高效液相色谱法测定人血浆3-氯酪氨酸浓度

Determination of 3-chlorotyrosine concentration in human plasma by HPLC with electrochemical detection

目的建立人血浆3-氯酪氨酸（3-CT）浓度的定量分析方法。方法用三氯乙酸沉淀血浆蛋白后，HPLC—ECD定量检测。结果3．氯酪氨酸血浆标准曲线的线性范围为0．0625—2．000μmol·L^-1，r〉0．9966，回收率〉71．8％。日内和日间的精密度（RSD％）分别〈8．8％和11．1％。3-氯酪氨酸的保留时间为（16．57±0．30）min，总分析时间32min。测得健康人和尿毒症患者（各6例）血浆3-CT浓度分别为（0．16±0．13）和（2．47±0．73）μmol·L^-1（P〈0．01）。结论本法所需样本量低，预处理步骤简便，适用于临床进行氧化应激、治疗、预后、诊断等研究工作之需。

AIM To develop a quantitative analysis method for the determination of 3-chlomtyrosine （3-CT） in human plasma. METHODS 3-CT in plasma was detennined using reversed-phase HPLC with electrochemical detection after precipitation all of the plasma proteins with trichloroacetic acid. RESULTS The linearity of the method was in the range of 0.062 5 to 2.000 μmol·L^- 1 for 3-CT with a correlation coefficient of more than 0. 996 6. Over 71.8% of the recovery of 3-CT in plasma was obtained. The relative standard deviation（RSD） of the method was less than 8.8% for within-day and 11.1% for between-day, respectively. The average retention time for 3-CT was （ 16.57 ± 0.30）rain. A single run for the measurement of 3-CT was 32 min. The average levels for measured 3-CT were （0.16 ± 0.13）μmol· L^- 1 for healthy population （ n = 6） and （2.47 ± 0.73） μmol· L- 1 for uremic patients （ n = 6）, respectively （ P 〈 0.01 ）. CONCLUSION The presented method is reliable and simple,and is applicable to those areas such as diagnosis, treatment and prognosis in clinic as well as the research work on oxidative stress and related diseases.