离子排阻色谱法快速测定益生菌发酵液中7种有机酸含量

Rapid Determination of Seven Organic Acids in Fermentation Broth of Probiotics by Ion-Exclusion Chromatography

运用离子排阻色谱法建立益生菌发酵液中主要有机酸的快速分离与检测方法。采用Aminex HPX-87H分析柱,选用3.5 mmol/L硫酸与乙腈混合液（体积比为99∶1）为流动相,在柱温50℃、流速0.6 m L/min及检测波长为210 nm条件下,对益生菌发酵产品中甲酸、乙酸、丙酸、乳酸、正丁酸、琥珀酸及柠檬酸7种有机酸进行分离与检测。结果表明：7种有机酸在0.02~22.34 g/L内线性关系较好,回归方程的线性相关系数为0.999 82~0.999 99。利用干酪乳杆菌（Lactobacillus casei）发酵液检测加标回收率为97.28%~107.33%,精密度为0.47%~2.07%（n=5）。利用该方法实现了对8株益生菌36 h发酵液中主要有机酸含量的快速分析比较。此法简单、快速、准确,适用于益生菌发酵液中主要有机酸的分离与检测。

An ion-exclusion chromatography method was established for the rapid separation and detection of the primary organic acids in fermentation broth of probiotics. Seven organic acids including formic,acetic,propionic,lactic,n-butyric,succinic and citric acids from fermentation products of probiotics were separated and measured under the condition of Aminex HPX-87 H column,the mixture mobile phase composed by3.5 mmol / L sulfuric acid and acetonitrile（ volume ratio 99 ∶ 1）,the column temperature at 50 ℃,the rate of mobile phase was 0.6 m L / min and the detection wavelength was 210 nm.The results demonstrated that the seven organic acids possessed good linear relationships in the range 0.02-22.34 g / L,and the liner correlation coefficients were 0.999 82-0.999 99. The recovery rates of standard addition were 97. 28%- 107. 33% for these seven organic acids by fermentation broth of Lactobacillus casei,and the precisions were 0.47%-2.07%（ n = 5）.The main organic acids in the 36 h fermentation liquids of eight strains of probiotics were analyzed and compared rapidly by the method.It was fairly simple,rapid,and accurate,and expected to be applied to the separation and detection of the primary organic acids in fermentation broth of probiotics.