|Harnly, James - Jim|
|Huang, Huilian - Jiangxi University Of Traditional Chinese Medicine|
|Colson, Kimberly - Bruker|
|Yu, Jimmy - Bruker|
|Mccoy, Joe-ann - North Carolina Arboretum|
|Harbaugh Reynaud, Danica - Authen Technologies|
|Harrington, Peter - Ohio University|
|Fletcher, Edward - Strategic Sourcing, Inc|
Submitted to: Planta Medica
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/3/2015
Publication Date: 12/21/2015
Citation: Harnly, J.M., Chen, P., Sun, J., Huang, H., Colson, K., Yu, J., Mccoy, J.H., Harbaugh Reynaud, D.T., Harrington, P.B., Fletcher, E. 2015. Comparison of Flow Injection MS, NMR, and DNA Sequencing: Methods for Identification and Authentication of Black Cohosh (Actaea racemosa). Planta Medica. 82:250-262.
Interpretive Summary: Plants used for botanical supplements have a complex chemical make-up. In this study, we examined the raw plants and commercial supplements of black cohosh (Actaea racemosa) and 4 other species of the Actaea genus using chemical fingerpringing (mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectrometry) and DNA sequencing. Authentic plant materials were collected from 4 reliable sources (American herbal Pharmacopoeia, North Carolina Arboretum, Strategic Sourcing, and National Institutes of Science and Technology). All 3 methods (MS, NMR, and DNA) can distinguish between the Actaea species. MS and NMR showed a considerable variation between the authentic materials suggesting location, harvesting, and/or processing differences. In every case, the commercial supplements were distinguishable from the raw plants, i.e. commercial processing produced a chemically different material. DNA sequencing was not successful in identifying the species of the commercial supplements due to the lack of quality DNA. Most commercial processing removes the DNA during the product preparation. This data shows that chemical fingerprinting (MS and NMR) and DNA sequencing are complimentary methods that can elucidate species differences.
Technical Abstract: Flow injection mass spectrometry (FIMS) and proton nuclear magnetic resonance spectrometry (1H-NMR), two metabolic fingerprinting methods, and DNA sequencing were used to identify and authenticate Actaea species. Initially, samples of Actaea racemosa L. from a single source were distinguished from other Actaea species based on principal component analysis (PCA) and soft independent modeling of class analogies (SIMCA) of FIMS and 1H-NMR metabolic fingerprints. The chemometric results for FIMS and 1H-NMR agreed well and showed similar agreement throughout the study. DNA sequencing using DNA sequence-data from two independent gene regions confirmed the metabolic fingerprinting results. Differences were observed between A. racemosa samples from four different sources although the variance within species was still significantly less than the variance between species. A model based on the combined A. racemosa samples from the four sources consistently permitted distinction between species. Additionally, the combined A. racemosa samples were distinguishable from commercial root samples and from commercial supplements in tablet, capsule, or liquid form. DNA sequencing verified the lack of authenticity of the commercial roots but was unsuccessful in characterizing many of the supplements due to the lack of available DNA.