高效液相色谱一步法快速测定10种有机酸

目的建立一种同时检测10种有机酸的高效液相色谱检测方法。方法采用Agilent ZORBAX SB-Aq液相色谱柱(250 mm×4.6 mm,5μm),优化分析条件:流动相KH₂PO₄溶液浓度、甲醇含量和柱温,同时检测草酸、酒石酸、丙酮酸、苹果酸、乳酸、乙酸、柠檬酸、丁二酸、富马酸和戊二酸的含量。结果流动相为40 mmol/L KH₂PO₄溶液(磷酸调pH 2.7)-甲醇(95:5),流速1.0 ml/min,柱温30℃,检测波长210 nm,10 min内10种有机酸达到完全分离,标准曲线的相关系数均大于0.999,方法精密度(n=6)的RSD为0.82%~2.6%,平均加标回收率(n=6)均在94%以上。结论该方法操作简便,省时快捷,灵敏度高,重复性好,可用于食品或药品中有机酸含量的检测。

Fast and Effective Determination of Ten Kinds of Organic Acids by HPLC

Objective To establish an HPLC method for the simultaneous determination of 10 organic acids.Methods The chromatographic separation was carried out on an Agilent ZORBAX SB-Aq liquid chromatography column(250 mm×4.6 mm,5μm)for the simultaneous determination of oxalic acid,tartaric acid,pyruvate,malic acid,lactic acid,Acetic acid,citric acid,succinic acid,fumaric acid and glutaric acid.The analytical conditions were optimized including the concentration of KH₂PO₄ solution,methanol content and column temperature.Results When the chromatographic conditions were as follows:40 mmol/L KH₂PO₄ solution with pH adjusted by phosphoric acid to 2.7 and methanol(95:5)as mobile phase,the flow rate of 1.0 ml/min,column temperature of 30℃and detection wavelength of 210 nm,ten kinds of organic acids could be separated fully in 10 min with the correlation coefficients all above 0.999.The precision of the method was tested.The relative standard deviations(RSDs)were in the range of 0.82%-2.6%and the average recoveries were all above 94%(n=6).Conclusion The method is simple,fast,sensitive and repeatable.It can be used to determine the organic acids in food and drug.