摘要
目的：建立高效液相色谱-二极管阵列法同时检测尿中内源性氢化可的松及其代谢产物的方法。方法：采用Kromasil C18反相色谱柱（250mm×4．6mm，5μm），柱温为40℃，流动相为水-乙腈（78：22，V／V，用磷酸调pH值至2．8，流速梯度洗脱），检测波长为238nm，尿液用固相萃取法处理。结果：6β-羟基氢化可的松和氢化可的松分别在20～1000、5～100μg／L浓度范围内线性关系良好，最低检测浓度分别为2、3μg／L，萃取回收率均大于80％，日内、日间相对标准差均小于5％。尿中6β-羟基氢化可的松与氢化可的松浓度的平均比值为0．66±0．37。结论：本方法专属性强、灵敏度高、简单可靠，可用于研究体内药物代谢机制及药物相互作用。
OBJECTIVE： To establish a HPLC- DAD method for simultaneous determination of endogenous cortisol and its metabolite in human urine. METHODS： The HPLC separation was performed on an Kromasil C18（250mm×4.6mm,5μm） reversion phase chromatography column at column temperature of 40℃. Water- acetonitrile （78 ： 22, pH value regulated to 2.8 with phosphoric acid, flow - rate gradient elution） was used as mobile phase. Wave length was 238nm. The urine was extracted with solid- phase extraction method. RESULTS： The calibration curves were linear in the range of 20- 1 000μg/L for 6β- hydroxycortisol and 5- 100μg/L for cortisol, respectively. The lowest detectable concentrations were 2μg/L and 3μg/ L respectively. The average extraction recoveries for the two compounds were above 80%. The intra - day and inter - day RSD were less than 5%. The average ratio of concentration of endogenous 6β- hydroxycortisol to cortisol in urine was 0.66 ± 0.37. CONCLUSION： The present method is highly specific, sensitive, simple and reliable, and can be used for the study of metabolic mechanism and interactions of drugs in vivo.
出处
《中国药房》
CAS
CSCD
北大核心
2006年第16期1235-1237,共3页
China Pharmacy
