학술논문
高效液相色谱法同时测定乳酸菌发酵液中L/D-乳酸方法的建立与验证 / Development and validation of HPLC for determination of L/D-lactic acid in culture supernatant of lactic acid bacteria
Document Type
Academic Journal
Source
食品与发酵工业 / Food and Fermentation Industries. 49(24):283-289
Subject
Language
Chinese
ISSN
0253-990X
Abstract
该研究采用高效液相色谱结合手性色谱柱的方法,建立了一种对乳酸菌发酵液中L/D-乳酸定性定量检测的分析方法.该方法中在1.0～400.0 μg/mL范围中,L/D-乳酸质量浓度与检测结果均呈良好线性关系,L/D-乳酸检出限为0.25 μg/mL,定量限为0.80 μg/mL.基于2株乳酸菌的发酵液对该方法进行验证,结果表明发酵液中L/D-乳酸得到良好分离,该方法精密度、重复性良好,回收率为90.0％～103.7％;在不同稀释度的发酵液中线性相关系数R2均在0.999以上;稳定性的相对标准偏差5％以下;对涵盖4属8种的18株可用于食品的乳酸菌菌株产生L/D-乳酸的情况进行分析,结果表明乳酸菌产L/D-乳酸含量具有菌株差异性,同种乳酸菌产D-乳酸比例具有相同趋势.利用该方法对乳酸菌发酵液中L/D-乳酸的定性定量检测,结果准确稳定.该研究为食品用菌种的安全性评价中菌株代谢产物D-乳酸的检测提供了有效数据支撑和实践参考.
In this study,an analytical method for the qualitative and quantitative determination of L/D-lactic acid in the fermentation broth of lactic acid bacteria was established by HPLC combined with chiral column.In this method,the concentration and detection results of L/D-lactic acid showed a good linear relationship in the range of 1.0-400.0 μg/mL.The limit of detection was 0.25 μg/mL,the lim-it of quantification was 0.80 μg/mL.The method was verified based on fermentation broth of two strains of lactic acid bacteria,the results showed that L/D-lactic acid in fermentation broth were well separated,and the method had good precision and repeatability,and the re-covery rate was between 90.0％and 103.7％.The linear correlation coefficient R2 were more than 0.999 in fermentation broth with dif-ferent dilution degrees,and relative standard deviation of stability less than 5％.The production of L/D-lactic acid by 18 strains of lactic acid bacteria that can be used in food,including 8 species and 4 genera,was analyzed,the results showed that the content of L/D-lactic acid produced by lactic acid bacteria was different among strains,and the proportion of D-lactic acid had the same trend in species.The qualitative and quantitative determination of L/D-lactic acid in lactic acid bacteria fermentation broth by this method was accurate and sta-ble.This study provides effective data support and practical reference for the detection of D-lactic acid,the metabolite of strains,in the safety evaluation of microbial food cultures.
In this study,an analytical method for the qualitative and quantitative determination of L/D-lactic acid in the fermentation broth of lactic acid bacteria was established by HPLC combined with chiral column.In this method,the concentration and detection results of L/D-lactic acid showed a good linear relationship in the range of 1.0-400.0 μg/mL.The limit of detection was 0.25 μg/mL,the lim-it of quantification was 0.80 μg/mL.The method was verified based on fermentation broth of two strains of lactic acid bacteria,the results showed that L/D-lactic acid in fermentation broth were well separated,and the method had good precision and repeatability,and the re-covery rate was between 90.0％and 103.7％.The linear correlation coefficient R2 were more than 0.999 in fermentation broth with dif-ferent dilution degrees,and relative standard deviation of stability less than 5％.The production of L/D-lactic acid by 18 strains of lactic acid bacteria that can be used in food,including 8 species and 4 genera,was analyzed,the results showed that the content of L/D-lactic acid produced by lactic acid bacteria was different among strains,and the proportion of D-lactic acid had the same trend in species.The qualitative and quantitative determination of L/D-lactic acid in lactic acid bacteria fermentation broth by this method was accurate and sta-ble.This study provides effective data support and practical reference for the detection of D-lactic acid,the metabolite of strains,in the safety evaluation of microbial food cultures.