Chromatographic Behaviour of Some Synthetic Dyes O!

Articles Indian Journal o f C hemical Techn ology Vo l. 9. Se pt ember 2002. pp. 432-437 . /

\.

Chromatographi c behaviour of some sy nthetic dyes O!1 unconventional layers

, I . " Ad il ya K Misra *, Pankaj SjSaxena & Upasana,Gupta _ ,_ Departmcnt ll r C hc lll istry. NMSN Dass (PC) Co ll ege. l3udalln 243 nO_I. In dia Receil'erl 20 Jill\' 200 1; reel'il'erl rel'ised 13 Mllrell 2002; lI("cepled 15 Mil\, 2002

tt:he thin layer chromatographic behaviour of some synthetic dyes has been studied on egg ~ h e ll layers in various organic solvents and inOJ-gani c salt solutions. The mobility of the dyes was found to depend on the polarity of the solvents. ~etones gave compact spots of dyes_

A large number of difficult separation of analytical interest have been achieved on the basis of R-r-RL value_ The results al-e compared with the chromatographic behaviour of phenols to establi sh the separation potential of the egg shell laye l- in pollution studies. \ -

'O Thin lay er chromatograph y (TLC) I S reliabl e, ammonium molybdate and copper sulphate , zinc ll l 2 inexpensive, portabl e and easil y performed by sulphate , nickel chloride , detergents') and nontec hnical operators. A great va ri ety of so l ve nts inorga nic ion exchanger l 4 ha ve been used to ach ieve can be used as mobile phases for TLC and several th e better and ef"fecti ve reso l uti on of sy ntheti c dyes . samples ca n be developed. TLC has grown rapidly in Micellar reverse phase TLC '5 has also bee n last two decades and is now widely accepted as a performed for th e same purpose. Some sta tionary rapid and efficient analytical tec hn ique, its sui tab i I ity phases like polyamide, ce llulose derivatives, ch itin, to th e analysis of organi c and inorganic substances chitocin, am inoplas t, polyacriloni trile, ion have been \vell documented in many papers and exchangers, chemically mod i fi ed and impreg nated 5 rev iews '- TLC in combinati on with modern hydrous so l ids have also been developed to ach ieve in strumentation such as spectrophotometry, the better separations and reproduci bi I i ty for va ri ous densi tometry , atomic absorption spectrophotometry organic and inorgani c compounds. Mohammad el 0/. (AAS) has been used for th e analysis of biologica l, have shown the high potential of th e chicken egg shell pharmaceutical, biomedical and environmental powder as TLC coating material for th e analysis o f l ' 7 samples . TLC has al so rep laced the paper metal chlorosu lphates (" amines and ph enols T hi s chromatography in dyes, amino acids and pes ti cide report is an attempt to use this material in th e anal ys is res idues analysis because of its higher reso lution and of synthetic dyes. The res ults have bee n compared shorter development time. with chromatographic behaviour of phenols, as The synthetic dyes are most widely used in toys, ph enols are also sel f visualized coloured compounds. food stuffs, cosmetics and drugs th e slight excess of the egg she ll is avai lable in abundance as waste which cau se acule hea lth hazard. So th eir separation, material after the use of protein matter all over th e identificati on and quantitation ha ve great importance world and being white in color has large number o f" in the fi eld of separation sc ience. The literature poss ibilities for Li se as an inexpensi ve adsorbent in avai lable up to June 2000 revea ls that the TLC. There is still a need to develop a simple, conve ntional layer materi als such as alumina, inex pensive, reproducible meth d to ac hieve better ce llulose, keiselguhr and silica gel have been widely se parations of synthetic dyes and th e proposed used as stationary phases . The separation of dyes is methods have better separation poss ibilities for difficult because of spots tailing and close IiRr va lue. sy ntheti c dyes. T o encounter this problem organic ion exc ange 6 materi als , silica gel impregnated with organic Experimental Procedure 7 compoun d , Sl'1 vel' nitr. ate,8 ca d'mlum acetate,9 Chemicals and Reagents Ethy l alcohol, methyl alcohol, propanol ethy l " For correspo nde nce: 20. Salldagaran, Bareill y 243 003, In dia acetate, acetone, hexane, ethyl methyl ketone and (E-mai l: akmisra 1234 @hotll1ail.eom; Fax: 550091) carbon tetrachloride, sodi um ch loride, sodium nitrate Mi sra el (fl.: Chrolllatog raph ic be ha viour of some sy nth etic dyes Articles were obtained from E. Merck (India), chicken egg usin g chicken egg shell powder as an effecti ve