Quantification of concentrated Chinese medicine granules by quantitative polymerase chain reaction

• Published in: Journal of pharmaceutical and biomedical analysis 2017-10, Vol.145, p.661-665
• Main Authors: Lo, Yat-Tung, Shaw, Pang-Chui
• Format: Article
• Language: English
• Subjects: Animal-derived drug; Concentrated Chinese medicine granules; DNA; DNA authentication; Medicine, East Asian Traditional; Polymerase Chain Reaction; qPCR; Species Specificity
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2017.07.047

[Display omitted] •DNA extracts from 5 to 50mg CCMG were preferred for qPCR.•Target DNA fragments in CCMG were amplified with short amplicon length.•Reproducible results were obtained for CCMG from the same company.•Constitute amount of CCMG in a mixture was determined by qPCR. Determination of the amount of constituent in a multi-herb product is important for quality control. In concentrated Chinese medicine granules (CCMG), no dregs are left after dissolution of the CCMG. This study is the first to examine the feasibility of using quantitative polymerase chain reaction (qPCR) to find the amount of CCMG in solution form. DNA was extracted from Hirudo and Zaocys CCMG mixed at different ratios and amplified in qPCR using species-specific primers. The threshold cycle (CT) obtained was compared with the respective standard curves. Results showed that reproducible quantification results could be obtained (1) for 5–50mg CCMG using a modified DNA extraction protocol, (2) amongst DNA extracted from the same batch of CCMG and (3) amongst different batches of CCMG from the same company. This study demonstrated the constitute amount of CCMG in a mixture could be determined using qPCR. This work has extended the application of DNA techniques for the quantification of herbal products and this approach may be developed for quality assurance in the CCMG industry.