酒石酸长春瑞滨长循环（热敏）脂质体在比格犬体内的药代动力学研究

摘要

目的：应用液相色谱质谱联用（ HPLC-MS／MS）建立长春瑞滨（ NVB）比格犬血浆检测方法并用于药代动力学研究。方法血浆样品采用蛋白沉淀和液－液萃取方法。色谱柱Venusil XBP C18（2．1 mm ×50 mm，3μm），柱温30℃，流动相∶水（10 mmol／L乙酸铵，1％乙腈）∶甲醇＝20∶80，流速0．3 ml／min，进样量5μl；采用正离子模式、电喷雾离子源（ESI）及多反应监测（MRM），NVB m／z 779．4～765．4，内标长春新碱m／z 825．4～122。结果 NVB在5～2500 ng／ml浓度范围线性良好（ r＝0．9994），准确度、精密度和提取回收率符合方法学验证要求，样品在－20℃和室温放置稳定性良好。比格犬给药后，Cmax分别为（833．51±150．42）和（1397．95±443．05） ng／ml、AUC（0－t）分别为（577．16±223．5）和（1059．82±408．27） ng／ml· h，Cl分别为（3014．64±1049．17）和（1633．10±551．77） ml／（h· kg）。显著性检验表明，两种制剂的Cmax、AUC（0－t）、AUC（0－∞）和Cl均有显著性差异（P≤0．05）。结论建立了准确、灵敏、可靠的定量检测比格犬血浆中NVB分析方法，可满足药代动力学研究。脂质体能提高药物达峰浓度，延长药物半衰期，增加药物暴露强度并降低毒性反应。%Objective To establish a quick method to analyze vinorelbine ( NVB) in plasma of beagle dogs and study its pharmacokinetics of long-circulation and thermosensitive liposome loaded vinorelbine bitartrate.Methods The plasma was treated with liquid-liquid extraction after precipitation in methanol.The analysis was perfomed on a Venusil XBP C18 column(2.1 mm ×50 mm, 3 μm) at 35℃,the mobile phase consisted of methanol and water( containing 10 mmol/L ammonium acetate, 1%acetonitrile) 80∶20 and injection volume was 10μl.The type of mass spectrum was multireactive monitoring(MRM) in a positive mode.The monitor transitions were m/z 779.4-765.4 for vinorelbine and m/z 825.4-122 for vincristine.Results The concentration range from 10 to 2500 ng/ml had a good linearity ( r=0.0994).The precision, accuracy and extraction efficiency were acceptable.The plasma samples were stable for 10 days at -20℃ and 24 h at room temperature.Pharmacokinetic study in beagle dogs showed that the main parameters for injection and liposome were Cmax(833.51 ±150.42) and (1397.95 ±443.05)ng/ml, AUC(0-t) (577.16 ±223.57) and (1059.82 ±408.27) ng/ml· h, Cl(3014.64 ±1049.17)and 1633.10 ±551.77 ml/(h· kg), respectively.Conclusion A reliable HPLC-MS/MS method for vinorelbine analysis is established and can be applied to the pharmacokinetics study of liposome.The results show that liposome has a higher AUC, Cmax and longer Cl than injection.Meanwhile, liposome has a lower irritability.