Molecular quantification, a new strategy for quality control of Chinese patent medicine containing animal-derived crude drug: Qi She in Jinlong capsule as an example
Authors: Haoting Ye
Fang Li
Chan Li
Zhi Chao
Enwei Tian
Yongshan Liang
Xuena Xie
Journal: Journal of Pharmaceutical and Biomedical Analysis (2022)
Discipline: PHARMACOLOGY & TOXICOLOGY
Keywords: Chinese patent medicine; Deinagkistrodon acutus; Jinlong capsule; Molecular quantification; Qi She
Affiliations: Department of Pharmacy, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China; Faculty of Medicinal Plants and Pharmacognosy, School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, China., Faculty of Medicinal Plants and Pharmacognosy, School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, China., Faculty of Medicinal Plants and Pharmacognosy, School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, China; Department of Pharmacy, Guangdong Second Provincial General Hospital, Guangdong Provincial Emergency Hospital, Guangzhou, China., Department of Pharmacy, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China; Faculty of Medicinal Plants and Pharmacognosy, School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, China; Guangdong Provincial Key Laboratory of Chinese Medicine Pharmaceutics, Guangzhou 510515, China. Electronic address: .

Abstract:  Quality control for Chinese patent medicine (CPM) containing animal-derived crude drug(s) is rather difficult. The methods based on chemical composition analysis, which are commonly used in CPM consisted of plant-derived crude drugs, are often not applicable for CPM containing animal-derived crude drug, because the effective constituents of most animal-derived crude drugs remain unknown. Even if there are such methods, they are usually qualitative rather than quantitative, and the specificity is generally poor. Here we proposed a molecular quantification method for CPM containing animal-derived crude drug, based upon the hypothesis that the amount of remnant DNA fragments could reflect feeding quantity of the crude drugs and thus ensure the quality of the CPM. Take Jinlong capsule [a hepatocellular carcinoma-resisting Chinese patent medicine comprising of three fresh animal drugs, i.e. Shougong (Peking gecko, Gekko swinhonis), Qi She (sharp-snouted pitviper, Deinagkistrodon acutus), and Jinqian Baihua She (many-banded krait, Bungarus multicinctus)] as an example, we established a qPCR assay for Qi She in the capsule, which verified the feasibility of the quality control method based on molecular quantification. Species-specific primers and TaqMan probe for Qi She were designed, and the qPCR assay system was then established. The assay exhibited a good specificity; there's a good linearity between Ct values and logarithm of the target amplicon copy numbers within the range of 8.8 × 101 to 8.8 × 106 copies/μL, and the limit of detection was 88 copies/μL. The method was validated through reproducibility, stability assessment. Recovery of spiked samples was between 91.59% and 101.69%. It was verified that the copy numbers reflected the original feeding amount of an animal-derived crude drug by self-made Jinlong capsules. The assay was successfully applied in Qi She-specific amplicon determination in 20 batches of Jinlong capsule. The study was expected to provide a new strategy for quality control of CPM containing animal-derived crude drug. 

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PubPeer comments:
 ## 4. Discussion There are various testing methods for quality control of Chinese patent medicines [11], [29], [30]. References 29. M.Q. Zhang, C. Zhao, Q.J. Shao, Z.D. Yang, X.F. Zhang, X.F. Xu, M. Hassan Determination of water content in corn stover silage using near-infrared spectroscopy Int. J. Agric. Biol. Eng., 12 (2019), pp. 143-148 It is puzzling to describe the subject of reference 29 as such. ,