AOAC 133rd Annual Meeting - Final Program

Scientific Sessions | Monday

SYMPOSIUM: Application of DNA Technologies and Standards in the Authentication of Botanicals for Quality Control of Botanical Dietary Supplements 3:30 PM – 5:00 PM Grand Ballroom 1 Chairs: Yanjun Zhang, Herbalife International of America Inc. Nandakumara Sarma, U.S. Pharmacopeia (USP) Silva Babajanian, Herbalife International of America Inc. 3:40 PM Validation Guidelines for PCR Based Botanical Species Identity Testing to Support Quality Control of Botanical Dietary Supplements Steven Newmaster, University of Guelph PCR-based molecular methods are routinely used to identify food borne pathogens and allergens in food. A survey of the literature indicates that some validation criteria are not addressed when developing PCR tests for botanicals. Here we provide guidelines with examples for validating qualitative real-time PCR methods for identity tests for botanical ingredients. This includes common criteria that underpins the development and validation of rigor- ous tests including 1) Aim of the test, 2) Applicability of different matrix variants, 3) Specificity in identifying the target species ingredient, 4) Sensitivity in detecting the smallest amount of the target material, 5) Repeatability of methods, 6) Reproducibility in detecting the target species in both raw and processed mate- rials, 7) Practicability of the test in a commercial laboratory, and 8) Comparison with alternative methods. In addition, we recommend additional criteria in which the practicability of the test method is evaluated by transferring the method to a second laboratory and a comparison with alternative methods. Properly validated PCR methods can be developed on small, real-time biotechnology that can be placed directly into the supply chain ledger in support of highly transparent data systems that supports quality control from farm to the fork of the consumer. 4:05 PM Developing Species-Specific DNA Testing Botanicals are a fast-growing segment of the dietary supplement industry. Besides being a regulatory requirement according to the dietary supplement GMPs (21 CFR 111), use of scientifically valid methods for establishing identity of botanicals is critical to avoid adulteration or substitution. USP botanical mono- graphs and General Chapter <563> Identification of Articles of Botanical Origin include appropriate methods for botanical identity based on macroscopic, microscopic, chromatographic and DNA barcode methods. Each of these methods has advantages and limitations. The current project examines the development and use of species-specific DNA-based methods as an orthogonal test method to the current compendial specifi- cations based on chromatographic methods (HPLC and HPTLC) Methods to Identify Botanicals Ning Zhang, U.S. Pharmacopeia (USP)

for establishing identify of botanical materials. Unlike the DNA barcoding methods that use universal primers which can give rise to false-positive results due to cross reactions, we have designed species-specific primers for four groups, e.g., ginseng (Asian ginseng, American ginseng and notoginseng), Echinacea ( E. purpurea , E. pallida , and E. angustifolia ), Ginkgo, and black cohosh ( A. racemosa , A. rubra , and A. podocarpa ). The data demonstrates that the optimized amplification conditions using the species-specific primers can differentiate closely related species with each other and can be used for orthogonal testing with other identification test methods. 4:30 PM Industry Experiences on Authentication of Botanical Materials using DNA-based Molecular Analysis Zhangfei Lu, Silva Babajanian, Yanjun Zhang, Peter Chang, Gary Swanson, Herbalife International of America Inc. Due to the diversity and complexity of botanical materials, indus- try botanical material authentication is frequently accomplished by multiple analytical methods. DNA-based molecular analytical methods, which target specific genetic information and provide objective result interpretation, are merged as an ideal orthogo- nal complement to the traditional morphological and chemical botanical authentication methods. In this presentation, we use industry examples to demonstrate how different DNA techniques, such as DNA barcoding, PCR and Next Generation Sequencing, can be utilized to trace and qualify botanical ingredients from starting botanical parts before manufacturing to finished product delivered to end consumers. Based on the types of botanical materials and processing stages, we also discuss the strengths and weaknesses of different DNA techniques in botanical authentication. Furthermore, compared to traditional analytical methods, we present data to show the correlation between results from DNA authentication and compendium chemical analytical methods and the benefit of DNA-based molecular analysis in reducing the risks of misiden- tification when applied to botanicals with limited reference materials and testing experiences. In conclusion, DNA-based molecular analysis has the potential to be applied at different stages of botanical manufacturing processes and is essential to the complete botanical authentica- tion toolbox for quality control of botanical products.

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