Quantitative Method for Simultaneous Analysis of a 5-Probe Cocktail for Cytochrome P450 Enzymes

Laureen A Lammers; Roos Achterbergh; Marcel C M Pistorius; Yuma Bijleveld; Emmely M de Vries; Anita Boelen; Heinz-Josef Klümpen; Johannes A Romijn; Ron A A Mathôt

doi:10.1097/FTD.0000000000000338

Quantitative Method for Simultaneous Analysis of a 5-Probe Cocktail for Cytochrome P450 Enzymes

Laureen A Lammers
, Roos Achterbergh
, Marcel C M Pistorius
, Yuma Bijleveld
, Emmely M de Vries
, Anita Boelen
, Heinz-Josef Klümpen
, Johannes A Romijn
, Ron A A Mathôt

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

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.

Original language	English
Pages (from-to)	761-768
Number of pages	8
Journal	Therapeutic drug monitoring
Volume	38
Issue number	6
DOIs	https://doi.org/10.1097/FTD.0000000000000338
Publication status	Published - Dec 2016
Externally published	Yes

Keywords

Chromatography, Liquid/methods
Cytochrome P-450 Enzyme System/metabolism
Drug Interactions
Humans
Pharmaceutical Preparations/blood
Sensitivity and Specificity
Tandem Mass Spectrometry/methods

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10.1097/FTD.0000000000000338

Cite this

@article{4a23e6464bfb4603bc24f34ebcbe4f94,

title = "Quantitative Method for Simultaneous Analysis of a 5-Probe Cocktail for Cytochrome P450 Enzymes",

abstract = "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.",

keywords = "Chromatography, Liquid/methods, Cytochrome P-450 Enzyme System/metabolism, Drug Interactions, Humans, Pharmaceutical Preparations/blood, Sensitivity and Specificity, Tandem Mass Spectrometry/methods",

author = "Lammers, \{Laureen A\} and Roos Achterbergh and Pistorius, \{Marcel C M\} and Yuma Bijleveld and \{de Vries\}, \{Emmely M\} and Anita Boelen and Heinz-Josef Kl{\"u}mpen and Romijn, \{Johannes A\} and Math{\^o}t, \{Ron A A\}",

year = "2016",

month = dec,

doi = "10.1097/FTD.0000000000000338",

language = "English",

volume = "38",

pages = "761--768",

journal = "Therapeutic drug monitoring",

issn = "0163-4356",

publisher = "Lippincott Williams \& Wilkins",

number = "6",

}

TY - JOUR

T1 - Quantitative Method for Simultaneous Analysis of a 5-Probe Cocktail for Cytochrome P450 Enzymes

AU - Lammers, Laureen A

AU - Achterbergh, Roos

AU - Pistorius, Marcel C M

AU - Bijleveld, Yuma

AU - de Vries, Emmely M

AU - Boelen, Anita

AU - Klümpen, Heinz-Josef

AU - Romijn, Johannes A

AU - Mathôt, Ron A A

PY - 2016/12

Y1 - 2016/12

N2 - 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.

AB - 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.

KW - Chromatography, Liquid/methods

KW - Cytochrome P-450 Enzyme System/metabolism

KW - Drug Interactions

KW - Humans

KW - Pharmaceutical Preparations/blood

KW - Sensitivity and Specificity

KW - Tandem Mass Spectrometry/methods

U2 - 10.1097/FTD.0000000000000338

DO - 10.1097/FTD.0000000000000338

M3 - Article

C2 - 27764027

SN - 0163-4356

VL - 38

SP - 761

EP - 768

JO - Therapeutic drug monitoring

JF - Therapeutic drug monitoring

IS - 6

ER -

Quantitative Method for Simultaneous Analysis of a 5-Probe Cocktail for Cytochrome P450 Enzymes

Abstract

Keywords

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