Discovery of Quality Markers in Hugan Qingzhi Formula by Integrating a Lipid‐Lowering Bioassay with UHPLC‐QQQ‐MS/MS

Abstract

Nonalcoholic fatty liver disease (NAFLD) is a prevalent chronic liver disease. The Hugan Qingzhi formula (HGQZ) has been proven effective in treating NAFLD through clinical and pharmacological mechanism studies. A screening study of the chemical components was carried out to better control the quality of this formula. Current research has combined biological activity assessment with chemical analysis to screen and identify the bioactive compounds in HGQZ for use as potential quality markers (Q-markers) to control the quality of this herbal product. The HGQZ extracted by three different solvents was evaluated in a free fatty acid-induced hepatic steatosis LO2 cell model. Simultaneously, the twelve major chemical constituents of these extracts were quantitatively measured by ultrahigh-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC-QQQ-MS/MS). Extraction with 50% ethanol showed the most potent lipid-lowering effect in steatosis LO2 cells and the highest extraction rate of major chemical constituents. Correlation analysis was used to establish the relationship between the biological activities and chemical characteristics of these extracts. The results showed that the contents of typhaneoside, hyperoside, isoquercitrin, isorhamnetin-3-O-neohesperidoside, notoginsenoside R1, and alisol B 23-acetate were positively correlated to the lipid-lowering effect. The subsequent bioassay confirmed that typhaneoside, isoquercitrin, and alisol B 23-acetate played the role of reducing the lipid effect. In conclusion, 50% of ethanol extraction produced the most active extract of HGQZ. Typhaneoside, isoquercitrin, and alisol B 23-acetate could be considered potential Q-markers for the quality control of HGQZ.

Pharmacological Properties and Molecular Targets of Alisol Triterpenoids from Alismatis Rhizoma

“…Ali-B and Ali-B 23-acetate are abundant in the plant roots, but also in the leaves, scapes, and inflorescences [ 138 ]. Ali-A is one of the quality markers which can be used to assess the quality of the TCM prescription Qiliqiangxin capsule (QLQX), used to treat chronic heart failure [ 18 ], whereas Ali-B 23-acetate is a quality marker for the Hugan Qingzhi formula used to treat nonalcoholic fatty liver disease [ 12 ]. However, the bioactivity not only depends on the quality of the phyto-preparation and the alisol content, but also on the respective proportions of the bioactive molecules, notably the ratio of Ali-A 24-acetate, Ali-B, and Ali-B 23-acetate [ 139 ].…”

“…Alismatis rhizoma is listed in the 2020 edition of the Chinese Pharmacopoeia, with specific recommendations on quality standards [ 8 ]. Those rhizoma are included in several TCMs, such as Ye Tianshi used for the treatment of dampness and heat [ 9 ], Zexie Tang used to treat metabolic diseases and liver injuries [ 10 ], Liuwei Dihuang decoction used to improve kidney yin deficiency [ 11 ], Hugan Qingzhi formula which is used to treat chronic liver diseases [ 12 ], Gandou decoction used to treat Wilson’s disease [ 13 , 14 ], and Dangguishaoyao-san (also called Toki-shakuyaku-san or TJ-23) used to reduce the symptoms of Alzheimer’s disease [ 15 , 16 ]. Alismatis rhizoma is also present in different TCM prescriptions, such as Qi-Li-Qiang-Xin capsule used for the treatment of chronic heart failure [ 17 , 18 ] and Qing-Fei-Pai-Du which is a lung cleansing and detoxifying decoction used historically for the treatment of exogenous fever, and recently proposed to reduce symptoms associated with coronavirus disease 2019 (COVID-19) [ 19 ].…”

Discovery of quality control ingredients in burdock root by combining anti‐tumor effects and UHPLC–QqQ–MS/MS

“…Therefore, the selection of quality control indicators is essential. These selected ingredients can achieve the general purpose of controlling the quality and efficacy of Chinese medicines (He et al, 2017a; He et al, 2020; Huang et al, 2020; Wang et al, 2020). At present, there is no pharmacopeia standard for burdock root, which lacks quality control.…”