl Adipate Sprague-Dawley not specified i.p. 1.8 ml/kg Singh et al 1973 Diethyl Adipate SSpragueprague-Dawle Dawleyy not sspecifiedpecified i.i.p. 22.5.5 ml/kml/kgg SinSinghgh et al 1973 Dipropyl Adipate Sprague-Dawley not specified i.p. 3.8 ml/kg Singh et al 1973 Dibutyl Adipate rats not specified i.p. 5.2 ml/kg Andersen 2006 Diisopropyl Adipate rats not specified i.v. 640 mg/kg Elder 1984 Diethylhexyl Adipate rats not specified i.v. 900 mg/kg Elder 1984 rabbits not specified i.v. 540 mg/kg Elder 1984 Sprague-Dawley not specified i.p. >50 ml/kg Singh et al 1973

CIR Panel Book Page 162 Table 9. Ocular Irritation - Esters of Dicarboxylic Acids Concentration Animals Procedure Results Reference Diethyl Malonate undiluted rabbits, no./gender not specified 0.1 ml slight to moderate irritation OECD 2005 Dimethyl Malonate undiluted rabbits, no./gender not specified 0.1 ml ,unrinsed slight to moderate irritation; OECD 2005 cleared by day 8 Dibutyl Adipate undiluted rabbits, no. not specified unrinsed minimally irritating Andersen 2006 undiluted 2 New Zealand rabbits unrinsed slight irritation Andersen 2006 0.1% in olive oil rabbits unrinsed non-irritating Andersen 2006 Diisopropyl Adipate undiluted 6 albino rabbits 0.1 ml, unrinsed negligible irritation Elder 1984 undiluted 6 albino rabbits 0.1 ml, unrinsed non-irritating Elder 1984 0.7% in formulation 9 albino rabbits 0.1 mi, undiluted, rinsed some corneal stippling Elder 1984

5% in formulation 6 albino rabbits not specified non-irritating Elder 1984 20.75% in formulation 6 albino rabbits not specified non-irritating Elder 1984 undiluted 3 albino rabbits 0.1 ml, unrinsed non-irritating EviC-CEBA 1998 Diethylhexyl Adipate undiluted 6 albino rabbits 0.1 ml, unrinsed non-irritating Elder 1984 0.01% in formulation 6 albino rabbits 0.1 ml, unrinsed non-irritating Elder 1984 0.175% in formulation 6 albino rabbits 0.1 ml, unrinsed mild transient irritant Elder 1984 Diisopropyl Sebacate 6 rabbits 0.1 ml, unrinsed minimally irritating Consumer Product Testing Co 1991 Diethylhexyl Sebacate 1.2% in formulation undiluted EpiOcular MTT viability non-irritating MB Research Laboratories 2003 assay Dioctyldodecyl Dodecanedioate undiluted 6 rabbits 0.1 ml, unrinsed MMTS = 0.0; non-irritating AMA 1990 Diisocetyl Dodecanedioate undiluteddil t d 6 rabbits bbit 0.101 ml, l unrinsed i d MMTS = 0.0; 0 0 non-irritatin i it ti g AMA 1990 Esterase Metabolites Ethylhexyl Alcohol (metabolite of diethylhexyl succinate, diethylhexyl adipate, and diethylhexyl sebacate) rabbits 20 μg moderately severe corneal ExxonMobil 2002 irritation Isopropyl Alcohol (metabolite of diisopropyl adipate and diisopropyl sebacate) rabbits severely irritatiting Inchem 2009 Hexyl Alcohol (metabolite of dihexyl adipate) rabbits highly irritative ExxonMobli 2002

CIR Panel Book Page 163 Table 10. Dermal irritation and sensitization - Esters of Dicarboxylic Acids Dose/Conc. Animals Procedure Results Reference DERMAL IRRITATION Diethyl Malonate not specified rabbits occlusive application; 4 h slightly irritating OECD 2005 Dimethyl Malonate not specified rabbits semi-occlusive application; 4 h not irritating; slight erythema at 30-60 min after patch removal OECD 2005 Dibutyl Adipate undiluted rabbits applied to belly PII of 2/8 Andersen 2006 undiluted 5 albino rabbits 0.1 ml, applied 8x in 4 h moderate erythema at 24 h undiluted 3 rabbits impregnated bands, 3 d application, 3 wks moderate erythema Andersen 2006 undiluted 5 rabbits impregnated bands, applied 2w/wk for 6 no progressive skin damage Andersen 2006 li ti undiluted 3 rabbits 0.025 ml to intact and abraded skin, 3 erythema and capillary injection during the study; desquamation was observed Andersen 2006 applications at 3 h intervals for 3 days 10% in acetone 5 hairless mice applied to ear, 1x/day, 10 days no adverse effect Andersen 2006 10% in acetone mice application to backs, 2x/day, 14 days no adverse effect Andersen 2006 Diisopropyl Adipate

CIR undiluted 9 albino rabbits 24 h, 0.1 ml, occlusive PII of 1.6/4; mild irritant Elder 1984

Panel undiluted 9 albino rabbits 24 h, 0.1 ml, occlusive PII of 1.3/4; mild irritant Elder 1984 undiluted 9 albino rabbits 24 h, 0.1 ml, occlusive PII of 0.06/4; minimally irritating Elder 1984

Book 5% in formulation 9 albino rabbits 24 h, 0.1 ml, occlusive PII of 0.33; minimally irritating Elder 1984 20.75% in formulation 9 albino rabbits 24 h, 0.1 ml, occlusive PII of 0.11; minimally irritating Elder 1984 Page undiluted 3 albino rabbits semi-occlusive application; 4 h, undiluted non-irritating EviC-CEBA 1998

164 DiethDiethylhexylylhexyl AdiAdipatepate undiluted 6 albino rabbits intact and abraded skin, 0.5 ml, 24 h, occlusive very mild irritant Elder 1984

0.175% in formulation 3 albino rabbits 4, 0.5 ml applications irritation index of 1.6/4 Elder 1984 Diisodecyl Adipate undiluted 3 albino rabbits semi-occlusive application, 4 h, undiluted non-irritating; scores of 0-1 for erythema and 0 for edema at 1-72 h; reversible EviC-CEBA 1994 Dioctyldodecyl Adipate undiluted 3 albino rabbits semi-occlusive application, 4 h, undiluted non-irritating; scores of 0-1 for erythema and 0 or 1 for edema at 24-72 h; reversible EviC-CEBA 1994 Diisocetyl Adipate undiluted 3 albino rabbits semi-occlusive application, 4 h, undiluted non-irritating; scores of 0-2 for erythema and 0 or 1 for edema at 1-72 h; reversible EviC-CEBA 1994 Diethyl Sebacate undiluted 8 rabbits intact and abraded skin, occlusive, 0.3 ml PII of 0.0 Council 1989 30% in ethanol 8 rabbits intact and abraded skin, occlusive, 0.3 ml PII of 0.3 Council 1989 Diisopropyl Sebacate undiluted 6 rabbits intact and abraded skin, occlusive, 0.5 ml PII of 2.88; not a primary irritant Consumer Product Testing 1991 undiluted 3 albino rabbits semi-occlusive application, 4 h, undiluted non-irritating; scores of 1 for erythema, with a 2 at 24 h, and 0 or 1 for edema at 1-72 h; EviC-CEBA 1994 reversible Table 10. Dermal irritation and sensitization - Esters of Dicarboxylic Acids Dose/Conc. Animals Procedure Results Reference Diethylhexyl Sebacate undiluted 3 albino rabbits semi-occlusive application, 4 h, undiluted non-irritating; scores of 1 for erythema and 0 for edema at 1-72 h; reversible EviC-CEBA 1994 undiluted 2-4 rabbits occlusive application; 48 h not irritating BIBRA 1996 Dioctyldodecyl Dodecanedioate undiluted 6 NZW rabbits occlusive application, 24 h, 0.5 ml PII = 0; not a primary irritant AMA 1990 Diisocetyl Dodecanedioate undiluted 6 NZW rabbits occlusive application, 24 h, 0.5 ml PII = 0; not a primary irritant AMA 1990 Esterase Metabolites Ethylhexyl Alcohol (metabolite of diethylhexyl adipate and diethylhexyl sebacate) 3 male rabbits occluision, 4 h irritating ExxonMobil 2002 rabbits occlusive, 0.5 ml highly irritating; not reversible ExxonMobil 2002 Caprylic Alcohol (metabolite of dicapryl succinate, dicapryl adipate, and dicaprylyl/capryl sebacate) undiluted rabbits mild irritation SENSITIZATION Dimethyl Malonate

CIR not specified guinea pigs Buehler method not sensitizing OECD 2005 Dibutyl Adipate Panel 25% 5 guinea pigs maximization test not sensitizing Andersen 2006 Diethylhexyl Adipate Book 0.1% in olive oil 10 male guinea induction: 10 injections; 2 wk non-treatment not sensitizing Elder 1984 pigs pd; challenge: 0.05 ml injection Page Diethylhexyl Sebacate undiluted rabbits occlusive patches, details not provided no reactions BIBRA 1996 165 DiDioct ty ldldodec dy l DDodecanedioate d di t 0.1 ml for intraderm 10 female guinea maximization test not sensitizing; slight erythema at induction AMA 1993 induction; 0.5 ml top. pigs induction /challenge Esterase Metabolites Hexyl Alcohol (metabolite of dihexyl adipate) 1% in petrolatum guinea pigs maximization test not sensitizing ExxonMobil 2002 Table 11. Genotoxicity studies - Esters of Dicarboxylic Acids

Concentration Vehicle Procedure Test System Results Reference IN VITRO Diethyl Malonate ≤5000 µg/plate not specified Ames test, +/- metabolic activation S. typhimurium TA1535, TA1537, TA98, TA100 negative OECD 2005 ≤5000 µg/plate not specified cytogenetic assay, +/- metabolic activation human peripheral lymphocytes negative; cytotoxic at 5000 OECD 2005 µg/plate Dimethyl Malonate ≤5000 µg/plate not specified Ames test, +/- metabolic activation S. typhimurium TA1535, TA1537, TA98, TA100 negative; cytotoxic at ≥1000 OECD 2005 µg/plate Dimethyl Succinate 20,000 µg/plate DMSO Ames test, +/- metabolic activation S. typhimurium TA1535, TA1537, TA98, TA100 negative Andersen and Jensen 1984 ≤10,000 µg/plate water NTP preincubation assay, +/- metabolic S. typhimurium TA100, TA1535, TA97, TA98 negative NTP1988 ID: 947738 activation Dimethyl Glutarate ≤10,000 µg/plate DMSO NTP preincubation assay, +/- metabolic S. typhimurium TA100, TA1535, TA97, TA98 negative NTP1995 ID: A20348 activation

CIR Dimethyl Adipate

Panel ≤10,000 µg/plate DMSO NTP preincubation assay, +/- metabolic S. typhimurium TA100, TA1535, TA97, TA98 negative NTP1994 ID: A45330 activation Dibutyl Adipate Book ≤5000 µg/plate Ames test, +/- metabolic activation S. typhimurium TA98, TA100, TA1535, TA1537, negative Andersen 2006

Page TA1538 Di-C7-9 Branched and Linear Alkyl Esters of Adipic Acid

166 ≤10.010.0 µµl/platel/plate not sspecifiedpecified Ames test, +/-+/ metabolic activation SS.. ttyphimuriumyphimurium TA1535, TA1537, TA98, TA100 nenegativegative EPA 2010 Ditridecyl Adipate ≤10 µl/plate DMSO Ames test, +/- metabolic activation S. typhimurium TA1535, TA1537, TA1538, TA98, negative EPA 2010 TA100 Diisobutyl Adipate ≤10,000 µg/plate DMSO Ames test, +/- metabolic activation S. typhimurium TA98, TA100, TA102, TA97, TA98, negative TOXNET 2001 E. coli wp2 Diethylhexyl Adipate ≤5 mg/plate not specified Ames test, +/- metabolic activation S. typhimurium TA1535, TA1537, TA1538, TA98, negative Elder 1984 TA100 5000 µg/plate not specified Ames test, +/- metabolic activation S. typhimurium TA1535, TA1537, TA1538, TA98, negative Simmon et al 1977 TA100 ≤0.01 M not specified Ames test, +/- metabolic activation S. typhimurium TA98, TA100 negative Warren et al 1982 10,000 µg/plate DMSO Ames test, +/- metabolic activation S. typhimurium TA1535, TA1537,. TA98, TA100 negative Zeiger et al 1985 10,000 µg/plate 95% ethanol Ames test, +/- metabolic activation S. typhimurium TA1535, TA1537,. TA98, TA100 negative Zeiger et al 1985 10,000 µg/plate acetone NTP preincubation assay, +/- metabolic S. typhimurium TA100, TA1535, TA97, TA98 negative NTP1988 ID: 963935 activation ≤150 µg/plate not specified Ames test, +/- metabolic activation S. typhimurium TA1535, TA1537, TA1538, TA98, negative EPA 2010 not specified DMSO liquid suspension assay S. typhimurium TA100 negative Seed 1982 Table 11. Genotoxicity studies - Esters of Dicarboxylic Acids

Concentration Vehicle Procedure Test System Results Reference ≤400 µg/ml not specified sister chromatid exchange assay, +/- Chinese hamster ovary cells negative w/out activation; Galloway et al 1987 metabolic activation equivocal w/activation ≤200µg/plate, 3 or 51 h DMSO sister chromatid exchange assay female F344 rat hepatocytes negative Reisenbichler and Eckl 1993 ≤400 µg/ml not specified chromosomal aberration assay, +/- metabolic Chinese hamster ovary cells some evidence w/out activation; Galloway et al 1987 activation negative w/activation ≤200µg/plate, 3 or 51 h DMSO chromosomal aberration assay female F344 rat hepatocytes negative Reisenbichler and Eckl 1993 ≤0.01 M not specified 3H-thymidine assay, +/- metabolic activation splenic lymphoid cells dose-dependent inhibition of 3H- Warren et al 1982 thymidine into replicating DNA, w/a dose-dependent increase in the ratio of acid- soluble to DNA-incorporated 3H- thymidine ≤1000 µg/plate forward mutation assay, +/- metabolic act. L5178Y cells negative McGregor et al 1988 urine of rats dosed with 2000 mg/kg diethylhexyl adipate

CIR corn oil Ames test negative DiVincenzo et al 1986 Diisononyl Adipate Panel ≤1000 µg/plate Ames test, +/- metabolic activation S. typhimurium TA98, TA100, TA1535, TA1537, negative McKee et al 1986 TA1538 Book ≤100 µg/ml lymphoma assay, +/- metabolic activation mouse lymphoma L5178Y cells negative McKee et al 1986

Page ≤1000 µg/ml transformation assay Syrian hamster embryo cells negative McKee et al 1986 ≤1.3 µ/ml BALB/3T3 assay negative McKee et al 1986

167 Diethyly Sebacate reversion assay E. coli Sd-4-73 negative Szybalski 1958 Dibutyl Sebacate not specified not specified Ames test S, typhimurium TA98, TA100, TA1535, TA1537, negative Wild et al 1983 TA1538 ≤10,000 µg/plate DMSO & Tween Ames test, +/- metabolic activation S. typhimurium TA98, TA100, TA1535, TA1537; negative TOXNET 2001 80 E. coli wp2 uvrA Diethylhexyl Sebacate ≤10,000 µg/plate DMSO Ames test, +/- metabolic activation S. typhimurium TA98, TA100, TA1535, TA1537 negative Zeiger et al 1985 ≤5000 µg/plate DMSO Ames test, +/- metabolic activation S. typhimurium TA98, TA100, TA1535, TA1537; negative TOXNET 2010 E. coli wp2 uvrA Esterase Metabolites Ethylhexyl Alcohol (metabolite of diethylhexyl succinate, diethylhexyl adipate, and diethylhexyl sebacate) 10,000 µg/plate Ames test, +/- metabolic activation negative Zeiger et al 1985 Ames test negative McKee et al 1986 ≤0.01 M not specified Ames test, +/- metabolic activation S. typhimurium TA98, TA100 negative Warren et al 1982 ≤5000 μg/plate not specified Ames test, +/- metabolic activation negative ExxonMobil 2002 0-1.5 mM DMSO liquid suspension assay S,. typhimurium TA100 negative Seed 1982 not specified not specified mouse lymphoma assay negative McKee et al 1986 not specified not specified unscheduled DNA synthesis negative McKee et al 1986 Table 11. Genotoxicity studies - Esters of Dicarboxylic Acids

Concentration Vehicle Procedure Test System Results Reference ≤0.01 M not specified 3H-thymidine assay, +/- metabolic activation splenic lymphoid cells dose-dependent inhibition of 3H- Warren et al 1982 thymidine into replicating DNA, w/a dose-dependent increase in the ratio of acid- soluble to DNA-incorporated 3H- thymidine 1000 mg/kg corn oil Ames test performed on urine from rats negative DiVincenzo et al 1986 dosed orally for 15 days MEHA (metabolite of diethylhexyl adipate) 10,000 µg/plate Ames test, +/- metabolic activation negative Zeiger et al 1985 ≤1000 µg/plate Ames test negative Dirven et al 1991 Mono-(2-Ethyl-5-Hydroxyhexyl)Adipate (metabolite of diethylhexyl adipate) ≤1000 µg/plate Ames test negative Dirven et al 1991 Mono-(2-Ethyl-5-Oxohexyl)Adipate (metabolite of diethylhexyl adipate) ≤1000 µg/plate Ames test negative Dirven et al 1991 Propyl and Isopropyl Alcohol (metabolite of dipropyl adipate, diisopropyl adipate, and diisopropyl sebacate) CIR bacterial and mammalian cell assays negative Heldreth 2010

Panel Isooctyl Alcohol (metabolite of diisooctyl adipate and diisooctyl sebacate) C7-9 branched alkyl bacterial and mammalian cell assays negative ExxonMobil 2002

Book IN VIVO Dimethyl Succinate

Page ≥1250 mg/kg corn oil micronucleus test, i.p. male F344 rats negative NTP 1996 ID:A71006 Dimethyl Glutarate

