高效液相色谱法定量尿酸、黄嘌呤和次黄嘌呤在人体血清中的浓度

目的:建立高效液相色谱法测定尿酸、黄嘌呤和次黄嘌呤在人体血清中的浓度.方法:用Agilent ZORBAX SB-Aq column色谱柱,ZORBAX-Extend C18预柱,以甲醇和47 mmol/LKH2PO4水溶液作为流动相,流速:1.0 mL/min,检测波长为260 nm.结果:血清中尿酸、黄嘌呤和次黄嘌呤的线性范围分别是(1.667～166.667)×103 ng/mL (r=0.9995)、(0.033～3.338)×103 ng/mL(r=0.9994)和(0.033～3.338)×103 ng/mL(r=0.9996),日间与日内精密度均＜15％.结论:该法灵敏度高,精密度和准确度佳,能满足非布司他体内药效学研究.

Quantification of uric acid,xanthine and hypoxanthine in human serum by HPLC

AIM： To develop a specific,sensitive,and accurate HPLC method to measure uric acid,xanthine and hypoxanthine concentrations in human serum.METHODS： Agilent ZORBAX SB-Aq column coupled with a ZORBAX-Extend C18 guard column was employed.The mobile phase consisted of methanol/ 47 mmol/L KH2PO4 solution and the flow rate was 1.0 mL/min.The detection wavelength was 260 nm.RESULTS：Good linearity of uric acid,xanthine,hypoxanthine in human serum were（1.667-166.667）×103 ng/mL（r=0.9995）,（0.033-3.338）×103 ng/mL（r=0.9994）,（0.033-3.338）×103 ng/mL（r=0.9996）,respectively.Intra-and inter-day validation were both less than 15%.CONCLUSION： A rapid,accurate and specific method was developed and validated for the determination of uric acid,xanthine and hypoxanthine in human serum.It was suitable for the pharmacodynamic studies of febuxostat.