UHPLC-Q-Exactive Orbitrap HRMS法同时测定当归配方颗粒中12种成分的含量

Content determination of twelve components in Angelica sinensis formula granules by UHPLC-Q-Exactive Orbitrap HRMS

目的:建立超高效液相色谱-四极杆/静电场轨道阱高分辨质谱(UHPLC-Q-Exactive Orbitrap HRMS)法同时测定当归配方颗粒中12种成分的含量。方法:采用ACQUITY UPLC^(■)BEH C_(18)(2.1 mm×100 mm, 1.7μm)色谱柱;以甲醇(A)-0.1%甲酸水溶液(B)为流动相,进行梯度洗脱;流速为0.3 mL·min^(-1);柱温为45℃;进样量为2μL。质谱采用电喷雾离子源,正负离子全扫描(Full MS/SIM)模式。对当归配方颗粒中的新绿原酸、绿原酸、隐绿原酸、咖啡酸、阿魏酸、洋川芎内酯I、洋川芎内酯H、丁烯基苯酞、洋川芎内酯A、正丁基苯酞、藁本内酯和欧当归内酯A共12种成分进行含量测定,并对含量测定结果进行化学计量学分析。结果:12种成分在各自范围内线性关系良好(r2≥0.997 9),精密度、重复性及稳定性良好(RSD≤9.22%);加样回收率在88.01%~108.77%,RSD均小于10%。聚类分析将18批当归配方颗粒聚为4类,主成分分析共筛选出6个差异标志物。结论:该方法可快速、灵敏测定当归配方颗粒中12种成分的含量,为当归配方颗粒质量评价提供部分新的科学依据和参考。

OBJECTIVE To establish an UHPLC-Q-Exactive Orbitrap HRMS method for the content determination of 12 components in Angelica sinensis formula granules.METHODS Using an ACQUITY UPLC^(■)BEH C_(18)(2.1 mm×100 mm,1.7μm)column,the mobile phase was methanol(A)-0.1%formic acid solution(B)with gradient elution;the flow rate was 0.3 mL·min^(-1);the column temperature was 45℃;the injection volume was 2μL.The mass spectrometry was performed by electrospray ion source with full MS/SIM mode of positive and negative ions.The contents of 12 components,neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,caffeic acid,ferulic acid,senkyunolide I,senkyunolide H,butenyl phthalide,senkyunolide A,n-butylphthalide,ligustilide and levistilide A in Angelica Sinensis formula granules were determined.The chemometrics analysis was performed for the content results of the 12 components.RESULTS The linear relationship of the 12 components was good in their respective ranges(r2≥0.9979).The precision,stability and repeatability were good(RSD≤9.22%).The average recoveries were from 88.01%to 108.77%,and the RSD was lower than 10%.The cluster analysis grouped 18 batches of Angelica sinensis formula granules into four categories,and six differential markers were screened by principal component analysis.CONCLUSION This method can quickly and sensitively determine the contents of the 12 components in Angelica sinensis formula granules,and provides new scientific basis and reference for the quality evaluation of Angelica sinensis formula granules.