Development and validation of DNA sequence-based methods of medicinal herb specie

DESCRIPTION (provided by applicant): In this proposal, we outline an innovative approach to accomplish at least two of NCCAMs priorities: validating DNA methods for botanical identification and developing complex DNA-authenticated botanical identity standard reference materials (BI-SRMs); this will have significant impact on the medicinal herb industry by helping to ensure the quality, safety, and efficacy of botanical medicines. There are many advantages to using DNA for botanical authentication: it can beused to identify any organism from a variety of material types, including raw, powder, and select processed products; it is extremely sensitive and powerful and can identify material from even a few molecules; it has ability to discriminate between closely related species, hybrids, and even lineages within species; and has the ability to identify mixtures of more than one primary or contaminating species, including toxic or chemically inert adulterants or additives. In Phase 1 we have two specific aims, orobjectives. In Objective 1, we will conduct a pre- validation study by identifying the two best candidate DNA regions for authentication of each of the following five species: licorice (Glycyrrhiza uralensis), goldenseal (Hydrastis canadensis), ginseng (Panax quinquefolius), rhubarb (Rheum palmatum), and skullcap (Scutellaria lateriflora); these five species were chosen because are widely used in herbal medicine, have known issues of adulteration, and taxonomically confusing, and could benefit from havingan alternative form of botanical identification. In Objective 2, we will conduc full single laboratory validation studies testing the specificity and performance for two gene regions for each of the five species on pure and impure samples following AOAC Internationals guidelines for qualitative method validation. This involves testing a total of 200 specimens representing the five species and their closely related adulterants, as well as 250 mixed samples, and calculating the Probability of Identification(POI). Through the products and services resulting from the research outlined in this proposal, we aim to bring DNA-authentication to the forefront of the medicinal herb industry. PUBLIC HEALTH RELEVANCE: In this proposal we will complete the first full-validation studies on DNA methods of identification for five commonly used medicinal herbs: ginseng, goldenseal, licorice, rhubarb, and skullcap. Through the data generated here, we aim to bring DNA methods to the forefront of medicinal herb authentication, by allowing us to offer not only the first validated DNA testing services to the public, but also develop the first DNA- authenticated complex botanical identity standard reference materials to use for in-house testing and validation studies of alternative methods. These products and services will undoubtedly help in ensuring the safety, quality, and efficacy of medicinal herb products.