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[目的]建立高效液相色谱法测定8种有机酸和双歧杆菌有机酸代谢产物的方法。[方法]优化色谱分离和检测条件,以甲醇-0.01mol/L磷酸二氢钠缓冲溶液(pH2.8)为流动相,梯度洗脱,用C18色谱柱分离,紫外检测器于215nm处检测8种有机酸及双歧杆菌培养基中的乙酸和乳酸。[结果]在优化的色谱条件下,草酸、抗坏血酸的线性范围为0.001～0.2g/L,酒石酸、苹果酸、乳酸、乙酸、柠檬酸和丁二酸的线性范围为0.01～2.5g/L(r﹥0.9996),细菌培养基中乳酸和乙酸的加标回收率分别为94.4%～104.3%,92.8%～98.1%,相对标准偏差分别为1.23%和1.00%。[结论]本方法灵敏快速,可为了解细菌的生长状况和优化培养条件提供参考,并能够通过检测培养基中有机酸的种类和相互比例,用于细菌的分类鉴定。
[Objective] The research aimed to establish a method for the determination of eight organic acids and Bifidobacterium organic acid metabolites by high performance liquid chromatography. [Method] The conditions of chromatographic separation and detection were optimized. The mobile phase consisted of methanol - 0.01 mol / L sodium dihydrogen phosphate buffer solution (pH2.8), gradient elution and separation by C18 column. The UV detector was used at 215 nm for 8 Acetic acid and lactic acid in organic acids and bifidobacteria medium. [Result] Under the optimized chromatographic conditions, the linear range of oxalic acid and ascorbic acid was 0.001-0.2 g / L, the linear range of tartaric acid, malic acid, lactic acid, acetic acid, citric acid and succinic acid was 0.01-2.5 g / L r> 0.9996). The spiked recoveries of lactic acid and acetic acid in bacterial culture medium were 94.4% -104.3% and 92.8% -98.1%, respectively, and the relative standard deviations were 1.23% and 1.00%, respectively. [Conclusion] The method was sensitive and rapid, which could provide a reference for understanding the growth status of bacteria and optimizing the culture conditions, and could be used for the classification and identification of bacteria by detecting the types and mutual proportions of organic acids in the culture medium.