摘要
目的建立并应用超高效液相色谱-串联质谱法（UPLC-MS／MS）测定人血浆中卡巴他赛与血浆蛋白的结合率。方法以拉洛他赛为内标，样品经叔丁基甲醚液液萃取，采用ACQUITYBEHC，。色谱柱（50mm×2．1mm，1．7μm）；流动相：2mmol·L^-1醋酸铵水溶液-乙腈；流速：0．2mL·min^-1；进样量：5μL；柱温：35℃；采用ESI正离子源，定量分析离子反应分别为m／z836．36→m／z555．26（卡巴他赛）和m／z832．25→m／z551．08（内标拉洛他赛）。结合平衡透析法，测定了卡巴他赛的人血浆蛋白结合率。结果卡巴他赛浓度在5～1000ng·mL^-1范围内线性关系良好，定量下限为5ng·mL^-1，日内和日间精密度均小于15．0％。提取回收率为70．0％。91．4％，基质效应为87．4％～100．9％，可用于卡巴他赛的血浆蛋白结合率研究。卡巴他赛在低、中、高三个浓度下的人血浆蛋白结合率为（87．8±4．0）％、（76．4±0．8）％和（73．5±6．4）％。结论本方法简单、快速、灵敏，能满足分析要求。卡巴他赛与人血浆蛋白结合率较高，与血浆蛋白结合能力在考察的浓度范围内无浓度依赖性。
AIM To establish a rapid and sensitive analytical method based on ultra performance liquid chromatography-tandem mass spectrometry （UPLC- MS/MS） for the determination of the human plasma protein binding rate of cabazitaxel. METHODS The equilibrium dialysis was utilized to imitate the binding process between cabazitaxel and plasma protein. Samples were extracted with methyl tert-butyl ether using larotaxel as internal standard. The analysis was conducted with a Cls column （50 mm × 2.1 mm, 1.7 μm） and the column temperature was kept at 35 ℃ with flow rate at 0.2 mL· min^-1. The mobile phase consisted of acetonitrile and 2 mmol·L^-1 ammonium acetate and aliquot of 5 μL was injected for the UPLC-MS/MS analysis. Detection was performed on a tandem mass spectrometry with electrospray ionization source in the positive ion mode. Multiple reactions monitoring mode was used to quantify m/z 836.36 → m/z 555.26 （cabazitaxel） and m/z 832.25 → m/z 551.08 （IS）, respectively. RESULTS The standard curves were linear over the range 5 - 1 000 ng·mL^-1 for plasma and dialysis samples with lower limit of quantification was 5 ng·mL^-1. The intra- and inter- day precision was less than 15.0%. Recoveries and matrix effects were satisfactory for all the samples, with 70.0% - 91.4%, and 87.4% - 100.9%, respectively. The validated method was successfully applied to the determination of binding rate of cabazitaxel in human plasma. The human plasma protein binding rate of cabazitaxel at low, middle and high concentrations were （87.8 ± 4.0） %, （76.4 ± 0.8） % and （73.5 ± 6.4） % respectively. CONCLUSION The method is simple, fast, sensitive and satisfied for the requirements. The binding power of cabazitaxel to human plasma is high without concentration dependence.
出处
《中国新药与临床杂志》
CAS
CSCD
北大核心
2014年第5期379-383,共5页
Chinese Journal of New Drugs and Clinical Remedies
关键词
卡巴他赛
蛋白质结合
色谱法
高压液相
串联质谱法
cabazitaxel
protein binding
chromatography, high performance liquid
tandem mass spectrometry
