Validation of Individual Quantitative Methods for Determination of Cytochrome P450 Probe Substrates in Human Dried Blood Spots with HPLC–MS/MS

Lad, Rakesh

doi:10.4155/bio.10.155

Cited by 16 publications

(13 citation statements)

References 24 publications

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“…The GC analysis reported by Brazier et al [11] required solvent extraction followed by derivatisation with methyxanthine prior to measurement. More recently a DBS method without the need for several work-up steps and compound derivatisation has been established [12]. This method, based on LC-MS/MS analysis, has a lower limit of quantification (LLOQ) of 250 ng/mL based on a 15 µL DBS sample.…”

Section: Introductionmentioning

confidence: 99%

Dried Blood Spot Sampling with LC-MS Analysis for Routine Therapeutic Caffeine Monitoring in Neonates

Lawson

Patel

Mulla

et al. 2012

ISRN Chromatography

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A liquid chromatography-mass spectrometry (LC-MS) method was developed and validated for the determination of therapeutic levels of caffeine in dried blood spot (DBS) samples. Caffeine is used in the treatment of Apnoea of Prematurity (AoP) in newborn children. Calibration DBS samples were prepared by spotting 15 µL of whole blood spiked with the analyte onto specimen collection cards. 3 mm disks cut from the centre of the DBS were extracted in methanol containing the internal standard. The extract was separated using a Zorbax Eclipse Plus C18 column and the MS, operated in electrospray positive ion mode, used single ion monitoring at m/z 195 for caffeine and m/z 198 for the IS. The overall extraction recovery of caffeine from spiked blood spots was demonstrated to be 44-47%. Validation of the microanalytical method showed good precision (coefficient of variation) and accuracy (relative error) and specificity and was linear within the tested calibration range 500-25000 ng/mL for caffeine. Investigation of different specimen collection papers revealed different matrix effects with significant ion suppression from the FTA Elute paper itself. Requiring only a microvolume (15 µL) blood sample for analysis, the developed DBS based microanalytical method has the potential to facilitate the routine monitoring of caffeine in neonates.

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Section: Introductionmentioning

confidence: 99%

Dried Blood Spot Sampling with LC-MS Analysis for Routine Therapeutic Caffeine Monitoring in Neonates

Lawson

Patel

Mulla

et al. 2012

ISRN Chromatography

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“…The developed UPLC-MS/MS, which was used for the subsequent quantitation of the six probe drugs, showed a faster analysis time than conventional HPLC (Jerdi et al, 2004;Yao et al, 2007;Johansson et al, 2012) or LC-MS (Scott et al, 1999;Zhang et al, 2002;Yin et al, 2004;He et al, 2007;Lad, 2010;Hu et al, 2012;Oh et al, 2012;Lee et al, 2013) and tremendously enhanced signal intensity. It took only 3 min to finish analyzing a plasma sample, which can save much time in experimental studies with hundreds of samples, and it was faster than the UPLC-MS/MS method with a run-time of 5 min for determination of the four probe drugs (caffeine, tolbutamide, metoprolol and dapsone) in rat plasma reported in the literature (Liu et al, 2013).…”

Section: Discussionmentioning

confidence: 99%

“…High‐throughput screening techniques have helped in the evaluation of potential CYP450 inhibition during drug discovery and development (Qin et al , ). Early publications have described methods for the simultaneous determination of multiple probe drugs using HPLC (Jerdi et al , ; Yao et al , ; Johansson et al , ), LC‐MS/MS (Scott et al , ; Zhang et al , ; Yin et al ., ; He et al , ; Lad, ; Hu et al , ; Oh et al , ; Lee et al , ; Wang et al , ) and UPLC‐MS/MS (De Bock et al , ; Liu et al , ). In this study, we developed an ultra‐performance liquid chromatography tandem mass spectrometry (UPLC‐MS/MS) method for analysis of the six probe drugs in a single‐run process.…”