168 ≥1250 mmg/kgg/kg corn oil micronucleus test, i.i.p. male F344 rats negativenegative NTP 1995 ID: A36863 Dibutyl Adipate ≤2000 mg/kg olive oil mouse micronucleus test mice negative Andersen 2006 ≥724 mg/kg corn oil micronucleus test, i.p. male F344 rats negative NTP 1994 ID: A86146 Ditridecyl Adipate ≤2000 mg/kg none micronucleus test; dosed dermally for 13 wks groups of 10 male and 10 female Sprague Dawley negative EPA 2010 rats Diethylhexyl Adipate

2000 mg/kg corn oil micronucleus test; dosed i.p. for 3 days 5 male B3C3F1 mice negative Shelby et al 1993

≤5000 mg/kg corn oil chromosomal aberration assay 8 male B3C3F1 mice negative NTP 1988 ID:959525

not specified corn oil chromosomal aberration assay 8 B6C3F1 mice negative Shelby and Witt 1995 5000 mg/kg corn oil micronucleus test single i.p. dose 6 male/6 female B3C3F1 mice negative EPA 2010 Dibutyl Sebacate 943-2829 mg/kg olive oil micronucleus test, i.p. micronucleus test negative Wild et al 1983 Diethylhexyl Sebacate 500 mg/kg not specified rat liver foci test single dose of known carcinogen, the dosing 3x/wk no activity Osterle and Deml 1988 for 11 wks Table 11. Genotoxicity studies - Esters of Dicarboxylic Acids

Concentration Vehicle Procedure Test System Results Reference Esterase Metabolites Ethylhexyl Alcohol (metabolite of diethylhexyl succinate, diethylhexyl adipate, and diethylhexyl sebacate) not specified not specified micronucleus test mice negative McKee et al 1986 not specified not specified transformation assay BALB/3T3 negative McKee et al 1986 Propyl and Isopropyl Alcohol (metabolite of dipropyl adipate, diisopropyl adipate, and diisopropyl sebacate) C7-9 branched alkyl micronucleus test negative Heldreth 2010 CIR Panel Book Page 169 Table 12. Clinical dermal irritation and sensitization - Esters of Dicarboxylic Acids

Test Material No. of Procedure Results Reference Subjects Dimethyl Malonate 8% in petrolatum 25 maximization test not a sensitizer OECD 2005 Dibutyl Adipate undiluted 10 24 h patch test no irritation at 24 or 48 h Andersen 2006 20% in alcohol 10 24 h occlusive patch test slight reactions in 4 subjects Andersen 2006 Diisopropyl Adipate undiluted 19 24 h occlusive patch, 0.1 ml no irritation Elder 1984 undiluted 19 24 h occlusive patch, 0.1 ml no irritation Elder 1984 undiluted 15 24 h occlusive patch, 0.1 ml no irritation Elder 1984 undiluted 15 24 h occlusive patch, 0.1 ml no irritation Elder 1984 undiluted 16 cumulative irritancy test moderately irritating; score of 395/945; irritation in 14/16 subjects Elder 1984 0.7% in formulation 13 cumulative irritancy test non-irritating; score of 2/630 Elder 1984 1.1% in formulation 17 cumulative irritancy test low potential for hazard to consumer; score of 0.29/84 Elder 1984 1.1% in formulation 17 cumulative irritancy test low potential for hazard to consumer; score of 0.24/84 Elder 1984 20.75% in a bath oil 7 cumulative irritancy test score of 8/84 Elder 1984 20.75% in formulation diluted to 1.25% 19 24 h occlusive patch, 0.1 ml minimal irritation Elder 1984

CIR 5.0% in formulation 19 24 h occlusive patch, 0.1 ml no irritation Elder 1984 1.08% in formulation 235 HRIPT no sensitization; slight hyperpigmentation Elder 1984 Panel 3.0% in formulation 50 HRIPT no irritation or sensitization Elder 1984 5.0% in formulation 108 HRIPT no irritation or sensitization Elder 1984 Book 5.0% aq. dispersion of a product 116 HRIPT minimal, faint erythema produced throughout the study Elder 1984 containing 20.75% Page 0.7% in formulation 25 maximization test no contact sensitization potential Elder 1984 DiDieth hy lhlhexy l Adipate 170 0.175% in formulation 11 cumulative irritancy test slightly irritating; score of 72/630 Elder 1984 0.01% in formulation 100 Schwartz-Peck prophetic patch test not an irritant or a sensitizer Elder 1984 0.01% in formulation 49 Shelanski and Shelanski HRIPT weak reactions in up to 4 subjects and strong reactions in 1 subject Elder 1984 9.0% in formulation 209 modified Draize-Shelanski patch test 3 strong reactions and 1 faint reaction at 2nd challenge Elder 1984 9.0% in formulation 151 modified Draize-Shelanski patch test irritant reactions in 2 subjects; no sensitization Elder 1984 product containing 0.7% of a 25% not given Shelanski-Jordan RIPT 1-2 subjects had reactions during the study Elder 1984 Diisostearyl Adipate undiluted 50 HRIPT not a primary irritant or sensitizer AMA Laboratories 1996 1.5% in formulation 20 SIOPT not irritating Anonymous 2003 1.5% in formulation 25 maximization test no contact sensitization potential KGL Inc 2003 Diisopropyl Sebacate 1.8% in formulation 20 SIOPT not irritating Anonymous 2000 undiluted 105 patch test no irritation or sensitization Product Investigations Inc. 2005 2.2% in formulation 27 maximization test no irritation or sensitization KGL Inc 2006 1% in formulation 110 modified HRIPT, semi-occlusive not an irritant or a sensitizer Reliance Clinical Testing 2007 1% in formulation 110 modified HRIPT, semi-occlusive not an irritant or a sensitizer Reliance Clinical Testing 2007 7.2% in formulation 51 HRIPT, semi-occlusive no skin reactivity observed Essex Testing Clinic 2007 Diethylhexyl Sebacate undiluted 15-30 occlusive patches no reactions BIBRA 1996 Dioctyldodecyl Dodecanedioate undiluted 50 HRIPT not a primary irritant or sensitizer AMA Laboratories 1996 Table 12. Clinical dermal irritation and sensitization - Esters of Dicarboxylic Acids

Test Material No. of Procedure Results Reference Subjects Diisocetyl Dodecanedioate undiluted 50 HRIPT not a primary irritant or sensitizer AMA Laboratories 1996 Esterase Metabolites Methanol (metabolite of dimethyl succinate, dimethyl glutarate, and dimethyl adipate primary irritation of the skin Andersen 2001 3.2% 274 provocative occupational study positive results Andersen 2001 5% closed patch test slight positive reaction (+) Andersen 2001 7 and 70% closed patch test +++ reactions Andersen 2001 Propyl Alcohol undiluted 20 24 h patch test no reactions Stillman et al. 1975 undiluted 116 48 h patch test no reactions Wahlberg and Maibach 1980 undiluted 16 24 h patch test no reactions Agner and Serup 1987 undiluted 42 48 h patch test no reactions Willis et al 1988 undiluted 16 24 h patch test no reactions Agner and Setup 1988 undiluted 7 24 h patch test no reactions Agner and Setup 2010 Isopropyl Alcohol (metabolite of diisopropyl alcohol and diisopropyl sebacate)

CIR 80.74% spray concentration 9 no sensitization potential Damato et al 1979 2.85% in formulation 109 HRIPT no sensitization Anonymous 2010 Panel undiluted 12 24 h patch test no reactions Suihko and Serup 2008 Cetyl Alcohol (metabolite of dicetyl succinate and dicetyl adipate)

Book 11.5% in formulation 80 topical tolerance study reaction in 1 subject Elder 1988 6.0% in formulation 12 cumulative irritancy test mild cumulative irritation Elder 1988

Page 8.4% in formulation 110 HRIPT not a primary irritant or sensitizer Elder 1988 30%informulation3.0% in formulation 25 HRIPT notnot a sensitizersensitizer ElderElder 1988

171 Myristyl Alcohol (metabolite of dimyristyl adipate) 0.80% in formulation 53 4 wk application no irritation Elder 1988 0.25% in formulation 51 4 wk application 1 reaction by 1 subject Elder 1988 0.25% in formulation 229 10 - 24 h occlusive patch not an irritant or an allergen Elder 1988 Stearyl Alcohol ((metabolite of distearyl succinate) undiluted SIOPT mild irritation Elder 1985 Isostearyl Alcohol (metabolite of diisostearyl glutarate, diisostearyl adipate, or diisostearyl sebacate) 25% in petrolatum 19 no irritation Elder 1988 25.0% in formulation no irritation Elder 1988 27.0% in formulation no irritation Elder 1988 28.0% in formulation no irritation Elder 1988 25% in 95% isopropyl alcohol 12 HRIPT 3 subject slight erythema at induction; no sensitization Elder 1988 5% in formulation 148 HRIPT, with rechallenge for reactors; 12 subjects had possible sensitization reactions at 1st challenge; 6 Elder 1988 add'l challenge with 5% in ethanol reacted at rechallenge; all 6 had positive reactions to 5% in alcohol 5% in formulation 60 HRIPT, rechallenge of 5% in ethanol 5 subjects reacted at 1 challenge1/5 rechallenged reacted Elder 1988 for reactors Caprylic Alcohol (metabolite of dicapryl succinate, dicapryl adipate, and dicaprylyl/capryl sebacate) 2% in petrolatum 25 48 h closed patch no irritation Opdyke 1973 Decyl Alcohol (metabolite of decyl succinate and didecyl sebacate) 3% in petrolatum 25 48 h closed patch no irritation Opdyke 1973 Table 13. Case report - Esters of Dicarboxylic Acids

Subject Presentation Follow-Up Testing/Discussion Reference Diethyl Sebacate 1 subject severe contact dermatitis after 7 days of treatment with a drug patch test with 20% diethyl sebacate in ethanol Schneider 1980 1 subject quick onset of contact dermatitis after use of drug ointment patch test with 20% diethyl sebacate in ethanol Schneider 1980 24 yr old female swelling and erythema with 3 mos of use of a cream; diethyl patch test results were positive to the cream containing diethyl sebacate and negative to the Sasaki et al 1997 sebacate was in the vehicle lotion that did not; patch testing with 30 add'l subjects was negative 28 yr old female erythema developed with use of an ointment; did not subside when positive parch test to 1st, but not 2nd ointment; upon patch testing with a panel of items, Kimura and Kawasa a new ointment was used including diethyl sebacate, only diethyl sebacate had a positive response at day 2, as 10% in 1999 pet, and on day 3, with 1 and d10 % in pet; further patch testing was with diethyl sebacate was positive for this patient but negative for others 39 yr old male 1 mo history of tinea cruris after use of a cream; he developed patch testing with the cream in 5% diethyl sebacate was positive Tanaka et al 2000 contact dermatitis 60 yr old male developed pruritic eczematoid eruptions after 1 yr use of a cream positive patch test with 5% diethyl sebacate in petrolatum, to the cream, and to cetyl alcohol Soga et al 2004 49 yr -old female contact dermatitis to a hand cream positive patch test t cream; further testing revealed only diethyl sebacate gave a positive Narita et al 2006 Diisopropyl Sebacate 22 yr old male reaction after 2 mos use of a cream containing 27% diisopropyl positive patch testing to diisopropyl sebacate, alone and in combination, was reported; an open Dooms-Goossen et al sebacate test on the forearm with diisopropyl sebacate produced red papules at 48 h 1996

CIR Diethylhexyl Sebacate patient was sensitized to 3 other sebacate esters 10% in petrolatum was not irritating deGroot et al 1991 Panel Esterase Metabolites Stearyl Alcohol (metabolite of distearyl succinate)

Book 3 individuals 2 had urticarial-type reactions 1 reaction was thought due to impurities in stearyl alcohol Elder 1985 Page 172 Data PersonalCare ProductsCouncil Committedto Safety, Quality& Innovation Memorandum

TO: F. Alan Andersen, Ph.D. Director - COSMETIC INGREDIENT REVIEW (CR)

FROM: John Bailey, Ph.D. Industry Liaison to the CR Expert Panel

DATE: February 25, 2010

SUBJECT: Concentration of Use Dicarboxylic Acid and their Salts and Esters

Updated concentration of use table on Sebacic Acid and related ingredients

Concentration of use additions to the Sebacic Acid report

11011 7th Street, N.W., Suite 300 Washington, D.C. 20036-4702 202.331.1770 202.331.1969 (fax) www.personalcarecouncil.org

CIR Panel Book Page 173 Concentration of Use - Additions to the Sebacic Acid and Related Ingredients Report Malonic Acid, Diethyl Malonate, Succinic Acid, Decyl Succinate, Dicapryl Succinate, Dicetearyl Succinate, Diethyihexyl Succinate, Diethyl Succinate, Dimethyl Succinate, Disodium Succinate, Sodium Succinate, Glutaric Acid, Diisostearyl Glutarate, Dimethyl Glutarate, Adipic Acid, Dibutyl Adipate, Di-C12-15 Alkyl Adipate, Dicapryl Adipate, Dicetyl Adipate, Diethyl Adipate, Diethyihexyl Adipate, Diheptylundecyl Adipate, Dihexyl Adipate, Dihexyldecyl Adipate, Dilsobutyl Adipate, Diisocetyl Adipate, Dhlsodecyl Adipate, Diisononyl Adipate, Dihsooctyl Adipate, Dhlsopropyl Adipate, Diisostearyl Adipate, Dimethyl Adipate, Dioctyldodecyl Adipate, Dipropyl Adipate and Ditridecyl Adipate*

Ingredient Product Category Concentration of Use

Diethyl Malonate Skin cleansing (cold creams, cleansing 0.0 1% lotions, liquids and pads)

Diethyl Malonate Night creams, lotions and powders 0.02% Diethyl Malonate Other skin care products 0.004%

Succinic Acid Bath oils, tablets and salts 26%

Succinic Acid Hair conditioners 0.2% Succinic Acid Hair straighteners 0.001%

Succinic Acid Rinses (noncoloring) 0.01% Succinic Acid Foundations 0.01% Succinic Acid Makeup bases 0.01% Succinic Acid Bath soaps and detergents 0.2%

Succinic Acid Face and neck creams, lotions and powders 0.2% Succinic Acid Moisturizing creams, lotions and powders 0.1%

Diethyihexyl Succinate Colognes and toilet waters 1%

Diethyihexyl Succinate Hair conditioners 5% Diethyihexyl Succinate Hair sprays (aerosol fixatives 0.02-0.3%

Diethyihexyl Succinate Permanent waves 3%

Diethylhexyl Succinate Tonics, dressings and other hair grooming 4% aids

Page 1 of 4

CIR Panel Book Page 174 Diethyihexyl Succinate Face powders 3%

Diethyihexyl Succinate Foundations 3%

Diethyihexyl Succinate Lipstick 3%

Diethyihexyl Succinate Preshave lotions (all types) 4%

Diethyihexyl Succinate Face and neck creams, lotions and powders 4-5%

Diethylhexyl Succinate Body and hand creams, lotions and 4-6% powders

Diethylhexyl Succinate Moisturizing creams, lotions and powders 3%

Diethylhexyl Succinate Other skincare preparations 4%

Diethylhexyl Succinate Suntan gels, creams and liquids 3-4%

Diethylhexyl Succinate Indoor tanning preparations 4%

Dimethyl Succinate Nail polish and enamel removers 0.2%

Dimethyl Succinate Skin cleansing (cold creams, cleansing 5% lotions, liquids and pads)

Dimethyl Succinate Eye lotion 0.002% Disodium Succinate Foundations 0.0005% Disodium Succinate Skin cleansing (cold creams, cleansing 0.0005% lotions liquids and pads)

Disodium Succinate Face and neck creams, lotions and powders 0.0 1-0.4%

Disodium Succinate Night creams, lotions and powders 0.02% Disodium Succinate Skin fresheners 0.0005%

Dimethyl Glutarate Nail polish and enamel removers 0.5%

Dimethyl Glutarate Skin cleansing (cold creams, cleansing 15% lotions, liquids and pads)

Adipic Acid Bath oils, tablets and salts 15-18% Adipic Acid Mascara 0.000001%

Adipic Acid Permanent waves 0.5% Adipic Acid Lipstick 0.000001%

Page 2 of 4

CIR Panel Book Page 175 Diethyihexyl Adipate Blushers (all types) 14%

Diethylhexyl Adipate Foundations 0.9%

Diethylhexyl Adipate Makeup bases 1%

Diethylhexyl Adipate Nail extenders 3%

Diethyihexyl Adipate Nail polish and enamel 2%

Diethyihexyl Adipate Deodorants (underarm) 0.9%

Diethyihexyl Adipate Aftershave lotions 4%

Diethylhexyl Adipate Shaving cream (aerosol, brushless and 0.6% lather)

Diethylhexyl Adipate Face and neck creams, lotions and powders 3%

Diheptylundecyl Adipate Moisturizing creams, lotions and powders 6%

Dihexyl Adipate Eye makeup remover 3%

Diisobutyl Adipate Bath oils, tablets and salts 0.5%

Diisobutyl Adipate Colognes and toilet waters 0.6%

Diisobutyl Adipate Perfumes 3%

Diisobutyl Adipate Hair conditioners 0.2% Diisobutyl Adipate Hair sprays (aerosol fixatives) 0.05-0.2%

Diisobutyl Adipate Tonics, dressings and other hair grooming 0.2% aids

Diisobutyl Adipate Basecoats and undercoats (manicuring 0.001-0.7% preparations)

Diisobutyl Adipate Cuticle softeners 0.5% Diisobutyl Adipate Nail polish and enamel 0.009-0.7%

Diisobutyl Adipate Bath soaps and detergents 0.009%

Diisobutyl Adipate Face and neck creams, lotions and powders 0.003%

Diisobutyl Adipate Body and hand creams, lotions and 0.8% powders

Diisobutyl Adipate Moisturizing creams, lotions and powders 0.03%

Page 3 of 4

CIR Panel Book Page 176 Diisobutyl Adipate Paste masks (mud packs) 0.002%

Diisobutyl Adipate Suntan gels, creams and liquids 0.003%

Diisopropyl Adipate Bubble baths 2%

Diisopropyl Adipate Mascara 1%

Diisopropyl Adipate Perfumes 0.005-8% Diisopropyl Adipate Other fragrance preparations 0.07-7%

Diisopropyl Adipate Tonics, dressings and other hair grooming 0.5-3% aids

Diisopropyl Adipate Blushers (all types) 0.02% Diisopropyl Adipate Foundations 0.02% Diisopropyl Adipate Aftershave lotions 0.009%

Diisopropyl Adipate Skin cleansing (cold creams, cleansing 7% lotions, liquids and pads)

Diisopropyl Adipate Face and neck creams, lotions and powders 3%

Diisopropyl Adipate Body and hand creams, lotions and 2% powders

Diisopropyl Adipate Body and hand sprays 2%

Diisopropyl Adipate Skin fresheners 1%

Diisostearyl Adipate Lipstick 10%

Diisostearyl Adipate Skin cleansing (cold creams, cleansing 3% lotions, liquids and pads)

Dimethyl Adipate Nail polish and enamel removers 0.2% *Ingredients found in the title of the table but not found in the table were included in the concentration of use survey, but no uses were reported.

