Advances in the application of mass spectrometry to studies of drug metabolism, pharmacokinetics and toxicology

Baillie, Thomas A.

doi:10.1016/0168-1176(92)85066-9

Cited by 45 publications

(25 citation statements)

References 123 publications

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“…Samples were subjected to chromatographic separation on a 150 × 2.0 mm Luna 5 µm C18 column (Phenomenex, Torrance, CA), and eluates were analyzed with a Thermo LCQ DecaXP ion trap mass spectrometer (ThermoElectron, San Jose, CA) operated in precursor ion monitoring mode to detect ions that fragment with loss of 129 Da, which corresponds to the loss of the glutamate residue (31). Elution solvents were 5% acetonitrile, 95% H 2 O, and 0.5% acetic acid (solvent A), and 95% acetonitrile, 5% H 2 O, and 0.5% acetic acid (solvent B).…”

Section: Lc-ms-ms Analyses and Identification Of Protein Adductsmentioning

confidence: 99%

Protein Targets of Reactive Electrophiles in Human Liver Microsomes

Shin

Liu

Stamer

et al. 2007

Chem. Res. Toxicol.

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Liver microsomes are widely used to study xenobiotic metabolism in Vitro, and covalent binding to microsomal proteins serves as a surrogate marker for toxicity mediated by reactive metabolites. We have applied liquid chromatography-tandem mass spectrometry (LC-MS-MS) to identify protein targets of the biotin-tagged model electrophiles 1-biotinamido-4-(4′-[maleimidoethylcyclohexane]-carboxamido)-butane (BMCC) and N-iodoacetyl-N-biotinylhexylenediamine (IAB) in human liver microsomes. The biotin-tagged peptides resulting from in-gel tryptic digestion were enriched by biotin-avidin chromatography and LC-MS-MS was used to identify 376 microsomal cysteine thiol targets of BMCC and IAB in 263 proteins. Protein adduction was selective and reproducible, and only 90 specific cysteine sites in 70 proteins (approximately 25% of the total) were adducted by both electrophiles. Differences in adduction selectivity correlated with different biological effects of the compounds, as IAB-but not BMCC-induced ER stress in HEK293 cells. Targeted LC-MS-MS analysis of microsomal glutathione-S-transferase cysteine 50, a target of both IAB and BMCC, detected time-dependent adduction by the reactive acetaminophen metabolite N-acetyl-p-benzoquinoneimine during microsomal incubations. The results indicate that electrophiles selectively adduct microsomal proteins, but display differing target selectivities that correlate with differences in toxicity. Analysis of selected microsomal protein adduction reactions thus could provide a more specific indication of potential toxicity than bulk covalent binding of radiolabeled compounds.

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Section: Lc-ms-ms Analyses and Identification Of Protein Adductsmentioning

confidence: 99%

Protein Targets of Reactive Electrophiles in Human Liver Microsomes

Shin

Liu

Stamer

et al. 2007

Chem. Res. Toxicol.

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“…Only precursor ion and neutral loss scans are usable in detection of unknown metabolites of a drug, while the product ion scan mode is more powerful in structure characterization of the detected metabolites. A unique feature of triple‐quadrupole mass spectrometers (QQQ) is its capability to identify families of metabolites by using neutral‐loss and precursor ion scans 33. However, the sensitivity in full‐scan MS/MS experiments is limited.…”

Section: Detection Of Metabolites and Determination Of The Site Of Bimentioning

confidence: 99%

“…Product ion scans are used for identification and multiple reaction monitoring (MRM) provides the high sensitivity required in quantitative analysis. The unique feature of QQQ is its capability to identify families of metabolites by using neutral‐loss and precursor ion scans 33. However, the sensitivity of identification of metabolites by the full‐scan mode may not always suffice and instead of QQQ the use of ion trap (IT) and time‐of‐flight (TOF) mass spectrometers has increased.…”

Section: Introductionmentioning

confidence: 99%

Liquid chromatography/atmospheric pressure ionization–mass spectrometry in drug metabolism studies

et al. 2003

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The study of the metabolic fate of drugs is an essential and important part of the drug development process. The analysis of metabolites is a challenging task and several different analytical methods have been used in these studies. However, after the introduction of the atmospheric pressure ionization (API) technique, electrospray and atmospheric pressure chemical ionization, liquid chromatography/mass spectrometry (LC/MS) has become an important and widely used method in the analysis of metabolites owing to its superior specificity, sensitivity and efficiency. In this paper the feasibility of LC/API-MS techniques in the identification, structure characterization and quantitation of drug metabolites is reviewed. Sample preparation, LC techniques, isotope labeling, suitability of different MS techniques, such as tandem mass spectrometry, and high-resolution MS in drug metabolite analysis, are summarized and discussed. Automation of data acquisition and interpretation, special techniques and possible future trends are also the topics of the review.

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“…The use of mass spectrometry has become vital to assessment of structure information in complex biologic matrices (Baillie 1992;Mutlib et al, 1995;Pochapsky and Pochapsky 2001;Nassar, 2003;Nassar and Adams, 2003;Watt et al, 2003). Time-of-flight mass spectrometry, a Fourier-transform ion cyclotron resonance-mass spectrometer (FTICR-MS), and Orbitrap mass spectrometers are among the most effective for this work.…”

Section: Abbreviations: Agt Omentioning

confidence: 99%

Biotransformation and Rearrangement of Laromustine

Nassar

Wisnewski

King

2016

Drug Metabolism and Disposition

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This review highlights the recent research into the biotransformations and rearrangement of the sulfonylhydrazine-alkylating agent laromustine. Incubation of [ 14 C]laromustine with rat, dog, monkey, and human liver microsomes produced eight radioactive components (C-1 to C-8). There was little difference in the metabolite profile among the species examined, partly because NADPH was not required for the formation of most components, which instead involved decomposition and/or hydrolysis. The exception was C-7, a hydroxylated metabolite, largely formed by CYP2B6 and CYP3A4/5. Liquid chromatography-multistage mass spectrometry (LC-MS n ) studies determined that collision-induced dissociation, and not biotransformation or enzyme catalysis, produced the unique mass spectral rearrangement. Accurate mass measurements performed with a Fourier-transform ion cyclotron resonance mass spectrometer (FTICR-MS) significantly aided determination of the elemental compositions of the fragments and in the case of laromustine revealed the possibility of rearrangement. Further, collision-induced dissociation produced the loss of nitrogen (N 2 ) and methylsulfonyl and methyl isocyanate moieties. The rearrangement, metabolite/decomposition products, and conjugation reactions were analyzed utilizing hydrogendeuterium exchange, exact mass, 13 C-labeled laromustine, nuclear magnetic resonance spectroscopy (NMR), and LC-MS n experiments to assist with the assignments of these fragments and possible mechanistic rearrangement. Such techniques produced valuable insights into these functions: 1) Cytochrome P450 is involved in C-7 formation but plays little or no role in the conversion of [ 14 C]laromustine to C-1 through C-6 and C-8; 2) the relative abundance of individual degradation/metabolite products was not species-dependent; and 3) laromustine produces several reactive intermediates that may produce the toxicities seen in the clinical trials.

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Advances in the application of mass spectrometry to studies of drug metabolism, pharmacokinetics and toxicology

Cited by 45 publications

References 123 publications

Protein Targets of Reactive Electrophiles in Human Liver Microsomes

Protein Targets of Reactive Electrophiles in Human Liver Microsomes

Liquid chromatography/atmospheric pressure ionization–mass spectrometry in drug metabolism studies

Biotransformation and Rearrangement of Laromustine

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