高效液相色谱-荧光检测法测定人体尿液中的蝶呤-6-羧酸  

Determination of pterin-6-carbonxylic acid in human urine by HPLC with fluorescence detection

作　　者：郭岚[1] 张云伟[2] 万益群[1] 鄢爱平[1] 

机构地区：[1]南昌大学分析测试中心,南昌330047 [2]江西省分析测试中心,南昌330029

出　　处：《分析试验室》2008年第5期17-19,共3页Chinese Journal of Analysis Laboratory

基　　金：国家自然科学基金(20765002);江西省自然科学基金(0520057,0620049)项目资助

摘　　要：利用蝶呤-6-羧酸具有天然荧光的特点,建立了人体尿液中蝶呤-6-羧酸的高效液相色谱-荧光分析方法。尿液经硫酸铵处理后,过0.45μm水相滤膜,直接进行液相色谱分析,色谱柱为NOVA-PAK C18柱,保护柱为RCSS Guard-PAKTMC18柱,流动相为V（5 mmol/L KH2P04-NaOH缓冲溶液,pH 7.3）∶V（甲醇）=99∶1）,流速为0.6 mL/min,荧光激发波长为338 nm,荧光发射波长为420nm。蝶呤-6-羧酸在0.05～1.2μg/mL范围内呈线性关系,相关系数为0.9974,检出限为0.010μg/mL。分别添加0.625、2.50和4.50μg蝶呤-6-羧酸标准品,平均回收率（n=5）在99.7%～105%之间,相对标准偏差小于3.6%。此法可用于临床尿样中蝶呤-6-羧酸的分析。The determination of pterin-6-carbonxylic acid in human urine by HPLC-FLD has been studied. The urine was filtered by 0.45 μm film after dealed with （NH4）2SO4, then the sample was analyzed by HPLC-FLD. The chromatographic column was NOVA-PAK C18, the guard column was RCSS Guard-PAKTM C18, the mobile phase was KH2PO4-NaOH （5 mmol/L, pH 7.3）-menthalol （ V： V= 99： 1）, the flow rate was 0.6 mL/min, λex = 338 nm, λem = 420 rim. Pterin-6-carbonxylic acid behaved linearly in the range of 0.05 - 1.2 μg/mL （r = 0.9974）, and the limit of detection was 0.010 μg/mL. The recoveries of urine samples spiked with 0.625, 2.50 and 4.50 μg pterin-6-car- bonxylic acid were 1.5%, 103%, 99.7% respectively with the RSD below 3.6%. The proposed method was applied to the determination of pterin-6-carbonxylic acid in human urine with satisfactory results.

关 键 词：蝶呤-6-羧酸 尿液 高效液相色谱 荧光检测 

分 类 号：O657.7[理学—分析化学]