HPLC-MS/MS法检测血根碱在鸡体内的药代动力学研究

The Pharmacokinetics Study of Sanguinarine in Broiler using HPLC-MS/MS

关键词:  血根碱  二氢血根碱  鸡  药代动力学

Key words:  sanguinarine  dihydrosanguinarine  broiler  pharmacokinetic

摘要:

为了建立给鸡灌胃血根碱后同时测定鸡血浆中血根碱及二氢血根碱浓度的HPLC-MS/MS检测方法，并评价血根碱在鸡体内的药代动力学特征，采用甲醇提取鸡血浆中的血根碱及代谢物二氢血根碱，以0.2 %甲酸水（A）-乙腈（B）为流动相，选用 Agilent Poroshell 120 EC C18（2.1 mm×150 mm，2.7 μm）色谱柱进行色谱分离，采用电喷雾离子源(ESI)三重四极杆串联质谱进行分析，多重反应监测 (MRM)方式进行检测。血根碱及代谢产物二氢血根碱均在0.1～50.0 ng/mL浓度范围内与色谱峰面积呈良好线性关系，提取回收率分别为88.95%～95.81%和82.00%～87.00%；日内及日间精密度（RSD）均小于15%；血根碱在鸡体内的药代动力学参数Cmax，Tmax，MRT，CL分别为0.90±1.053 ng/mL、0.38±0.30 h、5.11±0.74 h和44.09±3.05 h。血根碱在鸡体内血药达峰时间以及滞留在体内的平均时间较短，代谢物二氢血根碱的血药浓度是血根碱的血药浓度5.74倍，说明代谢物二氢血根碱的血药浓度远大于血根碱的血药浓度。该方法具有快速、灵敏度高、专属性强等特点，适用于同时测定鸡血浆中血根碱和二氢血根碱的血药浓度。

Abstract:

To establish a sensitive HPLC-MS/MS method for simultaneous determination the concentration of sanguinarine and dihydrosanguinarine in broiler plasma after oral administration of sanguinarine, and the pharmacokinetic characteristics of sanguinarine were evaluated. Sanguinarine and its metabolites dihydrosanguinarine were extracted from plasma with methanol, then separated on an Agilent Poroshell 120 EC C18 column using water-0.2% formic acid(A) and acetonitrile (B) as mobile phase. Electrospray ionization (ESI) source was applied and operated in multiple reaction monitoring (MRM) mode. The linear calibration curve was obtained in the concentration range of 0.1～50 ng/mL for sanguinarine and its metabolites dihydrosanguinarine, The extraction recoveries were 88.95%～95.81% and 82%～87% for sanguinarine and dihydrosanguinarine respectively. The inter-and intra-day precisions were less than 15%. The pharmacokinetic parameters of sanguinarine were as follows: Cmax=0.90±1.053ng/mL, Tmax=0.25±0.38h, MRT=5.11±0.74 h, CL=44.09±3.05 h. The time to peak of plasma and the average time of residence in the body were short; the plasma concentrations of metabolites dihydrosanguinarine is 5.74 times as high as that of sanguinarine; it shows that the plasma concentrations of metabolites dihydrosanguinarine is much higher than that of sanguinarine. The method was shown to be effective, convenient and suitable for simultaneous pharmacokinetic study of sanguinarine and dihydrosanguinarine in broiler plasma.