Database-aided UHPLC-Q-orbitrap MS/MS strategy putatively identifies 52 compounds from Wushicha Granule to propose anti-counterfeiting quality-markers for pharmacopoeia

Xican Li; Shaoman Chen; Jingyuan Zeng; Rongxin Cai; Yilan Liang; Chuanbin Chen; Ban Chen; Chunhou Li

doi:10.1186/s13020-023-00829-2

Database-aided UHPLC-Q-orbitrap MS/MS strategy putatively identifies 52 compounds from Wushicha Granule to propose anti-counterfeiting quality-markers for pharmacopoeia

Chin Med. 2023 Sep 9;18(1):116. doi: 10.1186/s13020-023-00829-2.

Authors

Xican Li¹, Shaoman Chen², Jingyuan Zeng², Rongxin Cai^{2

3}, Yilan Liang², Chuanbin Chen², Ban Chen⁴, Chunhou Li²

Affiliations

¹ School of Chinese Herbal Medicines, Guangzhou University of Chinese Medicine, Guangzhou, 510006, China. lixican@126.com.
² School of Chinese Herbal Medicines, Guangzhou University of Chinese Medicine, Guangzhou, 510006, China.
³ College of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu, 611137, China.
⁴ Key Laboratory of Fermentation Engineering (Ministry of Education), Cooperative Innovation Center of Industrial Fermentation (Ministry of Education and Hubei Province), Hubei University of Technology, Wuhan, 430068, China.

Abstract

Wushicha Granule, an over-the-counter-drug (OTC) prescription, consists of 19 traditional Chinese herbals medicines (CHMs), such as Chaihu, Hongcha, Chuanxiong, Houpo, and Gancao. The five however have not been effectively characterized by the quality-markers (Q-markers) system in current Pharmacopoeia. The study therefore established a novel database-aided ultra-high performance liquid chromatography-quadrupole-orbitrap mass spectrometry (UHPLC-Q-orbitrap MS/MS) strategy. The strategy has putatively identified 52 compounds from Wushicha Granule, mainly including flavonoids, saponins, alkaloid, lignins, and lactones. Especially, saponin "glycyrrhetinic acid" in the Granule was specifically identified as 18β-configuration (rather than 18α-configuration). Meanwhile, two pairs of isomers were fully discriminated, including vitexin vs isovitexin and daidzein vs 7,4'-dihydroxyflavone. 8β-Glycyrrhetinic acid, together with saponin saikosaponin A, alkaloid caffeine, lactone S-senkyunolide A, and lignin magnolol, were further studied using quantum chemical calculation, UV-vis spectra, and anti-counterfeiting validation experiment. In the validation experiment, they have successfully recognized 6 counterfeit Wushicha Granules, by means of a LC-MS equipped extraction software. Based on these results, 8β-glycyrrhetinic acid is recommended to replace the old Q-marker "glycyrrhetinic acid"; while saikosaponin A, caffeine, S-senkyunolide A, and magnolol are recommended as new Q-markers. These recommendations can not only recognize the counterfeits regarding Chaihu, Hongcha, Chuanxiong, Houpo, and Gancao, but also prevent the possible safety-incident. All these will greatly improve the efficiency and specificity of current Pharmacopoeia.

Keywords: Adulteration; Database UPLC-Q-orbitrap MS; Glycyrrhetinic acid; Quality-control; Quality-marker; Wushi Tea.