Information collected in 2010 Table prepared February 24, 2010

Page 4 of 4

CIR Panel Book Page 177

Confidential data removed PersonalCare ProductsCouncil Committedto Safety, Quality& Innovation Memorandum

TO: F. Alan Andersen, Ph.D. Director - COSMETIC INGREDIENT REVIEW (CIR)

FROM: John Bailey, Ph.D. Industry Liaison to the CIR Expert Panel

DATE: January 15, 2010 (revised summaries submitted July 26, 2010)

SUBJECT: Unpublished data on Diisostearyl Adipate, Diisocetyl Dodecandedioate and Dioctyldodecyl Dodecanedioate

For each ingredient there is a toxicological summary sheet (signed by Lisa Bouldin), followed by the studies on that compound. Please note that the AMA Laboratories, Inc. (1996) HRIPT includes all three compounds, and that a copy of this study is included following each summary sheet.

Bouldin L. Toxicological summary Liquiwax DISA (Diisostearyl Adipate)

AMA Laboratories, Inc. 1996. 50 human subject repeat insult patch test skin irritation/sensitization evaluation of several substances (including Liquiwax DISA [Diisostearyl Adipate]).

Bouldin L. Toxicological summary Liquiwax DICDD (Diisocetyl Dodecanedioate)

AMA Laboratories, Inc. 1996. 50 human subject repeat insult patch test skin irritation/sensitization evaluation of several substances (including Liquiwax DICDD [Diisocetyl Dodecanedioate]).

AMA Laboratories, Inc. 1991. Acute oral toxicity Liquiwax DICDD (Diisocetyl Dodecanedioate).

AMA Laboratories, Inc. 1990. Primary skin irritation Liquiwax DICDD (Diisocetyl Dodecanedioate).

AMA Laboratories, Inc. 1990. Primary eye irritation Liquiwax DICDD (Diisocetyl Dodecanedioate).

Bouldin L. Toxicological summary Liquiwax DIADD (Dioctyldodecyl Dodecanedioate)

AMA Laboratories, Inc. 1996. 50 human subject repeat insult patch test skin irritationlsensitization evaluation of several substances (including Liquiwax DIADD [Dioctyldodecyl Dodecanedioate]).

1101 N.W., Suite 3OO D.C. (fax) 17th Street, Washington, 20036-4702 202.331.1770 202.331.1969 www.personalcarecouncil.org I CIR Panel Book Page 197 AMA Laboratories, Inc. 1990. Acute oral toxicity Liquiwax DIADD (Dioctyldodecyl Dodecanedioate).

AMA Laboratories, Inc. 1990. Primary skin irritation Liquiwax DLkDD (Dioctyldodecyl Dodecanedioate).

AMA Laboratories, Inc. 1990. Primary eye irritation Liquiwax DJADD (Dioctyldodecyl Dodecandioate).

AMA Laboratories, Inc. 1993. Guinea pig maximization test Liquiwax DIADD (Dioctyldodecyl Dodecanedioate).

2

CIR Panel Book Page 198

CIR Panel Book Page 199

CIR Panel Book Page 200

CIR Panel Book Page 201

CIR Panel Book Page 202

CIR Panel Book Page 203

CIR Panel Book Page 204

CIR Panel Book Page 205

CIR Panel Book Page 206

CIR Panel Book Page 207

CIR Panel Book Page 208

CIR Panel Book Page 209

CIR Panel Book Page 210

CIR Panel Book Page 211

CIR Panel Book Page 212

CIR Panel Book Page 213

CIR Panel Book Page 214

CIR Panel Book Page 215

CIR Panel Book Page 216

CIR Panel Book Page 217

CIR Panel Book Page 218

CIR Panel Book Page 219

CIR Panel Book Page 220

CIR Panel Book Page 221

CIR Panel Book Page 222

CIR Panel Book Page 223

CIR Panel Book Page 224

CIR Panel Book Page 225

CIR Panel Book Page 226

CIR Panel Book Page 227

CIR Panel Book Page 228

CIR Panel Book Page 229

CIR Panel Book Page 230

CIR Panel Book Page 231

CIR Panel Book Page 232

CIR Panel Book Page 233

CIR Panel Book Page 234

CIR Panel Book Page 235

CIR Panel Book Page 236

CIR Panel Book Page 237

CIR Panel Book Page 238

CIR Panel Book Page 239

CIR Panel Book Page 240

CIR Panel Book Page 241

CIR Panel Book Page 242

CIR Panel Book Page 243

CIR Panel Book Page 244

CIR Panel Book Page 245

CIR Panel Book Page 246

CIR Panel Book Page 247

CIR Panel Book Page 248

CIR Panel Book Page 249

CIR Panel Book Page 250

CIR Panel Book Page 251

CIR Panel Book Page 252

CIR Panel Book Page 253

CIR Panel Book Page 254

CIR Panel Book Page 255

CIR Panel Book Page 256

CIR Panel Book Page 257

CIR Panel Book Page 258

CIR Panel Book Page 259 Persona Care ProductsCounci Committedto Safety, Quality& Innovation Memorandum

TO: F. Alan Andersen, Ph.D. Director - COSMETIC INGREDIENT REVIEW (CW)

FROM: John I}(O Industry Liaison to the CIR Expert Panel

DATE: February 15, 2010

SUBJECT: Physical Chemical Properties Sebacates

Hokoku Corporation. 2010. Physical/Chemical Properties Dibutyl Octyl Sebacate, Dihexyldecyl Sebacate and Dioctyldodecyl Sebacate.

Another supplier reports that the expected impurities in Diisopropyl Sebacate are the starting materials Sebacic Acid <0.3% and Isopropyl Alcohol <0.2%.

11011 7th Street, N.W., Suite 300 Washington, D.C. 20036-4702 202.331.1770 202.331.1969 (fax) www.personalcarecouncfl.org

CIR Panel Book Page 260 February 6, 2010. HOKOKU Corporation Physical/Chemical Properties

Chemical name IDibutyloctyl SebacatelDihexyldecyl SebacatelDioctyldodecyl Sebacate Phvsica oronerteis Density ( 20 C) g/ml 0. 90 0. 89 0. 87 Viscosity( 25 C) mm2/s 41 58 83 Flash point (C.0.C) C 236 260 270 Pour point C —62. 5 —42. 5 17. 5 Octanol/Water No data No data No data oart it ion Water solubility Insoluble Insoluble Insoluble

CIR Chemical properties

Panel Acid Value mgKOH/g 0. 1 0. 1 0. 1 Saponification Value mgKOH/g 209 173 147

Book Hydoroxyl Value mgKOH/g 0. 5 1. 0 2. 0 Iodine Value 12g/lOOg 0. 1 0. 4 0. 4 Page Irritation Data No data No data No data 261 SEBACIC ACID 11A - Aftershave Lotion 1 SEBACIC ACID 12A - Cleansing 3 SEBACIC ACID 12D - Body and Hand (exc shave) 1 SEBACIC ACID 12F - Moisturizing 7

DIETHYLHEXYL SEBACATE 03F - Mascara 4 DIETHYLHEXYL SEBACATE 05I - Other Hair Preparations 2 DIETHYLHEXYL SEBACATE 11A - Aftershave Lotion 7

DIMETHYL SEBACATE 10B - Deodorants (underarm) 1 ??

DIISOPROPYL SEBACATE 03D - Eye Lotion 1 DIISOPROPYL SEBACATE 04A - Cologne and Toilet waters 1 DIISOPROPYL SEBACATE 05A - Hair Conditioner 1 DIISOPROPYL SEBACATE 05G - Tonics, Dressings, and Other Hair Grooming5 Aids DIISOPROPYL SEBACATE 07I - Other Makeup Preparations 1 DIISOPROPYL SEBACATE 08A - Basecoats and Undercoats 1 DIISOPROPYL SEBACATE 10B - Deodorants (underarm) 4 DIISOPROPYL SEBACATE 11A - Aftershave Lotion 1 DIISOPROPYL SEBACATE 12C - Face and Neck (exc shave) 3 DIISOPROPYL SEBACATE 12D - Body and Hand (exc shave) 2 DIISOPROPYL SEBACATE 12F - Moisturizing 3 DIISOPROPYL SEBACATE 12J - Other Skin Care Preps 2 DIISOPROPYL SEBACATE 13B - Indoor Tanning Preparations 5

DIMETHYL SEBACATE 10B - Deodorants (underarm) 1 ??

SUCCINIC ACID 05C - Hair Straighteners 2 SUCCINIC ACID 12F - Moisturizing 2

SODIUM SUCCINATE 02B - Bubble Baths 1 SODIUM SUCCINATE 05F - Shampoos (non-coloring) 2 SODIUM SUCCINATE 10A - Bath Soaps and Detergents 1 SODIUM SUCCINATE 12F - Moisturizing 3

DISODIUM SUCCINATE 03D - Eye Lotion 2 DISODIUM SUCCINATE 03G - Other Eye Makeup Preparations 2 DISODIUM SUCCINATE 05A - Hair Conditioner 1 DISODIUM SUCCINATE 05E - Rinses (non-coloring) 1 DISODIUM SUCCINATE 05F - Shampoos (non-coloring) 1 DISODIUM SUCCINATE 05I - Other Hair Preparations 2 DISODIUM SUCCINATE 07C - Foundations 1 DISODIUM SUCCINATE 07F - Makeup Bases 1 DISODIUM SUCCINATE 12A - Cleansing 4 DISODIUM SUCCINATE 12C - Face and Neck (exc shave) 9 DISODIUM SUCCINATE 12D - Body and Hand (exc shave) 2 DISODIUM SUCCINATE 12F - Moisturizing 9 DISODIUM SUCCINATE 12G - Night 5 DISODIUM SUCCINATE 12I - Skin Fresheners 1 DISODIUM SUCCINATE 12J - Other Skin Care Preps 4

DIMETHYL SUCCINATE 08F - Nail Polish and Enamel Removers 12

DIETHYLHEXYL SUCCINATE 03D - Eye Lotion 1 DIETHYLHEXYL SUCCINATE 05A - Hair Conditioner 1 DIETHYLHEXYL SUCCINATE 05G - Tonics, Dressings, and Other Hair Grooming2 Aids

CIR Panel Book Page 262 DIETHYLHEXYL SUCCINATE 05I - Other Hair Preparations 1 DIETHYLHEXYL SUCCINATE 07A - Blushers (all types) 1 DIETHYLHEXYL SUCCINATE 07C - Foundations 1 DIETHYLHEXYL SUCCINATE 10E - Other Personal Cleanliness Products1 DIETHYLHEXYL SUCCINATE 11A - Aftershave Lotion 1 DIETHYLHEXYL SUCCINATE 12A - Cleansing 2 DIETHYLHEXYL SUCCINATE 12C - Face and Neck (exc shave) 2 DIETHYLHEXYL SUCCINATE 12D - Body and Hand (exc shave) 9 DIETHYLHEXYL SUCCINATE 12F - Moisturizing 8 DIETHYLHEXYL SUCCINATE 12G - Night 2 DIETHYLHEXYL SUCCINATE 12J - Other Skin Care Preps 3 DIETHYLHEXYL SUCCINATE 13B - Indoor Tanning Preparations 3

DIOCTYL SUCCINATE 05B - Hair Spray (aerosol fixatives) 1 actually Dicapryl DIOCTYL SUCCINATE 12C - Face and Neck (exc shave) 2 DIOCTYL SUCCINATE 12D - Body and Hand (exc shave) 1 DIOCTYL SUCCINATE 12F - Moisturizing 3 DIOCTYL SUCCINATE 12G - Night 1 DIOCTYL SUCCINATE 13C - Other Suntan Preparations 1

DIMETHYL GLUTARATE 08F - Nail Polish and Enamel Removers 13

ADIPIC ACID 02A - Bath Oils, Tablets, and Salts 1 ADIPIC ACID 05A - Hair Conditioner 8 ADIPIC ACID 05D - Permanent Waves 6 ADIPIC ACID 05F - Shampoos (non-coloring) 8 ADIPIC ACID 05G - Tonics, Dressings, and Other Hair Grooming2 Aids

DIMETHYL ADIPATE 08F - Nail Polish and Enamel Removers 12

DIETHYLHEXYL ADIPATE 01B - Baby Lotions, Oils, Powders, and Creams1 DIETHYLHEXYL ADIPATE 03C - Eye Shadow 1 DIETHYLHEXYL ADIPATE 03D - Eye Lotion 1 DIETHYLHEXYL ADIPATE 03G - Other Eye Makeup Preparations 1 DIETHYLHEXYL ADIPATE 04E - Other Fragrance Preparation 5 DIETHYLHEXYL ADIPATE 05G - Tonics, Dressings, and Other Hair Grooming1 Aids DIETHYLHEXYL ADIPATE 07A - Blushers (all types) 1 DIETHYLHEXYL ADIPATE 07C - Foundations 5 DIETHYLHEXYL ADIPATE 07E - Lipstick 1 DIETHYLHEXYL ADIPATE 07F - Makeup Bases 3 DIETHYLHEXYL ADIPATE 07I - Other Makeup Preparations 1 DIETHYLHEXYL ADIPATE 08A - Basecoats and Undercoats 1 DIETHYLHEXYL ADIPATE 08E - Nail Polish and Enamel 3 DIETHYLHEXYL ADIPATE 10E - Other Personal Cleanliness Products1 DIETHYLHEXYL ADIPATE 11E - Shaving Cream 4 DIETHYLHEXYL ADIPATE 12A - Cleansing 2 DIETHYLHEXYL ADIPATE 12C - Face and Neck (exc shave) 6 DIETHYLHEXYL ADIPATE 12D - Body and Hand (exc shave) 1 DIETHYLHEXYL ADIPATE 12F - Moisturizing 2 DIETHYLHEXYL ADIPATE 12G - Night 2 DIETHYLHEXYL ADIPATE 12H - Paste Masks (mud packs) 2 DIETHYLHEXYL ADIPATE 12J - Other Skin Care Preps 2 DIETHYLHEXYL ADIPATE 13A - Suntan Gels, Creams, and Liquids 1

DIBUTYL ADIPATE 03F - Mascara 2 DIBUTYL ADIPATE 04E - Other Fragrance Preparation 2 DIBUTYL ADIPATE 08E - Nail Polish and Enamel 1 DIBUTYL ADIPATE 12F - Moisturizing 1

CIR Panel Book Page 263 DICAPRYL ADIPATE 02A - Bath Oils, Tablets, and Salts 5 DICAPRYL ADIPATE 04E - Other Fragrance Preparation 1 DICAPRYL ADIPATE 10B - Deodorants (underarm) 30 DICAPRYL ADIPATE 12D - Body and Hand (exc shave) 7

DIHEXYL ADIPATE 12D - Body and Hand (exc shave) 1

DIISOBUTYL ADIPATE 05B - Hair Spray (aerosol fixatives) 5 DIISOBUTYL ADIPATE 08A - Basecoats and Undercoats 2 DIISOBUTYL ADIPATE 08E - Nail Polish and Enamel 7 DIISOBUTYL ADIPATE 11A - Aftershave Lotion 2 DIISOBUTYL ADIPATE 12D - Body and Hand (exc shave) 5 DIISOBUTYL ADIPATE 12F - Moisturizing 1

DIISODECYL ADIPATE 12G - Night 1

DIISOPROPYL ADIPATE 02A - Bath Oils, Tablets, and Salts 1 DIISOPROPYL ADIPATE 02D - Other Bath Preparations 1 DIISOPROPYL ADIPATE 03F - Mascara 2 DIISOPROPYL ADIPATE 04A - Cologne and Toilet waters 8 DIISOPROPYL ADIPATE 04B - Perfumes 6 DIISOPROPYL ADIPATE 04E - Other Fragrance Preparation 7 DIISOPROPYL ADIPATE 05G - Tonics, Dressings, and Other Hair 10Grooming Aids DIISOPROPYL ADIPATE 05H - Wave Sets 2 DIISOPROPYL ADIPATE 05I - Other Hair Preparations 5 DIISOPROPYL ADIPATE 07E - Lipstick 1 DIISOPROPYL ADIPATE 08F - Nail Polish and Enamel Removers 1 DIISOPROPYL ADIPATE 10B - Deodorants (underarm) 6 DIISOPROPYL ADIPATE 11A - Aftershave Lotion 11 DIISOPROPYL ADIPATE 12A - Cleansing 1 DIISOPROPYL ADIPATE 12D - Body and Hand (exc shave) 2 DIISOPROPYL ADIPATE 12F - Moisturizing 2 DIISOPROPYL ADIPATE 12J - Other Skin Care Preps 2 DIISOPROPYL ADIPATE 13A - Suntan Gels, Creams, and Liquids 1 DIISOPROPYL ADIPATE 13C - Other Suntan Preparations 1

DIISOSTEARYL ADIPATE 07E - Lipstick 4 DIISOSTEARYL ADIPATE 12A - Cleansing 2

DIOCTYLDODECYL ADIPATE 07E - Lipstick 3

DI-N-OCTYL ADIPATE 07C - Foundations 2 ??? DI-N-OCTYL ADIPATE 10E - Other Personal Cleanliness Products1

OCTYL ADIPATE 12C - Face and Neck (exc shave) 1 ??

