乃孜来颗粒中化学成分的UHPLC-Q-Orbitrap-MS分析

Identification and Attribution of Chemical Constituents in Naizilai Granules by UHPLC-Q-Orbitrap-MS

目的:采用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法(UHPLC-Q-Orbitrap-MS)对乃孜来颗粒中化学成分进行快速鉴定和归属。方法:选择ACQUITY UPLC BEH Shield RP C_(18)色谱柱(2.1 mm×100 mm,1.7μm),流动相选择0.1%甲酸水溶液(A)-乙腈(B)进行梯度洗脱(0~3 min,1%B;3~16 min,1%~11%B;16~30 min,11%~34%B;30~37 min,34%~52%B;37~42 min,52%~100%B;42~44 min,100%B),流速0.3 mL·min^(-1),柱温35℃;通过正、负离子模式采集质谱数据,根据精确相对分子质量、二级质谱裂解碎片等信息,结合对照品及相关文献信息比对,对乃孜来颗粒中化学成分进行鉴定。结果:共推测鉴别了175个化合物,包括72个黄酮类,77个有机酸类,15个倍半萜类,6个香豆素类和5个其他类;并对这些化合物的药材来源进行归属,其中归属于一枝蒿154个、神香草64个、破布木果33个、天山堇菜42个、莴苣子56个、薄荷65个、大枣28个、洋甘菊78个,其中7个为8味组方药材的共有成分。结论:建立的UHPLC-Q-Orbitrap-MS可实现对乃孜来颗粒中化学成分的快速、准确鉴定,基本涵盖了组方中各药材的成分信息,可为完善该制剂的质量评价体系和阐明其药效机制奠定基础。

Objective:Ultra-high performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry(UHPLC-Q-Orbitrap-MS)was used to rapidly analyze and assign the chemical constituents of Naizilai granules.Method:An ACQUITY UPLC BEH Shield RP C_(18) column(2.1 mm×100 mm,1.7μm)was selected for chromatographic analysis,the mobile phase was 0.1%formic acid aqueous solution(A)and acetonitrile(B)for gradient elution(0-3 min,1%B;3-16 min,1%-11%B;16-30 min,11%-34%B;30-37 min,34%-52%B;37-42 min,52%-100%B;42-44 min,100%B),flow rate was 0.3 mL·min^(-1) and the column temperature was 35℃.Mass spectrometry data of Naizilai granules were collected in positive and negative ion modes,the chemical constituents of this preparation were speculated and identified according to the precise molecular weight,secondary fragmentation and other information,combined with reference substance and literature data.Result:A total of 175 compounds were identified and speculated,including 72 flavonoids,77 organic acids,15 sesquiterpenes,6 coumarins and 5 other compounds.Among these identified chemical constituents,there were 154 from Artemisia rupestris,64 from Hyssopus cuspidatus,33 from Cordia dichotoma,42 from Viola tianshanica,56 from Lactuca sativa,65 from Mentha haplocalyx,78 from Matricaria chamomilla,28 from Ziziphus jujuba,7 of which were common components of these eight herbs.Conclusion:The established analytical method can realize the rapid and accurate identification of the chemical constituents in Naizilai granules,and basically covers the main constituents of each medicinal material in the formula,so as to provide a basis for improving the quality evaluation system of the preparation and lay a foundation for elucidating the pharmacodynamic mechanism.