UPLC法和HPLC法对比检测乳酸菌发酵液中的γ-氨基丁酸

Comparison of UPLC and HPLC for the determination of γ-aminobutyric acid in lactic acid bacteria fermentation broth

采用超高效液相色谱(UPLC)与高效液相色谱(HPLC)邻苯二甲醛(OPA)柱前衍生-紫外检测法对比分析乳酸菌发酵液中γ-氨基丁酸(GABA)含量,建立GABA质量浓度与吸光度值的线性回归方程。2种方法测得的GABA在质量浓度100.0~1000.0 mg/mL范围内均有良好的线性关系(相关系数R2>0.998)。UPLC法和HPLC法精密度试验结果相对标准偏差(RSD)均<1.5%,UPLC法的平均相对标准偏差(MRSD)为0.529%,平均回收率为99.59%;HPLC法的平均相对标准偏差为1.175%,平均回收率为98.06%。结果表明2种检测方法均可准确定量分析乳酸菌发酵液中GABA含量,二者相比较,UPLC法更快捷,更适合大量样品检测。

The content ofγ-aminobutyric acid(GABA)in the lactic acid bacteria fermentation broth was analyzed by ultra high performance liquid chromatography(UPLC)and high performance liquid chromatography(HPLC)with pre-column o-phthalaldehyde(OPA)derivatization-ultraviolet detection,and the linear regression equation of GABA mass concentration and absorbance value was established.The GABA determined by two kinds of methods had a good linear relationship(correlation coefficient R2>0.998)in the mass concentration range of 100.0-1000.0 mg/ml.The relative standard deviation(RSD)of the precision test results of UPLC and HPLC were less than 1.5%.The mean relative standard deviation(MRSD)of UPLC was 0.529%,and the average recovery rate was 99.59%.The MRSD of HPLC was 1.175%,and the average recovery rate was 98.06%.The results showed that the two kinds of methods could accurately and quantitatively analyze the GABA content in the lactic acid bacteria fermentation broth.Compared with the two kinds of methods,UPLC was faster and more suitable for the determination of a large number of samples.