AZELAIC ACID 11G - Other Shaving Preparation Products1 AZELAIC ACID 12C - Face and Neck (exc shave) 6 AZELAIC ACID 12H - Paste Masks (mud packs) 1 AZELAIC ACID 12J - Other Skin Care Preps 1

DIISOCETYL DODECANEDIOATE07I - Other Makeup Preparations 1 DIISOCETYL DODECANEDIOATE13B - Indoor Tanning Preparations 1

CIR Panel Book Page 264 DIOCTYLDODECYL DODECANEDIOATE04E - Other Fragrance Preparation 2 DIOCTYLDODECYL DODECANEDIOATE07E - Lipstick 1 DIOCTYLDODECYL DODECANEDIOATE12D - Body and Hand (exc shave) 2

CIR Panel Book Page 265 JOURNAL OF THE AMERICAN COLLEGE OF TOXICOLOGY Volume 3, Number 3, 19&Q Mary Ann Liebert, Inc., Publishers

3

Final Report on the Safety Assessment of Dioctyl Adipate and Diisopropyl Adipate

Dioctyl Adipate, the diester of octyl alcohol and adipic acid, and Diisopropyl Adipate, the diester of isopropyl alcohol, are used in cosmetics as emollients and bases. These two ingredients have a low acute oral and percutaneous tox- icity. Undiluted Dioctyl Adipate and Diisopropyl Adipate were, at most, only very mild, transient eye irritants. Primary dermal irritation tests indicated that Dioctyl Adipate was a very mild irritant and Diisopropyl Adipate was minimally irritating. Dioctyl Adipate was not a skin sensitizer in guinea pigs. An Ames test for the mutagenic potential of Dioctyl Adipate was negative. An assay of the carcinogenic potential of Dioctyl Adipate produced no un- toward effects and was noncarcinogenic to rats. Mice studies indicated a dose- related body weight reduction and a higher incidence of hepatocellular adenoma and carcinoma than controls. In a lifetime study Dioctyl Adipate caused no skin tumors when 10 mg was applied weekly to the back skin of mice. The teratogenicity potential of Dioctyl Adipate is reviewed. Clinical assessment of Dioctyl Adipate in formulations showed, at most, minimal erythema and papules when applied under occlusion. No UV sen- sitization occurred. Undiluted Diisopropyl Adipate produced no irritation in 24 h patch tests, but was moderately irritating in a 21-day cumulative irritancy test. Formulations containing up to 20°h Diisopropyl Adipate caused minimal to mild irritation, no sensitization and no photosensitization. On the basis of available data, it is concluded that Dioctyl Adipate and Diisopropyl Adipate are safe as presently used in cosmetics.

INTRODUCTION

ioctyl Adipate and Diisopropyl Adipate are common plasticizers and emol- D lient esters. In cosmetics they are used as emollients and as the base of many different types of products.“-3)

CHEMICAL AND PHYSICAL PROPERTIES

Structure

1. Dioctyl Adipate is the diester of octyl alcohol and adipic acid. It conforms generally to the formula: 101

CIR Panel Book Page 266 102 COSMETIC INGREDIENT REVIEW

0 II y2H5 HgCa-CH-CH2-0-C-(CH2)4-C-0-CH2-CH-C4Hg

CAS Number: 103-23-l Synonyms include: Di-(2-Ethylhexyl)Adipate and Wickenol 1 58.(3)

2. Diispropyl Adipate is the diester of isopropyl alcohol and adipic acid. It conforms generally to the formula:

0 0 II II (CHj)2CH-O-C-(CH2),-C-O-CH(CH3)2

CAS Number 6938-94-9 Other names include: Beta DIA Ceraphyl 230 Crodamol DA I so-Ad i pate 2/043 700 Prod i pate Schercemol DIA Standamul DIPA Tegester 504-D Wickenol 11 6.(3’

Production

Dioctyl Adipate is produced by the reaction of adipic acid and 2-ethylhexanol in the presence of an esterification catalyst such as sulfuric acid, p-toluenesulfonic acid or a proprietary catalyst. Purification of the reaction product includes re- moval of the catalyst, alkali refining and stripping. (4.5) Diisopropyl Adipate is pro- duced by esterification of adipic acid with an excess of isopropanol. The excess alcohol is removed by vacuum stripping and the ester is then alkali-refined and filtered.(6)

Properties Dioctyl Adipate and Diisopropyl Adipate are clear, colorless to light yellow viscous liquids with an aromatic odor. They are soluble in most organic solvents and insoluble in water. For other properties, see Table 1.

Analytical Methods Diisopropyl Adipate and Dioctyl Adipate can be identified through standard Infrared (IR) spectroscopy. (‘I Gas-liquid chromatography, liquid-liquid extrac- tion, mass spectrometry, and high-pressure liquid chromatography are also methods of analysis for the Adipates.“-lo)

Reactivity/Stability Dioctyl Adipate and Diisopropyl Adipate are considered stable; however,

CIR Panel Book Page 267 ASSESSMENT: DIOCTYL ADIPATE AND DIISOPROPYL ADIPATE 103

TABLE 1. Properties of Dioctyl and Diisopropyl Adipate.

Dioctyl Diisopropyl Property Adipate Ref. Adipate Ref.

Form Oily liquid 2 Oily liquid 7 Color Colorless to 2,4 Colorless, clear 7 light yellow Boiling point (“C) 417 2 - - Specific gravity 0.9268 2 0.950 to 0.965 7 (20/20%) Flash point, T, (“F) 204.4 (400) 11 - - Melting point (“C) -67.8 4 - - Molecular weight 370.56 11 230.34 4 Refractive index at 20°C 1.4474 11 1.4200-l .4245 7 Acid value 1 .O (max.) 5 2.0 (max.) 7 Saponification value 298-308 5 465-500 7 Iodine value 0.5 (max.) 5 1 .O (max.) 5 Viscosity (20°C) 13.7 cps - - Vapor pressure 0 200% 2.4 mm Hg - - - Soluble in: Alcohol 4 - - Ether Acetone Acetic acid Most organic solvents Insoluble in: Water 2,4 - - Glycerin and glycols

hydrolysis of the ester groupings may occur in the presence of aqueous acids or bases.‘5a6)

Impurities

No known minor impurities occur in either Dioctyl Adipate or Diisopropyl Adipate, although the acid values imply the presence of adipic acid or of the monoester in both.(5*6)

USE

Non-Cosmetic Uses

The Adipates are used primarily as plasticizers in food wraps, vinyl blood bags and hemodialysis bags. Adipates are also used as solvents and aircraft Iubes.(‘.2.4,‘2.13) Dioctyl Adipate has Indirect Food Additive (IFA) Status for use in food wrapping.(14)

Cosmetic Use The Adipates are used as components of cosmetic bases and as solvents and emollients in other cream-type skin preparations. (I) They are also used to modify the tactile and flow properties of emollient blends, especially in bath products.(‘5) The cosmetic product formulation computer printout which is made available by the Food and Drug Administration (FDA) is compiled through volun- tary filing of such data in accordance with Title 21 part 720.4 of the Code of

CIR Panel Book Page 268 104 COSMETIC INGREDIENT REVIEW

Federal Regulations. (14) Ingredients are listed in prescribed concentration ranges under specific product type categories. Since certain cosmetic ingredients are supplied by the manufacturer at less than 100% concentration, the value re- ported by the cosmetic formulator may not necessarily reflect the true, actual concentration found in the finished product; the actual concentration in such a case would be a fraction of that reported to the FDA. Since data are submitted only within the framework of preset concentration ranges; this presents the oppor- tunity for overestimation of the actual concentration of an ingredient in a par- ticular product. An entry at the lowest end of a concentration range is considered the same as one entered at the highest end of that range, thus, introducing the possibility of a two- to lo-fold error in the assumed ingredient concentration. According to the industry’s 1981 submission of product formulation data to the FDA, Dioctyl Adipate was used in 27 products in concentrations of I 0.1 Oh-1 % in some facial makeup, and up to lo%-25% in bath preparations(16) (see Table 2). Diisopropyl Adipate was used in 112 cosmetic formulations according to the 1981 FDA product formulation data. Its concentrations of use ranged from less than 0.1% up to 25%‘16) (see Table 2).

Surfaces to Which Commonly Applied and Frequency of Application

Dioctyl and Diisopropyl Adipates are found in cosmetics which may come in contact with the skin of the face, hands, and the general body surface, the mucous membranes, nails, scalp, and hair. Thus, cosmetics containing Adipates may be applied to the body once every few days to several times daily(16) (see Table 2).

TABLE 2. Product Formulation Data.a

No. of product formulations within each concentration Total no. range (% j b containing Product categoryb ingredient >JO-25 >5-10 >J-5 >O. J-J s0.J

Dioctyl Adipate Bath oils, tablets, and salts 4 - - - - Colognes and toilet waters - - 6 - - Blushers (all types) - - - - 1 Makeup foundations - 2 1 1 - Lipstick - - 5 - - Other makeup preparations (not eye) 1 - - 1 - - Nail polish and enamel remover 2 - - 2 - - Deodorants (underarm) 1 - - - 1 - Aftershave lotions 1 - - 1 - - Face, body, and hand skin care preparations (excluding shaving preparations) 1 - - 1 - - Other suntan preparations 1 - - - 1 -

198 1 TOTALS 27 4 2 17 3 1

CIR Panel Book Page 269 ASSESSMENT: DIOCTYL ADIPATE AND DIISOPROPYL ADIPATE 105

TABLE 2. (Continued.)

No. of product formulations within each concentration Total no. range (%) b containing Product categoryb ingredient > JO-25 >5-JO >J-5 >O.J-7 s0.J

Diisopropyl Adipate Bath oils, tablets, and salts 7 1 1 - Bubble baths 1 - 1 - - Eyeliner 1 - 1 - - Eye shadow 1 - - - - Colognes and toilet waters 15 12 3 - Perfumes 20 4 - - Sachets 1 - - - Other fragrance preparations 9 6 1 - Hair conditioners 3 - 1 2 Hair sprays (aerosol fixatives) 1 1 - - Tonics, dressings, and other hair grooming aids - - 4 - - Wave sets - 1 1 - Blushers (all types) - 1 - - Face powders 1 - - Makeup foundations - 1 - Other personal cleanliness products 1 - - - 1 - Aftershave lotions 16 - 9 7 - Preshave lotions (all types) 1 1 - - - Skin cleansing preparations (cold creams, lotions, liquids, and pads) 5 - - - 5 - Foot powders and sprays 1 - - - 1 - Moisturizing skin care preparations 2 - - 1 1 - Night skin care preparations 1 - 1 - - - Skin fresheners 11 3 - 6 2 Other skin care preparations 2 1 1 - - Suntan gels, creams, and liquids 2 - 2 - - - Indoor tanning preparations 2 - - 2 - -

1981 TOTALS 112 11 23 46 28 4

a Data from Ref. 16. bPreset product categories and concentration ranges in accordance with federal filing regulations (21 CFR 720.4).

BIOLOGICAL PROPERTIES

General Studies

Subcellular and Enzyme Effects

Dioctyl Adipate was fed to male rats in a dietary concentration of 2% for three weeks. Effects included hepatic peroxisome proliferation, increased size of the liver, and increase in the hepatic activities of the peroxisome-associated enzymes catalase and carnitine acetyl transferase. Hypolipidemia and a decrease

CIR Panel Book Page 270 106 COSMETIC INGREDIENT REVIEW in serum lipids were also observed in Dioctyl Adipate-treated animals. The authors postulated that the active portion of the compound, in the induction of hepatic peroxisome proliferation, may be the metabolite 2-ethylhexyl alcohol.(“)

Effect on Cultured Cells

When contracting chick embryo heart cells were maintained in tissue culture and exposed to Dioctyl Adipate at a level of 1.5 pglml (4 km), the number of car- diac contracting cells was reduced to 50% of control levels.(18) In cultures of human diploid cells of the WI38 strain, the IDso (dose that inhibits cell growth to 50% of the control culture) for Dioctyl Adipate was 32 pM.‘l’)

Animal Toxicology

Acute Toxicity

Oral Dioctyl Adipate was administered by gavage to nine groups of five male and five female F344 rats at doses of 0.08, 0.16, 0.31, 0.63, 1.25, 2.5, 5.0, 10, or 20 g/kg of the substance in corn oil. Two of five males of the 10 g/kg group died and one male and one female of the 20 g/kg group died.(‘O) Five groups of five male and five female mice were given Dioctyl Adipate in corn oil in single doses of 1.25, 2.5, 5.0, 10.0, or 20.0 g/kg. Mortality observations were: one male of the 1.25 g/kg group died “accidentally,” two males of the 10.0 g/kg group, three at the 20.0 g/kg group, one female of the 20 g/kg group. The estimated LDso for male mice was 15.0 g/kg, and for females it was 24.6 g/kg.(‘O) In an acute oral toxicity study using rats, Andreeva(z1) reported a no effect dose of Dioctyl Adipate in rats of 6 g/kg. Doses greater than this resulted in cen- tral nervous system (CNS) stimulation followed by depression which lasted for the five- to seven-day observation period. The single oral toxic dose of Dioctyl Adipate for the rat over an observation period of 14 days was 9.11 g/kg.(22) A group of five male and five female albino rats was fasted overnight, in- tubated with a dose of 7.4 g/kg Dioctyl Adipate, and then observed daily for 14 days. One animal died on Day 14, but other observations were not recorded.(23) A product containing 0.175% Dioctyl Adipate was administered in a single undiluted 6.5 g/kg dose to five male and five female Harlan Wistar rats. During the seven observation days, no signs of toxicity were observed and body weight gains were normal.(24) A face cream containing 0.7% Diisopropyl Adipate was intubated into groups of five male and five female Wistar rats per dose concentration. Animals were observed for 14 days; one male rat of the highest dose group (76.8 g/kg of the formulation) died on Day 4 of observation. At necropsy of this animal, the findings included urinary staining of the abdomen, prominent serosal blood vessels in the stomach, cecum and intestines, and red fluid in the intestines. No other deaths or abnormal findings were reported.‘25) A perfume containing 1.08% Diisopropyl Adipate was administered in a single 5 g/kg dose of the preparation to five male and five female Sprague- Dawley rats. The animals were observed for 14 days; one female died on Day 2.

CIR Panel Book Page 271 ASSESSMENT: DIOCTYLADIPATE AND DIISOPROPYLADIPATE 107

Necropsy findings of this animal were dark and mottled lungs and liver, reddened pylorus, and gas-filled GI tract. Other surviving animals showed signs of decreased activity, ataxia, diarrhea, gasping, and urinary incontinence.(26) In a similar study a perfume. containing 1.08% Diisopropyl Adipate was studied. Five male and five female Sprague-Dawley rats were given by oral in- tubation a single 5 g/kg dose of the formulation; animals were observed for 14 days. No animals died, but three males and five females had decreased activity and ataxia.r2” A product containing 5% Diisopropyl Adipate was administered as a single 5 g/kg dose of the formulation by intubation to five female albino rats; the animals were observed for seven days. No deaths or abnormal behavior were observed.‘2*) A dose of 15 g/kg of a product containing 20.75% Diisopropyl Adipate was administered orally by stomach tube to five female rats. After seven observation days, no deaths or abnormal responses were observed and the LDso was > 15 g/kg dose’29) (see Table 3).

74-day oral study A 14-day repeated dose study of Dioctyl Adipate was conducted using six groups of five male and five female F344 rats and of the same number of B6C3Fl mice. Dosages of 0 (control), 3,100, 6,300, 12,500, 25,000, and 50,000 ppm in the diet were fed to male rats and mice for 14 days. Female rats and mice were fed 0 (control), 6,300, 12,500, 25,000, 50,000, and 100,000 ppm Dioctyl Adipate in the diet. Weight gain was depressed in male rats fed 50,000 ppm and in female rats fed 25,000 ppm or more. Females fed 100,000 ppm lost weight and one died. All female mice receiving 100,000 ppm died, and males at 50,000 ppm and females at 25,000 or more lost weight.‘20’

Intravenous The intravenous LDso of Dioctyl Adipate to rats and rabbits was 900 mglkg and 540 mglkg, respectively.‘30) The acute intravenous LDso of Diisopropyl Adipate to rats was 640 mg/kg.‘30)

Percutaneous The acute dermal toxicity of Dioctyl Adipate was tested using eight albino rabbits. The trunk of each animal was clipped of all hair and half of the rabbits received longitudinal epidermal abrasions over the clipped area. The rabbits were immobilized and plastic sleeves were slipped over the shaved areas. The animals were placed into groups of two each and received doses of 0 (control), 3.6, 5.6, and 8.7 g/kg of pure Dioctyl Adipate under the sleeve. After 24 h, the sleeves were removed, the volume of unabsorbed material was cleaned from each animal and measured, and skin reactions were evaluated. The animals were observed for signs of toxicity for two weeks. Daily observation included body weights, food consumption, and behavior. Urinalysis, hematologic features, and skin changes were also observed and skin changes were rated according to stan- dard Draize scores. The animals had only slight erythema which increased in duration with increasing concentration. However, all irritation disappeared several days before the end of the observation period. Weight gain, feed con- sumption, urine and hematologic values, as well as behavior were normal in all animals. Dioctyl Adipate produced mild irritation, but no systemic toxic ef- fects. (23)

CIR Panel Book Page 272 - TABLE 3. Acute Oral Toxicity.

Species and No. Ing. Cont. Observation Ingredient of Animals 66) Dose/kg L Dsdkg period Comments Ref.

