提出了一种快速分析发酵液中9种有机酸的高效液相色谱方法。该方法利用RSpakKC-811色谱柱(300mm×8mmi.d.),采用超纯水-4mmol/L高氯酸为流动相进行梯度洗脱。流动相流速1.0mL/min,紫外检测波长210nm,柱温60℃时,能够快速、准确地分离测定发酵液中的丙酮酸、柠檬酸、苹果酸、延胡索酸、琥珀酸、乳酸、甲酸、乙酸等8种有机酸,分析时间只需15min。各种有机酸的线性相关系数R2≥0.9992,精密度的相对标准偏差为0.97%-2.72%,样品回收率为96.8%-102.5%,具有较高的精密度和准确度,可以用于1,3-丙二醇发酵液中有机酸的分析。

A high performance liquid chromatographic method was set up on a column of RSpak KC-811(300mm×8mm i.d.)for rapid measuring the concentrations of α-ketoglutaric acid,pyruvic acid,citric acid,malic acid,fumaric acid,succinic acid,lactic acid,formic acid and acetic acid.The mobile phase was water and perchloric acid solution(4 mmol/L,pH 2.4)at a flow rate of 1 mL/L.The ratio of perchloric acid to water was controlled by a gradient elution program.The assay was carried out at a column temperature of 60℃ and detection wavelength of 210 nm.The method could effectively separate and detect 8 organic acids in 1,3-propanediol fermentation broths in 15 min.All the correlation coefficients(R2)were no less than 0.999 2.The recovery and precision(RSD)were 96.8%-102.5%,0.97%-2.72%,respectively.This precise and accurate method could be used to analyze the organic acids in 1,3-propanediol fermentation broths.