AOAC SMPR® 2017.015 Standard Method Performance Requirements

Home , Caftaric acid, Chlorogenic acid

AOAC SMPR® 2017.015 Raw materials.—Fresh, dried, or cut plant material. Recovery.—The fraction or percentage of spiked analyte that is recovered when the test sample is analyzed using the entire method. Standard Method Performance Requirements Repeatability.—Variation arising when all efforts are made (SMPRs®) for Determination of Phenolic Compounds in Dietary Supplements and Dietary Ingredients to keep conditions constant by using the same instrument and Containing Echinacea operator and repeating during a short time period. Expressed as the

repeatability standard deviation (SDr); or % repeatability relative standard deviation (%RSD ). Intended Use: Quality Assurance and Compliance to r Reproducibility.—The standard deviation or relative standard Current Good Manufacturing Practices (CGMPs) deviation calculated from among-laboratory data. Expressed as 1 Purpose the reproducibility standard deviation (SDR); or % reproducibility

AOAC SMPRs describe the minimum recommended relative standard deviation (%RSDR). performance characteristics to be used during the evaluation of a 5 Method Performance Requirements method. The evaluation may be an on-site verification, a single- laboratory validation, or a multi-site collaborative study. SMPRs See Tables 4 and 5. are written and adopted by AOAC stakeholder panels composed of representatives from industry, regulatory organizations, contract 6 System Suitability Tests and/or Analytical Quality Control laboratories, test kit manufacturers, and academic institutions. Suitable methods will include blank check samples, and check AOAC SMPRs are used by AOAC expert review panels in their standards at the lowest point and midrange point of the analytical evaluation of validation study data for method being considered for Performance Tested MethodsSM or AOAC Official Methods of range. A control sample must be included. SM Analysis , and can be used as acceptance criteria for verification 7 Reference Material(s) at user laboratories. 2 Applicability See Tables 2 and 6 for a list and sources of reference and testing materials. Quantitative determination of individual phenolic compounds, including caftaric acid, chlorogenic acid, cichoric acid, cynarin, and Refer to Annex F: Development and Use of In-House Reference echinacoside (Table 1 and Figure 1) in Echinacea angustifolia DC., Materials in Appendix F: Guidelines for Standard Method E. pallida (Nutt.) Nutt., and E. purpurea (L.) Moench (Table 2) Performance Requirements, Official Methods of Analysis of AOAC in raw materials, and dietary ingredients and dietary supplement INTERNATIONAL (2016) 20th Ed., AOAC INTERNATIONAL, products listed in Table 3. Rockville, MD, USA (http://www.eoma.aoac.org/app_f.pdf) 3 Analytical Technique 8 Validation Guidance Any analytical technique that meets the method performance requirements in this SMPR is acceptable. All target analytes and examples of claimed matrixes listed in 4 Definitions Tables 1–3 shall be considered. Data for at least one analyte per Dietary ingredients.—A vitamin; a mineral; an herb or other claimed matrix/plant is acceptable provided that all analytes and botanical; an amino acid; a dietary substance for use by man claimed matrices/plants are represented in the complete evaluation. to supplement the diet by increasing total dietary intake; or a Appendix D: Guidelines for Collaborative Study Procedures to concentrate, metabolite, constituent, extract, or combination of any of the above dietary ingredients. {United States Federal Food Drug Validate Characteristics of a Method of Analysis, Official Methods and Cosmetic Act §201(ff) [U.S.C. 321 (ff)]} of Analysis of AOAC INTERNATIONAL (2016) 20th Ed., AOAC Dietary supplements.—A product intended for ingestion that INTERNATIONAL, Rockville, MD, USA (http://www.eoma.aoac. contains a “dietary ingredient” intended to add further nutritional org/app_d.pdf) value to (supplement) the diet. Dietary supplements may be found Appendix K: Guidelines for Dietary Supplements and Botanicals, in many forms, such as tablets, capsules, softgels, gelcaps, liquids, or powders. Official Methods of Analysis of AOAC INTERNATIONAL (2016) Echinacea angustifolia.—See USP 40 Dietary Supplements, 20th Ed., AOAC INTERNATIONAL, Rockville, MD, USA ® Echinacea 6923; or American Herbal Pharmacopoeia (AHP) (http://www.eoma.aoac.org/app_k.pdf). Also at: J. AOAC Int. 95, Monograph: Echinacea angustifolia root. 268(2016); DOI: 10.5740/jaoacint.11-447 Echinacea pallida.—See USP 40 Dietary Supplements, Echinacea 6931; or AHP Monograph: Echinacea pallida root. 9 Maximum Time-to-Determination Echinacea purpurea.—USP 40 Dietary Supplements, Echinacea 6937; or AHP Monograph: Echinacea purpurea aerial and AHP No maximum time. Monograph: Echinacea purpurea root. Approved by the AOAC Stakeholder Panel on Dietary Limit of quantitation (LOQ).—Minimum concentration or mass of analyte in a given matrix that can be reported as a quantitative Supplements (SPDS) on September 22, 2017. Final Version Date: result. September 22, 2017.