Dioctyl Adipate Ingredient F344 rats - 0.08-20.0 g 45.0 g for 14 days 2 of 5 males died in the 10 g/kg dose 20 45 male, male9 group; 1 of 5 males and 1 of 45 female 26.0 g for 5 females died in the 20 g/kg dose female? group. B6C3Fl mice - 1.25-20.0 g 15.0 g for 15 days 2 of 5 males died in the 10 g/kg dose 20 25 male, male9 group; 3 males and 1 of 5 females 25 female 24.6 g for died in the 20 g/kg group. female9 CIR rats - 6g - 7 days 6 g/kg = “No effect level.” Greater 21 doses cause CNS disturbance. Panel 5 male rats - - 9.11 g 14 days - 22 (7.28-l 1.4) Book 5 male, 5 female - 7.4 g - 14 days One animal died on Day 14. 23 rats Page

Formulation 5 male, 5 female (0.175) 6.5 g - 7 days No signs of toxicity; weight gains 24

273 Wistar rats normal.

Diisopropyl Adipate Formulation 5 male, 5 female 0.7 up to 76.8 g - 14 days One male rat at the 80 ml/kg level 25 Wistar albino died on Day 4 of observation. rats per dosage 5 male, 5 female 1.08 5.0 g - 14 days On Day 2 of observation, one 26 Sprague-Dawley animal died. Necropsy showed rats red, mottled lungs and liver, gas- filled C.1. tract. 5 male, 5 female 1.08 5.0 g 14 days 3 males and 5 females showed 27 Sprague-Dawley decreased activity and ataxia. rats No deaths. 5 female albino 5.0 15.0 g - 7 days No deaths occurred and all animals 28 rats appeared normal. 5 female albino 20.75 15.0 g - 7 days No deaths occurred and all animals 29 rats appeared normal.

a Extrapolated by author. ASSESSMENT: DIOCTYL ADIPATE AND DIISOPROPYL ADIPATE 109

Immersion test A product containing 20.75% Diisopropyl Adipate was tested for dermal ir- ritation and percutaneous toxicity- in a whole-body immersion test using six albino guinea pigs. The product was diluted to 0.5% w/v with water so that the actual concentration of the Adipate was 0.10%. The animals were clipped of all abdominal hair and placed in restraining cylinders. The lower parts of the body were immersed in the 37°C test solution for 4 h per day for three consecutive days. Forty-eight hours after the last exposure, the skin of the abdomen was graded according to a scale from 10 (normal) to 1 (moribund as determined by skin in- juries). Clinical signs were recorded daily, and products with a score less than seven are considered potential irritants. On observation, four animals were nor- mal (score = 10) and two had a “first hint of scaling” (score = 9). There were no signs of systemic toxicity and the degree of skin irritation was considered minimal.(31r

Ocular Undiluted Dioctyl Adipate (0.1 ml) was instilled into one eye of each of six albino rabbits. The untreated eye served as control. The eyes were graded at 24, 48, and 72 h on a scale of 0 (normal) to 4 (cornea1 opacity, iridial destruction, red conjunctivae, and swelling). No irritation (all scores = 0) was found at any of the observation periods.(23) Each of six albino rabbits was treated in one eye with 0.1 ml of a cosmetic moisturizer containing 0.175% Dioctyl Adipate. The animals were observed up to seven days following instillation. After 1 h, slight conjunctival redness was observed, but it had disappeared after 24 h. No other effects were noted.‘24) A 0.1 ml sample of a rouge product containing 0.01% Dioctyl Adipate was instilled into one eye of each of six albino rabbits. The untreated eye served as the control and all eyes were graded after 24,48, and 72 h. This product produced no conjunctival redness or chemosis, keratitis, or iritis and it was considered nonirritating.(32) Two lots of undiluted Diisopropyl Adipate were tested for ocular irritation us- ing six albino rabbits per lot. One eye of each animal received 0.1 ml of the ingre- dient and examinations for irritation were made daily until all scores were negative or up to seven days. One lot caused neligible irritation on Day 1, which disappeared by Day 2. No irritation was caused by the second lot.(33’ A face cream formulation containing 0.7% Diisopropyl Adipate was tested on nine albino rabbits for ocular irritation. One-tenth milliliter of the undiluted test material was placed in one eye of each animal; the other eye served as the control. Thirty seconds after instillation, the treated eyes of three rabbits were rinsed with 20 ml of deionized water. Observations for ocular reactions were made at 24, 48, and 72 h, and four and seven days after administration. In rabbits with unwashed eyes, two had conjunctival redness for 72 h and one had some presence of cornea1 stippling for 48 h. No other reactions were noted. The washed eyes of two rabbits had some cornea1 stippling up to Day 4; no other reactions were noted.(34) Two products, one containing 5.0% Diisopropyl Adipate and one containing 20.75%, were each tested for ocular irritation using six albino rabbits. The prod- ucts were instilled into one eye of each animal; the untreated eye served as the control. Observations were made until all eyes were negative for up to seven

CIR Panel Book Page 274 110 COSMETIC INGREDIENT REVIEW days. The 5.0% product produced minimal irritation (score = 6 out of 110) on Day 1 and the irritation had disappeared by Day 2.(35’ The second product pro- duced minimal irritation (score = 2 out of 110) on Day 1 and the irritation had disappeared by Day 2(36) (see Table 4).

Primary skin irritation The primary cutaneous irritation of undiluted Dioctyl Adipate was studied using six albino rabbits. An intact and an abraded site on each rabbit received 0.5 ml of the Adipate under an occluded patch. After 24 h of exposure, the patches were removed and the sites evaluated for irritation according to the Draize method. A second observation was made 48 h after patch removal. Only very slight, barely perceptible erythema was observed in all animals at 24 h. After 72 h, the irritation had decreased in severity in all animals and had disappeared in one. The Primary Irritation Index (PII) was 0.83, indicating that Dioctyl Adipate was a very mild irritant.‘13’ The primary skin irritation of a moisturizing cream containing 0.175% Dioc- tyl Adipate was tested using three albino rabbits. The formulation was applied in four single daily 0.5 ml applications to the shaved backs of the animals and obser- vations were made for seven days. After 24 h, slight erythema was observed which persisted throughout the seven-day period. One animal had well defined erythema with edema, and mild desquamation was seen on day seven. The irrita- tion index was 1 .6.(24) The primary skin irritation of three lots of Diisopropyl Adipate was in- vestigated according to the Draize method. In each experiment, 0.1 ml of the un- diluted product was applied under occlusion to the clipped back skin of nine albino rabbits. After 24 h of contact, the dressing was removed and the sites scored on a Primary Skin Irritation (PSI) scale of 0 (no effect) to 4 (severe erythema with or without edema). The PII was the average score of the total number of test subjects. The first lot had a PII of 1.6 and the second, 1.3. These scores indicated that the material was a mild irritant.“‘) The third lot caused no irritation in eight rabbits and only barely perceptible erythema in one. The PII score of 0.06 indicated that this compound was minimally irritating.(3B1 Two products containing Diisopropyl Adipate at 5.0% and 20.75% were tested by the Draize technique. The undiluted product (0.1 ml) was applied under occlusion to the shaved skin of nine albino rabbits for 24 h. Observations were made 24 and 72 h after contact. The product with 5.0% Diisopropyl Adipate had a PII of 0.33, indicating that the product was minimally irritating.(39) The product with 20.75% Diisopropyl Adipate had a PII of 0.11 and was minimally ir- ritatingC40) (see Table 5).

Sensitization The skin sensitizing potential of Dioctyl Adipate was studied using 10 white male guinea pigs. An area on the backs and flanks, clipped free from hair, was in- jected intracutaneously with 0.1% Dioctyl Adipate in olive oil. Injections were made every other day, three times weekly, until 10 had been given. The first in- jection was 0.05 ml and all subsequent ones were 0.1 ml each. Two weeks after the last injection, a challenge dose of 0.05 ml was injected. Observations were made 24 h after each injection as to area, height, and color of reaction. The retest or challenge injection reaction was compared with an average of-the scores taken after the original 10 doses. The area and height of the retest area was

CIR Panel Book Page 275 TABLE 4. Ocular Irritation.

Applied irritation score Species and no. Cont. amount Observation ingredient of animals (%) (ml) period Max. score Comments Ref.

Dioctyl Adipate Ingredient 6 albino rabbits 100 0.1 72 h o/4 Nonirritating to rabbit eves. 23

Formulation 6 albino rabbits 0.175 0.1 7 days - Slight conjunctival redness after 1 h; 24

CIR cleared after 24 h. Formulation 6 albino rabbits 0.01 0.1 72 h - No irritation. 32 Panel

Diisopropyl Adipate

Book Ingredient Lot 75 6 albino rabbits 100 0.1 72 h l/110 Negligibly irritating on Day 1. Irritation 33 disappeared on Day 2. Page Lot 76 6 albino rabbits 100 0.1 72 h O/l 10 Nonirritating. 33 276 Formulation 9 albino rabbits 0.7 0.1 7 days - Unwashed eyes; 2 showed conjunctival 34 (6 unwashed eyes) redness for 72 h, 1 had cornea1 (3 washed eyes) stippling for 48 h. Washed eyes; 2 showed cornea1 stippling to Day 4. 6 albino rabbits 5.0 0.1 2 days 6/l 10 Minimal irritation occurred on Day 1 35 and disappeared on Day 2. 6 albino rabbits 20.75 0.1 2 days 2/l 10 Minimal irritation occurred on Day 1 36 and disappeared on Day 2. TABLE 5. Primary Dermal Irritation.

Applied Time Ingr. Cont. amount Species and no.

CIR Ingredient 6) (ml) of animals Contact Observ. P///Max. Comments Ref.

Panel Dioctyl Adipate Ingredient 100 0.5 6 albino rabbits 24 h 48 h 0.83ia.o Very slight, barely perceptible 23 erythema in all animals. Book ------Product-moisturizer 0.175 0.5 3 albino rabbits 4 days 7 days 1.6140 After 24 h, slight erythema 24 Page persisting to Day 7 with desquamation. 277

Diisopropyl Adipate Ingredient 100 0.1 9 albino rabbits 24h - 1.6l4.0 Mild irritant. 37 100 0.1 9 albino rabbits 24h - 1.314.0 Mild irritant. 37 loo 0.1 9 albino rabbits 24h - 0.06/4.0 Minimally irritating. 38

Products 5.0 0.1 9 albino rabbits 24 h 72 h 0.3314.0 Minimally irritating. 39 20.75 0.1 9 albino rabbits 24 h 72 h 0.11/4.0 Minimally irritating. 40 ASSESSMENT: DIOCTYL ADIPATE AND DIISOPROPYL ADIPATE 113 smaller and lower than the average induction reactions; therefore, Dioctyl Adipate was not a sensitizer.(23)

P hototoxici ty Primary dermal phototoxic irritation studies were conducted on two per- fumes both containing 1.1% Diisopropyl Adipate. Four male and three female New Zealand white rabbits were clipped of all back hair and 200 mg of the un- diluted product was applied to gauze patches which were then affixed to the shaved areas. Six patches were applied to the back of each test rabbit and one rabbit received two positive control patches. After a 2 h exposure, the patches on the right-hand side of each animal were removed and the skin irradiated for 15 min with four F40BLB bulbs (wavelength of 320-420 nm, peaking at approx- imately 360 nm), 24 in from the skin. The left-hand side was not irradiated. The patches were replaced on the right side and sealed with an occlusive wrap. All patches were removed 48 h after the initial application and 1 h after removal, sites were scored according to the Draize criteria. Scores were again recorded 72 and 96 h postdose. Both perfumes scored 0 (no irritation) for primary dermal ir- ritation as well as primary dermal phototoxic irritation.(41’42)

Mucous membrane irritation Six female albino rabbits were used to test the mucous membrane irritancy of a product containing 0.175% Dioctyl Adipate. The animals were given a single 0.1 ml topical application of the product to the genital mucosa. During the seven- day observation period, no irritation was noted.(24’

Subchronic Toxicity

Oral Diets containing 0, 1,600, 3,100, 6,300, 12,500, or 25,000 ppm Dioctyl Adipate were fed for 13 weeks to six groups of ten F344 rats and ten B6C3Fl mice of both sexes. Observations were made twice daily and animals were weighed weekly. After 91 days, all survivors were sacrificed, necropsy was performed, and tissues were examined histopathologically. Weight gain was depressed for male rats at the 12,500 and 25,000 ppm dosage levels. No other compound-related abnor- malities were found. Weight gain depression occurred in male mice fed 3,100 ppm or more and in female mice fed 6,000 or 25,000 ppm. No other compound- related abnormalities occurred.(20)

Chronic Toxicity

Oral lntragastric doses of Dioctyl Adipate of 0.4, 1 .O or 2.0 g/kg given for six months to rats caused no enzymatic changes, but did increase the level of sulphydryl compounds in the blood. Hepatic detoxification appeared depressed at the onset of the study, but it was accelerated after six months. Administration of 0.1 g/kg for 10 months decreased CNS excitability. (21) (See Carcinogenesis Section of this report for additional chronic test results.)(20)

CIR Panel Book Page 278 114 COSMETIC INGREDIENT REVIEW

Special Studies

Mutagenesis

Dioctyl Adipate (5 mg/plate or the dose which gave a toxic response, whichever was lower), was tested in the Ames Salmone//a/microsome assay. The compound was nonmutagenic when S. typhimurium strains TA1535, TA1537, TA1538, TA98, and TAlOO were exposed to the chemical with and without metabolic activation systems from rat livers.‘43)

Carcinogenesis

Oral administration Groups of 50 male and 50 female F344 rats and 50 male and 50 female B6C3Fl mice were fed diets containing 12,000 or 25,000 ppm Dioctyl Adipate for 103 weeks. Fifty untreated rats and mice of both sexes were used as controls, and all surviving animals were sacrificed at 104-107 weeks. In rats, mean body weights of the 25,000 ppm group were lower than those of the controls. Males had survival rates of 68% in both the control and low-dose (12,000 ppm) group and 80% in the high-dosed (25,000 ppm) group. In females, 58% of controls, 78% of the low-dose group, and 88% of the high-dose group survived the study. Neoplastic and nonneoplastic lesions were seen with equal frequency in treated and control groups and none appeared related to administration of the com- pound. Dioctyl Adipate was not carcinogenic in F344 rats.(20) In the prechronic studies a dose level of 12,500 ppm and 25,000 ppm caused a weight change in male mice, relative to controls, of minus 15% and minus 25%, respectively. In female mice, the same low and high dose concentrations caused a plus 5.6% and minus 13% weight loss, respectively. In three of these four dose concentrations, the weight loss exceeded the criteria for selecting the Maximum Tolerated Dose.(44) In treated mice, mean body weights of either sex were lower than those of controls. Males had survival rates of 72% in controls, 64% in the low-dose group, and 82% in the high-dose group. In females, 84% of controls, 78% of the low- dose group, and 73% of the high-dose group survived. Incidence of hepatocellular adenomas in male mice were dose-related and statistically significant in the high- dose group. The incidence of hepatocellular carcinomas in male mice was higher in dosed groups, but was not statistically significantly increased. In female mice, there was a significant, dose-related trend and significantly higher incidence of hepatocellular adenomas or carcinomas in each of the dosed groups than in the control group (see Table 6). Since hepatocellular tumors were induced in this bioassay, Dioctyl Adipate was considered carcinogenic in B6C3Fl mice.(20) Hodge and associates (45) fed rats a diet containing O%, O.l%, 0.5%, or 2.5% Dioctyl Adipate for two years. A total of 33 tumors were found which were mainly lymphomas and adenomas; one fibroma occurred. Also two carcinomas of the mammary gland and one carcinoma of the kidney were found, but the incidence of these tumors was not different from controls and not related to dietary treat- ment. They concluded the compound was not carcinogenic. No tumors were found when dogs were maintained for one year on diets containing O%, 0.07%, 0.15%, or 0.2% Dioctyl Adipate.r45)

CIR Panel Book Page 279 ASSESSMENT: DIOCTYL ADIPATE AND DIISOPROPYL ADIPATE 115

TABLE 6. Hepatocelhlar Tumors in Mice.a

Time to first observed tumor (weeks) incidence (%)

Tumor Dose @pm) F M F M

Adenoma 0 106 46 4 12 12,000 103 37 10 16 25,000 a4 101 12 31 Carcinoma 0 106 86 2 14 12,000 a5 68 28 24 25,000 79 65 24 24

a Data from Ref. 20.

Subcutaneous and skin application Dioctyl Adipate was included in a study which tested the carcinogenic potency of six chemicals by subcutaneous implantation and by repeated skin ap- plication. Three compounds, aminotriazole, Aramite [2-(p-tert-butylphenoxy) isopropyl 2-chloroethyl oulfite] and Flectol H (a polymer of 1,2,dihydro-2, 2,4trimethylquinoline) were chosen as carcinogens or suspected carcinogens. Two additional reported noncarcinogenic compounds, butylated hydroxyanisole and dioctyl adipate, were included in the study. Groups of 50 male and 50 female C3HIAnF strain mice were used for each chemical and for each dosing method and dose concentration. A single 10 mg subcutaneous injection dose was used for one group of animals. A weekly application of either 0.1 or 10 mg of each chemical in acetone was applied for life to the clipped skin of the back in two separate groups of animals. All animals were observed for life. No significant adverse treatment-related effects were reported, nor were any of the compounds tested considered to be carcinogenic by the test methods used. The authors con- cluded that methods used are not substitutes for tests by other routes of ad- ministration.(45)

Teratogenesis/Dominant lethal Study

Dioctyl Adipate was injected i.p. to each of 10 male albino Swiss strain mice at doses of 0.47, 0.93, 4.7, or 9.3 g/kg. Two groups of controls were injected with distilled water. Immediately after injection, two virgin female mice were caged with each male mouse. Females were replaced weekly for eight weeks. Pregnant mice were sacrificed on Day 15 ( f 2) of gestation and necropsy was performed to determine the number of corpora lutea, implantations, preimplantation losses, early and late fetal deaths, and viable fetuses. The antifertility effect was con- sidered a function of the reduction in the number of pregnancies; the dominant lethal mutation was determined directly from the number of early fetal deaths in individual females and indirectly from the number of implantations. The results indicated no compound-related changes in the incidence of late fetal deaths. The 10 ml/kg dose of Dioctyl Adipate reduced the number of pregnancies, but the lower doses had values comparable to controls. The compound caused a dose- dependent and time-dependent decrease in implants per pregnancy, and there was a dose-related increase in early fetal death, a direct measure of dominant lethal mutation. A dose- and time-related decrease in the number of live fetuses

CIR Panel Book Page 280 116 COSMETIC INGREDIENT REVIEW

TABLE 7. Embryonic-Fetal Toxicity of Dioctyl Adipate on Rat Fetuses.=fb

Number Dose of Number of live in jetted corpora resorptions Dead fetuses Mean weight Treatment groups b’k) lutea (percent) fetuses Cpercen t) of fetuses (g)’

Blunt needle (injection) - 69 4t6.0) 63 (94.0) 3.91 f 0.02 Distilled water 10.00 59 4C6.8) 55 (93.2) 4.40 f 0.33 Normal saline Control 10.00 62 7f11.5) 54 (88.5) 4.10 f 0.13 Cottonseed oil 9.2 71 S(7.5) 62 (92.5) 3.89 f 0.09

Di-2-ethylhexyl adipate 0.93 62 3f5.3) 54 (94.7) 3.90 f 0.09 4.7 65 2f3.1) 63 (96.9) 3.83 f 0.03~ 9.3 60 4f7.0) 53 (93.0) 3.49 f 0.14d

a Data from Ref. 48. b Five pregnant female rats were injected in each group on Days 5, 10, and 15 of pregnancy. CNumbers represent the average values (g) f the standard error of the mean for each group. dp S 0.05.

occurred in groups treated with Dioctyl Adipate. The authors concluded that the mutational effects occurred mainly during the postmeiotic stage of sper- matogenesis. (46) A comment received on this study questioned the author’s con- clusions, noting that additional data on the number of pregnancies per treated male were required. It was also suggested the number of corpora lutea was necessary if one is to determine whether the differences in implantations per pregnancy are associated with male infertility or are a dominant lethal effect. The comment also stressed the need for historical control data on the test species, as well as the need to have included a positive control in the experiment.(47’ Singh et al.“” studied the embryonic-fetal toxicity and teratogenic effect of Dioctyl Adipate in rats. The compound was administered i.p. at 0.93, 4.7, and 9.3 g/kg to pregnant rats on the 5th, lOth, and 15th days of gestation. The diluents at each dose level were water, saline and cottonseed oil, respectively. Animals

were sacrificed on Day 20. Resorption rates were 5.3%, 3.1 O/O, and 7.0% for each increasing dose; each control had similar or greater rates (Tables 7 and 8). One

TABLE 8. Gross, Skeletal, and Visceral Malformationsa

Dose injected Resorptionsb Abnormalities Treatment groups (g/kg) f%o) CrossC ske/etald Viscera/e

Blunt needle (injection) - 4f6.0) 0 1(3.0%) 0 Distilled Water 10.00 4f6.8) 0 0 - Normal saline 10.00 7Ul.5) 1(1.9%) 4(14.3%) - Cottonseed oil 10.00 5f7.5) 1(1.6%) 2(6.3%) 0

Di-2-ethylhexyl adipate 0.93 3f5.3) 0 1(3.6%) 0 4.7 2(3.1) 1(1.6%) 3(9.4%) 1(3.2%) 9.3 4(7.0) 2(3.8%)’ 2(7.1%) 1(4.0%)

a Data from Ref. 48. bPercent resorptions are based on total number of resorptions and dead and live fetuses. c Percent gross abnormalities are based on total number of fetuses. d Percent skeletal abnormalities are based on total number of stained fetuses (50% of total fetuses). ePercent visceral abnormalities are based on total number of unstained fetuses, ‘Values greater than the 95% confidence interval of the “pooled volume control.”

