茵陈蒿中芹菜素6,8-二-C-β-D-葡萄糖苷的确认及含量测定  

Identification and Content Determination of Apigenin 6,8-di-C-β-D-Glucoside in Artemisia Capillaris

作　　者：黄志永 周荣荣 王璐瑶 窦志华[1,2,3] HUANG Zhiyong;ZHOU Rongrong;WANG Luyao;DOU Zhihua(Nantong University,Nantong,Jiangsu,China 226019;Affiliated Nantong Hospital 3 of Nantong University,Nantong,Jiangsu,China 226006;Nanjing University of Chinese Medicine,Nanjing,Jiangsu,China 210023)

机构地区：[1]南通大学,江苏南通226019 [2]南通大学附属南通第三医院,江苏南通226006 [3]南京中医药大学,江苏南京210023

出　　处：《中国药业》2023年第7期72-77,共6页China Pharmaceuticals

基　　金：江苏省重点研发计划(社会发展)项目[BE2018674];江苏省南通市科技计划项目[JCZ20168];江苏省药学会-天晴医院药学基金课题[Q202131];南通市药学会-常州四药医院药学基金项目[ntyx2012]。

摘　　要：目的 确认我国内地产茵陈蒿中存在芹菜素6,8-二-C-β-D-葡萄糖苷,并建立测定其含量的高效液相色谱(HPLC)法。方法 采用高效液相色谱-二极管阵列检测器(HPLC-DAD)法鉴定,色谱柱为Symmetry C18柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.1%磷酸水溶液(梯度洗脱),流速为1.0 mL/min,柱温为30℃,检测波长为200~400 nm;采用超快液相色谱-三重四极杆飞行时间串联质谱(UFLC-Triple-Q-TOF-MS/MS)法鉴别,电喷雾电离负离子模式下采集数据,PeakView 1.6软件提取总离子流图;采用对照品比对,比较对照品溶液和供试品溶液色谱图中色谱峰的保留时间、紫外吸收光谱及总离子流图中相同保留时间分子离子峰的一级和二级质谱数据,鉴定茵陈蒿中的芹菜素6,8-二-C-β-D-葡萄糖苷;同时,采用HPLC法,于270 nm波长处测定35批茵陈蒿样品中芹菜素6,8-二-C-β-D-葡萄糖苷的含量。结果 对照品溶液和供试品溶液色谱图中相同保留时间色谱峰的紫外吸收光谱基本相同,总离子流图中相同保留时间分子离子峰的一级和二级质谱数据基本一致。芹菜素6,8-二-C-β-D-葡萄糖苷质量浓度在0.082 3~16.640μg/mL范围内与峰面积线性关系良好(r=0.999 9,n=8);定量限和检测限分别为36.67 ng/mL和20 ng/mL;日内精密度、日间精密度、稳定性和重复性试验的RSD均小于4.10%(n=6);平均加样回收率为96.37%,RSD为1.40%(n=6)。35批茵陈蒿样品中芹菜素6,8-二-C-β-D-葡萄糖苷含量在0.037~0.339 mg/g之间,其中产地为河南的平均含量最高(0.144 mg/g)。结论 确认了我国内地产茵陈蒿中存在黄酮二糖碳苷类成分芹菜素6,8-二-C-β-D-葡萄糖苷。所建立方法操作简单、重复性好,可用于茵陈蒿中芹菜素6,8-二-C-β-D-葡萄糖苷的含量测定。Objective To identify the apigenin 6,8-di-C-β-D-glucoside in Artemisia capillaris produced in Chinese mainland,and to establish the high-performance liquid chromatography(HPLC)method for the determination of its content.Methods High-performance liquid chromatography-diode array detector(HPLC-DAD)method was used for identification.The chromatographic column was Symmetry C18 column(250 mm×4.6 mm,5μm),the mobile phase was acetonitrile-0.1%phosphoric acid aqueous solution(gradient elution),the flow rate was 1.0 mL/min,the column temperature was 30℃,and the detection wavelength was in the range of 200-400 nm.The ultra-fast liquid chromatography-triple quadrupole-time of flight-tandem mass spectrometry(UFLC-Triple-Q-TOF-MS/MS)method was used for identification,the data were collected in the negative ion mode of electrospray ionization,and the total ion chromatogram was recorded by the PeakView 1.6 software.The retention time and the UV absorption spectrum of the chromatographic peaks in the chromatogram of the reference solution and the test solution,and the primary and secondary mass spectrometry data of the molecular ion peak with the same retention time in the total ion chromatogram were compared and analyzed to identify apigenin 6,8-di-C-β-D-glucoside in Artemisia capillaris.At the same time,the content of apigenin 6,8-di-C-β-D-glucoside in 35 batches of Artemisia capillaris samples was determined by HPLC at the wavelength of 270 nm.Results The UV absorption spectrums of the chromatographic peaks with the same retention time in the chromatograms of the reference solution and the test solution were similar,and the primary and secondary mass spectrometry data of the molecular ion peaks with the same retention time in the total ion chromatogram were similar.The linear range of apigenin 6,8-di-C-β-D-glucoside was 0.0823-16.640μg/mL(r=0.9999,n=6).The limit of quantitation(LOQ)and the limit of detection(LOD)were 36.67 ng/mL and 20 ng/mL respectively.The RSDs of intra-day precision,inter-day precision,stability a

关 键 词：茵陈蒿 高效液相色谱-二极管阵列检测器法 超快液相色谱-三重四极杆飞行时间串联质谱法 黄酮碳苷 芹菜素6 8-二-C-β-D-葡萄糖苷 含量测定 

分 类 号：R932[医药卫生—生药学] R284.1[医药卫生—药学]