Establishment of a new approach for quality evaluation of Pseudobulbus Cremastrae seu Pleiones (Shancigu) based on multicomponent analysis and anti-liver cancer pharmacological effect

Yuxin CaoZhuangzhuang HaoMengmeng LiuJingwen XueYuqing WangTingyue JiangGe ZhangWenxin FanChunguo WangJinli Shi^*

• Beijing University of Chinese Medicine, Beijing, China

Background: Pseudobulbus Cremastrae seu Pleiones (Shancigu), a traditional Chinese medicine (TCM), has been extensively used in clinical practice for the treatment of various tumors, particularly liver cancer. Shancigu is classified into two commercial specifications -"Maocigu" and "Bingqiuzi"-which exhibit significant differences in appearance, chemical composition, and price, posing challenges for quality control of medicinal materials.The aim of this study was to clarify the quality evaluation indicators based on the anti-liver cancer active components of Shancigu, and to establish a reliable quality evaluation method to preliminarily assess the quality of Shancigu from different commercial specifications and producing areas.Methods: Twenty-six batches of Shancigu samples were collected. High-performance liquid chromatography (HPLC) was employed to establish fingerprint spectra. In vitro anti-liver cancer pharmacological effects indicators were determined using the CCK-8 assay and scratch woundhealing assay. Through spectrum-effect relationship analysis, serum pharmacochemistry analysis, and in vitro/in vivo anti-liver cancer activity evaluation, the effective component combinations of Bingqiuzi and Maocigu were identified and validated. Gray relational analysis (GRA) and the TOPSIS method were subsequently applied to assess the quality of Shancigu from different specifications and origins. Results: Eleven key anti-liver cancer active components from Shancigu were screened and confirmed, including malic acid, citric acid, 2-isobutylmalic acid, gastrodin, batatasin III, 2-phydroxybenzyl-5,3'-dihydroxy-3-methoxybibenzyl, coelonin, 1-p-hydroxybenzyl-2,7-dihydroxy-4methoxyphenanthrene, blestriarene A, blestriarene B, and monbarbatain A. These components exist in Bingqiuzi and Maocigu in different proportions, and the anti-liver cancer pharmacological effects of the effective component combinations was equivalent to that of the original materials both in vitro and in vivo. These eleven components can be used as indicators for evaluating the quality of Shancigu. The quality evaluation revealed no significant difference between Bingqiuzi and Maocigu.For Bingqiuzi, medicinal materials produced in Guizhou and Yunnan demonstrated better quality; for Maocigu, those from Guizhou and Sichuan were superior.This study established quality evaluation criteria for Shancigu and developed an innovative method to comprehensively assess the quality of Shancigu from different commercial specifications and producing regions. By integrating component analysis with anti-liver cancer activity assessment, this research provides a valuable reference for the quality evaluation of other Chinese medicinal materials.