基于UHPLC-QTOF-MS/MS辨识的侧柏叶炒炭前后化学成分分析  

Chemical Composition Analysis of Platycladi Cacumen Before and After Being Carbonized Based on Identification by UHPLC-QTOF-MS/MS

作　　者：石典花[1,2] 戴衍朋[1] 王丽凤 周倩[1] 张学兰[2] 张军[1] SHI Dian-hua;DAI Yan-peng;WANG Li-feng;ZHOU Qian;ZHANG Xue-lan;ZHANG Jun(Shandong Academy of Chinese Medicine,Ji'nan 250014,China;Shandong University of Traditional Chinese Medicine,Ji'nan 250355,China;Shandong Maternal and Child Health Care Hospital,Ji'nan 250014,China)

机构地区：[1]山东省中医药研究院,济南250014 [2]山东中医药大学,济南250355 [3]山东省妇幼保健院,济南250014

出　　处：《中国实验方剂学杂志》2021年第8期107-116,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家科技基础性工作专项(2014FY111100);全国中药特色技术传承人才培训项目(国中医药人教函[2019]43号);国家中医药管理局中药炮制技术传承基地建设项目;山东省中医药科技发展计划项目(2015-173,2017-130);山东省职业教育技艺技能传承创新平台项目(201712)。

摘　　要：目的:对侧柏叶炒炭前后化学成分进行辨识。方法:采用超高效液相色谱串联四极杆飞行时间质谱法(UHPLCQTOF-MS/MS)对侧柏叶及其制备的侧柏炭进行化学成分辨识和比较,色谱条件为ACQUITY UPLC BEH C18色谱柱(2.1 mm×100 mm,1.7μm),以0.1%甲酸水溶液(A)-乙腈(B)为流动相进行梯度洗脱(0~3.5 min,5%~15%B;3.5~6 min,15%~30%B;6~6.5 min,30%B;6.5~12 min,30%~70%B;12~12.5 min,70%B;12.5~18 min,70%~100%B;18~22 min,100%B),进样量5μL,流速0.4 mL·min^(-1);质谱条件为电喷雾离子源,正、负离子全扫描模式采集一、二级质谱数据。利用自建二级质谱数据库及相应裂解规律匹配法对含有MS/MS数据的色谱峰进行物质鉴定。结果:正、负离子模式分别辨识出了77,76种变化趋势一致的物质,侧柏叶炒炭后消失的成分基本为氨基酸类、酮醛等易挥发性成分,而新产生成分包括6种黄酮苷元类(鼠李素,3,6,3’-三羟基黄酮,6,7,3’-三羟基黄酮,4’-羟基-2’-甲基-3,4,5-三甲氧基查尔酮,草质素,3’,5’-二甲氧基-3,5,7,4’-四羟基黄酮),3种香豆素类(7-羟基香豆素,7,8-二羟基香豆素和8-乙酰基-7-羟基香豆素)及3种苯甲酸类(3-甲基苯邻二酚、邻苯二酚、色酮-3-甲酸)。其中辨识出的化学物质中共计黄酮类成分40种(槲皮苷、槲皮素、山柰酚等)。结论:侧柏叶炒炭后化学成分发生了明显的量变和质变,辨识出的主要有效成分黄酮类可为侧柏叶炒炭前后功效变化物质基础的确定提供依据。Objective:To identify the chemical constituents of Platycladi Cacumen before and after being carbonized.Method:Chemical constituents in 3 batches of Platycladi Cacumen and its carbonized products were identified and compared by ultra performance liquid chromatography-quadrupole-time of flightmass spectrometry(UHPLC-QTOF-MS/MS).Chromatographic separation was performed on an ACQUITY UPLC BEH C18 column(2.1 mm×100 mm,1.7μm)with 0.1%formic acid aqueous solution(A)-acetonitrile(B)as mobile phase for gradient elution(0-3.5 min,5%-15%B;3.5-6 min,15%-30%B;6-6.5 min,30%B;6.5-12 min,30%-70%B;12-12.5 min,70%B;12.5-18 min,70%-100%B;18-22 min,100%B).The flow rate was 0.4 mL·min^(-1)and the injection volume was 5μL.Mass spectrometry was performed by an electrospray ionization,and the primary and secondary mass spectrometry data were collected with the full scan mode of positive and negative ions,the peaks containing MS/MS data were identified by self-established secondary mass spectrometry database and corresponding fragmentation law matching method.Result:A total of 77 and 76 substances with the same change trend were identified under positive and negative ion modes.After being carbonized,the disappeared components of Platycladi Cacumen were mainly amino acids,ketone aldehydes and other volatile components.Among newly produced components,there were 6 kinds of flavonoid aglycones(rhamnetin,6,7,3’-trihydroxyflavone,3,6,3’-trihydroxyflavone,4’-hydroxy-2’-methyl-3,4,5-trimethoxychalcone,herbacetin and 3’,5’-dimethoxy-3,5,7,4’-tetrahydroxyflavone),3 kinds of coumarins(7-hydroxycoumarin,7,8-dihydroxycoumarin and 8-acetyl-7-hydroxycoum-arin)and 3 kinds of benzoic acids(3-methylcatechol,pyrocatechol and chromone-3-carboxylic acid).There were a total of 40 flavonoids(quercitrin,quercetin,kaempferol,etc.)among these identified chemical constituents.Conclusion:There are significant quantitative and qualitative changes in the chemical compositions of Platycladi Cacumen after being carbonized.The flavonoids,the identifie

关 键 词：侧柏叶 炒炭品 超高效液相色谱串联四极杆飞行时间质谱法(UHPLC-QTOF-MS/MS) 化学成分 炮制 黄酮类 物质基础 

分 类 号：R22[医药卫生—中医基础理论] O657[医药卫生—中医学]