二维液相色谱法同时测定人血浆中伊马替尼及去甲伊马替尼药物浓度及临床应用

Simultaneous Determination of Imatinib and N-desmethyl Imatinib in Human Plasma by Two-Dimensional Liquid Chromatography and Its Clinical Application

关键词:  二维液相色谱  伊马替尼  N-去甲伊马替尼  治疗药物监测

Key words:  two-dimensional liquid chromatography  imatinib  N-desmethyl imatinib  therapeutic drug monitoring

摘要:

目的 基于全自动二维液相色谱技术，建立一种能同时测定人血浆中伊马替尼及活性代谢物N-去甲伊马替尼简单而灵敏的方法，并应用于临床检测。方法 标本经简单去蛋白前处理后直接进样分析，待测物在一维色谱柱Aston SX1(3.5 mm×25 mm，5 μm)上进行初步萃取分离，通过中间柱Aston SCB(4.6 mm×10 mm，3.5 μm)转移至二维色谱柱Aston SCB(4.6 mm×125 mm，5 μm)进行二次分离分析，一维流动相为CAA-1D移动相，流速0.8 mL·min^–1；二维流动相为伊马替尼-2D移动相，流速1.2 mL·min^–1，柱温40 ℃，检测波长264/235 nm。结果 伊马替尼与血液中其他杂质分离良好，伊马替尼及N-去甲伊马替尼浓度分别在13.44~1 400 ng·mL^–1及13.17~1 372 ng·mL^–1内与峰面积呈良好的线性关系(r²=0.999 9，r²=0.999 9)，伊马替尼3个浓度水平(33.6，896，1 120 ng·mL^–1)及N-去甲伊马替尼3个浓度水平(32.93，878.08，1 097.60 ng·mL^–1)回收率分别为>96%及98%，浓度日内、日间RSD均<5.0%。结论 该方法准确度高，重现性好，自动化程度高，相比传统HPLC更具优越性，适用于临床伊马替尼血药浓度检测及其药动学研究。

Abstract:

OBJECTIVE To establish a simple and sensitive method based on two-dimensional liquid chromatography for the determination of imatinib and N-desmethyl imatinib in human plasma. METHODS Samples were directly injected and analyzed after simple deproteinization pretreatment. Samples were preliminarily extracted and separated with on the one-dimensional chromatographic column Aston SX1(3.5 mm×25 mm, 5 μm), and transferred from the one-dimension chromatographic column to the two-dimension chromatographic column as Aston SCB(4.6 mm×125 mm, 5 μm) through the intermediate column Aston SCB(4.6 mm×10 mm, 3.5 μm) for secondary separation analysis. The one-dimensional mobile phase was CAA-1D extractant with the flow rate of 0.8 mL·min^-1 and the two-dimensional mobile phase was imatinib-2D flowing phase with the flow rate of 1.2 mL·min^-1, the column temperature was 40℃. The concentration of imatinib and N-desmethyl imatinib were tested under the the wavelength of 264/235 nm. RESULTS It was found that the plasma concentration of imatinib and N-desmethyl imatinib showed a good linear relationship with their peak areas over the concentration range of 13.44-1 400 ng·mL^-1 and 13.17-1 372 ng·mL^-1, respectively(r²=0.999 9, r²=0.999 9). The mean recoveries obtained from three concentrations of imatinib(33.60, 896, 1 120 ng·mL^-1) and N-desmethyl imatinib(32.93, 878.08, 1 097.6 ng·mL^-1) were higher than 96% and 98%, respectively. The intra-day and inter-day RSD for imatinib and N-desmethyl imatinib were both <5.0%. CONCLUSION The method showed good accuracy, good reproducibility and high degree of automation. This method is more superior than traditional HPLC and can be applied to determine the clinical plasma concentration of imatinib and its pharmacokinetic studies.