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/L KH₂PO₄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.