基于LC-MS/MS测定大鼠血清中大麻二酚及其药动学研究

Determination of cannabidiol in rat serum and its pharmacokinetic effects based on LC-MS/MS

目的:建立SD大鼠血清中大麻二酚(CBD)的LC-MS测定方法,研究CBD在正常大鼠体内药动学特征。方法:SD大鼠灌胃给予CBD溶液(20、50、100 mg·kg^(-1))及CBD油(50 mg·kg^(-1)),在给药前及给药后0.083、0.25、0.5、1、1.5、2、3、4、5、6、7、8、10、12、24、48 h眼眶采血,采用LC-MS技术,以Waters Atlantis T3色谱柱,以0.1%甲酸-水为流动相A,0.1%甲酸-乙腈为流动相B,梯度洗脱,多反应监测(MRM),正离子模式检测,测定大鼠体内CBD的浓度,采用Winnonlin软件计算药动学相关参数。结果:CBD在血清中线性关系良好;提取回收率在103.3%~115.7%;低、中、高3个浓度CBD日内RSD分别为3.3%、1.5%、0.29%,日间RSD分别为3.8%、2.7%、5.8%;提取回收率为(104.9±2.4)%、(115.7±0.4)%、(103.3±3.6)%;样品于-80℃放置稳定。CBD溶液T_(max)为2 h,CBD油T_(max)为9 h,不同剂量CBD给药血药浓度差异较大,C_(max)、AUC随剂量而升高,MRT在7~13 h。结论:相较于已有文献该分析方法耗时短,进样量少,专属性、灵敏度均能满足大鼠血清药动学测定。CBD溶液在血中分布迅速,T_(max)仅为2 h,CBD溶液与CBD油在给药后30 min内快速通过血脑屏障入脑。不同剂量CBD给药血药浓度差异较大,C_(max)、AUC随剂量而升高。

Objective:To establish an LC-MS method for determination cannabidiol(CBD)in rat to verify the methodology and to study the influence of CBD on pharmacokinetics in normal rats.Methods:CBD solution(20,50,100 mg·kg^(-1))and CBD oil(50 mg·kg^(-1))were administrated intragastricly.The blood samples were collected before administration and at 0.083,0.25,0.5,1,1.5,2,3,4,5,6,7,8,10,12,24 and 48 h after administration.The concentrations of CBD were determined by LC-MS method,which using Waters Atlantis T3 column,0.1%formic acid-water as mobile phase A,0.1%formic acid-acetonitrile as mobile phase B,gradient elution,multiple reaction monitoring(MRM)andpositive ion mode detection.Winnonlin software package was used to analyze the calculations of pharmacokinetic parameters.Results:Excellent linear relationships of CBD were obtained in the serum.The recoveries of the analytes were 103.3%-115.7%.The intra-day RSDs of low,medium and high CBD concentrations were 3.3%,1.5%and 0.29%,and the intra-day RSDs were 3.8%,2.7%,and 5.8%,respectively.The extraction recoveries were(104.9±2.4)%,(115.7±0.4)%and(103.3±3.6)%.The sample was stable at-80℃.The T_(max)of CBD solution was 2 h,and the T_(max)of CBD oil was 9 h.The plasma concentrations of CBD in different doses were significantly different.C_(max)and AUC increased with the dose,and MRT was between 7 and 13 h.Conclusion:The proposed method consumed shorter analysis time and less injection than investigation reported before.The specificity and sensitivity can meet requirements of serum pharmacokinetic assay.The CBD solution distributes rapidly in the blood,with a T_(max)of only 2 h.The CBD solution and CBD oil quickly pass through the blood-brain barrier and enter the brain within 30 min after administration.The blood concentration of CBD administered with different doses is quite different,and C_(max)and AUC increase with the dose.