Caftaric acid MW, MW, g/mol 354.311 312.230 474.374 786.733 516.455

6440397 5281764 1794427 5281771 5281769 PubChem ID Chicoric (cichoric) acid

Chlorogenic acid

InChI Key FSBUXLDOLNLABB-ISAKITKMSA-N CWVRJTMFETXNAD-JUHZACGLSA-N YDDGKXBLOXEEMN-IABMMNSOSA-N SWGKAHCIOQPKFW-JTNORFRNSA-N YDDUMTOHNYZQPO-RVXRWRFUSA-N

Echinacoside I04O1DT48T 318ADP12RI 85D81U9JAV FDA UNII No. FDA S4YY3V8YHD WCV7W3174L 9 9 12 20 12 O O O O O 12 18 18 46 24

H H H H H 13 16 22 35 25 Formula C C C C C

1,3-Dicaffeoylquinic CAS No. 6537-80-0 Alternative

30964-13-7 acid (cynarin)

Figure 1. Molecular structures. CAS No. 327-97-9 84696-11-7 67879-58-7 70831-56-0 82854-37-3 97281-15-7 90028-20-9 212891-05-9 extract extract extract Caftaric acid Echinacoside Echinacea pallida Chlorogenic acid Echinacea purpurea Echinacea angustifolia Reference standard Chicoric (cichoric) acid 1,3-Dicaffeoylquinic acid (cynarin) 1,3-Dicaffeoylquinic Polyphenols Powdered Powdered Powdered 1. Table No. 1 2 3 4 5 6 7 8

© 2017 AOAC INTERNATIONAL Table 2. Echinacea Table 5. Method performance requirements as a function of range Source Echinacea CAS No. Form (ID No.) Range, % Echinacea angustifolia 84696-11-7 Powdered USP Parameter 0.01–1 >1 extract (1231706) Recovery, % 90–107 95–105 Root Alkemist

Labs RSDr, % ≤6 ≤3

Herb Alkemist RSDR, % ≤8 ≤4 Labs Echinacea pallida 97281-15-7 Powdered USP extract (1231717) Alkemist Root Labs Echinacea purpurea 90028-20-9 Powdered USP extract (1231728) Aerial Alkemist Table 6. Sources of phenolic compound reference materials parts Labs CAS No. Root Alkemist Polyphenol (alternative) Source Product No. Labs Caftaric acid 67879-58-7 Chromadex ASB-00003028 Alkemist Seed Labs PhytoLab 89170 Sigma 15029

USP 1086039

Chicoric (cichoric) acid 70831-56-0 Chromadex ASB-00003640 (6537-80-0) Extrasynthese 4987S Table 3. Examples of dietary ingredient and dietary supplement matrixes PhytoLab 89177 Ingredients Sigma C7243 Extracts (including encapsulated) USP 1105315 Herb powders Chlorogenic acid 327-97-9 Acro 109240000 Juice Alfa Asean J60457 Supplements Chromadex ASB-00003450 Powders Extrasynthese 4991S Tablets LKT Labs C2943 Gummies MP Biomedicals 0215061801 Liquids PhytoLabs 89175 Capsules Sigma C3878 Softgel capsules USP 1115545 Tinctures Echinacoside 82854-37-3 Chromadex ASB-00005020 Gelcaps Extrasynthese 4988S Chewables LKT Labs E0929 Juice PhytoLab 89188

Sigma 01710580

USP 1231750

1,3-Dicaffeoylquinic acid 212891-05-9 Chromadex ASB-00003990 (cynarin) (30964-13-7) Table 4. LOQ and analytical range Extrasynthese 4995S Parameter PhytoLab 89179 Analytical range, % 0.01 to 5 Sigma 91801 LOQ, % ≤0.01 USP