Abstract
BackgroundDNA in herbal decoctions is usually fragmented by extensive boiling and is usually regarded as unsuitable for molecular authentication. This study aims to evaluate the feasibility for molecular authentication methods by multiplex polymerase chain reaction (PCR) and DNA sequencing for decoctions.MethodsThe DNA extraction procedure, sample pulverization and boiling time were examined in (1) single herb decoction with Panax ginseng (“ginseng”) or P. quinquefolius (“American ginseng”), (2) decoctions of two classical ginseng prescriptions of five herbal materials (Aconitum carmichaeli, Atractylodes macrocephala, P. ginseng, Glycyrrhiza uralensis and Zingiber officinale) and (3) a commercial ready-to-serve ginseng soup. Primers were designed from 26S-18S, ITS2 and trnH-psbA region, with DNA sequences obtained from GenBank. Multiplex PCR was also employed in ginseng or American ginseng decoctions to differentiate between these two herbs.ResultsAll six herbal species tested in this study could be identified in decoctions. We had four main observations: (1) sample pulverization before boiling improved PCR detection; (2) prolonged boiling increased the DNA concentration but decreased the intactness of DNA fragments; (3) ginseng could be differentiated from American ginseng by multiplex PCR; (4) identification of individual herbs in multi-herb decoctions with prolonged boiling time of 180 min could be achieved.ConclusionsDNA could be amplified from extensively boiled ginseng decoctions, multi-herb decoctions and commercial soup although DNA degradation was critical to successful PCR.Electronic supplementary materialThe online version of this article (doi:10.1186/s13020-015-0029-x) contains supplementary material, which is available to authorized users.
Highlights
Deoxyribonucleic acid (DNA) in herbal decoctions is usually fragmented by extensive boiling and is usually regarded as unsuitable for molecular authentication
polymerase chain reaction (PCR) products and DNA concentration in single herb decoction We investigated if a 191 bp DNA of the P. ginseng 26S18S intergenic spacer region could be amplified from the single herb P. ginseng decoction prepared by boiling sliced or pulverized samples for 30, 60 and 120 min
With the concentration of the crude and extracted DNA adjusted to that in the original decoction, visible PCR products were found in crude and extracted DNA with samples boiled for 30 min
Summary
DNA in herbal decoctions is usually fragmented by extensive boiling and is usually regarded as unsuitable for molecular authentication. Intentional or inadvertent adulteration of herbal products, including health food and dietary supplements, can cause adverse effects or even poisoning [1,2,3,4,5,6]. The chemical constituents in decoction may be altered by physiological conditions, harvesting period, storage and boiling conditions of the herbs involved. Such methods make differentiation between related species quite difficult owing to their similar chemical profiles and demanding sophisticated instrumentation [14]
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