高效液相色谱-柱后化学发光法检测人体尿液中的卡托普利

在酸性条件下,Ce?氧化Ru(bipy)32+生成Ru(bipy)33+,同时氧化卡托普利生成二硫化物中间活性态(RS-SR]*),Ru(bipy)33+和二硫化物中间活性态之间相互反应产生强烈的化学发光。基于此,根据发光试剂Ru(bipy)32+水溶性好、试剂稳定等特点,将其加入到流动相中,通过高效液相色谱分离,建立了柱后化学发光快速灵敏检测卡托普利的方法。在以甲醇-0.01mol/L KH2PO4-1g/L Ru(bipy)32+(80∶20∶2,V/V)为流动相,流速为0.9mL/min,8.0×10-4 mol/L Ce?的优化实验条件下,方法的线性范围为2.0×10-7～1.0×10-4 mol/L(R2=0.9988),检出限为6.0×10-8 mol/L(S/N=3),并对1×10-5 mol/L卡托普利平行测定11次,相对标准偏差(RSD)为1.8%。将本方法用于人体尿液中卡托普利含量的测定,结果令人满意。结合化学发光光谱,对该体系发光机理进行了探讨。

Determination of Captopril in Human Urine Samples by High Performance Liquid Chromatography Coupled to Post-column Chemiluminesence Detection

An improving approach for the determination of captopril in human urine samples by high performance liquid chromatography(HPLC) with chemiluminescence(CL) detection was developed.The method was based on the CL reaction of captoril with Cerium sulfate(Ce(SO 4) 2) and tris(2,2′bipyridyl) ruthenium?(Ru(bipy) 3 2+),which was added in the mobile phase.The chromatographic separation was performed on a Kromasil TM RP-C 18 column(150mm×4.6mm i.d.,particle size : 5"m,pore size : 10nm) with a mobile phase consisting of methanol-0.01 mol / L KH 2 PO 4-1g / L Ru(bipy) 3 2+(80∶20∶2,V / V) at a flow rate of 0.9 mL / min,the total analysis time was within 10min.Under the optimal conditions of 0.1mol / L H 2 SO 4,8.0×10-4 mol / L Ce(SO 4) 2,the linear range was 2.0×10 -7-1.0×10-4 mol / L(R 2 =0.9988) with the detection limit of 6.0×10-8 mol / L,and the relative standard detection(RSD) was 1.8% for 1.0×10-5 mol / L(n=11).The possible mechanism of the CL reaction was also discussed briefly.