CIR Panel Book Page 281 ASSESSMENT: DIOCTYL ADIPATE AND DIISOPROPYL ADIPATE 117 malformed fetus occurred at the 4.7 g/kg dose and two at the 9.3 g/kg dose. The equivalent control had one abnormality. Skeletal abnormalities occurred at rates of 3.6%, 9.4%, and 7.1% as compared with 6.3% in the control. Visceral abnor- malities of O%, 3.2%, and 4.0% were observed for each increasing dose. No similar results were seen in the control. The investigator concluded that Dioctyl Adipate depressed the mean body weight of the developing fetus. Further, there was a significant increase of gross fetal abnormalities in the high-dose group as compared with the pooled controls. However, the available data did not indicate teratogenic effect when the same results were compared to control groups for each dose concentration. The lack of data on historical controls, and the failure to include a positive control in the study, make it difficult to accept the validity of the statistical procedures used and the conclusions made by the investigator.

Clinical Assessment of Safety

Dioctyl Adipate

Patch tests A Schwartz-Peck prophetic patch test was used to assess the irritation and sensitization potential of a rouge product containing 0.01% Dioctyl Adipate. A 48 h patch impregnated with the formulation was applied under occlusion to the cleansed upper backs of 100 panelists. Simultaneously, an open patch was affixed for 48 h to the inside of the right upper arm and after the allotted time, the sites were scored. After a 1Cday rest, a second open and closed insult was applied and graded 48 h later. The sites on the backs were then irradiated for 1 min at a distance of 12 in with a UV source (Hanovia Tanette Mark I Lamp) at a wave- length of 360 nm. These sites were read 48 h after irradiation. Two of the 100 sub- jects had a weak erythematous reaction at the open patch site after the first patch and one individual had a strong edematous or vesicular reaction after the second open patch. No reactions occurred after the UV exposure. The investigators con- cluded the product was nonirritating, nonsensitizing and nonphotosensitizing.(49) A Shelanski and Shelanski repeated insult patch test was conducted on the same rouge product discussed above. A series of 10 successive 24 h open and closed patches was applied to the skin of 49 panelists and each site was graded after patch removal. After a two- to three-week rest, an 11 th challenge patch was applied for 48 h and read after patch removal. Ultraviolet light sensitization was evaluated after removal of patch numbers 1, 4, 7, 10, and 11 by irradiating the sites for 1 min at a distance of 12 in with a Hanovia Tanette Mark I Lamp. No photosensitivity was indicated by this test, but weak reactions were produced in three, one, and four panelists, after the fourth, fifth, and tenth open patch ex- posures, respectively. Strong reactions occurred in one panelist after the sixth open patch, and in another after the 1 lth (challenge) open patch.(49) A liquid makeup product containing 9.0% Dioctyl Adipate was assayed in a Modified Draize-Shelanski patch test on 209 men and women. The undiluted product was applied under occlusion to sites on the upper back on Monday, Wednesday, and Friday for three consecutive weeks. Patches were removed and sites scored on the next patch replacement day. After a two-week rest, two con- secutive 48 h challenge patches were applied to adjacent sites on the back and these areas were scored 48 and 96 h after application. Three subjects had moderate to strong erythematous reactions, with or without infiltration and

CIR Panel Book Page 282 118 COSMETIC INGREDIENT REVIEW vesicles, and one subject had a macular faint erythema over 25% of the test area after the second challenge.‘50’ Another makeup product containing 9.0% Dioctyl Adipate was tested as above in a Modified Draize-Shelanski patch test. The product caused irritant reactions in two of the 151 men and women tested, but no significant sensitiza- tion or primary irritation occurred.(S” A Shelanski-Jordan repeated insult procedure was used to evaluate primary irritation and allergic sensitivity of a moisturizing product containing 0.7% of a 25% solution of Dioctyl Adipate (0.175% actual concentration). Patches contain- ing the material were affixed to the cleansed back for 24 h on each Monday, Wednesday, and Friday for 3% consecutive weeks for 10 insults. A lo- to 14-day rest followed removal of the 10th insult, at which time a 48 h challenge patch was applied. The challenge site was scored and seven to 10 days later, a second 48 h challenge patch was applied and graded immediately and 24 h after patch removal. One subject had erythema and papules on the test site after the ninth and tenth inductions. The second challenge patch caused erythema and papules in one subject. No other reactions were noted.(52)

Cumulative irritancy test A similar moisturizing product containing 0.175% Dioctyl Adipate was tested in a 21-day cumulative irritation assay. The product, 0.2 ml, was applied under cotton patches to the backs of 11 female panelists for 21 consecutive days. The patches were removed 23 h after application and the sites were scored 1 h after patch removal. New patches were applied immediately. The cumulative irrita- tion score for this product was 72 out of a possible 630. This product was slightly irritating.(53)

Photopatch test A photopatch test was conducted using a formulation containing 9.0% Dioc- tyl Adipate. Each of 25 panelists received patches containing 0.1 ml of the prod- uct. Twenty-four hours later, the patches were removed and the sites were ir- radiated with a Xenon Arc Solar Simulator (150 W) with a continuous emission in the UVA and UVB range (290-400 nm). Forty-eight hours later, the irradiated sites were scored for irritation. This entire procedure was repeated twice weekly for a total of six exposures. After a lo-day rest, a challenge patch was applied for 24 h and then irradiated for 3 min. This site was then scored 0.25, 24, 48, and 72 h after irradiation. Two control sites, one with the test product with no irradia- tion, and a second receiving irradiation but no product, were included in the test program. None of the 25 individuals had phototoxic or photoallergic reac- tions(54) (see Table 9).

Diispropyl Adipate

24-hour patch tests Diisopropyl Adipate, alone and in a formulation, was assayed for skin irrita- tion potential in 24 h patch tests. Occlusive patches containing 0.1 ml of the substance were affixed to the volar surface of the forearm and/or the medial aspect of arm. The patches were removed 24 h later, and the sites read 2 and 24 h later. The sites were scored on a scale of 0 (no irritation) to 4 (severe deep red

CIR Panel Book Page 283 TABLE 9. Clinical Assessment of Safety.

Applied Time Irrit. score No. of Cont. amount ingredient Test subjects (%) (ml) Contact Observ. Max. Comments Ref.

Dioctyl Adipate Formulation Schwartz-Peck 100 M,F 0.01 - 2 24-h 48 h - Two panelists showed a mild reaction 49

CIR prophetic after the first open patch. 1 panelist patch showed a strong reaction after the Panel 2nd open patch. Formulation Shelanski- 49 M,F 0.01 - 11 24-h - - Ten induction patches; 14-day rest; 49

Book Shelanski RIPT + uv challenge patch (1 lth). UV irradiation after patch nos. 1,4,7,10,11.

Page Weak reaction in 3/49 after patch 4; in l/49 after patch 5; in 4/49 after

284 patch 10. Strong reaction in l/49 after patch 6; in l/49 after patch 11. No UV reaction. Formulation Modified Draize- 209 M,F 9.0 - See 48, 96 h - Nine 48 h inductions; 14-day rest; 50 Shelanski RIPT comments one 48 h challenge. Three moderate to strong erythematous reactions during induction. One faint erythematous reaction from challenge patch. Not a sensitizer or irritant. Modified Draize- 151 M,F 9.0 - See 48, 96 h Nine 48 h inductions; 14-day rest; 51 Shelanski RIPT comments one 48 h challenge. Two subjects had irritant reactions; no sensitization. TABLE 9. (Continued.)

Applied Time Init. score No. of Cont. amount hgredient Test subjects (%) (ml) Contact Observ. Max. Comments Ref.

Formulation Shelanski-Jordan 210 M,F 0.225 - See See - Ten 24 h inductions; 14-day rest; one 52 RIPT comments comments 48 h challenge; 7- to lo-day rest and 2nd 48 h challenge. Results: Insults l-8 caused no reaction.

CIR Insults 9,10 caused erythema, papules in 2/210. Panel Challenge 1 caused no reaction. Challenge 2 caused erythema, Book papules in l/210 after 72 h. Formulation 21-day cumulative 11 F 0.175 0.2 21 23-h - 721630 Sites scored 1 h after patch removal. 53 Page irritant test Maximum irritation occurred in 3 panelists after insults 3,7,18; slight

285 irritation occurred in 1 after patch 16. Formulation Photopatch test 25 M,F 9.0 0.1 See - - Six 24 h patches (twice weekly for 54 comments 3 weeks). Sites irradiated (Xenon UV lamp) on patch removal and read 48 h later; lo-day rest; one 24 h patch to new site; irradiation after patch removal; readings taken 0.25, 24, 48, and 72 h after irradiation. Results: No phototoxicity or photo- allergenicity.

Diisopropyl Adipate Ingredient 24 h patch 19 M,F 100 0.1 24 h 2, 24 h o/4 No irritation. 55 24 h patch 19 M,F 100 0.1 24 h 2, 24 h o/4 No irritation. 55 24 h patch 15 M,F 100 0.1 24 h 2, 24 h o/4 No irritation 56 24 h patch 15 M,F 100 0.1 24 h 2, 24 h o/4 No irritation. 56 Formulation 24 h patch 19 M,F 0.26 0.1 24 h 2. 24 h 0.1614.0 13 subjects had no irritation; 6 had 57 i?i barely perceptible erythema. i4 Minimal irritation. w 24 h patch 19 M,F 5.0 0.1 24 h 2, 24 h o/4 No irritation. 58 3 Formulation Maibach-Marzulli 235 M,F 1.08 0.5 See - - Ten 48 h patches; 14-day rest; One 59 RIPT comments 48 h patch. Erythema occurred in T one person after induction. Hyper- 6 pigmentation occurred in 171235, K but no sensitization reactions occurred. 2 Formulation RIPT 50 M,F 3.0 - See See - Ten 24 h patches, 7-day rest, one 24 h 60 comments comments challenge and observation after 24 and 48 h. No irritation or sensitization reactions. Modified Draize 108 M,F 5.0 0.4 See See - Nine 24 h induction patches; 14-day 61 F RIPT comments comments rest; one 24 h challenge read 48 $ CIR and 96 h after application. No 0 irritation was seen in any panelist.

Panel x RIPT 116 M.F 1.04 0.1 See See - Nine 24 h inductions, 3-week rest, 62 0 comments comments one 24 h challenge scored 24 and 2

Book 48 h after removal. If a challenge 0 reaction occurred, a 2nd 24 h s

Page challenge was applied. Results: Induction No. l-faint 6

286 erythema in 2/l 16; mild z erythema in l/l 16. 4 Induction No. 2-faint erythema in 41116; inductions 3 and 5- faint erythema in l/l 16; and induction 4-mild erythema in l/l 16. Challenge produced faint erythema in 21116. No potential for allergic sensitization. Formulation Kligman 25 M,F 0.7 - See See - Five 48 h induction patches; eight 63 maximization comments comments lo-day rest; 1 h pretreatment with test SLS; 48 h challenge patch read at patch removal and after 24 and 48 h. No contact sensitization d occurred. I: TABLE 9. (Continued.) CIR Applied Time Irrit. score

Panel No. of Cont. amount ingredient Test subjects (%) (ml) Contact Observ. Max. Comments Ref. Book Ingredient 21-day cumulative 16M,F 100 0.3 21 22-h - 26.33/630 All but one panelist showed erythema. 64 irritation test patches Ingredient is “moderately irritating.” Page

Formulation 21-day cumulative 10 F 0.7 0.3 21 23-h - 21630 One panelist showed minimal 65

287 irritation test patches erythema after patch no. 18. Product is “essentially nonirritating.” 21-day cumulative 17F 1.08 0.5 See - 0.29184 Application of material was made on 66 irritation test comments 5 consecutive days during 3 consecutive weeks. Continuous contact was made on Sat./Sun. to allow for 21-day continuous exposure. Two panelists had questionable erythema, one had vesiculation. Low irritation. 21-day cumulative 17 F 1.08 0.5 See - 0.24184 Procedure as above. One panelist had 66 irritation test comments questionable erythema, 2 had definite erythema. Low irritation. 21 -day cumulative 7 M,F 20.75 - 21 24-h - 8184 - 67 irritation test patches Formulation Schwartz-Peck 98 M,F 0.7 - 2 24-h 48 h - All patch tests were negative. UV test 68 prophetic patch + uv was negative. + uv Formulation Draize-Shelanski 49 F 0.7 - See 48 h - Ten 48 h inductions; 14-day rest; one 68 > RIPT + UV comments 48 h challenge. UV irradiation after M patch nos. 1,4,7,10,11. All patch Ii tests + UV tests were negative. 52 Formulation Modified 50 M,F 3.0 - See - Patch test sites pretreated with 5% aq. 69 T maximization comments SLS for 30 min. Six to 8 h later, the + uv first test patch was applied and read 48 h later. A second 48 h patch was then applied. This procedure was repeated twice and followed by a 5-day rest. An SLS pretreatment preceded the 48 h challenge patch. Duplicate sites were exposed to a Hanovia UV Lamp after patches 1,3,5, and 7 (challenge) and read 48 h later. No photoallergic

CIR responses occurred. 0 Modified 49 M,F 3.0 - See - - Test performed as directly above. 70 = Panel maximization comments Product is not a photosensitizer 5 + uv or phototoxic. 2

Book Modified 50 M,F 17.0 See - - Test performed as directly above. 71 0 maximization comments Product is not a photoallergic 3 Page + uv sensitizer. Modified 50 M,F 17.0 See - - Test performed as directly above 72 G

288 maximization comments Product is not a photoallergic zj + uv sensitizer or a primary irritant. m 124 COSMETIC INGREDIENT REVIEW

erythema, vesiculation). Two different lots of the undiluted ingredient were tested on 15 men and women. Neither product caused irritation.(55) Additionally, 19 individuals were tested with two different lots of the undiluted ingredient. None of the 19 had signs of irritation.‘55*56) Product formulations containing Diisopropyl Adipate were tested as discussed in the preceding paragraph. One formulation containing the ingredient at 20.75% was diluted to 1.25% in water (actual ingredient concentration = 0.26%) and tested on 19 individuals. Thirteen subjects had no irritation and six had minimal faint erythema. The PII was 0.16 (possible score of 4.0).‘57’ Another for- mulation containing 5.0% Diisopropyl Adipate was tested undiluted on 19 panelists and produced no irritation.(5B)

Repeated insult patch tests A Marzulli-Maibach repeat insult sensitization study was performed using a perfume containing 1.08% Diisopropyl Adipate (18% of a 6% solution). The per- fume (0.5 ml) was applied under occlusion to the skin of the upper backs of 235 women for 48 h (72 h on weekends). The sites. were scored on a scale of 0 (no reaction) to 5 (erythema with induration and bullae). New patches were applied to the same sites; this procedure was repeated for a total of 10 applications. A two-week rest was followed by a challenge patch applied to an adjacent, un- treated site for 48 h. During the induction series, one individual had erythema covering the entire test site and one had erythema with induration and vesicula- tion. This patient had no reaction when challenged. Seventeen subjects had slight hyperpigmentation, but no sensitization reaction occurred in any of the 235 volunteers.r59) A similar test was conducted using a suntan lotion containing 3.0% Diisopropyl Adipate. No reactions were produced in the 50 men and women panelists. The product was neither a sensitizer nor a contact irritant.‘60’ A hair grooming preparation containing 5.0% Diisopropyl Adipate was tested as above on 108 men and women. This product caused no reactions and gave no evidence of sensitization.‘61) A 5.0% aqueous dispersion of a bath oil containing 20.75% Diisopropyl Adipate was tested as above on 116 men and women. The first insult produced minimal faint to pink erythema in four panelists. Minimal faint erythema occurred in four panelists after insult 2, in one after insult 3, and in one after insult 5. Pink, uniform erythema occurred in one person after insult 4. The challenge patch produced minimal, faint erythema in two persons after 24 h, and no reactions occurred after 48 h.(62)

Maximization test A facial cream containing 0.7% Diisopropyl Adipate was evaluated for contact-sensitization potential in a maximization test. The material was applied under occlusion to the skin of the volar forearm or back of 25 subjects for five consecutive 48 h periods. The patch site was then treated with 2.5% sodium lauryl sulfate for 24 h under occlusion. A challenge.patch was then applied for 48 h and the site read immediately after patch removal and 24 h later. The product produced no reactions indicative of contact-sensitization.‘6J,

