尿中苯乙醛酸和苯乙醇酸测定的高效液相色谱法

Determination of phenylglyoxylic acid and mandelic acid in urine by high performance liquid chromatography method

目的对尿中苯乙醛酸和苯乙醇酸的高效液相检测标准方法进行修订。方法通过试验对原高效液相色谱方法进行评估，比较ACQUITYUPLCBEHC8,C18、Phenyl（50mm×2．1mm×1．7μm）色谱柱，并对定量方法、方法检出限、定量下限及方法稳定性进行研究。结果采用苯基（BEHPhenyl）色谱柱分离，内标工作曲线定量，苯乙醇酸和苯乙醛酸内标工作曲线回归方程分别为y=3．6607x＋0．0663和y=5．1612x-0．0073；线性相关系数分别为0．9993和0．9991，线性范围分别为0．10～1．00mg／ml和0．04～0．40mg／ml；苯乙醛酸回收率为91．6％～97．1％，批内和批间精密度分别为1．7％-2．4％、0．9％～4．6％，方法检出限1．1mg／L，定量下限为3．7mg／L；苯乙醇酸回收率为84．3％-99．0％，批内和批间精密度分别为0．5％-1．9％、1．1％-1．8％，方法检出限5．4mg／L，定量下限为17．9mg／L。结论该方法采用苯基修饰色谱柱，明确了定量下限，提高了方法稳定性、检测准确度和检出限，满足职业人群生物样品的检测。

Objective To-revise the standard method for the determination of phenylglyoxylic acid （PGA） and mandelic acid （MA） in urine by uhra-performance liquid chromatography. Methods The original standard method was evaluated by experiment, and the chromatographic column, the detection limit, quantitation limit and stabilityof the method were studied. Results The samples were separated by BEH Phenyl（50 mm×2.1 mm×1.7 μm） column and the internal standard working curve method was used. The regression equations were y=3.660 7x＋0.066 3 and y=5.161 2x-0.007 3 for MA and PGA respectively. Linear correlation coefficients were 0.999 3 and 0.999 1. Linearity ranges were 0.10-1.00 mg/ml, 0.04-0.40 mg/ml. The recoveries of PGA and MA were 91.6%-97.1% and 84.3%-99.0%, the precision were 0.9%-4.6% and 0.5%-1.9%. The detection limit and quantitation limit of the method were 1.1 mg/L and 3.7 mg/L for PGA, 5.4 mg/L and 17.9 mg/L for MA. Conclusion The method uses the phenyh modified chromatographic column, determines the detection limit. The method can improve quantitation limit, the detection accuracy and meet the detection of occupational population samples.