HPLC-FD测定大鼠尿液中儿茶酚胺类化合物

Determination of Catecholamines Excretion in Rats Urinary by HPLC-FD

摘要:

目的: 建立HPLC-FD测定大鼠尿液中儿茶酚胺类化合物(CAs)的含量,为虚热证的药理学研究提供参考. 方法: 采用酸性氧化铝吸附法制备大鼠尿样;使用HiQSil C₁₈V色谱柱,流动相0.02 mol·L^-1 KH₂PO₄水溶液,流速0.5 mL·min^-1,激发波长(E_X)280 nm,发射波长(E_M)316 nm,柱温40℃. 结果: 大鼠尿样供试液中的儿茶酚胺类化合物可以与各干扰组分达到基线分离;CAs各组分线性、精密度和重复性良好;该法制备的样品溶液在16 h内测定,RSD<2%;CAs的回收率在65%~71%,试验中测得适应期后大鼠尿液中CAs含量稳定. 结论: 该文采用的供试品溶液制备方法简便,建立的色谱分析方法稳定、可靠,适合于大鼠尿液中CAs的含量测定.

Abstract:

Objective: To establish an HPLC-FD method for the determination of catecholamines excretion in rats urinary, and hence providing a reference for the pharmacological research of the hyperactivity of Yin deficiency and Huo exuberance. Method: Catecholamines was concentrated by acid alumina adsorption from rats urinary, followed by the analysis on a HiQ-Sil C₁₈V column at 40℃ with the 0.02 mol· L^-1 KH₂PO₄solution as mobile phases. FD excitation wavelength was set a 280 nm and emission wavelength was set a 316 nm, flow rate was 0.5 mL· min^-1 throughout the analysis. Result: Catecholamines could be satisfactorily separated from the interferential components in the sample. The linearity of the calibration curve, precision and repeatability was good with the average recovery reached 65% to 71%. In addition, the sample solution prepared was stable within 16 h(RSD<2%). And, the catecholamines excretion in rats urinary after adjustment raise was stable. Conclusion: The method for sample solution preparation was convenient. Moreover, the HPLC-FD method developed in the study was stable and reliable, and hence can be used to determine the catecholamines excretion in rats urinary.