Cumulative irritancy test Twenty-one day cumulative irritancy tests were performed on Diisopropyl Adipate alone and in a formulation. The undiluted ingredient was tested on 16 men and women. No irritation was observed until after the sixth patch was ap-

CIR Panel Book Page 289 ASSESSMENT: DlOCTYL ADIPATE AND DIISOPROPYL ADIPATE 125 plied (sixth day). After this time, irritation was reported in 14 of 16 panelists where erythema and papules were the most severe reaction. The undiluted in- gredient had a total irritation score of 395 out of a possible 945; it was classified by the authors as “moderately irritating.” The formulation, a face cream product containing 0.7% Diisopropyl Adipate, was tested on 13 individuals. Minimal erythema occurred in one person after the third patch (third day), and in one per- son after patch 18 (18th day). This product had a score of 2 out of a possible 630; the product was classified as nonirritating.‘64*65) A cumulative irritancy test of two products containing 1.1% Diisopropyl Adipate was tested on 17 subjects using procedures similar to that previously noted for Dioctyl Adipate. cs3) The first product had a score of 0.5 (questionable erythema) in two subjects, T.0 (definite erythema) in one, and 3.0 (vesiculation) in one. The mean score was 0.29 (possible 84). The second product had a score of 0.5 in one person, 1 .O in two people, and 1.5 (definite erythema and possible induration) in one person. The mean score for this product was 0.24 (possible 84). These product scores indicate a low potential for hazard to the consumer.(66) In a similar test, a bath oil containing 20.75% Diisopropyl Adipate was tested on seven patients. The bath oil caused an average score of 8 (possible 84).(57’

Photopatch tests A face cream containing 0.7% Diisopropyl Adipate was evaluated for irrita- tion potential by a Schwartz-Peck prophetic patch test followed by UV exposure and by the Draize-Shelanski repeated insult patch test which was also followed by UV exposure. In the Schwartz-Peck Procedure, 98 panelists were patch tested with the product on the back and on the volar surface of the right arm for 48 h. A second patch was applied 12-14 days later and graded 48 h after application. After the patch site was scored, the same site was irradiated with a UV source (Hanovia Tanette Mark I Lamp) at 12 in for 1 min. The site was graded 48 h after exposure. Reactions were not observed at the induction patch, the challenge patch, or the irradiation sites.(6s) In the Draize-Shelanski test, 10 consecutive 48 h inductions were applied for 48 h to each of 49 panelists and an 11th challenge patch was ap- plied for 48 h approximately 14 days later. Skin sites which received patches 1, 4, 7, 10, and 11 also were exposed to UV radiation (Hanovia Tanette Mark I Lamp) at 12 in for 1 min. These light-exposed sites were graded 48 h after irradiation. No reactions occurred after any induction patch, challenge patch, or UV exposure. The product was neither a primary irritant nor a sensitizer.‘68) Several products were used in modified maximization tests with UV ex- posure. The procedures followed were: on Day 1 of the test (Monday), patch test sites were pretreated for 30 min with 0.5 ml of 5.0% aqueous sodium lauryl sulfate. The test material was then applied to the test site 6-8 h later for a period of 48 h. On Wednesday, the sites were graded immediately after patch removal, and a new patch was then applied for 48 h. On Friday, this site was graded and left untreated over the weekend. This regimen was repeated for two more weeks. After the last induction patch was graded (Friday of the third week), a five-day nontreatment period followed and a patch of sodium lauryl sulfate was again ap- plied for 30 min. After 6-8 h, a challenge test patch was applied and graded 48 h later. A control site was treated with sodium lauryl sulfate, but no test material was applied to it. To assess UV sensitization, skin sites which received patches 1, 3, 5, and 7 were also exposed to a UV source (Hanovia Tanette Mark I Lamp) at 12 in for 1 min. Sites were graded 48 h later. A suntan product containing 3.0%

CIR Panel Book Page 290 126 COSMETIC INGREDIENT REVIEW

Diisopropyl Adipate was tested on 50 people. There were no reactions and the product was not a photoallergic sensitizer. (6g) A sunburn lotion containing 3.0% Diisopropyl Adipate was similarly tested on 49 individuals. No reaction occurred and the product was not a photosensitizer or a phototoxic agent.“‘) Two sun- burn foam bases, each containing 17.0% Diisopropyl Adipate, were tested on two panels of 50 subjects. Neither material caused any reaction and the products were not photoallergic sensitizers or primary irritants’71.72’ (see Table 9).

SUMMARY

Dioctyl Adipate, the diester of octyl alcohol and adipic acid, and Diisopropyl Adipate, the diester of isopropyl alcohol and adipic acid, are plasticizers and emollients. They are produced by the esterification of adipic acid and the ap- propriate alcohol in the presence of an esterification catalyst. Both Adipates are clear, colorless to light yellow viscous liquids, with an aromatic odor. In noncosmetic products, the two Adipates are used in plastic food wraps, blood and hemodialysis bags, solvents, and lubricants. Dioctyl Adipate has In- direct Food Additive status for use in food wrapping materials. The Adipates are used in cosmetics as emollients and bases. Dioctyl Adipate is used in 27 products in concentrations of I O.l%-25%, and Diisopropyl Adipate is used in 112 for- mulations, ranging in concentration from I O.l%-25%. Dioctyl Adipate had low acute oral toxicity, with the LDso ranging from 9.11 g/kg to 45.0 g/kg (estimated). Likewise, Diisopropyl Adipate had low oral toxicity. Estimated LDSOs ranged from greater than 5 g/kg to greater than 76.8 g/kg. In a 14-day study, rats and mice fed up to 50,000 ppm (for males) and 100,000 ppm (females) had weight loss and weight gain reduction at the highest concentra- tions. Females fed the 100,000 ppm diet died. The intravenous LDso of Dioctyl Adipate to rats and rabbits was 900 mglkg and 540 mg/kg, respectively. Diisopropyl Adipate had an intravenous LDso of 640 mg/kg for rats. A per- cutaneous absorption test on rabbits showed that Dioctyl Adipate had an LDso of 16 g/kg, but up to 8.7 g/kg for 24 h was not toxic to rabbits in another test. An im- mersion test of a formulation containing Diisopropyl Adipate (20.75%) indicated no toxicity to guinea pigs. The intraperitoneal LDso of Dioctyl Adipate in mice was 1 .O g/kg; in rats, 47.0 g/kg; and 38.0 g/kg in rabbits. In ocular irritation studies, undiluted Dioctyl Adipate was nonirritating; for- mulations containing up to 0.175% of the ingredient were, at most, mild, tran- sient irritants. Undiluted Diisopropyl Adipate was a very mild, transient irritant; formulations containing the ingredient produced minimal irritation. The results of primary dermal irritation tests indicated that Dioctyl Adipate, when ad- ministered alone and in formulations, was a very mild irritant and Diisopropyl Adipate was minimally irritating. Dioctyl Adipate was not a skin sensitizer in guinea pigs. Two perfumes containing 0.108% Diisopropyl Adipate were neither irritating nor phototoxic to rabbits and a product with 0.175% Dioctyl Adipate caused no mucous membrane irritation in rabbits. Mice and rats fed up to 25,000 ppm Dioctyl Adipate for 91 days had weight gain depression, but no other abnormalities. An Ames test for the mutagenic potential of Dioctyl Adipate was negative. An assay of the carcinogenic potential of Dioctyl Adipate showed that administration

CIR Panel Book Page 291 ASSESSMENT: 9lOCTYL ADIPATE AND DIISOPROPYL ADIPATE 127 of up to 25,000 ppm of the compound for 103 weeks produced no untoward ef- fects and was noncarcinogenic to rats. Mice fed the same amount for 103 weeks had dose-related body-weight reductions and a higher incidence of hepatocellular adenoma and carcinoma than controls. Hodge and associates reported that rats fed up to 2.5 Dioctyl Adipate for two years had a tumor incidence similar to that of the control group. They also found no tumors in dogs fed up to 0.2% Dioctyl Adipate for one year. A single 10 mg dose of Dioctyl Adipate given by subcutaneous injection was not carcinogenic in mice. In a lifetime study Dioctyl Adipate caused no skin tumors when 10 mg was applied weekly to the back skin of mice. The teratogenicity of Dioctyl Adipate was studied in mice. According to the author, intraperitoneal injection of up to 9.3 g/kg of the ingredient to male mice caused antifertility effects in females to which they were mated. lntraperitoneal injections of up to 9.3 g/kg Dioctyl Adipate were administered to pregnant rats on the 5th, lOth, and 15th days of gestation. The investigator reported that resorp- tion rates were similar; however, there was a greater incidence of skeletal and visceral abnormalities. The experimental design and interpretation have been questioned by some. Clinical assessment of Dioctyl Adipate at concentrations of O.Ol%-9.0% in formulation showed, at most, erythema and papules when applied under occlu- sion for extended periods of time. No UV sensitization occurred. Undiluted Diisopropyl Adipate produced no irritation in 24 h patch tests, but was mod- erately irritating in a 21-day cumulative irritancy test. Formulations containing concentrations of 0.26%-20.75% Diisopropyl Adipate caused minimal to mild ir- ritation, no sensitization and no photosensitization.

DISCUSSION

The Expert Panel, in reviewing the animal and human test data on Dioctyl Adipate and Diisopropyl Adipate, found them adequate to evaluate the safety of these ingredients as used in cosmetic products. No human data were available for Dioctyl Adipate as a pure ingredient; however, data were available on for- mulations up to a concentration of 9.0%. These data, plus animal test data at a concentration of 100% of the ingredient, indicated Dioctyl Adipate is, at most, a weak irritant. Sensitization and phototoxicity tests were negative. In a formula- tion Diisopropyl Adipate at a concentration of 0.1% was neither an irritant or phototoxic agent to rabbits. Two studies by the same investigator, one which reported on fetal toxicity and teratogenic effects, and the second on mutation and antifertility effects of Dioctyl Adipate, were reviewed. The author concluded a statistically significant effect in each study, but there were several deficiencies noted in each study which made the author’s conclusions questionable. Several carcinogenic studies have been reported. All but one were negative; one oral feeding study conducted by the National Toxicology Program indicated that Dioctyl Adipate was carcinogenic in female mice and was probably car- cinogenic in male mice. The Expert Panel noted that the Maximum Tolerated Dose was significantly exceeded in this chronic study; thus, these test data may not be relevant in a safety assessment for humans.

CIR Panel Book Page 292 128 COSMETIC INGREDIENT REVIEW

CONCLUSION

On the basis of available data, the Panel concludes that Dioctyl Adipate and Diisopropyl Adipate are safe as presently used in cosmetics.

ACKNOWLEDGMENT

Ms. Anne Moore, Scientific Analyst and writer, prepared the literature review used by the Expert Panel in developing this report.

REFERENCES

1. BALSAM, M.S. and SAGARIN, E. (ed.). (1972). Cosmetics: Science and Technology. New York, NY: Wiley Interscience. 2. HAWLEY, G.G. (ed.). (1971). The Condensed Chemical Dictionary, 8th ed. New York, NY: Van Nostrand Reinhold Co. 3. ESTRIN, N.F., CROSLEY, P., and HAYNES, C. (ed.). (1982). CTFA Cosmetic Ingredient Dictionary, 3rd ed. Washington, DC: Cosmetic, Toiletry and Fragrance Assoc. 4. NATIONAL LIBRARY OF MEDICINE (NLM). (1981). Toxicology database; computer database. 5. CTFA. (Oct. 9, 1981). Submission of data of CTFA. Cosmetic ingredient chemical description for Dioctyl Adipate.* 6. CTFA. (Oct. 9, 1981). Submission of data by CTFA. Cosmetic ingredient chemical description for Diisopropyl Adipate.’ 7. CTFA. (1972). Specifications. Washington, DC. 8. RAMSEY, JO., LEE, T.D., OSSELTON, M.D., and MOFFAT, A.C. (1980). Gas liquid chromatographic reten- tion indices of 296 nondrug substances on SE-30 or OV-1 likely to be encountered in toxicological analyses. J. Chromatogr. 184(2), 185-206. 9. JANARDAN, K.G., SCHAEFFER, D.)., and SOMANI, S.M. (1980). Efficiencies of liquid-liquid extraction car- bon and XAD-2 absorption in isolating organic compounds from environmental sources. Bull. Environ, Contam. Toxicol. 24(l), 145-51. 10. THRUSTON, Jr., A.D. (1978). High pressure liquid chromatography techniques for the isolation and iden- tification of organics in drinking water extracts. 1. Chromatogr. Sci. 16(6), 254-9. 11. CLAYTON, C.D. and CLAYTON, F.E. (eds.). (1978). Patty’s fndustrial Hygiene and Toxicology, 3rd rev. ed., vol. ZA, Toxicology. New York, NY: Wiley-lnterscience Publication. 12. VANDERVORT, R. and BROOKS, S.M. (1977). Polyvinyl chloride fils thermal decomposition products as an occupational illness: I. Environmental exposures and toxicology. 1. Occup. Med. 19(3), 188-91. 13. BAXTER TRAVENOL LABS. (May 7, 1980). Japan Kokai Tokkyo Koho Patent No. 80 60459. 14. CODE OF FEDERAL REGULATIONS (CFR). Title 21, Parts 175.105; 177.1200; 177.1210; 177.2600; 178.3740. 15. SOCIETY OF COSMETIC CHEMISTS (SCC). (March 5, 1982). Submission of data by CTFA. Personal com- munication. 16. FOOD AND DRUG ADMINISTRATION (FDA). (Dec. 22, 1981). Cosmetic product formulation data. Com- puter printout. Washington, DC. 17. MOODY, D.E. and REDDY, J.K. (1978). Hepatic peroxisome (microbody) proliferation in rats fed plasticizers and related compounds. Toxicol. Appl. Pharmacol. 45(2), 497-504. 18. RUBIN, R.]. and JAEGER, R.J. (1973). Some pharmacologic and toxicologic effects of di-2-ethylhexyl phthalate (DEHP) and other plasticizers. Environ. Health Perspect. 1973(3), 53-9. 19. NAPIER, Jr., E.A. (1976). Accumulation, toxicity and metabolism of common plasticizers in humans, U.S.N.T.I.S. PB 260406. 20. NATIONAL TOXICOLOGY PROGRAM (NTP). (March 1982). Carcinogenesis bioassay of Di(2&hylhexyl)- adipate. NTP report series No. 212, NIH Publication No. 81-l 768.

*Available on request: Administrator, Cosmetic Ingredient Review, Suite 810, 11 10 Vermont Ave., NW, Washington, DC 20005.

CIR Panel Book Page 293 ASSESSMENT: DIOCTYL ADIPATE AND DIISOPROPYL ADIPATE 129

21. ANDREEVA, G.A. (1971). Gig. Primen. Toksikol. Pestits. Klin. 9, 373. 22. SMYTH, H.F., CARPENTER, C.P., and WEIL, C.S. (1951). Range-findingtoxicitydata: list IV. Arch. Ind. Hyg. 4,119-22. 23. CTFA. (Aug. 1967). Submission of data by CTFA. Unpublished primary skin irritation, primary irritation of eye mucous membrane, acute oral toxicity, acute dermal toxicity, and skin sensitization study of Dioctyl Adipate.* 24. CTFA. (March 18, 1982). Submission of data by CTFA. Unpublished acute oral, dermal, ocular, and mucous membrane testing of a moisturizing product containing 0.7 percent of a 25 percent solution of Dioctyl Adipate (0.175 percent).* 25. BIODYNAMICS, INC. (BI). (Jan. 16, 1976). Submission of data by CTFA. Unpublished acute oral toxicity in rats of a face cream containing 0.7 percent Diisopropyl Adipate.’ 26. FOOD AND DRUG RESEARCH LABS (FDRL). (June 20, 1980). Submission of data by CTFA. Unpublished acute oral toxicity in rats of Diisopropyl Adipate (18 percent of a 6 percent solution in perfume [1.08 per- cent]).* 27. FDRL (May 1, 1979). Submission of data by CTFA. Unpublished acute oral toxicity in rats of Diisopropyl Adipate (18 percent of a 6 percent solution in perfume [1.08 percent]).* 28. CTFA. (Feb. 21, 1975). Submission of data by CTFA. Unpublished acute oral toxicity test of a product con- taining 5.0 percent Diisopropyl Adipate.* 29. CTFA. (Nov. 9, 1978). Submission of data by CTFA. Unpublished acute oral toxicity test of a product con- taining 20.75 percent Diisopropyl Adipate.* 30. NATIONAL INSTITUTE FOR OCCUPATIONAL SAFETY AND HEALTH (NIOSH). (1977). Registry of Toxic Effects and Chemical Substances, vols, I and II. Cincinnati, OH. 31. CTFA. (Nov. 9, 1978). Submission of data by CTFA. Unpublished immersion test of a product containing 20.75 percent Diisopropyl Adipate.’ 32. CTFA (1975). Submission of data by CTFA. Unpublished ocular irritation test summary of a product contain- ing 0.01 percent Dioctyl Adipate.* 33. CTFA. (May 28, 1973). Submission of data by CTFA. Unpublished eye irritation test of Diisopropyl Adipate.’ 34. BI. (Dec. 29, 1975). Submission of data by CTFA. Unpublished rabbit eye irritation study of a face cream containing 0.7 percent Diisopropyl Adipate.’ 35. CTFA. (Feb. 21, 1975). Submission of data by CTFA. Unpublished eye irritation test of a product containing 5.0 percent Diisopropyl Adipate.’ 36. CTFA. (Nov. 9, 1978). Submission of data by CTFA. Unpublished eye irritation test of a product containing 20.75 percent Diisopropyl Adipate.* 37. CTFA. (Feb. 2, 1973). Submission of data by CTFA. Unpublished primary skin irritation study of Diisopropyl Adipate.* 38. CTFA. (Nov. 2, 1978). Submission of data by CTFA. Unpublished primary skin irritation test of Diisopropyl Adipate.’ 39. CTFA. (Feb. 21, 1975). Submission of data by CTFA. Unpublished primary skin irritation study of a product containing 5 percent Diisopropyl Adipate.* 40. CTFA. (Nov. 9, 1978). Submission of data by CTFA. Unpublished primary skin irritation test of a product containing 20.75 percent Diisopropyl Adipate.* 41. FDRL. (July 21, 1980). Submission of data by CTFA. Unpublished primary dermal phototoxic irritation study of Diisopropyl Adipate (18 percent of a 6 percent solution in perfume [1.08 percent]).* 42. FDRL. (July 21, 1980). Submission of data by CTFA. Unpublished primary dermal phototoxic irritation study with Diisopropyl Adipate (18 percent of a 6 percent solution in perfume [1.08 percent]).* 43. SIMMON, V.F., KAUHANEN, K., and TARDIFF, R.G. (1977). Mutagenic activity of chemicals identified in drinking water. Dev. Toxicol. Environ. Sci. Prog. Genet. Toxicol. 2, 249-58. 44. SONTAG, J.M., PAGE, N.P., and SAFFIOTTI. (Feb. 1976). National Cancer Institute. PB-264-061. 45. HODGE, H.C., MAYNARD, E.A., DOWNS, W.L., ASHTON, J.K., and SALERNO, L.L. (1966). Tests on mice for evaluating carcinogenicity. Toxicol. Appl. Pharmacol. 9(3), 583-96. 46. SINGH, A.R., LAWRENCE, W.H., and AUTIAN, J. (1975). Dominant lethal mutations and antifertility effect of di-2-ethylhexyl adipate and diethyl adipate in male mice. Toxicol. Appl. Pharmacol. 32(3), 566-76. 47. NORTHUP, S. (March 9, 1982). Comment received on the CIR Scientific Literature Review on Dioctyl Adipate and Diisopropyl Adipate.* 48. SINGH, A.R., LAWRENCE, W.H., and AUTIAN, J. (1973). Embryonic fetal toxicity and teratogenic effects of adipic acid esters in rats. J. Pharm. Sci. 62(10), 1596-600. 49. CTFA. (1977). Submission of data by CTFA. Unpublished human prophetic patch test and repeated insult patch test of a product containing 0.01 percent Dioctyl Adipate.* 50. CTFA. (March 10, 1978). Submission of data by CTFA. Unpublished human modified Draize-Shelanski-Jor- dan patch test of a product containing 9 percent Dioctyl Adipate.’

