生物合成咖啡酸苯乙酯体系的液相色谱-串联质谱快速分析

Rapid Determination of Enzymatic Synthesis Caffeic Acid Phenethyl Ester in an Enzymecatalytic and Synthetic System by Liquid Chromatography-Tandem Mass Spectrometry

建立液相色谱-串联质谱(LC-MS/MS)检测生物合成咖啡酸苯乙酯体系中咖啡酸(Caffeic acid,CA)和咖啡酸苯乙酯(Caffeic acid phenethylester,CAPE)的方法。采用LC-MS/MS电喷雾电离(ESI),负离子选择反应监测(SRM)模式检测。以V(乙腈)∶V(水)∶V(冰醋酸)=55∶45∶0.5为流动相,流速1.0mL/min,Hypersil C18色谱柱分离并检测生物合成咖啡酸苯乙酯体系中的咖啡酸以及咖啡酸苯乙酯的含量,并对生物合成咖啡酸苯乙酯的收率进行了动力学分析。本方法在进样量为0.2～20μg时具有良好的线性关系,咖啡酸和咖啡酸苯乙酯样品的加标回收率分别为93.4%～98.2%和90.3%～97.8%,相对标准偏差分别为1.79%～2.56%和1.82%～3.67%,咖啡酸苯乙酯收率在3d内可以达到15.54%,表明本方法简便、快速、可靠。

A liquid chromatography-tandem mass spectrometric method was established for the simultaneous determination of caffeic acid （ CA） and caffeic acid phenethyl ester （ CAPE） in enzymatic synthesis,and the biosynthesis kinetics of caffeic acid phenethyl ester was analyzed. Identification was carried out by electrospray ionization （ ESI） in negative mode using selected reaction monitoring. CA and CAPE in enzymatic synthesis system were separated on a hypersil C18 column. The mobile phase consisted of acetonitrile-deionized wateracetic acid （ 55∶45∶0. 5,V/V） at a flow-rate of 1. 0 mL/min. The linear ranges of CA and CAPE were both between 0. 2 and 20 μg. The recoveries of CA and CAPE were 93. 4% 98. 2% and 90. 3% 97. 8% with relative standard deviations of 1. 8% 2. 6% and 1. 8% 3. 7% respectively. The yield of CAPE was up to 15. 54% in three days. The results indicated that the developed method is rapid,simple and reliable.