用于细胞色素P450酶5探针混合物同步分析的定量方法。
Quantitative Method for Simultaneous Analysis of a 5-Probe Cocktail for Cytochrome P450 Enzymes.
作者信息
Lammers Laureen A, Achterbergh Roos, Pistorius Marcel C M, Bijleveld Yuma, de Vries Emmely M, Boelen Anita, Klümpen Heinz-Josef, Romijn Johannes A, Mathôt Ron A A
机构信息
Departments of *Hospital Pharmacy, †Medicine, ‡Endocrinology and Metabolism, and §Medical Oncology, Academic Medical Center, Amsterdam, the Netherlands.
出版信息
Ther Drug Monit. 2016 Dec;38(6):761-768. doi: 10.1097/FTD.0000000000000338.
BACKGROUND
The metabolic activity of P450 enzymes in vivo can be determined using selective probe drugs. The simultaneous administration of multiple CYP-specific probe drugs is commonly known as the "cocktail approach." Disadvantages of a cocktail are large volumes of samples required for analysis and time-consuming analyses. The aim of this study was to develop and validate a simplified but sensitive method for the simultaneous quantification of 5 probe drugs [caffeine (CYP1A2), metoprolol (CYP2D6), midazolam (CYP3A4), omeprazole (CYP2C19), and S-warfarin (CYP2C9)] in a previously validated cocktail using a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method.
METHODS
The method entailed a single method for sample preparation that enables quick processing of the samples containing all 5 probe drugs in a small volume of blood (≥10 μL) followed by a chiral and nonchiral LC-MS/MS method. The method was validated for selectivity, specificity, resolution of racemic warfarin, linearity, accuracy, imprecision, recovery, process efficiency, ionization efficiency, and carryover effect.
RESULTS
The method showed good selectivity without matrix interferences and differentiated S- and R-warfarin enantiomers with adequate resolution (Rs = 1.55). For all analytes, the mean process efficiency was >95%, and the mean ionization efficiency was >97%. Furthermore, the accuracy was between 94.9% and 108% for all analytes, and the within- and between-run imprecision were <11.7% for the lower limit of quantification and <12.6% for the middle level and upper limit of quantification.
CONCLUSIONS
The method presented here enables the simultaneous quantification of the 5 probes in a very small blood volume (≥10 μL). Furthermore, it is less time consuming than previously reported methods because it requires only 1 simple method for sample preparation followed by a nonchiral and chiral LC-MS/MS method that can be performed sequentially.
背景
体内P450酶的代谢活性可通过使用选择性探针药物来确定。同时给予多种CYP特异性探针药物通常被称为“鸡尾酒法”。鸡尾酒法的缺点是分析所需的样本量较大且分析耗时。本研究的目的是开发并验证一种简化但灵敏的方法，用于使用液相色谱 - 串联质谱（LC-MS/MS）法同时定量先前已验证的鸡尾酒中的5种探针药物[咖啡因（CYP1A2）、美托洛尔（CYP2D6）、咪达唑仑（CYP3A4）、奥美拉唑（CYP2C19）和S-华法林（CYP2C9）]。
方法
该方法需要一种单一的样品制备方法，能够在少量血液（≥10μL）中快速处理包含所有5种探针药物的样品，随后采用手性和非手性LC-MS/MS方法。该方法针对选择性、特异性、外消旋华法林的拆分、线性、准确性、不精密度、回收率、处理效率、电离效率和残留效应进行了验证。
结果
该方法显示出良好的选择性，无基质干扰，并且以足够的分辨率（Rs = 1.55）区分了S-和R-华法林对映体。对于所有分析物，平均处理效率>95%，平均电离效率>97%。此外，所有分析物的准确度在94.9%至108%之间，定量下限的批内和批间不精密度<11.7%，定量中间水平和上限的批内和批间不精密度<12.
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