HPLC法同时测定尿样中苯、甲苯的4种代谢产物  

Simultaneous determination of four urinary metabolites of Hippuric acid andMethylHippuric acid by HPLC

作　　者：肖润梅[1,2] 孙敬智[2] 梅勇[2] 刘婕[2] 陈勇[1] 

机构地区：[1]湖北大学生命科学院湖北省生物技术省重点实验室,湖北武汉430062 [2]湖北省新华医院理化实验室,湖北武汉430015

出　　处：《中国卫生检验杂志》2013年第12期2559-2561,2565,共4页Chinese Journal of Health Laboratory Technology

摘　　要：目的建立高效液相色谱/二极管阵列检测法同时测定尿样中马尿酸、(邻,间,对)-甲基马尿酸含量的方法。方法以0.5 ml乙酸乙酯为萃取溶剂,样品用量0.5 ml,NaCl加入量75 mg,萃取时间3 min,以1000 r/min速度离心5 min,取0.4 ml萃取剂于具塞试管在低于70℃水浴中挥发至干,用1.0 ml流动相溶解残留物,分析条件:柱温35℃,流动相(甲醇:水:冰乙酸=30:70:0.1)波长235 nm,实现了4种代谢物的有效分离。结果在优化条件下,4种待测组分在20 mg/L^500 mg/L范围内呈线性,相关系数(r)均不低于0.9997,检出限(S/N=3)为21μg/L^200μg/L;样品的平均加标回收率为97%~104%,相对标准偏差(RSD)小于5%。结论该方法确定了最佳样品预处理条件,优化了色谱分离及检测条件,灵敏、准确,能够满足有害物质代谢产物的监测要求。Objective A high performance liquid chromatographic（HPLC） method with diode array detection was established for simultaneous determination of hippuric acid （ HA）, methyl hippuric acid（2 - MHA, 3 - MHA, 4 - MHA） in urine. Methods The target analytes in 0.5 ml urine sample were extracted with 0.5 ml ethyl acetate as extraction solvent in 3 min after the addition of 75 mg NaCL. After centrifugation, the four metabolites in sample were well separated on symmetry C18（ 150 mm× 4.6 mm,5 μm） column with mobile phase consisting of methanol： water： glacial acetic acid = 30：70：0.1, column temperature at 35 ℃, the wavelength was set at 235 nm. Results Under the optimal conditions, the calibration curves of the four metabolites were linear in the range of 20 mg/L - 500 mg/L and the correlation coefficients were not less than 0. 9997. The detection limits （ S/N = 3 ） were in the range of 21 μg/L- 200μg/L. The spiked recoveries were in the range of 97% - 104% with relative standard deviations（RSD） less than 5%. Conclusion The method was sensitive, accurate and it could meet the needs of biological monitoring on the metabolites of harmful substances in urine.

关 键 词：高效液相色谱法 代谢产物 马尿酸 甲基马尿酸 

分 类 号：O657.72[理学—分析化学]