shell were collected from a natural source. All the sorbent layer for th e separati on of transitional metals, l 6 ' 7 dyes were of analytical grade from BDH (Poole) or E. rare earths , phenols and amines . The results Merck (Damstred). obtained were promisin g despite the limitation that egg shell powder is un stable under acidic medium . Test soiutiolls The spots at more than 3 cm has been treated as tai led The soluti ons of sy nthetic dyes ( I 0-3M) were spots. Atleast 1.5 cm di stance between th e RI. of one prepared in ethanol-water (4: I) ratio (v/v). Dyes are spot and RT of th e oth er spot has been used to resol ve sel f vis ual ized compounds. the dye. RT and RI. are th e rear and front limits of th e spot of dye respec ti vely. Preparatioll of tile chicken egg-shell powder Broken pi eces of egg shell were washed wi th RI. + RT 12 boiling water th en di still ed water and th en soaked in ----'-'------'----- and 10% aOH so lution for 20-30 h to remove th e protein 10 content. The cleaned egg shell pieces we re then washed several times with distilled water to ensure IiRr= Rrx lOO co mpl ete remova l of NaOH , dri ed at 100°C, ground and sieved to furni sh 250- 1SO mes h powder. The Table I represents the abbreviations and stru ctural res ulting egg she ll power was used to prepare slurry formu la of th e dyes. The results obtained have been for coatin g on to TLC plates with out addition of summari zed in Table 2 and Fi g.l. Table 2 shows th e bi nder. IiR r va lu es of 24 syn th etic dyes in different mobi le phases. Resu lts have been compared with th ose Preparation ofTLC plates obtained on conventi onal sili ca gel G layers Crable 3). Th e so rbent was mixed with demin erali zed water in very few separations are possible on such layers while ( I :3) rat io (11'Il\' ) with constant sha kin g to furnish on haematoxy Ii ne, bi smark brown, resorci nol bl ue. homogenous slurry. This slurry was app li ed to 3. 0x 1S cllrclllllin S, eri ochrome black T, indi gocarmine and CI11 glass plates as 0.25 mm layer by means of Sudan black show streak in both eth anol and Toshniwal (India) TLC app li cator. The plates we re methanol mobi le phases. Spots were large and dri ed at room temperature and than activated at hindered in th e separation on si li ca gel G layers. The IOS±2°C by hea tin g in an electricall y controll ed oven same problem has been seen on aluminium oxide for C(l I h. The activated plates were stored in a closed layers. On the basis of Rr - RL va lues, se veral chamber at room te mperature until used. important , clear and di llicult separations of analytical Procedure interest have been obtain ed as fo ll ows: The test so lu tions (500 ng) were appl ied to TLC The binary separa ti on of amid oswartz, curcu min S. plates C(f 1.5 cm above th e lower edge by means of a ali zarin red S and naphthol green B has been ac hi eved mi croppipette. The spots we re ai r dri ed and plates indi vidua ll y from crys tal violet, gen tia nt viol et. were developed with suitable mobiie phases by fusc hin basic, bri lliant gree n, safranill , methyl red and ascenc in g tech nique in Sx20 cm glass jars sudan blac k and ternary separation of naphthol green presaturatecl with th e mobile phase. The solve nt B - sudan black - and one or more of th e following ascent was fixed to 10 cm above th e po int of dyes: genti ant violet, fll sch in basic, brilli ant green. application. Arter development the plates were safranin , methyl red has been achieved in methan ol. removed from the jars, dri ed at room temperature The binary separati l) ns of resorcino l blu e and (25 °C) and rela tive humidity (65 %). The spots were amidoswartz from crys tal violet, genti ant vio let. se lf visualized. bismark brown, briliiant green. safrani n and The fo ll owing solvent systems were used: rhodamine B; eri ochrome black T and indi gocarmi ne (i) Meth ano l. (ii ) Ethanol, (iii ) Propanol, (iv) Acetone, from cry stal vio let, gentiant violet, bismark brown. (v) Ethyl meth yl ke tone, (v i) Eth yl acetate, (v ii ) 0.1 M brilliant breep. safranin. rh odamin e B and sudan black NaC!, (v iii ) 0. 1M Na 0 ]. and ternary separations of indi gocamine, resorcinol blue and amidoswartz (indi vidua ll y) fro m sud an black Results and Discussion and one or more of th e foll owing dyes: crystal violet. Studi es to develop new sorbent phases usin g gentian violet, bi smark brown, brilliant green. unconventional material s revealed th e possibility of safranin , rhodamin e B; and ternary separati on of.

