RP-HPLC法测定人血液和尿液中草酸  

Determination of oxalate in human plasma and urine by RP-HPLC

作　　者：刘宏胜[1] 吴胜群[1] 

机构地区：[1]天津市第一中心医院卫生部危重病急救医学重点实验室,天津300192

出　　处：《中草药》2014年第20期2935-2938,共4页Chinese Traditional and Herbal Drugs

摘　　要：目的建立RP-HPLC法测定人血液与尿液中草酸的质量浓度,监测原发性高草酸盐尿症（PH）患者,实施肝肾联合移植术前后草酸质量浓度的变化。方法采用Agilent XDB C18（150 mm×4.6 mm,5μm）色谱柱;Agilent Zorbax extend-C18（12.5 mm×4.6 mm,5μm）为预柱;流动相为甲醇-0.1 mol/L醋酸胺水溶液（15∶85）,体积流量1.2 mL/min;紫外检测波长314 nm;柱温26.3℃;进样量50μL;以邻苯二胺为衍生剂,与血液与尿液中草酸反应,生成具有强紫外吸收化合物2,3-二羟基喹喔啉。结果血液中最低检测草酸质量浓度为0.3 mg/L,线性范围为1.953-125 mg/L,平均回收率为94.89%,精密度RSD为4.1%;尿液中草酸最低检测质量浓度为0.5 mg/L,线性范围为1.953-125 mg/L,平均回收率为94.31%,精密度RSD为3.2%。结论该方法简便,灵敏度高,重复性好,回收率高,是临床检测草酸质量浓度的可信方法。Objective To establish a method of reversal phase-high performance liquid chromatography（RP-HPLC） for determining the mass concentration of oxalate in human plasma and urine and to monitor the variation of mass concentration of oxalate in the patients with primary hyperoxaluria（PH） before and after combined liver-kidney transplantation. Methods Agilent XDBC18（150 mm × 4.6 mm, 5 μm） column and Agilent Zorbax extend-C18（12.5 mm × 4.6 mm, 5 μm） guard column were used. Methyl alcohol and aqueous solution containing 0.1 mol/L ammoniom acetate（15∶85） were used as mobile phase. The flow rate was at 1.2 mL/min, ultraviolent determination wavelength was 314 nm, column temperature was at 26.3 ℃, and injection volume was 50 μL. o-phenylenediamine was used as derivating agent, reacted with oxalate in human plasma and urine so as to obtain the compound with better ultraviolet absorption—2, 3-dyhydroxy quinoxaline. Results The detection limit in human plasma was 0.3 mg/L, the linear range was 1.953—125 mg/L, the average recovery was 94.89%, and its RSD was 4.1%; The detection limit in urine was 0.5 mg/L, the linear range was 1.953—125 mg/L, the average recovery was 94.31%, and its RSD was 3.2%. Conclusion The method is believable for determining the mass concentration of oxalate with its simplicity, sensibility, repeatability, and better recovery rate.

关 键 词：尿草酸 血草酸 原发性高草酸盐尿症(PH) 肝肾联合移植术 邻苯二胺 2 3-二羟基喹喔啉 RP-HPLC 

分 类 号：R286.02[医药卫生—中药学]