CIR Panel Book Page 294 130 COSMETIC INGREDIENT REVIEW

51. CTFA. (Sept. 22, 1976). Submission of data by CTFA. Unpublished human modified Draize-Shelanski test of a product containing 9 percent Dioctyl Adipate.* 52. LEO WINTER ASSOCIATES (LWA). (Sept. 1978). Submission of data by CTFA. Unpublished human Shelan- ski-Jordan repeated insult procedure of a product containing 0.175 percent Dioctyl Adipate.* 53. HILL TOP RESEARCH (HTR). (Oct. 6, 1978). Submission of data by CTFA. Unpublished human cumulative irritancy test of a product containing 0.175 percent Dioctyl Adipate.* 54. CTFA. (March 10, 1978). Submissiion of data by CTFA. Unpublished human photopatch test of a product containing 9 percent Dioctyl Adipate.* 55. CTFA. (Jan. 24, 1973). Submission of data by CTFA. Unpublished human skin irritation test of Diisopropyl Adipate.* 56. CTFA. (May 18, 1973). Submission of data by CTFA. Unpublished human skin irritation of Diisopropyl Adipate.* 57. CTFA. (Oct. 26, 1978). Submission of data by CTFA. Unpublished human skin irritancy test of a product containing 20.75 percent Diisopropyl Adipate.* 58. CTFA. (Feb. 12, 1975). Submission of data by CTFA. Unpublished human skin irritation of a product con- taining 5 percent Diisopropyl Adipate.* 59. CONCORDIA RESEARCH LABORATORIES (CRL). (Oct. 29, 1980). Submission of data by CTFA. Unpublished human repeated insult patch test of a product containing 1.08 percent Diisopropyl Adipate.* 60. CTFA. (March 5, 1974). Submission of data by CTFA. Unpublished human repeated insult patch test of a product containing 3.0 percent Diisopropyl Adipate.* 61. HTR. (Nov. 3, 1976). Submission of data by CTFA. Unpublished human repeated insult patch test of a product containing 5 percent Diisopropyl Adipate.* 62. CTFA. (Feb. 1, 1982). Submission of data by CTFA. Unpublished human allergic contact sensitizatron test of a product containing 20.75 percent Diisopropyl Adipate.* 63. IVY RESEARCH LABORATORIES (IRL). (Feb. 17, 1976). Submission of data by CTFA. Unpublished human maximization test of a product containing 0.7 percent Diisopropyl Adipate.’ 64. HTR. (March 30, 1976). Submission of data by CTFA. Unpublished human cumulative irritancy test of Diisopropyl Adipate.* 65. HTR. (Feb. 18, 1976). Submission of data by CTFA. Unpublished human cumulative irritancy test of a product containing 0.7 percent Diisopropyl Adipate.’ 66. CRL. (June 9, 1980). Submission of data by CTFA. Human 21-day cumulative irritation test of a product con- taining 1.08 percent Diisopropyl Adipate.* 67. CTFA. I(May 11, 1978). Submission of data by CTFA. Unpublished human 21.day cumulative irritancy test of a product containing 20.75 percent Diisopropyl Adipate.* 68. RESEARCH TESTING LABORATORIES (RTL). (March 4, 1976). Submission of data by CTFA. Unpublished human prophetic patch test of a product containing 0.7 percent Diisopropyl Adipate.’ 69. CTFA. (June 21, 1974). Submission of data by CTFA. Unpublished human modified maximization test of a product containing 3.0 percent Diisopropyl Adipate.’ 70. CTFA. (March 3, 1975). Submission of data by CTFA. Unpublished human modified maximization test of a product containing 3.0 percent Diisopropyl Adipate.* 71. CTFA. (June 21, 1974). Submission of data by CTFA. Unpublished human photopatch test of a product con- taining 17.0 percent Diisopropyl Adipate.* 72. CTFA. (June 21, 1974). Submission of data by CTFA. Unpublished human modified maximization test of a product containing 17.0 percent Diisopropyl Adipate.’

CIR Panel Book Page 295

CIR Panel Book Page 296

CIR Panel Book Page 297

CIR Panel Book Page 298

CIR Panel Book Page 299

CIR Panel Book Page 293

CIR Panel Book Page 294 Persona Care’ ProductsCounci Committedto Safety, Quaity & nnovation Memorandum

TO: F. Alan Andersen, Ph.D. Director - COSMETIC INGREDIENT REVIEW (C]R)

FROM: John Bailey, Ph.DZ E__3 I. I2 Industry Liaison to the Cifi Expert Panel

DATE: December 1, 2009

SUBJECT: Comments on the Draft Report on the Dicarboxylic Acids and their Salts and Esters prepared for the December 7-8, 2009 CIR Expert Panel meeting

General Comment - This report is still very incomplete. Diethyihexyl Adipate, Dibutyl Adipate and Diisopropyl Adipate, have already been reviewed by CIR and found safe as used. This report fails to mention these earlier CIR reviews, and this report does not include the information that was summarized in these original CW reports, including an NTP bioassay on Diethyhexyl Adipate.

Looking only at the Safety/Regulatory Information for the added ingredients on the On-Line, the following items are missing from this report. Oxalic Acid: listed in EU Annex ifi; NTP continuous breeding study; NTP tested negative in Salmonella (Haworth et al. 1983) Malonic Acid: NTP tested negative in Salmonella Dimethyl Succinate: NTP micronucleus test in male rats, negative; NTP tested negative in Salmonella (Zeiger et al. 1992) Glutaric Acid: NTP tested negative in Salmonella Dimethyl Glutarate: NTP micronucleus test in male rats, equivocal; NTP tested negative in Salmonella Adipic Acid: NTP tested negative in Salmonella Dimethyl Adipate: NTP has selected Dimethyl Adipate for further study; there is a nomination background report that includes information that is not in the CIR report; NTP tested negative in Salmonella Diethylhexyl Adipate - IARC unclassifiable; CIR report; NTP carcinogenicity study (includes 14-day, 90-day and 2-year studies); NTP battery of genotoxicity assays Report organization - Generally CW reports are organized by duration of exposure, route of exposure, then compound. It is not clear how this report is organized. There are very few route of exposure section headings. Inhalation exposure is its own section, not under any duration of exposure. The Introduction states “the data are presented by order of carbon chain length, beginning with Oxalic Acid.” This is a good idea, but it is not how the report is organized.

11011 7th Street, N.W., Suite 300 Washington, D.C. 20036-4702 202.331.1770 202.331 .1969 (fax) www.personalcarecouncil.org CIR Panel Book Page 295 Most sections start with information on Sebacic Acid and the Sebacates. Both the text (each section) and all the tables should be presented by order of carbon chain length as indicated in the Introduction. p.1 - Only the esters of the dicarboxylic acids have side chains between 1-18 carbons (not the salts). p.1 - In the introduction, it should be mentioned that Diethyihexyl Adipate, Dibutyl Adipate and Diisopropyl Adipate have previously been reviewed by CW and found safe for use. p.2 - Please delete the sentence concerning the name of Sodium Oxalate. If you search on ChemlD for Sodium Oxalate, the compound found has two sodium atoms, a compound with one sodium is not found. p.2 - In the second paragraph under the heading Dicarboxylic Acids - General, it would be helpful to add some of the additional ingredients that have been added to this report. p.3, 4, 49 - Anonymous (2000b) should be changed to IARC (2000). p.4 - Ingredients in addition to the sebacates should be mentioned in the first paragraph of the Cosmetic use section. p.5 - As the ingredients added to this report since the September meeting have not yet been included in a Council concentration of use survey, it is likely that the results will not be available until sometime in March. Please look at the VCRP again to be sure all of the ingredients included in the report were found. Searching the VCRP for adipate revealed 2 uses of Dimethyl Adipate, 2 uses of Dibutyl Adipate and 41 uses of Diethylhexyl Adipate that are not included in the Cifi report. The uses of Diethyihexyl Adipate, Dibutyl Adipate and Diisopropyl Adipate from the old reports (and re-reviews) should also be mentioned in this report p.6 - Please change “setting” to “settling”. How did Lehman-McKeeman (2008) define nanoparticles? p.6 - Information about how these ingredients are regulated in Europe should be added to the cosmetic use section. p.9 - In the last paragraph of this page, what is meant by “both substances”? p.10, 25, 44, 45 - The INCI names for diethylmalonate and dimethylmalonate includes a space (Diethyl Malonate, Dimethyl Malonate). p.10 - The following is not a complete sentence: “Tyrode’s solution as receptor fluid.” p.12 - Please delete the last sentence on p.12 (“According to Lubrizol (2000), Diisopropyl Sebacate is rapidly absorbed and is fast-spreading, but no supporting data were provided.”), as it does not add any useful information. p.13 - What is meant by “the same oxidative stress parameters”? Please define TAR the first time it is used. p.13 - It is not clear what is meant by “the study supported a NOAEL of 4000 mg/kg for the oral route and 8000 mg/kg”. Was 4000 mg/kg the highest dose tested and some effects were observed, and was 8000 mg/kg the highest dose tested and no effects observed? p.14 - What were the i.v. doses given to rabbits? Currently, it states “from 300 to 1800 mg” should this be mg/kg b.w.? p.14 - The information on Diethylhexyl Sebacate in Table 5 is also presented in Table 7. p.15 - Please delete “in repeat dose studies, and no effects in teratologic evaluations” from the Acute Exposure section. There is no information about acute studies in Table 9. Please provide a reference for “A similar relationship existed for ocular irritation, where severity of response decreases with increasing carbon number.”

2

CIR Panel Book Page 296 p.15 - If route subheadings are going to be used, they should be used throughout the report. p.15 - It is not clear what is meant by “Smyth et al. (1969) reported an acute oral toxicity in rats of DEM of 15,794 mg/kg bw (SIDS 2005)”. If Smyth et al. (1969) did the study, why is there also a citation to SIDS (2005)? Smyth et al. (1969) is not in the reference section. p.15 - What is “Gemini surfactant (AGS) mixture”? p.16 - Please delete the second paragraph under Short-Term Exposure as it presents the same information as in the first paragraph of this section. p.17 - The 3-day rat study should not be presented in the Subchronic Exposure section. p.17-18 - The protocol of the Mingrone et al. (1983) study of Azelaic Acid is not clear. Animals were fed for 90 days. Was there also a recovery group that was kept for about 90 days after exposure ended? p.18 - What compound were the beagle dogs fed for 90 days (American Chemistry Council, 2003)? p.19 - The three week study of Diethyihexyl Sebacate should not be in the Subchronic Exposure section. p.19, 45 - Why is 2-ethylhexyl adipate considered a surrogate diester? 2-Ethyihexyl adipate is Diethylhexyl Adipate, a cosmetic ingredient being reviewed in this report, and previously reviewed by CIR and found safe for use in cosmetics. p.19, 31 - Why is Ditridecyl Adipate included in the Diesters subsection? As it is an ingredient included in this report, it should have its own subsection. p.20 - The 90-day studies should not be discussed in the Chronic Exposure section. p.21, 26 - It is not necessary to present the information on butyl stearate for the Smith (1953) study. p.22 - The inhalation exposure studies are usually presented under the various duration subheadings. p.25 - The Magnussen Kligman test of Dodecanedioic Acid (SIDS 1994) should be moved to the Dermal Sensitization section. p.26 - There are no additional details of the BIBRA (1996) reproductive study in TableS (the same information is presented in Table 7). p.27 - The acute exposure studies cited to Greco et al. (1990) should not be in the Reproductive and Developmental Toxicity section. p.27 - Please clarify what days of gestation the ewes were treated in the Scheifer et al. (1976) study. p.27 - Were reproductive outcomes studied in the Goldman et al. (1977) study of Oxalic Acid? Did they look for testicular effects? p.27-28 - The protocol of the Lamb et al. (1997) study is not clear. Is this the NTP continuous breeding study? p.28 - Please change “life” to “live” p.29 - What route of exposure was used in the Bradford et a!. (1984) study? When during gestation were the rats and rabbits treated? p.29 - When during gestation were the rats treated with the seven adipates? p.29 - When during gestation were the rats and rabbits treated in the Mingrone et al. (1983) study? Were the rabbits really sacrificed on gestation day 19? p.30 - In the last paragraph of the Azelaic Acid subsection, please change “rams” to “dams”. p.31 - Please delete the following sentence, as this study has already been presented (more than once). “No adverse reproductive , suckling and growth affects were evident in a four-generation study in rats fed a diet containing 200 ppm Diethylhexyl Sebacate (BIBRA 1996).”

3

CIR Panel Book Page 297 p.32 - The BII3RA (1996) study is mentioned again at the top of p.32 and should be deleted. p.33 - In the first sentence under the heading Bacterial Gene Mutation Assay, what is meant by “in this assessment:? p.40 - Did De Groot et al. (1991) really use PEG-300? Or is this PEG-6 (a PEG with an average molecular weight of 300)? p.43 - Please check the VCRP again for additional adipate ingredients. p.4-7 - The following sentence should be moved with the rest of the genotoxicity information. “In the HPV report on diesters, mutagenicity nor clastogenicity were exhibited by those diesters cited in vivo or in vitro tests, with or without metabolic activation.” What is meant by those diesters? Please change “cited” to “studied”. p.48 - There is no mention of the cancer studies in the Summary. Table section - The table section is missing page numbers. Table 1- This table should be organized as stated on the first page of the report (from smallest to largest dicarboxylic acid). The title of the table needs to be change to reflect all of the ingredients included in the report. For those ingredients for which the definition is based on the structure, the table should refer to the figure in which the structure is shown. Please delete “q.v.” from the table or define it at the end of the table. Other names that are the same as the INCI name, with another group designation, e.g., Diisobutyl Adipate (RIFM) should be deleted from the report. If these names are left in the report the group, e.g., RIFM, should be defined. Table 2 - The title of Table 2 should indicate the types of compounds included in the table. The table should be organized as stated on the first page of the report (from smallest to largest dicarboxylic acid). On the second page of the table, what does “*“ and “**“ mean? Please use the INCI name, Diethylhexyl Adipate rather than Di(2-Ethylhexyl) Adipate. Please use the INCI names for Dimethylmalonate and Dimethylmalonate. Some units in this table are presented in the first column, some are presented in the second column; please be consistent. Table 3 - The title of the table should include more that just Sebacates. Please organize this table as stated on the first page of the report (from smallest to largest dicarboxylic acid). Table 4 - The title of the table needs to be changed to include the ingredients that have been added to the report. When the Council concentration of use survey for the other ingredients is complete, the concentration of use for Diethyl Sebacate will be revised to 1.5% (it was rounded to 2%). Please check the VCRP to be sure all the ingredients in this report were identified. The VCRP also has 2 uses for Dimethyl Adipate, 2 uses for Dibutyl Adipate and 41 uses for Diethylhexyl Adipate. Consider adding old concentration of use information for the ingredients already reviewed (and re-reviewed) by CIR. The total uses for Diisobutyl Adipate is listed as Dicapryl Adipate. The total uses for Diisopropyl Adipate is listed as Diisostearyl Adipate. The footnote “Ingredients not found in the table were included in the concentration of use survey, but no uses were reported.” is not correct as the Council concentration of use survey on the ingredients added to this report has not yet been sent out. Table 5- Since Diethyihexyl Sebacate is the only ingredient included in this table, the first column can be deleted. Please consider deleting this complete table as all this information is also presented in the last row of Table 7. Table 6 - The heading of column 3 needs to be changed as it includes more than just 50LD values. In the route column, please change “skin” to “dermal”. It is not clear what is meant by the NOELs

4

CIR Panel Book Page 298 of 100 mg/kg (males), 500 mg/kg (females) and then in the next row a NOEL for males and females of 1000 mg/kg is given. Are the first values adult NOELs and the last value a NOEL for developmental effects? Table 7 -The Summary table of Diesters Toxicity is misleading as it does not include all of the data on each ingredient. If it is left in the report, it should include al the data on each ingredient with appropriate references, not just the information as presented by the American Chemistry Council (2003). Table 8- Please identify what was in the Dicarboxylic Acid Mixture. In what species were the dermal and eye irritation tests completed? Table 9 - What route of exposure was used for these studies? The doses provided are actually concentrations. What do they represent - concentration in the diet? What species were tested in the developmental and reproductive toxicity studies? What doses were used in the developmental and reproductive toxicity studies? Table 10 - In the title of this table, please indicate which ingredient was studied.

5

CIR Panel Book Page 299