433 Indian J. Cheill. Techn o!., Septclll bcr 2002

Table I- The naPles, abbrev,lIi ons and st ructura l formulae of the dyes in vesti gated- Collfd ~ ", a, :((H,H,X;( (CHJ. ~ 1 ...... -9 h CHICHJ. 19 Bromolhymol blue BIB aSQ,H<;

0 H NiJO,S V ~ ..... /' ~ ?" 20. Indigo carmine Ie C:.-.... N...... c=( ...... C C~ so Na ~ g , IC,H,I,NV:°ONlc,H",

21 Rhodamno O ~ " C """

Oc~'" I ICH,I,N'O o NteHJ, " c """ 22. Methyl..,iolet MV ¢ NHCIl ,

0 OH

23 Alizarin Red S A,S ,.. I" $~'::::-... I , ...-::: so , N.l 0 H 24 Sudan Black SO O-N- = oN_ = ~_ ~ 'c ...... CH , ~ '" ~ h N/ ' Ctl, ~ curcumin S - bromothymol blue - reso rcinol blue, bri II iant green, bi smark brown have bee n achi eved in ethanol. Binary se parati on 01' amidoswartz, cllrcumin S, eri oc hrome black T , naphthol green B, indigocarmine, orange G, bromoph enol blue, haematoxyline, eos in from crystal violet , genti ant violet, reso rcinol blue, fu schin basic, bri lliant green , sa fran in, ponceu S, meth yl red, rh odamine B, meth y l violet and sudan black has been ach ieved in propanol. Bin;} ry separations of reso rcinol blue, amidoswartz, curclImin S, naphthol green B, orange G, alizarin "eel S from crystal violet, gentiant violet, bi smark brown, lac moid, fusc hin basic, brilliant gree n, sarranin, meth yl red , rh odamine B, meth yl violet ha ve been achieved in acetone . Binary separati ons of erioc hrome black T, indigocarmine from crystal violet, gentian violet, ponceu S, sud an black and rh odam ine B have also been achieved in eth ylmeth y l ketone. The tern ary separati ons of resorci nol bl ue, arni dos wartz, curcu mi n S, erioc hrome black T, bromoph eno l blue, brilliant green, naphthol green B, orange G, indigocarmine - crystal violet - ponceu S, meth y l red and sudan black individually have been ob tain ed in eth y l acetate. Methyl red has been separated from all oth er dyes except ponceu S in hexane. The quarternary separation of crystal vi olet, haemat oxy Ii ne, reso rci nol blue, amidoswartz, curcumin S, eri ochrome blac k T, bromophenol blue, safranin, oran ge G, nap hthol gree n B, eos in, bromothymol blue, indigocarmine, ali za rin Misra el at.: Chromatographic behaviour of some synthetic dyes Articles

