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高效液相色谱法测定米根霉乳酸发酵液中乳酸的光学纯度
引用本文:白冬梅,赵学明,胡宗定. 高效液相色谱法测定米根霉乳酸发酵液中乳酸的光学纯度[J]. 色谱, 2001, 19(1): 13-15
作者姓名:白冬梅  赵学明  胡宗定
作者单位:天津大学化工学院生物工程系,
基金项目:国家自然科学基金资助项目(No.29776035)
摘    要: 以 2 ,3,6 三甲基 β 环糊精作手性流动相添加剂 ,C18柱为固定相 ,考察了流动相的pH和手性流动相添加剂浓度对乳酸对映体分离度的影响 ,建立了DL 乳酸的拆分定量分析方法。用归一化法分析DL 乳酸时引入了定量校正因子 ,确定了外标法测定L 乳酸的回归方程、精密度和回收率。测定了米根霉L 乳酸发酵液中D 型异构体的质量分数及其随放置时间的变化情况。

关 键 词:高效液相色谱法  乳酸  手性分离
文章编号:1000-8713(2001)01-0013-03
修稿时间:2000-05-15

Determination ofEnantiomeric Purity for Lactic Acid in Fermentation Broth by Rhizopus Oryzae with HighPerformance Liquid Chromatography
BAI Dong mei, ZHAO Xue ming, HU Zong ding. Determination ofEnantiomeric Purity for Lactic Acid in Fermentation Broth by Rhizopus Oryzae with HighPerformance Liquid Chromatography[J]. Chinese journal of chromatography, 2001, 19(1): 13-15
Authors:BAI Dong mei   ZHAO Xue ming   HU Zong ding
Affiliation:Department of Biochemical Engineering, School of Chemical Engineering & Technology, Tianjin University, Tianjin 300072, China.
Abstract:A procedure for the resolution of DL-lactic acid and the determination of D-isomer ratio in L-lactic acid fermentation broth by Rhizopus oryzae is described. The effects of pH of mobile phase and concentration of chiral mobile phase additives on resolution of DL-lactic acid were investigated. The optical isomers of lactic acid were resolved by RP-HPLC with 2,3,6-tri-O-beta-cyclodextrin(TM-beta-CD) as a chiral mobile phase additive, and C18 column as stationary phase, and detected at wavelength 210 nm. The results showed that a correction factor should be introduced into the equation for calculation of the percentage of D-lactic acid, because the UV absorption of D-lactic acid and L-lactic acid might not be the same when TM-beta-CD was present. Quantitation was achieved with external standard method, the average recovery was 100.4%, and the relative standard deviation was 0.82%. This method can be used for the determination of the percentage of D-isomer in L-lactic acid fermentation broth by Rhizopus oryzae, and it is simple, rapid and accurate. The results showed that the mass fraction of D-isomer in the fermented broth increased during the period of storage.
Keywords:high performance liquid chromatography  lactic acid  chiral resolution
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