SMPR 2018.XXX; Version 2; January 25, 2018
1 DRAFT AOAC SMPR 2018.XXX; Version 2; January 25, 2018. 2 3 Method Name: Limit Test for Determination of Selected Compounds from Teucrium 4 spp. in Skullcap Materials in Commerce 5 6 Approved by: Stakeholder Panel on Dietary Supplements (SPDS) 7 8 Intended Use: For quality assurance and compliance to current good manufacturing practices. 9 10 1. Purpose 11 12 AOAC SMPRs describe the minimum recommended performance characteristics to be used 13 during the evaluation of a method. The evaluation may be an on-site verification, a single- 14 laboratory validation, or a multi-site collaborative study. SMPRs are written and adopted by 15 AOAC Stakeholder Panels composed of representatives from the industry, regulatory 16 organizations, contract laboratories, test kit manufacturers, and academic institutions. 17 AOAC SMPRs are used by AOAC Expert Review Panels in their evaluation of validation study 18 data for method being considered for Performance Tested Methods or AOAC Official 19 Methods of Analysis, and can be used as acceptance criteria for verification at user 20 laboratories. [Refer to Appendix F: Guidelines for Standard Method Performance 21 Requirements, Official Methods of Analysis of AOAC INTERNATIONAL (2012) 20th Ed., AOAC 22 INTERNATIONAL, Gaithersburg, MD, USA.] 23 24 2. Applicability: 25 Detection of Teucrin A and/or Teucrioside, and/or Teuflin, and/or Verbascoside (Acteoside, 26 Kusaginin, Russetinol, Stereospermin) in raw materials and dietary ingredients labelled as 27 skullcap (Scutellaria lateriflora) to ensure that raw materials and dietary ingredients labelled 28 as skullcap do not contain compounds indicative of the presence of Teucrium. 29 30 See Table 2 for additional information on analytes and figure 1 for molecular structures. 31 32 3. Analytical Technique: 33 Any analytical technique that meets the following method performance requirements is 34 acceptable. 35 36 4. Definitions: 37 38 Analytical range – Includes all steps of the analytical procedure including sample 39 preparation and further dilutions. 40 41 Dietary ingredient.— A vitamin; a mineral; an herb or other botanical; an amino acid; a 42 dietary substance for use by man to supplement the diet by increasing total dietary intake; 43 or a concentrate, metabolite, constituent, extract, or combination of any of the above 44 dietary ingredients. {United States Federal Food Drug and Cosmetic Act §201(ff) [U.S.C. 321 45 (ff)]} 46 47 Dietary supplement.— A product intended for ingestion that contains a “dietary ingredient” 48 intended to add further nutritional value to (supplement) the diet. Dietary supplements may 49 be found in many forms such as tablets, capsules, softgels, gelcaps, liquids, or powders. 50 51 Limit of Quantitation (LOQ) .— The minimum concentration or mass of analyte in a given 52 matrix that can be reported as a quantitative result 53 54 Plant Materials – Fresh, dried or cut plant materials 55 56 Repeatability .— Variation arising when all efforts are made to keep conditions constant by 57 using the same instrument and operator and repeating during a short time period. 58 Expressed as the repeatability standard deviation (SDr); or % repeatability relative standard 59 deviation (%RSDr). 60 61 Reproducibility.— The standard deviation or relative standard deviation calculated from 62 among-laboratory data. Expressed as the reproducibility relative standard deviation (SDR); or 63 % reproducibility relative standard deviation (% RSDR). 64 65 Recovery.