Table 2-hRr values of synthetic dyes in various solvent systems on egg shell layers SI. Dyes Mobile Qhases No. A B C D E F G H CrY 77 88 94 98 98 26 00 00 2 GnY 90 88 94 96 97 28 20 00 3 Hm 36 27 10 42 42 17 00 20 4 Bb 66 82 15 95 91 17 6 00 5 La 47 24 8 93 12 36 00 00 6 Rb 00 6 97 00 00 00 00 7 7 FB 87 80 97 98 89 13 10 10 8 AS 00 6 00 00 12 00 00 00 9 CrS 00 00 00 00 00 00 00 5 10 EBT 17 8 00 20 9 00 00 49 11 BrB 12 11 5 10 8 00 00 17 12. BrG 94 76 97 96 77 00 50 00 13 S 95 87 96 93 75 9 00 10 14 BtB 38 25 12 70 78 15 00 8 15 PS 59 45 97 57 85 94 57 00 16 MR 90 96 96 91 97 97 20 49 17 NgB 9 21 00 00 00 00 00 16 18 E 12 6 15 7 9 00 00 7 19 OrG 25 30 00 4 00 6 00 38 20 IC 12 8 00 24 00 00 00 21 21 Rh 87 90 84 92 95 00 8 12 22 MY 85 85 97 90 98 25 II 00 23 ArS 00 00 00 6 8 8 00 00 24 SB 60 40 97 85 76 95 91 00 A-Methanol, B-Ethanol, C-Propanol, D-Acetone, E-Eth ylmethylketone, F-Ethyl acetate, G-Carbon tetrachloride, H-O. I M Sodium chloride

.60 red S - gentiant violet - brilliant green or ponceu S - sudan black and ternary separation of bismark .40 brown, fuschin basic, rhodamine B - brilliant green or ponceu S - sudan black have been achieved in .20 carbon tetrachloride. The binary separation of eriochrome black T and methyl red from crystal violet, gentiant violet, bismark brown, lacmoid, amidoswartz, ponceu S, methyl violet, sudan black, .60 alizarin red S, resorcinol blue, fuschin basic, curcumin S, safranin, bromothymol blue and eosin .40 have actually been achieved in aqueous 0.1 M sodium chloride . . 20 The studies revealed that the polarity index of the solvent has great influence on the mobility of dyes. .. en.t en m '" As the molecular weights of alcohol and ketones iD '" Dyes ------> increase (in mobile phases) the Rr values of most of the dyes increase slightly although some dyes show decrease in Rr values like bromothymol blue. In . . . general, the spots of dyes were more compact in r ::a ... •

