基于UPLC-Q-TOF-MS^E技术分析麸炒前后枳实及其辅料麦麸化学成分的差异

Analysis of Differential Chemical Compositions of Aurantii Fructus Immaturus Before and After Stir-frying with Bran and Chemical Compositions of Wheat Bran After Processing by UPLC-Q-TOF-MS^E

目的:应用超高效液相色谱-四级杆-飞行时间串联质谱法(UPLC-Q-TOF-MS^E)技术结合UNIFI数据库筛查方法快速分析和鉴定枳实麸炒前后的差异性化学成分以及辅料麦麸的化学成分。方法:采用ACQUITY UPLC BEH C18色谱柱(2.1 mm×100 mm,1.7μm),流速0.3 mL·min^-1,进样量2μL,流动相0.1%甲酸水溶液(A)-乙腈(B)梯度洗脱(0~11 min,98%~70%B;11~15 min,70%~55%B;15~16 min,55%~35%B;16~20 min,35%~5%B;20~20.5 min,5%~98%B;20.5~22 min,98%B);选择电喷雾离子源(ESI),正离子检测模式,质量扫描范围m/z 50~1 500;结合主成分分析(PCA)和正交偏最小二乘法-判别分析(OPLS-DA)寻找枳实炮制前后的差异性成分,并检测炮制后麦麸的化学成分。结果:初步鉴定枳实中含有64种化学成分,麸炒枳实58种化学成分,炮制后麦麸18种化学成分,枳实与麸炒枳实差异性成分19个,主要为挥发油、黄酮类、酚酸类、香豆素类和皂苷类成分。结论:从麸炒前后差异性成分和辅料麦麸携带的化学成分结果分析,麸炒能够缓和枳实的峻烈之性,证实了麸炒对于枳实炮制工艺的必要性,可为枳实的炮制机制研究提供较为全面的实验依据。

Objective: To quickly analyze and identify the differential chemical compositions of Aurantii Fructus Immaturus before and after stir-frying with bran and chemical compositions of wheat bran after processing by ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MSE)combined with UNIFI database screening method. Method:ACQUITY UPLC BEH C18 column(2.1 mm×100 mm,1.7 μm)was used for chromatographic separation with 0.1% formic acid solution(A)-acetonitrile(B)as the mobile phase for gradient elution(0-11 min,98%-70%B;11-15 min,70%-55%B;15-16 min,55%-35%B;16-20 min,35%-5%B;20-20.5 min,5%-98%B;20.5-22 min,98%B)at the flow rate of 0.3 mL·min^-1 and the injection volume of 2 μL. The analytes were determined in positive ion mode with electrospray ionization(ESI)and data collection range of m/z 50-1 500. Principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA)were used to find the component differences between raw and processed products of Aurantii Fructus Immaturus,and the chemical compositions of wheat bran after processing were determined. Result: There were 64 compounds in raw products,58 compounds in bran-fried products,and 18 compounds in wheat bran. There were 19 different components between raw and processed products of Aurantii Fructus Immaturus,mainly volatile oil,flavonoids,phenolic acid,coumarins and saponins. Conclusion:Based on the analysis of these different components before and after stir-frying with bran and the chemical compositions carried by wheat bran,the stir-frying with bran can alleviate the intensity of Aurantii Fructus Immaturus, which proves the necessity of stir-frying with bran for the processing technology of this herb,and provides a comprehensive experimental basis for research on processing mechanism of Aurantii Fructus Immaturus.