摘要
同时检测黄花蒿Artemisia annua中多个化合物,以对黄花蒿的品质进行多角度评价。建立超高效液相色谱-三重四极杆串联质谱(UPLC-QQQ-MS/MS)同时检测黄花蒿中紫穗槐-4,11-二烯、青蒿醛、双氢青蒿酸、青蒿酸、青蒿素B、青蒿烯和青蒿素7个青蒿素类相关化合物的方法,并对黄花蒿不同组织部位(根、茎、叶和侧枝)中这7个化合物含量进行差异比较,检测其在黄花蒿中的累积情况。采集生长盛期4月龄黄花蒿的根、茎、叶和侧枝,检测黄花蒿不同组织部位7个青蒿素类相关化合物的含量。采用UPLC-QQQ-MS/MS的多反应监测(MRM)采集模式,大气压力化学离子源(APCI)正离子模式,色谱柱为Eclipse Plus RRHD C_(18)(2.1 mm×50 mm,1.8μm),流动相为含甲酸(0.1%)-甲酸铵(5 mmol·L^(-1))水溶液(A)和甲醇(B),梯度洗脱(0~8 min,55%~100%B;8~11 min,100%B;平衡3 min)。流速0.6 mL·min^(-1),柱温40℃,进样量5μL,检测时间8 min。通过方法学考察,建立了基于UPLC-QQQ-MS/MS同时定量检测青蒿素生物合成途径中7个代表性化合物含量的方法。对黄花蒿根、茎、叶和侧枝的含量比较分析结果表明:黄花蒿中青蒿素含量高于青蒿素B,且青蒿素和双氢青蒿酸在黄花蒿各部位的含量相对较高;7个化合物在各部位的含量存在较大差异,均在叶片中最高,且青蒿烯和青蒿醛在根中未检测到。该研究建立了基于UPLC-QQQ-MS/MS同时检测青蒿素生物合成途径上7个代表性化合物的定量方法,准确、灵敏且高效,可用于黄花蒿中青蒿素类相关化合物的含量测定、新品种选育和中药材质量控制等。
A variety of compounds in Artemisia annua were simultaneously determined to evaluate the quality of A.annua from multiple perspectives.A method based on ultra-high performance liquid chromatography-triple quadrupole tandem mass spectrometry(UPLC-QQQ-MS/MS)was established for the simultaneous determination of seven compounds:amorpha-4,11-diene,artemisinic aldehyde,dihydroartemisinic acid,artemisinic acid,artemisinin B,artemisitene,and artemisinin,in A.annua.The content of the seven compounds in different tissues(roots,stems,leaves,and lateral branches)of A.annua were compared.The roots,stems,leaves,and lateral branches of four-month-old A.annua were collected and the content of seven artemisinin-related compounds in different tissues was determined.A multi-reaction monitoring(MRM)acquisition mode of UPLC-QQQ-MS/MS was used,with a positive ion mode of atmospheric pressure chemical ion source(APCI).Chromatographic separation was achieved on an Eclipse Plus RRHD C_(18) column(2.1 mm×50 mm,1.8μm).The gradient elution was performed with the mobile phase consisted of formic acid(0.1%)-ammonium formate(5 mmol·L^(-1))(A)and the methanol(B)gradient program of 0-8 min,55%-100%B,8-11 min,100%B,and equilibrium for 3 min,the flow rate of 0.6 mL·min^(-1),the column temperature of 40℃,the injection volume of 5μL,and the detection time of 8 min.Through methodological investigation,a method based on UPLC-QQQ-MS/MS was established for the simultaneous quantitative determination of seven representative compounds involved in the biosynthesis of artemisinin.The content of artemisinin in A.annua was higher than that of artemisinin B,and the content of artemisinin and dihydroartemisinic acid were high in all the tissues of A.annua.The content of the seven compounds varied considerably in different tissues,with the highest levels in the leaves and neither artemisinene nor artemisinic aldehyde was detected in the roots.In this study,a quantitative method based on UPLC-QQQ-MS/MS for the simultaneous determination of seven representa
作者
王梦月
万会花
向丽
蒲雨婷
尹青岗
高冉冉
师玉华
邬兰
WANG Meng-yue;WAN Hui-hua;XIANG Li;PU Yu-ting;YIN Qing-gang;GAO Ran-ran;SHI Yu-hua;WU Lan(Key Laboratory of Beijing for Identification and Safety Evaluation of Chinese Medicine,Artemisinin Research Center,Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2024年第5期1260-1265,共6页
China Journal of Chinese Materia Medica
基金
国家重点研发计划专项“政府间国际科技创新合作”项目(2019YFE0108700)
中央级公益性科研院所基本科研业务费专项(ZZ16-ND-10-28,ZZ16-ND-10-09,ZZ16-ND-10-10,ZZ13-YQ-106)
海南省院士创新平台科研专项(SQ2021PTZ0052,YSPTZX202137)。
