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Detection of quaternary ammonium drugs in equine urine by liquid chromatography-mass spectrometry

机译：液相色谱-质谱法检测马尿中季铵盐类药物

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Quaternary ammonium drugs are anticholinergic agents and some of which have been known to be abused in equine sports. A general screening method for this class of drugs in equine urine by liquid chromatography-mass spectrometry (LC-MS) has not been reported. The paper describes an efficient LC-MS-MS method for the simultaneous detection and confirmation of twenty quaternary ammonium drugs at low ng mL(-1) in equine urine after solid-phase extraction. Quaternary ammonium drugs were extracted from equine urine by solid-phase extraction using ISOLUTE(R) CBA SPE Columns and analysed by LC-MS-MS in the positive electrospray ionisation mode. Separation of twenty quaternary ammonium drugs (the quaternary ammonium ions of edrophonium chloride, pyridostigmine bromide, neostigmine bromide, bretylium tosylate, ipratropium bromide, tubocurarine chloride, N-butylscopolammonium bromide, mepenzolate bromide, rocuronium bromide, clidinium bromide, pipenzolate bromide, isopropamide iodide, glycopyrronium bromide, alcuronium chloride, oxyphenonium bromide, propantheline bromide, tridihexethyl chloride, vecuronium bromide, mivacurium chloride and poncuronium bromide) was achieved in a reversed phase column with a mixture of aqueous ammonium formate, (pH 3.0, 10 mM) and acetonitrile as the mobile phase. Detection and confirmation of the twenty quaternary ammonium drugs at about 5 ng mL-1 in equine urine could be achieved within 22 min using product-ion scan MS-MS. The target analytes were detected by examination of extracted-ion chromatograms of their product ions. Drugs spiked in different equine urine (n = 15) were consistently detected. Negative samples (n = 30) of normal post-race equine urine have also been analysed, no matrix interference at the targeted masses and retention times was observed. The method was successfully applied to the analyses of drug-administration samples. Other method validation data including reproducibility and recovery will also be presented. An LC-MS-MS method for the simultaneous detection and confirmation of twenty quaternary ammonium drugs in equine urine was developed. The methodology should be applicable to other biological matrices suck as human urine.
机译：季铵药物是抗胆碱能药，其中一些已知在马运动中被滥用。尚未通过液相色谱-质谱法（LC-MS）对马尿中此类药物进行一般筛选。本文描述了一种有效的LC-MS-MS方法，用于固相萃取后同时检测和确认马尿中低ng mL（-1）的二十种季铵药物。使用ISOLUTE CBA SPE色谱柱通过固相萃取从马尿中提取季铵药物，并在正电喷雾电离模式下通过LC-MS-MS分析。分离二十种季铵盐药物（四氢呋喃鎓，溴化吡啶​​斯的明，溴化新斯的明，溴化甲苯磺酸bre，溴化异丙托溴铵，氯化铜尿嘧啶，氯化正丁基溴化铵，溴化溴甲烷，溴化间苯二甲酰亚胺，溴化异丙基溴化物，溴化吡啶​​鎓，溴化吡啶​​鎓，溴化吡啶​​鎓，溴化吡啶​​鎓在反相色谱柱中用甲酸铵水溶液（pH 3.0，10 mM）和乙腈的混合物制得了格隆溴铵，氯化铝，溴化苯氧铵，溴化苯乙氧铵，溴化丙胺，三二乙基氯乙烷，溴化维库溴铵，氯化米伐库铵和溴化ponpononium。流动相。使用产物离子扫描MS-MS可以在22分钟内检测和确认马尿中约5 ng mL-1的二十种季铵药物。通过检查其产物离子的提取离子色谱图来检测目标分析物。始终检测到加标在不同马尿（n = 15）中的药物。还分析了赛马后正常尿液的阴性样本（n = 30），未观察到目标质量和保留时间的基质干扰。该方法已成功应用于药物管理样品的分析。也将介绍其他方法验证数据，包括重现性和回收率。建立了一种同时检测和确认马尿中20种季铵盐药物的LC-MS-MS方法。该方法应适用于像人类尿液一样吮吸的其他生物基质。

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