雄黄饮片ICP-MS无机元素分析及其抗结肠癌谱效关系研究

Inorganic element analysis of realgar decoction pieces by ICP-MS and its spectrum-effect relationship against colon cancer

目的:建立雄黄饮片的无机元素指纹图谱并对其进行分析与评价,同时进行体外抗结肠癌活性研究,探讨多种无机元素与抑制结肠癌细胞增殖的谱效关系。方法:采用ICP-MS测定12个不同批次雄黄饮片中30种无机元素的含量,建立无机元素指纹图谱,进行相似度评价,运用SPSS 26.0、SMICA 14.1软件进行聚类分析、主成分分析(PCA);采用MTT法测定雄黄对结肠癌HCT116细胞的增殖抑制率,运用SMICA 14.1、DPS 7.5软件分别进行偏最小二乘回归法和灰色关联度法分析其谱效关系。结果:检测的30种元素中,As、Fe、Sb、Al、Mg含量较高,Na、Zn和重金属Cu、Cd均未检出,Pb、Hg部分检出。建立了12种无机元素特征指纹图谱,相似度(夹角余弦值)均大于0.900 0。12批次雄黄分为了2类,综合评价得分产地贵州的雄黄饮片排在前列。MTT实验表明雄黄能抑制结肠癌HCT116细胞增殖,偏最小二乘回归和灰色关联法分析谱效关系表明,VIP>1且关联度>0.7的元素有As、Sr、Ga、Ti、Al、Fe、V、Sb、Co、Mg、Mn。结论:不同批次雄黄饮片无机元素的种类和含量存在一定差异,该研究建立了准确高效分析评价雄黄饮片无机元素的方法。雄黄抗结肠癌作用是多种元素协同作用的结果,主要与As、Sr、Ga、Ti、Al、Fe、V、Sb、Co、Mg、Mn 11种元素密切相关。

OBJECTIVE To establish,analyze and evaluate the inorganic element fingerprint of mineral medicine realgar prepared by water grind processing,and to conduct in vitro anti-colon cancer activity research to explore the spectrum-effect relationship between various inorganic elements and the inhibition of colon cancer cell proliferation.METHODS Inductively coupled plasma-mass spectrometry(ICP-MS)was used to determine the contents of 30 inorganic elements in 12 different batches of realgar prepared by water grind processing to establish inorganic element fingerprints,and the similarity was evaluated.MTT method was used to determine the proliferation inhibition rate of realgar on colon cancer HCT116 cells,and partial least squares regression(PLSR)and gray relational degree(GRA)were combined to analyze the spectrum-effect relationship.RESULTS Among the 30 elements detected,the contents of As,Fe,Sb,Al and Mg were relatively high,while Na,Zn and heavy metals Cu and Cd were not detected,while Pb and Hg were partially detected.The characteristic fingerprints of 12 inorganic elements were established,and the similarity(cosine value of the included angle)was all greater than 0.9000.The 12 batches of realgar were divided into 2 categories,and the comprehensive evaluation score of realgar from Guizhou ranked in the forefront.MTT experiments showed that realgar inhibited the proliferation of colon cancer HCT116 cells.The partial least squares regression analysis and gray correlation analysis showed that the elements with VIP>1 and correlation degree>0.7 were As,Sr,Ga,Ti,Al,Fe,V,Sb,Co,Mg and Mn.CONCLUSION There are certain differences in the types and contents of inorganic elements in different batches of realgar prepared by water grind processing.This study established an accurate and efficient method for analyzing and evaluating inorganic elements in realgar prepared by water grind processing,and the anti-colon cancer effect of realgar is the result of the synergistic action of various elements,mainly closely related to the 11 elements of As,Sr,Ga,Ti,Al,Fe,V,Sb,Co,Mg and Mn.