435 Articles Indian 1. Chern. Technol., September 2002

Table 3--hRf values of syntheti c dyes on sili ca gel G layers and their possible separati ons in alcohols S.No. Dyes Methanol Ethanol Separati ons (R,--Rd R,--RL hRr R,--RL hRr I CrY 0.0-2.4 12 2. 1-4.0 3 1 In methanol 2 GnV 1.2-2.9 20 2.4-3.6 30 Binary separations 3 Hm 0.0-7.5 37T 0.0- 10.0 50s OrG/BrB/AS/Rb ---- CrV/GnVIFBlBrG/S s 4 Bb 0.0- 10.0 50 0.0- 10.0 50s 5 LA 8.0-10.0 90 8.6-10.0 93 Ternary separations 6 Rb 8. 5-10.0 93 0.0-10.0 50s E/B rB/BtB ---- PS ---- CrV/GnV/MV 7 FB 1.7-2.7 22 4.5-5 .6 51 8 AS 6.7-8.5 76 9.0-10.0 95 In ethanol 9 CrS 0.0-10.0 50s 0.0- 10.0 50s Ternary separations 10 EBT 0.0- 10.0 50s 0.0- 10.0 50s AS/LalBrB/O rGIMRlNgB/E/BtB ---- FB/BrG/S/Rh ---- ArS I I BrB 9.0-10.0 95 8.0-10.0 90 12 BrG 1.4-2.2 18 3.2-4.6 39 13 S 1.7-3.1 24 3.6-4.7 44 14 OrG 9.0- 10.0 95 8.3-9.5 89 15 PS 5. 8-7.4 66 6.7-8.0 74 16 MR 8.2-9.4 88 8. 1-8.9 85 17 NgB 8.0-9.2 86 7. 8-10.0 89 18 E 9.0-10.0 95 9.0- 10.0 95 19 BtB 9.4- 10.0 97 9. 1-10.0 95 s 20 IC 0.0-10.0 50s 0.0- 10.0 50 2 1 Rh 4.5-6.3 54 3.2-7.4 63T 22 MV I. Ci-2.4 17 2.8-4.8 38 23 ArS 0.0-3.0 15 0.0-2.2 II 5 s 24 SB 0.0-8.9 42 0.0-10.0 50 Abbrev iat ions as per Table I. S = Streak; T = Tailing containing one hydroxyl group shows low Rr value Gallic acid (GA), Hydroquinone (Hqn), 2-Naphthol while the dye having two or three benzene rings or (2-Nol), 2-Nitrophenol (2-Nphl), p-nitrophenol di azo groups shows differential behaviour. In some (P-NPhl), Orcinol (001), Phloroglucinol (Phglol), cases adsorption behaviour of the dye on egg shell Picric acid (PA), Pyrogallol (Pol), Resorci nol (Rol), layer differential solubilities of dyes, intermolecular Thymol crystal (Tol) and alpha-Naphthol (alpha-Nol). hydrogen bonding aI1l d steric effect are responsible. A The results are compared in Fig. 1 which shows that large number of dyes show either no movement or the dyes and phenol have similar behaviour. It is well small movement in ethyl acetate and carbon a known fact that some dyes have phenoli c tetrachloride due to their low polarity index. No components. separation is possible in 0.1M sodium nitrate solution All the dyes can be easily extracted from the plates perhaps due to the salt formation. Although few with the help of absolute alcohol and water (9: 1) ratio separations have been achieved in 0.1 M sodium (vlv) or 90% alcohol and can be quantitized after pH chloride solution. adjustment. The method is useful for separating the The studies on calcium carbonate have also been dyes from food stuffs, cosmetics and drugs. can·ied out in hexane to compare the results obtained on pure egg shell layers due to the fact that calcium Acknowledgement carbonate is a maj or component of egg shell layers'B. The fin ancial assistance from UPCST (Luck now), The comparison has also been made with the India is gratefully acknowledged. 17 chromatographic behaviour of phenols • The References fo llowing phenols were taken for the comparison 1 Sherma J & Fried 8 (Eds), Handbook of th ill layer purpose: p-Aminophenol (P-Aph), m-Cresol (m-Crol), chromatography (Marcel Dekker. NY). 1991 & 1996.

436 Misra et al.: Chromatographic behaviour of some synthetic dyes Articles

2 Touchstone JC, Practice of thin layer chromatography, 3rd 10 Srivastava S P, Bhushan R & Chauhan R S, J Liq ed (Wi ley I nterscience, NY), 1992. Chromatogr, 8 (1985), 1255. 3 Sherma J, Anal Chem, 70 (1998) 7R. II Gupta V K, J Liq Chromatogr, 9 (1986) 3489. 4 Sherma J, Anal Chem, 72 (2000) 9R. 12 Gupta V K, Ali I & Joshi A, J ind Chem Soc, 68 (1991) 3 11. 5 Misra A K, Agrawal S K, Kumar S & Rajput RPS, ind J Chern Technol, 5 (1998) 383. 13 Van Peteghem C & Bizl J, J Chromatogr, 210 (1980) 113. 6 Lepri L, Desideri P G & Coas V, J Chromatogr, 16 (1978) 14 Mi sraAK, JLiqChromatogr, 13 ( 1991 ) 1461. 279. 15 Shtykov S N, Sumina, E G, Smushkina EV & Tyunina NV, J 7 Vcnkataraman K. The analytical chemistry of synthetic dyes, Planar Chromatogr, 12 (1999) 129. (A Wiley Interscience Publication, USA), Chap. " (1977) 16 Mohammad A, Naseem KT & Mohammad Nazar PA , J 25-55. Planar Chromatogr, II (1998) 127. 8 Copius-Peereboom J W & Beakes H W, J Chromatogr. 20 (1965) 43. 17 Mohammad A & Iraqi E, J Planar Chromatogr, 12 ( 1999) 9 Srivastava S P, Chauhan L S & Dua VK, J Liq Chromatogr. 288. 3 (1980) 1929. 18 Guiying L & Shanguam Z, Huaxlle Shijia, 33 (1992) 522.