HPLC法测定人尿液中的3-甲基组氨酸

Determination of human urinary 3-methylhistidine by HPLC

目的采用HPLC法测定人尿液中3-甲基组氨酸(3-MH)的含量。方法尿样经乙腈沉淀蛋白和邻苯二甲醛柱前衍生化,60s内进样,以乙醇胺为内标,测定尿液中的3-MH的含量。结果3-MH的衍生物与尿液中其他杂质的衍生物完全分离。线性范围为10~400μmol·L^(-1)(r=0.9993),平均回收率为94.03%~97.13%(n=5),日内和日间RSD分别<7.80%和8.25%,最低检出浓度达5μmol·L^(-1)。结论该方法灵敏度高,可用于人尿液中3-MH含量的测定。

AIM To develop a HPLC method to determine the concentration of 3-methylhistidine in human urine. METHODS The concentration of 3-methylhistidine in human urine was determined by precipitating the urine with acetonitrile, precolunm deriving with o-pathaldialdehyde and injecting within 60 s, using ethanolamine as internal standard. RESULTS The derivative of 3-methylhistidine was clearly separated from other derivatives in urine. The linear range was 10 - 400μmol·L^-1 （ r = 0.999 3）, the average recovery was between 94.03 % - 97.13 % （ n = 5）. The intra-day and inter-day precision were less than 7.80% and 8.25%, respectively, and the detection limit reached 5 μmol·L^-1. CONCLUSION This is a high sensitive and reliable method to determine the urinary 3-methylhistidine with a widerange.