液相色谱-串联质谱法用于酶法制备L-苯丙氨酸生产过程的监测

LC-MS analysis of four compounds in preparation of L-phenylalanine by enzymatic synthesis procedure

针对苯丙酮酸酶法制备L-苯丙氨酸体系,建立了液相色谱-串联质谱法定性定量检测丙酮酸、L-天氡氨酸、苯丙酮酸、L-苯丙氨酸的方法,并对质谱检测条件进行了详细探讨。色谱条件如下:流动相为乙腈、乙酸铵溶液(10mmol/L,乙酸pH5.5),等度洗脱,Eclipse CB-C8色谱柱,流速0.2mL/min,室温;质谱条件如下:电喷雾离子源,负离子MRM扫描模式,气帘气流量0.138MPa,电喷雾电压-4200V,辅助雾化气流量0.380MPa;温度500℃,干燥气流量0.380MPa。实验表明,丙酮酸、L-天冬氨酸、苯丙酮酸、L-苯丙氨酸的线性相关系数分别为0.9995、0.9984、0.9999和0.9991,平均回收率在98.8%～103%之间,相对标准偏差为4.3%～5.0%。该方法简便、快速、准确,可同时定量测定体系中4种成分的含量,为研究代谢过程和优化生产工艺提供了科学依据。

An LC-MS method was developed for the analysis of four compounds produced in the preparation of L- phenylalanine detail. Under pyruvic acid, by enzymatic synthesis procedure. The mass spectrometric conditions were investigated optimal conditions, linear relationships existed between peak areas and concentrations L-aspartic acid, phenylpuruvic acid, and L-phenylalanine with correlation coefficients 0. 9995, 0. 9984, 0. 9999, and 0. 9991 in of of respectively. The average recoveries were 98.8%-103%, and the RSDs were 4.3%-5.0%. The method is easy to use, fast, and accurate, and can provide scientific foun dation to optimize production technology.