不同蟾皮提取物中蟾毒灵、华蟾酥毒基和脂蟾毒配基血浆蛋白结合率的研究

Study on plasma protein binding rates of bufalin,cinobufagin and resibufogenin in different kinds of toad skin extracts

目的研究不同蟾皮提取物中蟾毒灵、华蟾酥毒基和脂蟾毒配基的血浆蛋白结合率。方法采用平衡透析法,以高效液相色谱-串联质谱(LC-MS/MS)法测定,比较两种蟾皮提取物及不同浓度的蟾毒灵、华蟾酥毒基和脂蟾毒配基对照品溶液的血浆蛋白结合率。结果建立的LC-MS/MS法线性良好,精密度、准确度、回收率以及稳定性均符合生物样品分析要求。3种不同浓度蟾毒灵的血浆蛋白结合率分别为(75.03±1.97)%、(72.33±2.51)%、(71.80±1.53)%;蟾皮药材提取物1中的为(-16.67±1.58)%,蟾皮药材提取物2中的为(71.93±1.48)%。3种不同浓度的华蟾酥毒基血浆蛋白结合率分别为(52.33±4.13)%、(52.47±3.84)%、(47.60±2.46)%;蟾皮药材提取物1中的为(-51.23±3.53)%,蟾皮药材提取物2中的为(40.93±1.29)%。3种不同浓度的脂蟾毒配基血浆蛋白结合率分别为(91.27±1.09)%、(93.60±0.85)%、(88.13±0.76)%;蟾皮药材提取物1中的为(52.67±3.58)%,蟾皮药材提取物2中的为(80.13±2.38)%。结论蟾毒灵、华蟾酥毒基和脂蟾毒配基的血浆蛋白结合率跟药物浓度不呈依赖性,蟾皮药材提取物1中蟾毒灵、华蟾酥毒基和脂蟾毒配基的血浆蛋白结合率显著低于对照品蟾毒灵、华蟾酥毒基和脂蟾毒配基,而蟾皮药材提取物2的血浆蛋白结合率基本与对照品一致;提示蟾皮药材提取物1中混合物体系导致蟾毒灵、华蟾酥毒基和脂蟾毒配基的血浆蛋白结合率的改变。

Objective To study the plasma protein binding rates of bufalin,cinobufagin and resibufogenin in different kinds of toad skin extracts.Methods The plasma protein binding rates of bufalin,cinobufagin and resibufogenin in two extracts of toad skin and control solution with different concentrations of bufogenin,cinobufagin and cinobufagin were determined by equilibrium dialysis and LC-MS/MS method.Results The LC-MS/MS method showed good linearity,the accuracy,precision,recovery and stability of the analytes all met the methodology requirements.The plasma protein binding rates of bufalin in three different concentrations of control solution and in two kinds of toad skin extracts were(75.03±1.97)%,(72.33±2.51)%,(71.80±1.53)%,(-16.67±1.58)%and(71.93±1.48)%,respectively.The plasma protein binding rates of cinobufagin in three different concentrations of control solutionand in two kinds of toad skin extracts were(52.33±4.13)%,(52.47±3.84)%,(47.60±2.46)%,(-51.23±3.53)%and(40.93±1.29)%,respectively.The plasma protein binding rates of resibufogenin in three concentrations of control solution and in two kinds of toad skin extracts were(91.27±1.09)%,(93.60±0.85)%,(88.13±0.76)%,(52.67±3.58)%and(80.13±2.38)%,respectively.Conclusions The results show that the plasma protein binding rates of bufalin,cinobufagin and resibufogenin are not dependent on the drug concentration.The plasma protein binding rates of them in extract 1 are significantly lower than those of control solution,while the plasma protein binding rates of them in toad skin extract 2 are basically the same as those of the control solution.It is suggested the mixture system of toad skin extract 1 leads to the change of plasma protein binding rates of bufalin,cinobufagin and resibufogenin.