[ Objective ] The aim was to improve the measuring method of 1 -hydroxypyrene in urine by high performance liquid chromatography with fluorescence detection by comparing concentrated and non-concentrated eluate of C18 solid phase extraction. [ Method] Take two same urine samples, hydrolyzed by enzyme first, then enriched by C18 solid phase extraction, elution by methanol, one nitrogen dry to 1.5 ml, another direct volume to 1.5 ml, then were determined with HPLC/FLD. With Vacetnnitnle: Vwater =75 = 25 as mobile phase and 1 ml/min of flow rate, the excitation(λexI) and emission wavelengths( λem ) were 275 and 430 nm, respectively. [ Result ] The determined values of urinary 1-hydroxypyrene which after concentrating were 72.9% - 80. 6% of which non-concentrating. When use the method of non-condensed enzymatic hydrolysis high performance liquid chromatography with fluorescence detection, the limit of detection was 0.02 ng/ml, the calibration curve was linear at a given concentration range, the correlation coefficient of linear regression equation was y = 2. 404 la; - 0. 083 ( r = 0.999 9) , the recoveries were from 94.8% to 108.7% , and the average RSD was 2.7%. [ Conclusion] The method of non-condensed enzymatic hydrolysis high performance liquid chromatography with fluorescence detection can be used to determine the urine of 1-hydroyxpyrene, and it is simple, have high sensitivity and precision, good recoveries and reproducibility.%[目的]通过C18固相萃取小柱洗脱液浓缩与不浓缩试验对比,改进尿样中1-羟基芘的高效液相色谱-荧光检测器测定方法.[方法]取2组相同尿样,经酶避光水解后,通过C18固相萃取小柱进行富集,甲醇洗脱.一组对洗脱液进行氮吹浓缩后定容至1.5ml,另一组直接洗脱定容至1.5ml,然后,使用高效液相色谱-荧光检测器检测.以 V乙腈∶V水 =75∶25为流动相,等度洗脱,流速为1 ml/min,荧光检测器 λex=275 nm,λem=430nm.[结果]浓缩后测定的1-羟基芘的值为不浓缩测定值的72.9％～80.6％.利用不浓缩的酶水解高效液相色谱-荧光检测器法测定尿样中的1-羟基芘,栓出限为0.02ng/ml,在给定浓度范围内,1-羟基芘的峰面积与浓度有良好的线性关系,线性回归方程为y =2.404 1x -0.083(r =0.999 9),加标回收率为94.8％ ～ 108.7％,平均相对标准偏差为2.7％.[结论]不浓缩的酶水解高效液相色谱- 荧光检测器法可用于测定尿样中的1-羟基芘,且该方法具有操作简单,灵敏度、精密度高,重现性、回收率好的特点.
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