He Jin, Chen Shouwen, Yu Ziniu
College of Life Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, China.
J Chromatogr A. 2002 Oct 11;973(1-2):197-202. doi: 10.1016/s0021-9673(02)01215-3.
A new capillary electrophoresis method for determining poly-beta-hydroxybutyric acid (PHB) in Bacillus thuringiensis was established. Poly-beta-hydroxybutyric acid in samples was hydrolyzed by sulphuric acid and neutralized by Ba(OH)2. The content of produced beta-hydroxybutyrate was then determined by capillary zone electrophoresis (CZE) with indirect UV detection at 254 nm. With 5 mM p-hydroxybenzoate and 0.5 mM tetradecyltrimethylammonium bromide (TTAB) at pH 8.0 as carrier electrolyte, beta-hydroxybutyrate can be determined within 6 min. Standard regression equation was made by beta-hydroxybutyrate, and the linear range was 2-1000 microg/ml. The relative standard deviations (RSDs) for migration time and peak area are both less than 1.0%. The detection limit for beta-hydroxybutyrate was 0.2 microg/ml, which is two to three orders of magnitude lower than that of the gas chromatography (GC) method. The capillary electrophoresis method was successfully applied to determine poly-beta-hydroxybutyric acid in fermentation broth and single colony. The added standard recovery was 96%.
建立了一种测定苏云金芽孢杆菌中聚-β-羟基丁酸（PHB）的新型毛细管电泳方法。样品中的聚-β-羟基丁酸经硫酸水解，并用Ba(OH)₂中和。然后通过在254 nm处进行间接紫外检测的毛细管区带电泳（CZE）测定生成的β-羟基丁酸盐的含量。以5 mM对羟基苯甲酸和0.5 mM十四烷基三甲基溴化铵（TTAB）在pH 8.0作为载体电解质，β-羟基丁酸盐可在6分钟内测定。由β-羟基丁酸盐建立标准回归方程，线性范围为2 - 1000 μg/ml。迁移时间和峰面积的相对标准偏差（RSD）均小于1.0%。β-羟基丁酸盐的检测限为0.2 μg/ml，比气相色谱（GC）法低两到三个数量级。该毛细管电泳方法成功应用于测定发酵液和单菌落中的聚-β-羟基丁酸。加标回收率为96%。
