摘要
建立了高效液相色谱-荧光检测法同时测定人体尿液中的蝶呤、新蝶呤、生物蝶呤和墨蝶呤。尿液在酸性条件下经碘-碘化钾溶液氧化30 min,滴加抗坏血酸还原液后,可进行液相色谱分析。色谱柱为Diamonsil C18柱,用体积比9∶10的水-甲醇为流动相,流速为1.0 mL/min,荧光检测波长为λex=360 nm,λem=445 nm。新蝶呤、生物蝶呤、蝶呤和墨蝶呤的线性范围分别是0.005～1.5μg/mL、0.05～2.0μg/mL、0.01～1.0μg/mL和0.3～10.0μg/mL,检出限依次为0.003μg/mL、0.002μg/mL、0.001μg/mL和0.15μg/mL。加标平均回收率在84.8%～120.0%之间。用该方法测定了健康人、普通病人和癌症病人尿样中的蝶呤、新蝶呤、生物蝶呤和墨蝶呤,发现癌症病人尿样中的新蝶呤水平与健康人和普通病人相比有所升高,生物蝶呤水平有所降低。该方法准确,可行,适用于临床上尿样中蝶呤类化合物的检测。
The pteridines in human urine have been determined by high performance liquid chromatography（HPLC） fluorescence detection.The urine was oxidized by I2-KI under acidic condition for 30 min and mixed with ascorbic acid,then the sample was diluted 5 times,filtered by 0.45 μm membrane and analyzed by HPLC-fluorescence detection.The chromatographic column was Diamonsil C18,the mobile phase was H2O-CH3OH（90∶10,V/V）,the flow rate was 1.0 mL/min,the excitation and emission wavelengths were 360 nm and 445 nm,respectively.The linear ranges of neopterin,biopterin,pterin and sepiapterin were 0.005～1.5 μg/mL,0.05～2.0 μg/mL,0.01～1.0 μg/mL and 0.3～10.0 μg/mL,respectively,and the limits of detection were 0.003 μg/mL,0.002 μg/mL,0.001 μg/mL,0.15 μg/mL.The recoveries of four target analytes in real urine samples were in the range of 84.8%～120.0%.The concentrations of neopterin,biopterin,pterin and sepiapterin in urines were determined and a comparison of the analytical results for healthy people,normal patients（control groups） and cancer patients was presented.The concentration of neopterin in cancer group was higher than that in control groups,while the concentration of biopterin in cancer group was lower.The method is accurate and can be used to detect pteridines in clinical urine samples.
出处
《分析科学学报》
CAS
CSCD
北大核心
2011年第1期21-25,共5页
Journal of Analytical Science
基金
国家自然科学基金(No.20765002)
江西省自然科学基金(No.0620049)
关键词
蝶呤类化合物
人体尿液
高效液相色谱
荧光检测
Pteridines
Urine
High performance liquid chromatography
Fluorescence detection
