A quantitative LC-MS/MS method for determination of a small molecule agonist of EphA2 in mouse plasma and brain tissue.
A quantitative LC-MS/MS method for determination of a small molecule agonist of EphA2 in mouse plasma and brain tissue.
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DOI：
10.1002/bmc.4461
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发表时间：
2018-12
期刊：
影响因子：
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通讯作者：
Bo Zhong;Yaxin Li;Nethrie D. Idippily;A. Petty;Bin Su;B. Wang
中科院分区：
文献类型：
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作者：
Bo Zhong;Yaxin Li;Nethrie D. Idippily;A. Petty;Bin Su;B. Wang
化合物27{1，12-bis[4-(4-amino-6，7-dimethoxyquinazolin-2-yl)piperazin-1-yl]dodecane-1，12-dione}是一种新型的EphA2受体酪氨酸激酶小分子激动剂。与亲本化合物多沙唑嗪相比，其激活EphA2受体的活性显著提高。为进一步研究该化合物的药理和毒理作用，建立了高效液相色谱-电喷雾电离串联质谱仪(LC-MS/MS)定量测定该化合物的方法。采用液-液萃取法从小鼠血浆和脑组织匀浆中提取该化合物。采用反相色谱梯度洗脱将化合物27从基质中的内源分子中分离出来，然后采用正离子多反应监测模式进行MS检测。选择多重反应监测峰m/z 387.3→290.1和m/z 384.1→247.1分别用于化合物27和内标哌唑嗪的监测。线性范围为2~200 ng/mL，日内、日间精密度和准确度均在可接受的范围内。该方法成功地应用于不同给药途径的小鼠血浆和脑组织中化合物27的定量分析。
Compound 27 {1, 12-bis[4-(4-amino-6,7-dimethoxyquinazolin-2-yl)piperazin-1-yl]dodecane-1,12-dione} is a novel small molecule agonist of EphA2 receptor tyrosine kinase. It showed much improved activity for the activation of EphA2 receptor compared with the parental compound doxazosin. To support further pharmacological and toxicological studies of the compound, a method using liquid chromatography and electrospray ionization tandem mass spectrometry (LC-MS/MS) has been developed for the quantification of this compound. Liquid-liquid extraction was used to extract the compound from mouse plasma and brain tissue homogenate. Reverse-phase chromatography with gradient elution was performed to separate compound 27 from the endogenous molecules in the matrix, followed by MS detection using positive ion multiple reaction monitoring mode. Multiple reaction monitoring transitions m/z 387.3 → 290.1 and m/z 384.1 → 247.1 were selected for monitoring compound 27 and internal standard prazosin, respectively. The linear calibration range was 2-200 ng/mL with the intra- and inter-day precision and accuracy within the acceptable range. This method was successfully applied to the quantitative analysis of compound 27 in mouse plasma and brain tissue with different drug administration routes.