Cassette analysis of eight beta-blockers in bovine eye sclera, choroid-RPE, retina, and vitreous by liquid chromatography-tandem mass spectrometry
Cassette analysis of eight beta-blockers in bovine eye sclera, choroid-RPE, retina, and vitreous by liquid chromatography-tandem mass spectrometry
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DOI：
10.1016/j.jchromb.2008.12.027
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发表时间：
2009-01-15
影响因子：
3
通讯作者：
Kompella, Uday B.
中科院分区：
文献类型：
--
作者：
Kadam, Rajendra S.;Kompella, Uday B.
建立了一种简单、选择性和灵敏的LC-MS/MS方法，用于同时提取和测定各种牛眼组织（包括巩膜、脉络膜-RPE、视网膜和玻璃体）中的8种β-受体阻滞剂（阿替洛尔、索他洛尔、纳多洛尔、吲哚洛尔、噻吗洛尔、美托洛尔、倍他洛尔和普萘洛尔）。样品经2%NaOH水溶液碱化后，采用液-液萃取法进行提取。采用Hypersil-ODS C18柱进行色谱分离（100 mm × 2.1 mm，3.9 μ m），使用（A）5 mM甲酸铵在水中的梯度混合物（用甲酸调节pH 3.5）和（B）含0.02%三乙胺的乙腈：甲醇（75：25）（pH4.0;用甲酸调节）作为移动的相，流速为0.4 ml/min。化合物在正电喷雾电离（ESI）模式下电离，并在多反应监测（MRM）中检测。模式除索他洛尔（>51%）外，所有4种眼组织中所有β受体阻滞剂的平均回收率均> 82%。在四种眼组织中，β受体阻滞剂的基质效应范围为81 - 110%。该分析方法经过验证，并成功应用于组织暴露后使用盒式给药方法同时定量巩膜中的β-受体阻滞剂。所有分析物的校准曲线在10-2000 ng/ml范围内呈线性，相关系数>0.996。所有分析物的日内和日间精密度（% CV）均小于15%，准确度范围为85%至110%。巩膜吸收最低的是索他洛尔和阿替洛尔，两种亲水性β受体阻滞剂，最高的是普萘洛尔，亲脂性β受体阻滞剂。由爱思唯尔公司出版
A simple, selective, and sensitive LC-MS/MS method was developed for the simultaneous extraction and determination of eight beta-blockers (atenolol, sotalol, nadolol, pindolol, timolol, metoprolol, betaxolol and propranolol) in various bovine eye tissues including sclera, choroid-RPE, retina, and vitreous. The analytes were extracted by liquid-liquid extraction after samples were alkalinized with 2% NaOH solution in water. The chromatographic separation was performed on a Hypersil-ODS C18 column (100 mm x 2.1 mm, 3.9 mu m) using a gradient mixture of (A) 5 mM ammonium formate in water (pH 3.5 adjusted with formic acid) and (B) acetonitrile:methanol (75:25) containing 0.02% triethyl amine (pH 4.0; adjusted with formic acid) as mobile phase at a flow rate of 0.4 ml/min. The compounds were ionized in the positive electrospray ionization (ESI) mode and detected in the multiple reaction monitoring (MRM) mode. The average recoveries in all four eye tissues for all beta-blockers were >82%, except for sotalol (>51%). The matrix effect for beta-blockers ranged from 81 to 110% in the four eye tissues. This analytical method was validated and applied successfully for simultaneous quantification of the beta-blockers in sclera after tissue exposure using cassette dosing method. The calibration curve was linear in the range of 10-2000 ng/ml for all analytes, with the correlation coefficient >0.996. Intra-day and inter-day precision (% CV) was less than 15%, and accuracy ranged from 85 to 110% for all analytes. Scleral uptake was the lowest for sotalol and atenolol, two hydrophilic beta-blockers, and the highest for propranolol, a lipophilic beta-blocker. Published by Elsevier B.V.