Determination of poly-beta-hydroxybutyric acid in Bacillus thuringiensis by capillary zone electrophoresis with indirect ultraviolet absorbance detection.
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%.
He
Jin
J
College of Life Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, China.
Chen
Shouwen
S
Yu
Ziniu
Z
D001413:Bacillus thuringiensis / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D006885:Hydroxybutyrates / Q000032:analysis*; D011091:Polyesters / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*; D000094122:Polyhydroxybutyrates
D006885:Hydroxybutyrates; D011091:Polyesters; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates