朱砂莲中马兜铃酸类成分的UPLC-QTOF-MS/MS定性与定量分析  

Qualitative and Quantitative Analysis of Aristolochic Acids in Aristolochia cinnabarina Dried Root Tubers by UPLC-QTOF-MS/MS

作　　者：郭宁 赵雍[2] 孙奕[2] 王安琪 徐凌川[1] 梁爱华[2] GUO Ning;ZHAO Yong;SUN Yi;WANG An-qi;XU Ling-chuan;LIANG Ai-hua(College of Pharmacy,Shandong University of Traditional Chinese Medicine,Ji'nan 250355,China;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]山东中医药大学药学院,济南250355 [2]中国中医科学院中药研究所,北京100700

出　　处：《中国实验方剂学杂志》2021年第11期162-170,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家“重大新药创制”科技重大专项(2015ZX09501004,2018ZX09101-002);中国中医科学院自主选题(ZZ12-001)。

摘　　要：目的:建立朱砂莲块根中马兜铃酸类成分的定性与定量测定方法。方法:采用超高效液相色谱-四极杆飞行时间串联质谱法(UPLC-QTOF-MS/MS)对朱砂莲块根进行定性与定量分析,检测条件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)为流动相梯度洗脱(0~1 min,10%B;1~9 min,10%~30%B;9~11 min,30%~50%B;11~15 min,50%~90%B),流速0.45 mL·min^(-1),柱温35℃,检测波长250 nm;电喷雾电离源(ESI),正离子模式检测。同时,建立了21批朱砂莲样品中马兜铃酸类成分的UPLC指纹图谱,根据指纹图谱分析结果,测定了不同产地和不同采收期朱砂莲样品中5个马兜铃酸类成分的含量。结果:结合文献信息和质谱数据,一共鉴定了朱砂莲中17个马兜铃酸类化合物,包括8个马兜铃酸,7个马兜铃内酰胺和2个4,5-二羰基阿朴菲类生物碱。UPLC指纹图谱中指认了10个共有峰,分别为朱砂莲苷,马兜铃内酰胺Ⅰa-N-β-D-葡萄糖苷,马兜铃酸Ⅳa-O-β-D-葡萄糖苷,马兜铃内酰胺Ⅲa-N-β-D-葡萄糖苷,马兜铃内酰胺Ⅰ-N-β-D-葡萄糖苷,马兜铃酸C,马兜铃酸D,马兜铃酸Ⅱ,马兜铃内酰胺Ⅰ,马兜铃酸Ⅰ。定量分析结果显示,马兜铃酸Ⅰ,Ⅱ,C,D及马兜铃内酰胺Ⅰ在线性范围内呈良好的线性关系,精密度、稳定性、重复性试验的RSD均<3.0%,加样回收率范围在97.06%~101.84%(RSD<3.0%)。21批朱砂莲中马兜铃酸Ⅰ,马兜铃酸Ⅱ,马兜铃酸C,马兜铃酸D,马兜铃内酰胺Ⅰ的质量分数范围分别为0.938 6~3.567 5,1.377 6~3.688 1,0.056 3~0.527 7,0.108 8~0.305 5,0.021 0~0.081 7 mg·g-1。结论:朱砂莲中马兜铃酸类成分的含量存在一定差异,马兜铃酸Ⅰ和Ⅱ的含量要高于其他马兜铃酸类成分。建立的分析方法快速简便、准确可靠,可为朱砂莲的质量控制与评价提供参考。Objective: To establish a qualitative and quantitative method for the determination of aristolochic acids in Aristolochia cinnabarina dried root tubers. Method:The dried root tubers of A. cinnabarina was qualitative and quantitative analysis by ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry(UPLC-QTOF-MS/MS). The analysis was performed on Waters ACQUITY UPLCBEH C18 column( 2.1 mm×100 mm,1.7 μm)with the mobile phase of 0.1% formic acid aqueous solution(A)-acetonitrile(B)for gradient elution(0-1 min,10%B;1-9 min,10%-30%B;9-11 min,30%-50%B;11-15 min,50%-90%B). The flow rate was 0.45 mL·min^(-1),column temperature was 35 ℃,and the detection wavelength was 250 nm. Mass spectral data was acquired in positive mode of electrospray ionization(ESI). At the same time,the UPLC fingerprints of aristolochic acids in 21 batches of A. cinnabarina dried root tubers were established,and the contents of 5 aristolochic acids in A. cinnabarina dried root tubers from different producing areas and different harvesting periods were determined. Result: A total of 17 compounds, including 8 aristolochic acids,7 aristololactams and 2 4,5-dioxoaporphine alkaloids,were identified from A. cinnabarina dried root tubers by mass spectrometry data and bibliographic information. Ten common peaks were identified in the UPLC fingerprint, and they were tuberosinone-N-β-D-glucoside, aristolactam Ⅰ a-N-β-D-glucoside,aristolochic acid Ⅳ a-O-β-D-glucoside,aristolactam Ⅲ a-N-β-D-glucoside,aristolactam Ⅰ-N-β-D-glucoside,aristolochic acid Ⅲ a,aristolochic acid Ⅳ a,aristolochic acid Ⅱ,aristolactam Ⅰ and aristolochic acid Ⅰ.According to the quantitative analysis,the results exhibited that aristolochic acid Ⅲ a,aristolochic acid Ⅳ a,aristolochic acid Ⅱ,aristolactam Ⅰ and aristolochic acid Ⅰ had good linear relationships in the linear range.The relative standard deviations(RSDs)of precision,stability and reproducibility tests were all less than 3.0%,the recovery was 97.06%-101.84%(R

关 键 词：朱砂莲 马兜铃酸类成分 超高效液相色谱-四极杆飞行时间串联质谱法(UPLC-QTOF-MS/MS) 指纹图谱 马兜铃内酰胺 4 5-二羰基阿朴菲类生物碱 

分 类 号：R22[医药卫生—中医基础理论] R917[医药卫生—中医学]