24 h盐酸吗啡缓释片与市售品在比格犬体内的药代动力学对比研究

Comparative Study on Pharmacokinetics of Morphine Hydrochloride 24-Hour Sustained-Release Tablets and Its Commercial Products in Beagle Dogs

目的建立测定比格犬血浆中盐酸吗啡浓度的液相色谱串联质谱(LC-MS/MS)法,比较自制24 h盐酸码啡缓释片与市售品的药代动力学行为和缓释特征。方法以磷酸可待因为内标,用乙腈沉淀分离蛋白;色谱柱为ACE Phenyl柱(100 mm×2.1 mm,5μm),流动相为0.1%甲酸水溶液-0.1%甲酸乙腈溶液(梯度洗脱),流速为500μL/min;采用电喷雾离子源、多反应监测模式,监测离子对分别为质荷比(m/z)286.1~165.3(吗啡)和m/z 300.2~165.1(可待因)。比格犬单剂量、三周期交叉口服方式给予24 h盐酸吗啡缓释片和2种盐酸吗啡市售品,采用LC-MS/MS法测定血药浓度,由Phoenix WinNonlin 7.0软件计算药代动力学参数。结果吗啡质量浓度在5~2000 ng/mL范围内与待测物和内标峰面积比值的线性关系良好(r=0.9984,n=8);准确度、精密度、稳定性试验结果的RSD和RE均低于20%。自制品与2种市售品的血药浓度时间曲线下面积(AUC_(0-t))、最大血药浓度(C_(max))无显著差异(P>0.05),达峰时间(t_(max))分别为市售品1的13.9倍、市售品2的7.1倍。结论自制24 h盐酸吗啡缓释片较2种市售品有明显的缓释特性。

Objective To establish a high-performance liquid chromatography-tandem mass spectrometry(LC-MS/MS)method for the determination of morphine hydrochloride concentration in the plasma of Beagle dogs,and to compare the pharmacokinetic behavior and sustained-release characteristics of self-made morphine hydrochloride 24-hour sustained-release tablets with commercial products.Methods Protein was isolated by precipitation with acetonitrile with phosphate as an internal standard.The chromatographic column was ACE Phenol column(100 mm×2.1 mm,5μm),the mobile phase was 0.1%formic acid aqueous solution-0.1%formic acid acetonitrile solution(gradient elution),the flow rate was 500μL/min.The quantitation analysis was performed by electrospray ionization source under multiple-reaction monitoring mode at mass charge ratio(m/z)of 286.1-165.3 for morphine and m/z of 300.2-165.1 for codeine.A single and three-cycle cross administration of the morphine hydrochloride 24-hour sustained-release tablets and the two commercial products were given in Beagle dogs,and the morphine hydrochloride concentration in plasma was determined by the LC-MS/MS method.The relevant pharmacokinetic parameters were calculated by the Phoenix WinNonlin 7.0 software.Results The linear range of morphine was 5-2000 ng/mL(r=0.9984,n=8).The RSD and RE of accuracy,precision,and stability test results were all lower than 20%.There was no significant difference in the area under the blood drug concentration-time curve(AUC_(0-t))and maximum blood drug concentration(C_(max))between self-made product and two commercial products(P>0.05).The peak time(t_(max))of the self-made product was 13.9 times that of commercial product 1 and 7.1 times that of commercial product 2,respectively.Conclusion Self-made morphine hydrochloride 24-hour sustained-release tablets have more significant sustained-release characteristics than the two commercial products.