反相高压液相色谱法测定恶臭假单胞菌腈水合酶活力

采用高压液相色谱法（ＨＰＬＣ）分析丙烯腈及其微生物酶法转化产物丙烯酰胺，配制两者的混合溶液做标准曲线，通过外标法定量，测定反应液中腈水合酶活力．实验结果表明，采用Ｎｏｖａ－ＰａｋＣ１８柱、以流速为１．６ｍＬ／ｍｉｎ的甲醇－水溶液（体积比２０：８０）作流动相、采用２１０ｕｍ紫外检测时两者分离度好．ＨＰＬＣ法与化学法相比具有准确、简单、快速、重现性好且灵敏度高等优点，适于批量样品的连续测定．

Determination of Nitrile Hydratases Activities of Pseudomonas Putida Cells by Reversed-Phase High Performance Liquid Chromatograph

By using RP-HPLC, the conditions of the determination of nitrile hydratases activities produced by micro-oganism have been studied. Nova-Pah C18 column is used; the optimal wavelength is 210urn ; CH3OH-H2O(20: 80 V/V) fluent at 1. 6ml/min is the most suitable mobile phase ; a linear relationship between peak area and content of acrylamide is obtained. Under these conditions, the acrylonitrile and microbial transformed product of acrylonitrile-acrylamide are separated completely. The results show that compared with the chemical method. RP-HPLC is more precise, simple, quicke and sensitive and has good reproducibility. It can be applied to the continuous determination for a lot of samples.