高效液相色谱法同时测定尿中新喋呤和生物喋呤

 建立了同时分离检测尿中新喋呤（neopterin，NP）和生物喋呤(biopterin,BP)的高效液相色谱（HPLC）-荧光检测方法。采用Hypersil BDS C18柱、甲醇-水(体积比为10∶90)流动相(流速0.5 mL/min)、激发波长360 nm、发射波长 440 nm、柱温20 ℃的色谱条件，同时分离测定了尿中的NP和BP。尿标本经三氯乙酸处理，在4 ℃下，以12000 r/min的速率离心15 min，上清液用碱中和后，取30　μL直接进样。研究结果表明，NP的线性范围为0.12～10

Simultaneous Determination of Neopterin and Biopterin in Urine by High Performance Liquid Chromatography

A method was established for simultaneous determination of neopterin (NP) and biopterin (BP) in urine by high performance liquid chromatography-fluorescence detection. Conditions: column, Hypersil BDS C18 (4.6 mm i. d. x 200 mm, 5 microm); mobile phase, methanol-water (10:90, v/v); flow rate, 0.5 mL/min; column temperature, 20 degrees C; excitation wavelength, 360 nm; emission wavelength, 440 nm. Urine samples were first precipitated with trichloroacetic acid, then centrifuged at 12 000 r/min for 15 min at 4 degrees C, and finally added with NaOH to neutralize the supernatant. Injection volume was 30 microL. The linear ranges of determination of the standards NP and BP were 0.12 - 1000 microg/L and 0.48 -1000 microg/L, with the detection limits of 0.01 microg/L and 0.1 mictog/L, the relative standard deviations (RSDs) in urine of 1.09% and 3.39%, and average recoveries of 100.7% and 100.3%, respectively. The concentrations of NP and BP, and concentration ratio of NP and BP in urines were detected and compared between controls and uremia patients. The concentration of creatinine (Cr) in urines was detected with picric acid method. The concentration ratios of NP and Cr, NP and BP in uremia group increased more significantly than that in normal group (P < 0.05). The method is simple, fast, accurate and convenient. It can be used to detect NP and BP for clinical urine samples and provide a more convenient and accurate tool for the diagnosis of patients.