— The fraction or percentage of spiked analyte that is recovered when the test 66 sample is analyzed using the entire method. 67 68 Skullcap.- For the purposes of this SMPR, the term skullcap refers to the species Scutellaria 69 lateriflora. 70 71 72 5. Method Performance Requirements: 73 74 See table 3 and 4. 75 76 6. System suitability tests and/or analytical quality control: 77 Suitable methods will include blank check samples, and check standards at the lowest point 78 and midrange point of the analytical range. A control sample must be included. 79 80 7. Reference Material(s): 81 82 See table 5 for sources of flavonoids reference materials. See table 6 for sources of 83 Scutellaria and Teucrium species. 84 85 Refer to Annex F: Development and Use of In-House Reference Materials in Appendix F: 86 Guidelines for Standard Method Performance Requirements, 19th Edition of the AOAC 87 INTERNATIONAL Official Methods of Analysis (2012). Available at: 88 http://www.eoma.aoac.org/app_f.pdf 89 90 8. Validation Guidance: 91 92 Appendix D: Guidelines for Collaborative Study Procedures To Validate Characteristics of a 93 Method of Analysis; 19th Edition of the AOAC INTERNATIONAL Official Methods of Analysis 94 (2012). Available at: http://www.eoma.aoac.org/app_d.pdf 95 96 Appendix K: Guidelines for Dietary Supplements and Botanicals 19th Edition of the AOAC 97 INTERNATIONAL Official Methods of Analysis (2012). Also at: . AOAC Int. 95, 268(2012); DOI: 98 10.5740/jaoacint.11-447 and available at: http://www.eoma.aoac.org/app_k.pdf 99 100 9. Maximum Time-To-Determination: No maximum time. 101 102 103 104 105 106 Table 1: Examples of Plant Material, Dietary Supplements and Dietary Ingredients 107 108 Dried plant material 109 Liquid extracts (including tinctures) 110 Dry extracts 111 Tablets 112 Capsules 113 Multi-botanical finished product 114 115 116 117 Table 2: Additional Information on Select Flavanoids 118 Compound Name CAS # UNII InChI Key PubChem
5,6,7-Trihydroxy-2-phenyl-4H-1- FXNFHKRTJBSTCS-
Baicalein (Noroxylin) 491-67-8 benzopyran-4-one 49QAH60606 UHFFFAOYSA-N 5281605
5,6-Dihydroxy-4-oxo-2-phenyl-4H-1- Baicalin (Baicalein-7-O- IKIIZLYTISPENI-
benzopyran-7-yl β-D- 21967-41-9 glucuronide) 347Q89U4M5 ZFORQUDYSA-N 64982 glucopyranosiduronic acid
Chrysin (5,7- 5,7-Dihydroxy-2-phenyl-4H-1-benzopyran- RTIXKCRFFJGDFG-
480-40-0 Dihydroxyflavone) 4-one 3CN01F5ZJ5 UHFFFAOYSA-N 5281607
Dihydrobaicalin (5,6,7- (2S)-3,4-Dihydro-5,6-dihydroxy-4-oxo-2- UVNUGBQJLDGZKE-
Trihydroxyflavanone 7-O-β- phenyl-2H-1-benzopyran-7-yl β-D- 56226-98-3 NA XDZPIWCFSA-N 14135325 D-glucuronide) glucopyranosiduronic acid
5,6-Dihydroxy-2-(2-hydroxyphenyl)-4-oxo- ARCCSELFQKSKDR-
Ikonnikoside I 4H-1-benzopyran-7-yl β-D- 119152-49-7 NA ZFORQUDYSA-N 10183148 glucopyranosiduronic acid
Lateriflorein (5,6,7- 5,6,7-Trihydroxy-2-(2-methoxyphenyl)- OHWUYMZEFLQFQB- Trihydroxy-2'- 1329-06-2 NA 10913608 4H-1-benzopyran-4-one UHFFFAOYSA-N methoxyflavone)
Lateriflorin (Lateriflorein 7- 5,6-Dihydroxy-2-(2-methoxyphenyl)-4- 521066-22-8 NA NA NA O-β-D-glucuronide) oxo-4H-1-benzopyran-7-yl
119 120 Table 2: Additional Information on Select Flavanoids (cont’d.) Compound Name CAS # UNII InChI Key PubChem Oroxylin A 7-O-β-D- 5-Hydroxy-6-methoxy-4-oxo-2-phenyl-4H- glucuronide (6- QXIPXNZUEQYPLZ-
1-benzopyran-7-yl β-D- 36948-76-2 Methoxybaicalein 7-O-β-D- O84RM2NAEQ QSUZLTIMSA-N 14655552 glucopyranosiduronic acid glucuronide)
Scutellarein (6- 5,6,7-Trihydroxy-2-(4-hydroxyphenyl)-4H- JVXZRQGOGOXCEC-
529-53-3 Hydroxyapigenin) 1-benzopyran-4-one P460GTI853 UHFFFAOYSA-N 5281697
