长春新碱血药浓度测定方法的建立及其与肿瘤患儿细胞色素P450 3A酶活性的相关性

Determination of vincristine concentration in human plasma and its correlation analysis with cytochrome P450 3A activity in pediatric oncology patients

目的建立一种测定人血浆中长春新碱浓度的HPLC-MS/MS方法 ,并对肿瘤患儿中长春新碱血药浓度与细胞色素P450 3A(CYP3A)酶活性相关性进行探讨。方法用液液萃取法处理血浆样本。色谱柱:Shim-pack XR-ODS(2. 0mm×100. 0 mm,2. 2μm),流动相:0. 1%乙酸溶液-甲醇,梯度洗脱。用正离子模式,多重反应监测进行定量分析。考察该方法的专属性、标准曲线与定量下限、精密度与回收率、基质效应和稳定性,并用SPSS 13. 0对长春新碱血药浓度与CYP3A酶活性进行相关性分析。结果长春新碱在0. 5~100. 0μg m L-1内,线性关系良好,质控样品批内和批间RSD均小于10%,提取回收率和基质效应分别在95. 7%~105. 9%和95. 7%~105. 9%。在肿瘤患儿体内CYP3A酶活性与长春新碱血药浓度呈负相关。结论本研究建立的长春新碱血药浓度测定方法准确、灵敏、高效,适用于临床上血药浓度监测和体内药代动力学的研究。在肿瘤患儿体内长春新碱血药浓度与CYP3A酶活性有一定的相关性。

Objective To establish a HPLC-MS/MS method for the determination of vincristine and explore the correlation between vincristine concentration and cytochrome P450 3 A(CYP3 A)activity in pediatric oncology patients.Methods The plasma samples were prepared using liquid-liquid extraction.The chromatographic separation was carried out on Shim-pack XR-ODS column(2.0 mm×100.0 mm,2.2μm)with0.1%acetic acid-methanol and gradient elution.The compounds were quantified by positive ion multiple-reaction monitoring(MRM).The correlation analysis between vincristine concentration and CYP3 A activity was processed using SPSS 13.0 software.Results The calibration curves of vincristine were linear over the range 0.5-100.0μg·m L-1.The intra-day and inter-day precisions(RSD%)of quality control samples were within±10%.The extraction recoveries and matrix effects were 95.7%-105.9%,95.7%-105.9%,respectively.Moreover,the analysis results showed that the logarithm of CYP3 A activity was negatively correlated with the vincristine concentration in pediatric oncology patients.Conclusion This method possesses the profiles of accuracy,sensitivity and efficiency.It was suitable for therapeutic drug monitoring in clinical and pharmacokinetic studies.Furthermore,there was a correlation between the vincristine concentration and the activity of CYP3 A in pediatric oncology patients.