基于UHPLC-Q-Orbitrap HRMS分析桃仁化学成分及快速测定苦杏仁苷和野黑樱苷的含量  

Identification of chemical compositions and rapid quantification of amygdalin and prunasin in Prunus persica by UHPLC-Q-Orbitrap high resolution mass spectrometry

作　　者：张聪聪 王长虹[2] 李兴佳 程雪梅[2] 王永丽[2] 戚胜兰 张定棋 刘平 岳鹏飞 刘伟 ZHANG Cong-cong;WANG Chang-hong;LI Xing-jia;CHENG Xue-mei;WANG Yong-li;QI Sheng-lan;ZHANG Ding-qi;LIU Ping;YUE Peng-fei;LIU Wei(Key Laboratory of Modern Prepara-tion of TCM,Ministry of Education,Jiangxi University of Traditional Chinese Medicine,Jiangxi Nanchang 201203,China;In-stitute of Chinese Materia Medica,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Institute of Liver Diseases,Key Laboratory of Liver and Kidney Diseases,Ministry of Education,Shanghai Key Laboratory of Traditional Chinese Clinical Medicine,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]江西中医药大学现代中药制剂教育部重点实验室,江西南昌330004 [2]上海中医药大学中药研究所,上海201203 [3]上海中医药大学附属曙光医院肝病研究所,肝肾疾病病证教育部重点实验室,上海市中医临床重点实验室,上海201203

出　　处：《中国医院药学杂志》2022年第4期347-355,共9页Chinese Journal of Hospital Pharmacy

基　　金：江西中医药大学现代中药制剂教育部重点实验室开放基金项目(编号:TCM-202008);国家自然科学基金项目(编号:81703681,81530101);上海市教育委员会和上海市教育发展基金会“晨光计划”项目(编号:20CG50);中国科协青年托举工程项目(编号:2020QNRC001)。

摘　　要：目的:采用UHPLC-Q-Orbitrap HRMS法对桃仁化学成分进行分析鉴定,并建立快速测定桃仁中苦杏仁苷和野黑樱苷含量的方法。方法:采用ACQUITY UPLC HSS T3(100 mm×2.1 mm,1.8μm)色谱柱,以0.1%甲酸水-乙腈为流动相,梯度洗脱;采用正负离子Full MS/dd-MS^(2)模式,通过化合物的一级、二级质谱信息,结合对照品比对确认,进行桃仁主要化学成分的快速、精准识别。采用负离子Full MS-SIM模式,建立快速测定桃仁中苦杏仁苷和野黑樱苷含量的UHPLC-Q-Orbitrap HRMS方法,并测定10批次燀桃仁中苦杏仁苷和野黑樱苷的含量。结果:共鉴定出桃仁中47种化学成分,包括14种氨基酸类成分、12种核苷酸类成分、12种糖苷类成分、3种氰苷类成分以及6种其他成分。成功建立一种4 min完成桃仁中苦杏仁苷和野黑樱苷含量测定的UHPLC-Q-Orbitrap HRMS方法,2种成分在3.28~2000.00ng·mL^(-1)范围内均线性关系良好(r≥0.9994);精密度、重复性及稳定性良好(RSD≤9.57%);加样回收率在104.01%~104.64%,RSD≤4.52%。10批次样品中苦杏仁苷含量范围为15.62~46.92 mg·g^(-1)、野黑樱苷含量范围为0.18~1.66 mg·g^(-1)。结论:运用UHPLC-Q-Orbitrap HRMS方法实现了桃仁化学成分的快速、准确分析。建立的定量方法分析时间短、灵敏度高、准确性好,方法学考察结果均符合含量测定的要求,可用于桃仁中苦杏仁苷和野黑樱苷含量的快速测定,可为其质量评价提供部分科学参考。OBJECTIVE To establish UHPLC-Q-Orbitrap HRMS method for the identification of the chemical components and simultaneous determination of amygdalin and prunasin in Prunus persica.METHODS ACQUITY UPLC HSS T3(100 mm×2.1 mm,1.8μm)was used for analytes separation with gradient elution of 0.1%formic acid water-acetonitrile as mobile phase.The positive and negative ion Full MS/dd-MS^(2) mode was used for the identification of chemical constituents of P.persica.The main constituents of P.persica were identified quickly and accurately,via comparison with the retention times and MS/MS spectra of the standards.UHPLC-Q-Orbitrap HRMS method for the rapid determination of amygdalin and prunasin in P.persica was established by negative ion Full MS-SIM mode,and the contents of amygdalin and prunasin in10 batches of P.persica were determined.RESULTS Total 47 chemical compositions were identified,including 14 amino acids,12 nucleotides,12 glycosides,3 cyanosides and6 other components.UHPLC-Q-Orbitrap HRMS method was successfully established for the deter-mination of amygdalin and prunasin in peach kernel in 4 min.The 2 constituents showed a good linear relationship in the range of 3.28~2000.00ng·mL^(-1)(r≥0.9994).The precision,repeatability and stability were good(RSD≤9.57%).The recovery was 104.01%~104.64%with the RSD≤4.52%.The content range of amygdalin in10 batches of samples was 15.62~46.92 mg·g^(-1),and the content range of prunasin was 0.18~1.66 mg·g^(-1).CONCLUSION The rapid and accurate analysis of chemical constituents in P.persica was established by UHPLC-Q-Orbitrap HRMS.The established quantitative method has the advantages of short analysis time,high sensitivity and good accuracy.The methodological investigation results meet the requirements of constituents determination.It can be used for the rapid determination of amygdalin and prunasin in P.persica,and provide some scientific references for its quality evaluation.

关 键 词：桃仁 超高效液相色谱-四极杆/静电场轨道阱高分辨质谱 化学成分 苦杏仁苷 野黑樱苷 

分 类 号：R282[医药卫生—中药学]