发酵液中克拉维酸含量测定方法地研究.pdfVIP

M061 发酵液中克拉维酸含量测定方法的研究 左志晗 韩英素 张建 于国营 高辉 王艳萍* （天津科技大学食品科学与生物技术学院，天津市食品营养与安全重点实验室，天津 300222 ） 摘 要 本文介绍了三种测定克拉维酸含量的方法，即高效液相色谱法、紫外分光光度法与生 物效价法三种方法。克拉维酸与咪唑反应衍生后在 312nm 处有特征吸收，衍生后的色谱图在反 相高效液相色谱柱上可有效避免发酵液中其它组分的干扰；紫外分光光度法也基于克拉维酸与 咪唑反应的衍生物在 312nm 具有特征吸收；生物效价法采用Klebsiella pneumoniae ATCC29665 作为指示菌。并对三种方法测定同样发酵样品克拉维酸含量的结果进行了比较，得出高效液相 色谱法具有操作简便、快速、色谱系统简单等特点，不仅适用于成品及半成品的检测，也适用 于发酵液中克拉维酸含量的常规检测。生物效价测定法与高效液相色谱法所测结果存在一定差 异，但它是生物活性最直接的反映，是评价发酵和产品质量的重要参数，当发酵液克拉维酸含 量较高时二者的测定结果较为接近，但其测定方法复杂，难以掌握且测定时间长。紫外分光光 度法是检测发酵液中最简便快速的方法，与高效液相色谱法所测结果存在一定偏差，适用于发 酵液相对含量的测定，因此可以根据不同的实验目的选用不同的检测方法。 STUDY ON DETERMINATION METHODS OF CLAVULANIC ACID CONCENTRATION IN FERMENTATION LIQUOR ZUO Zhihan ， HAN Yingsu ， ZHANG Jian ，YU Guoying ， GAO Hui and WANG Yanping (College of Food Science and Bioengineering, Tianjin University of Science Technology, Tianjin 300222) Abstract The use of highperformance liquid chromatography (HPLC), spectrophotometry and bioassay methods to determine the concentration of clavulanic acid in fermentation liquor are described in the present paper. Clavulanic acid was assayed by reacting the sample with imidazole, which rapidly produces a derivative absorbing at 312nm. This derivative chromatographs on reverse-phase HPLC columns clear of interfering components in fermentation liquor. Spectro photometry method is also based on the absorbance at 312nm of the clavulanic acid of reaction with imidazole. The bioassay of clavulanic acid method was used with Klebsiella pneumoniae ATCC29665 as the indicator organism.Comparing the clavulanic acid concentration of the same cultures which were assayed by the there methods, result is concluded that HPLC is easy, rapid and simple,