Comparison of Flow Injection MS, NMR, and DNA Sequencing: Methods for Identification and Authentication of Black Cohosh (Actaea racemosa)

Abstract

Flow injection mass spectrometry and proton nuclear magnetic resonance spectrometry, two metabolic fingerprinting methods, and DNA sequencing were used to identify and authenticate Actaea species. Initially, samples of Actaea racemosa from a single source were distinguished from other Actaea species based on principal component analysis and soft independent modeling of class analogies of flow injection mass spectrometry and proton nuclear magnetic resonance spectrometry metabolic fingerprints. The chemometric results for flow injection mass spectrometry and proton nuclear magnetic resonance spectrometry 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.

Profiles of Phenolic Acids and Triterpene Glycosides in Commercial and Cultivated Black Cohosh

“…Our findings support the results of Harnly et al [32]. The authors investigated black cohosh samples from different sources, next to other Actaea spp., qualitatively by NMR and flow injection mass spectrometry (FIMS).…”

DNA Barcoding for the Identification of Botanicals in Herbal Medicine and Dietary Supplements: Strengths and Limitations

“…However, with slight modifications in the DNA extraction protocol, it is sometimes possible to obtain useful DNA as a template for PCR from a powder, capsules, or tablets [51]. In contrast to this, the processes by which tinctures and extracts are made, which can include extensive heat treatment, filtration, extractive distillation, or supercritical fluid extraction [52], thus degrading or removing the plant DNA completely, often make these materials unsuitable for DNA barcoding analysis [53].…”

Selection and characterization of botanical natural products for research studies: a NaPDI center recommended approach

“…However, it is important to note that the "ngerprint" of which metabolites are present differs depending on the extraction procedure as well as analytical technique used to collect the data and the parameters of the specic analysis. 68 Thus, it is necessary to include authenticated reference material for comparison in the same analysis with the sample under question and processed by the same extraction procedure and solvent system (see Purchasing authenticated reference materials, Section 5.5) ( Fig. 2).…”