高效液相色谱-荧光分析法测定人体尿液中的墨蝶呤

Determination of sepiapterin in human urine by HPLC with fluorescence detection

建立了人体尿液中墨蝶呤的高效液相色谱-荧光分析方法。尿液经乙腈处理,过0.45μm水相滤膜后定容,可进行液相色谱分析。色谱柱为Diamonsil C18柱,以水-甲醇(80:20,v/v)为流动相,流速为1.0 mL/min,荧光检测波长为λex=425 nm,λem=530 nm。墨蝶呤含量在0.005～1.0μg/mL范围内与色谱峰面积呈良好的线性关系,线性回归方程为y=106x-328.02(r=0.999 0),检测限是0.002μg/mL。其加标平均回收率在98.7%～106.8%之间,相对标准偏差小于7.34%。该方法简便,应用于胃癌病人和健康人尿样中墨蝶呤测定,结果较好。

The determination of sepiapterin in human urine by HPLC-FLD has been studied.The urine was filtered by 0.45 μm membrane after dealed with CH3CN,then the sample was analyzed by HPLC-FLD.The chromatographic column was Diamonsil C18,the moble phase was H2O-methanol（80：20,v/v）,the flow rate was 1.0 mL/min,the excitation and emission wavelengths were 425 nm and 530 nm,respectively.The linear range of sepiapterin was 0.005-1.0 μg/mL.The linear regression equation was y=106x-328.02（r=0.999 0）,and the limit of detection was 0.002 μg/mL.The average recoveries were in the range of 98.7%～106.8% with the RSD below 7.34%.The proposed method was applied to the determination of sepiapterin in both the gastric cancer patients＇urine samples and healthy human urine with satisfactory results.