Bio-generation of stable isotope-labeled internal standards for absolute and relative quantitation of phase II drug metabolites in plasma samples using LC-MS/MS

Pei Li, Zi Li, Wayne D. Beck, Patrick Michael Callahan, Alvin V Terry, Maor Bar-Peled, Michael G. Bartlett

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Quantification of drug metabolites in biological samples has been of great interest in current pharmaceutical research, since metabolite concentrations and pharmacokinetics can contribute to a better understanding of the toxicity of drug candidates. Two major categories of Phase II metabolites, glucuronide conjugates and glutathione conjugates, may cause significant drug toxicity and therefore require close monitoring at early stages of drug development. In order to achieve high precision, accuracy, and robustness, stable isotope-labeled (SIL) internal standards (IS) are widely used in quantitative bioanalytical methods using liquid chromatography and tandem mass spectrometry (LC-MS/MS), due to their capability of compensating for matrix effects, extraction variations and instrument response fluctuations. However, chemical synthesis of SIL analogues of Phase II metabolites can often be very difficult and require extensive exploratory research, leading to higher cost and significant delays in drug research and development. To overcome these challenges, we have developed a generic method which can synthesize SIL analogues of Phase II metabolites from more available SIL parent drugs or SIL conjugation co-factors, using in vitro biotransformation. This methodology was successfully applied to the bio-generation of SIL glucuronide conjugates and glutathione conjugates. The method demonstrated satisfactory performance in both absolute quantitation and assessment of relative exposure coverage across species in safety tests of drug metabolites (MIST). This generic technique can be utilized as an alternative to chemical synthesis and potentially save time and cost for drug research and development.

Original languageEnglish (US)
Pages (from-to)4053-4063
Number of pages11
JournalAnalytical and Bioanalytical Chemistry
Volume407
Issue number14
DOIs
StatePublished - May 1 2015
Externally publishedYes

Fingerprint

Metabolites
Isotopes
Plasmas
Pharmaceutical Preparations
Glucuronides
Drug-Related Side Effects and Adverse Reactions
Glutathione
Research
Drug Costs
Toxicity
Biotransformation
Tandem Mass Spectrometry
Liquid Chromatography
Pharmacokinetics
Liquid chromatography
Safety
Costs and Cost Analysis
Mass spectrometry
Costs
Monitoring

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

Cite this

Bio-generation of stable isotope-labeled internal standards for absolute and relative quantitation of phase II drug metabolites in plasma samples using LC-MS/MS. / Li, Pei; Li, Zi; Beck, Wayne D.; Callahan, Patrick Michael; Terry, Alvin V; Bar-Peled, Maor; Bartlett, Michael G.

In: Analytical and Bioanalytical Chemistry, Vol. 407, No. 14, 01.05.2015, p. 4053-4063.

Research output: Contribution to journalArticle

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