Development and validation of an UPLC-MS/MS assay for quantitative analysis of the ghrelin receptor inverse agonist PF-5190457 in human or rat plasma and rat brain.
Development and validation of an UPLC-MS/MS assay for quantitative analysis of the ghrelin receptor inverse agonist PF-5190457 in human or rat plasma and rat brain.
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DOI：
10.1007/s00216-015-8730-2
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发表时间：
2015-07
影响因子：
4.3
通讯作者：
Akhlaghi, Fatemeh
中科院分区：
文献类型：
--
作者：
Ghareeb, Mwlod;Leggio, Lorenzo;El-Kattan, Ayman;Akhlaghi, Fatemeh
PF-5190457是一种生长激素释放肽受体反向激动剂，目前正在进行临床开发，用于治疗酒精中毒。我们的目的是开发并验证一种简单、灵敏的方法，用于采用液相色谱-串联质谱法定量分析人或大鼠血浆和大鼠脑中的PF-5190457。使用0.1%甲酸的乙腈溶液，通过蛋白质沉淀从50 μL血浆或大鼠脑匀浆中提取分析物和稳定同位素内标物。色谱在具有1.7 μm粒度和130 μ m孔径的Acquity UPLC BEH C18（2.1 mm X 50 mm）上进行。流速为0.5 mL/min，总色谱运行时间为2.2分钟。移动的相由含有0.1%甲酸的水：乙腈95：5%（v/v）（溶剂A）和含有0.1%甲酸的100%乙腈（溶剂B）的梯度混合物组成。采用PF-5190457的m/z 513.35 → 209.30和内标物的m/z 518.47 → 214.43，在正电喷雾电离模式下进行多反应监测。所有基质的回收率范围为102-118%，CV小于6%。在研究的浓度范围内，所有基质的校准曲线均呈线性（R2 ≥ 0.998，n = 3）。大鼠或人血浆中的定量下限为1 ng/mL，大鼠脑中的定量下限为0.75 ng/g。运行内和运行间平均准确度百分比在85-115%之间，不精密度百分比≤ 15%。在该化合物的临床前和临床药理学研究中，该试验成功用于测定PF-5190457的浓度。
PF-5190457 is a ghrelin receptor inverse agonist that is currently undergoing clinical development for the treatment of alcoholism. Our aim was to develop and validate a simple and sensitive assay for quantitative analysis of PF-5190457 in human or rat plasma and rat brain using liquid chromatography-tandem mass spectrometry. The analyte and stable isotope internal standard were extracted from 50 μL plasma or rat brain homogenate by protein precipitation using 0.1% formic acid in acetonitrile. Chromatography was carried on an Acquity UPLC BEH C18 (2.1 mm X 50 mm) with 1.7 μm particle size and 130Å pore size. Flow rate was 0.5 mL/min and total chromatographic run time was 2.2 minutes. Mobile phase consisted of gradient mixture of water: acetonitrile 95:5% (v/v) containing 0.1% formic acid (Solvent A), and 100% acetonitrile containing 0.1% formic acid (Solvent B). Multiple reaction monitoring was carried out in positive electro-spray ionization mode using m/z 513.35 → 209.30 for PF-5190457 and m/z 518.47 → 214.43 for the internal standard. The recovery ranged from 102-118% with CV less than 6% for all matrices. The calibration curves for all matrices were linear over the studied concentration range (R2 ≥ 0.998, n = 3). Lower limit of quantification was 1 ng/mL in rat or human plasma and 0.75 ng/g in rat brain. Intra- and inter-run mean percent accuracy were between 85–115% and percent imprecision was ≤ 15%. The assays were successfully utilized to measure the concentration of PF-5190457 in pre-clinical and clinical pharmacology studies of the compound.
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影响因子：
3.4
作者：
Leggio L;Ferrulli A;Cardone S;Nesci A;Miceli A;Malandrino N;Capristo E;Canestrelli B;Monteleone P;Kenna GA;Swift RM;Addolorato G
通讯作者：
Addolorato G
影响因子：
7.7
作者：
Tong J;Prigeon RL;Davis HW;Bidlingmaier M;Kahn SE;Cummings DE;Tschöp MH;D'Alessio D
通讯作者：
D'Alessio D
DOI：
10.1016/s1044-0305(03)00574-9
发表时间：
2003-11-01
影响因子：
3.2
作者：
Dams, R;Huestis, MA;Murphy, CM
通讯作者：
Murphy, CM
DOI：
10.1016/j.jchromb.2009.05.031
发表时间：
2009-07-15
影响因子：
3
作者：
Zhang, Guodong;Wujcik, Chad E.
通讯作者：
Wujcik, Chad E.
影响因子：
--
作者：
Leggio, Lorenzo
通讯作者：
Leggio, Lorenzo