5,6-Dihydroxy-2-(4-hydroxyphenyl)-4-oxo- Scutellarin (Scutellarein 7-O- 27740-01-8 DJSISFGPUUYILV-
4H-1-benzopyran-7-yl β-D- β-D-glucuronide) (1329-06-2) 16IGP0ML9A ZFORQUDYSA-N 185617 glucopyranosiduronic acid
(3S,5S,5'aS,7'S,8'S,8'aR)- 5-(3-Furanyl)-3', 4,5,5',5'a,7',8',8'a-octahydro-8'-hydroxy- AONLJCCUYGGOSW-
Teucrin A 12798-51-5 NA 7'-methyl-spiro[furan-3(2H),6'-[6H] PJERILTQSA-N 159529 naphtho[1,8-bc]furan]-2,2'(4'H)-dione (E)-4-[3-(3,4-Dihydroxyphenyl)-2- propenoate]-2-(3,4-dihydroxyphenyl)ethyl UDHCHDJLZGYDDM-
Teucrioside O-α-L-lyxopyranosyl-(1→2)-O-6-deoxy-α- 115872-98-5 NA JPCQEIDESA-N 14034194 L-mannopyranosyl-(1→3)-β-D- glucopyranoside (E)-4-[3-(3,4-Dihydroxyphenyl)-2- Verbascoside (Acteoside, propenoate]-2-(3,4-dihydroxyphenyl)ethyl FBSKJMQYURKNSU-
Kusaginin, Russetinol, 61276-17-3 O-α-rhamnopyranosyl(1→3)-4-O-caffeoyl- 3TGX09BD5B ZLSOWSIRSA-N 5281800 Stereospermin) β-D glucopyranoside
5,7-Dihydroxy-8-methoxy-2-phenyl-4H-1- XLTFNNCXVBYBSX-
Wogonin 632-85-9 benzopyran-4-one POK93PO28W UHFFFAOYSA-N 5281703
Wogonoside (Oroxinidin, 5-Hydroxy-8-methoxy-4-oxo-2-phenyl-4H- LNOHXHDWGCMVCO-
Glychionide B, Wogonin 7-O- 1-benzopyran-7-yl β-D- 51059-44-0 ETX4944Z3R NTKSAMNMSA-N 3084961 β-D-glucuronide) glucopyranosiduronic acid 121 122 Table 3: LOD 123 Parameter Minimum Requirement LOD 10 ppm
124 125 126 Table 4: Method Performance Requirements 127
Minimum Requirement Parameter (at detection limit)
% Recovery 80 - 110
RSDr 7
RSDR 11 128 129 130 131 132 133 134 135 Table 5: Sources of Selected Flavanoids 136 Compound Phytolab Extrasynthese Sigma EP USP
Baicalein (Noroxylin) 89577 1171 92081 1048357
Baicalin (Baicalein-7-O-glucuronide) 89322 1280 S 94121 Y0001273 1048368
Chrysin (5,7-Dihydroxyflavone) 80382 1362 S 95082 NA NA
Dihydrobaicalin (5,6,7-Trihydroxyflavanone 7-O-β- NA NA NA NA NA D-glucuronide)
Ikonnikoside I NA NA NA NA NA
Lateriflorein (5,6,7-Trihydroxy-2'-methoxyflavone) NA NA NA NA NA
Lateriflorin (Lateriflorein 7-O-β-D-glucuronide) NA NA NA NA NA
Oroxylin A 7-O-β-D-glucuronide (6- NA NA NA NA NA Methoxybaicalein 7-O-β-D-glucuronide)
Scutellarein (6-Hydroxyapigenin) 83283 1334 S S0327 NA NA
Scutellarin (Scutellarein 7-O-β-D-glucuronide) 89788 NA 73577 NA NA
Teucrin A 89285 NA NA NA NA
NA NA NA NA NA Teucrioside
Verbascoside (Acteoside, Kusaginin, Russetinol, NA Stereospermin) 89289 4994 S v4015 1711455
Wogonin 89825 1304 S 681670 NA NA
Wogonoside (Oroxinidin, Glychionide B, Wogonin NA NA NA 7-O-β-D-glucuronide) 82684 SMB00081 137 138 139 Table 6: Sources of Scutellaria species. 140 141 Skullcap Aerial Parts: American Herbal Pharmacopoeia, Product Code 524. 142 Scutellaria baicalensis, root. American Herbal Pharmacopoeia, Product Code 563474. 143 Scutellaria lateriflora, herb. Alkemist Labs, Product Code 942495. 144 Scutellaria lateriflora, herb. American Herbal Pharmacopoeia, Product Code 533097. 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 Figure 1: Molecular Structures of Selected Flavanoids 164 165 166 O OH O OH
S OH S OH HO S S HO S S
O R O R S OH S OH
O O HO O O O
S
HO HO HO
OH O OH O OH O
Baicalin Baicalein Dihydrobaicalin O OH
S OH HO S S
R O H3CO H3CO S OH
O O HO O
HO HO
OH O OH O
Lateriflorin Lateriflorein
O OCH3 OCH3 O O O HO O
HO S S
S R S HO OH
OH OH O OH O
167 Wogonoside Wogonin 168 169 Figure 1: Cont’d 170 171 O OH
S OH HO S S
R H3CO O O S OH
O O O HO O O O
HO S S
S R S HO OH H3CO OH OH O OH O OH O
2'-Methoxy-chrysin 7-O-Glucoronide Chrysin Oroxylin A 7-O-Glucoronide
O OH O OH
OH OH S S HO S S HO S S
O R OH OH O R HO S OH S OH
O O HO O O O
HO HO HO
OH O OH O OH O
172 Scutellarin Scutellarein Ikonnikoside I