大鼠皮下氢化可的松浓度在线检测方法的建立

On-line detection of hydrocortisone concentration in rat subcutaneous adipose with liquid chromatography-mass spectrometry/mass spectrometry

目的利用微透析采样技术和高效液相色谱-质谱(LC-MS/MS)检测方法,建立大鼠皮下局部药物浓度氢化可的松(HC)的检测方法。方法采用微透析方法对大鼠皮下进行透析采样,以5%乙醇-Ringer’s液作为灌流液,并建立LC-MS/MS对样品中HC浓度进行检测。结果采用离子对m/z 363.2→121.1进行HC定量分析,在0.5～1000 ng/ml范围内,浓度峰面积呈现良好的线性关系,日间及日内精密度、准确度均在15%以内,无明显的基质效应。微透析实验方法的探针回收率较高,达到59%。结论本研究所建立的实验方法,专属性好,灵敏度高,可成功用于大鼠皮下HC含量测定。

Objective To develop a method for detecting the topical concentration of hydrocortisone（HC） in the subcutaneous adipose of rats using microdialysis sampling technique and liquid chromatography-mass spectrometry/mass spectrometry（LC-MS/MS）.Methods Topical samples were collected by applying the probe into the subcutaneous adipose of rats,with alcohol（5%）-ringer＇s solution as the perfusion solution.A LC-MS/MS method was established for detecting the HC concentration in the dialysates.Results The protonated precursor to produce ion transitions monitored for HC was m/z 363.2→121.1.The calibration curve was linear over the range of 0.5-1000 ng/ml,with the intra-and inter-day precision and accuracy within ±15%,and no significant matrix effect was noted.The in vivo recovery of the probe was about 59%.Conclusion A selective and sensitive method has been successfully established for the on-line HC detection in the subcutaneous adipose of rats.