Section: Introductionmentioning

confidence: 99%

Simultaneous determination of bupropion, metroprolol, midazolam, phenacetin, omeprazole and tolbutamide in rat plasma by UPLC‐MS/MS and its application to cytochrome P450 activity study in rats

Wang

Zhang

et al. 2015

Biomedical Chromatography

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A specific ultra-performance liquid chromatography tandem mass spectrometry method is described for the simultaneous determination of bupropion, metroprolol, midazolam, phenacetin, omeprazole and tolbutamide in rat plasma with diazepam as internal standard, which are the six probe drugs of the six cytochrome P450 isoforms CYP2B6, CYP2D6, CYP3A4, CYP1A2, CYP2C19 and CYP2C9. Plasma samples were protein precipitated with acetonitrile. The chromatographic separation was achieved using a UPLC® BEH C18 column (2.1 × 100 mm, 1.7 µm). The mobile phase consisted of acetonitrile and water (containing 0.1% formic acid) with gradient elution. The triple quadrupole mass spectrometric detection was operated by multiple reaction monitoring in positive electrospray ionization. The precisions were <13%, and the accuracy ranged from 93.3 to 110.4%. The extraction efficiency was >90.5%, and the matrix effects ranged from 84.3 to 114.2%. The calibration curves in plasma were linear in the range of 2-2000 ng/mL, with correlation coefficient (r(2) ) >0.995. The method was successfully applied to pharmacokinetic studies of the six probe drugs of the six CYP450 isoforms and used to evaluate the effects of erlotinib on the activities of CYP2B6, CYP2D6, CYP3A4, CYP1A2, CYP2C19 and CYP2C9 in rats. Erlotinib may inhibit the activity of CYP2B6 and CYP3A4, and may induce CYP2C9 of rats.

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“…Busulfan TDM LC-MS/MS [71] 11. Caffeine PK studies LC-MS/MS [72] 12. Canrenone Neonatal screening LC-MS [73] 13.…”

Section: Amodiaquine Chloroquine and Chlorthalidonementioning

confidence: 99%

Dried blood spots: Concepts, present status, and future perspectives in bioanalysis

Sharma

Jaiswal

Shukla

et al. 2014

Drug Testing and Analysis

144

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Over the past several years, dried blood spot (DBS) sampling technique has emerged as a pertinent method in both qualitative and quantitative bioanalysis context. In the DBS method, the blood sample is directly soaked on to a paper (with or without treatment). After drying it can be analyzed by modern analytical, immunological, or genomic detection systems. Several advantages of the DBS technique such as low blood volume requirement, transportation and storage without special treatment, better analytes stability, enhanced clinical cooperation in clinical trials, and reduced unforeseeable exposure of analysts to biohazards, make it the most appropriate blood sampling technique. This review illustrates the information available on the DBS method which may serve as a single window for investigators in the field of bioanalysis. Also, it explores the proficiency and appliance of the DBS method in pharmacokinetic (PK), therapeutic drug monitoring (TDM), toxicokinetic (TK), metabolomic, and disease diagnosis.

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Validation of Individual Quantitative Methods for Determination of Cytochrome P450 Probe Substrates in Human Dried Blood Spots with HPLC–MS/MS

Abstract: This work demonstrates that quantitative analysis of a drug extracted from dried blood spots can provide high-quality data while minimizing the volume of blood withdrawn from volunteers.

Cited by 16 publications

References 24 publications

Dried Blood Spot Sampling with LC-MS Analysis for Routine Therapeutic Caffeine Monitoring in Neonates

Dried Blood Spot Sampling with LC-MS Analysis for Routine Therapeutic Caffeine Monitoring in Neonates

Simultaneous determination of bupropion, metroprolol, midazolam, phenacetin, omeprazole and tolbutamide in rat plasma by UPLC‐MS/MS and its application to cytochrome P450 activity study in rats

Dried blood spots: Concepts, present status, and future perspectives in bioanalysis

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