加压毛细管电色谱法测定肥胖大鼠尿液中内源性代谢物

利用氯甲酸乙酯作为衍生化试剂,建立了测定尿样中内源性代谢物的加压毛细管电色谱方法。采用毛细管色谱柱EP-150-30/50-5-C18,以0.01%TFA水溶液(A)与0.01%TFA的95%乙腈水溶液(B)组成流动相,加压13MPa;工作电压为2kV;流速0.08mL/min;检测波长214nm;室温;柱上检测;进样体积5μL,同时测定了正常大鼠尿液与肥胖大鼠尿液中几种代谢物的含量。在最佳条件下,6种内源性代谢物在1.2～500mg/L范围内线性良好,相关系数为0.9988～0.9999;日间和日内精密度小于5%;平均加样回收率在95.9%～103.2%之间。本方法用于大鼠尿液中内源性代谢物含量的测定,简单、灵敏、准确。

Determination of Endogenous Metabolites in Obesity Rat Urine by Pressurized Capillary Electrochromatography with Ethyl Chloroformate Derivatization

Pressurized capillary electrochromatographic(pCEC) method was developed to analyze metabolites in normal and obesity rat urines with ethyl chloroformate derivatization.The separation was performed on a capillary column(EP-150-30/50-5-C18),the good resolution was obtained with the mobile phase of water(0.01% trifluoroacetic acid,TFA)-95% acetonitrile(0.01% TFA) by using the following gradient:0-30 min,0-50% B;30-40 min,50%-75% B.Under the optimized conditions,such as pressure of 13 MPa,applied voltage of 2 kV,flow rate of 0.08 mL/min and room temperature,the contents of metabolites in the urine of normal and obesity rats were obtained,and the correction coefficients were 0.9988-0.9999 in the range of 1.25-500 g/L.The intra-and inter-day precisions were less than 5%.The average recoveries were ranged from 95.9% to 103.2%.The method is simple,sensitive and accurate for the determination of saponins in rat urine sample.