高效液相色谱荧光法测定尿中二元有机酸研究

Determination of dicarboxylic acids in urine by high-performance liquid chromatography with fluorescence detection

目的：建立尿中丙二酸、丁二酸、戊二酸3种二元有机酸的高效液相色谱荧光测定方法。方法：采用荧光检测器，利用Waters C18（250mm×4．6mmi．d．，5μm）色谱柱，以70％（v／v）乙腈为淋洗液，流速1．0ml／min，采用激发波长345nm、发射波长480nm进行检测，外标法定量。结果：丙二酸、丁二酸、戊二酸在相应的试验浓度范围内均具有良好的线性关系，其相关系数均大于0．999，最低检出限在0．02～0．2mg／L范围，平均回收率在80．2％～109．6％之间，RSD均小于5．0％。结论：该法具有简单、准确、灵敏等优点，适合尿中丙二酸、丁二酸、戊二酸含量的检测。

Objective： To establish an analytical method for the determination of diearboxylic acids including malonie acid, succinic acid and glutaric acid in urine by by high - performance liquid chromatography with fluorescence detection. Methods：The dicarboxylic acids in urine were separated on sunfire C18 column （i50 mm × 4.6 mm i. d. , 5 μm）by using 70% acetonitrile as an elnent at a flow rate of 1.0 ml/min. Detection was performed on an fluorescence detector. The excitation and emission wavelengths were 345 nm and 480 nm. Quantification was calculated by an external standard method. Results：The linearity of each organic acid in the tested concentration range was good and the correlation coefficients were all over 0. 9990. The limits of quantification were found to be 0. 02 - 0. 2 mg/L for the target compounds. The extraction recoveries were between 80. 2% - 109.6% and the RSD all less than 5.0%. Conclusion： This method is simple, sensitive and accurate for the determination of the dicarboxylic acids in urine and it can be satisfactorily applied to the routine analysis.