基于超高效液相色谱-质谱联用技术和化学计量学方法比较白芥子炒制前后的化学成分

Comparison of Chemical Constituents of Sinapis alba before and after Stir-frying Based on UPLC-MS and Chemometrics Methods

目的:比较白芥子炒制前后的化学成分。方法:采用超高效液相色谱-四极杆-静电场轨道阱质谱(UPLC-Q-Exactive Obitrap MS)联用技术对白芥子炒制前后的化学成分进行分析。色谱柱为Waters CORTECS T3,流动相为甲醇-0.1%甲酸溶液(梯度洗脱),流速为0.25 mL/min,柱温为30℃,进样量为2μL;高分辨MS采用加热电喷雾离子源,于正离子扫描模式下扫描,扫描范围为m/z 120~1000。采用Compound Discover 3.2软件结合相关数据库等对白芥子炒制前后的化学成分进行鉴定,以峰面积对成分含量变化进行初步评价,再以峰面积为变量对成分含量变化进行化学计量学分析。结果:从白芥子中共鉴定出54种化学成分,主要为脂肪酸类(以芥子酸为代表)、生物碱类(以芥子碱为代表)、黄酮类等化合物。经炒制后共有19种化学成分的含量发生了明显变化,其中10种显著降低、9种显著升高(P<0.05)。主成分分析和正交偏最小二乘法判别分析均能将白芥子与炒白芥子明显区分开。结论:白芥子经炒制后,其部分化学成分含量发生了明显变化,这可能是该药材炒制后功效发生改变的重要原因之一。

OBJECTIVE:To compare the chemical components in Sinapis alba before and after stir-frying.METHODS:UPLC-Q-Exactive Obitrap MS was adopted to analyze chemical constituents of S.alba before and after stir-frying.The determination was performed on Waters CORTECS T3 column with mobile phase consisted of methanol-0.1%formic acid solution(gradient elution)at the flow rate of 0.25 mL/min.The column temperature was 30℃and the sample size was 2μL.High resolution MS adopted heating electrospray electron source,positive ion scanning mode,scanning range m/z 120-1000.The chemical constituents of S.alba before and after stir-frying were identified by Compound Discover 3.2 software combined with relevant database,and the content changes of chemical constituents were analyzed by using peak area.Chemometrics analysis was performed for the content changes of chemical constituents using peak area as variable.RESULTS:A total of 54 chemical components were identified in S.alba,mainly fatty acids(represented by erucic acid),alkaloids(represented by sinapine),flavonoids.After stir-frying,the contents of 19 chemical components changed significantly,of which the contents of 10 components decreased significantly and those of 9 components increased significantly(P<0.05).Principal component analysis and orthogonal partial least squares discriminant analysis could clearly distinguish S.alba from stir-fried S.alba.CONCLUSIONS:The contents of some chemical components of S.alba change significantly after stir-frying,which may be one of the important reasons for the change of efficacy after stir-frying.