A robust and rapid liquid chromatography tandem mass spectrometric method for the quantitative analysis of 5-azacytidine

Nicole Anders, Teresia M. Wanjiku, Ping He, Nilofer Azad, Michelle Rudek-Renaut

Research output: Contribution to journalArticle

Abstract

The DNA methyltransferase inhibitor 5-azacytidine is being evaluated clinically as an oral formulation to treat various solid tumors. A sensitive, reliable method was developed to quantitate 5-azacytidine using LC-MS/MS to perform detailed pharmacokinetic studies. The drug of interest was extracted from plasma using Oasis MCX ion exchange solid-phase extraction 96-well plates. Chromatographic separation was achieved with a YMC J'sphere M80 C18 column and isocratic elution with a methanol-water-formic acid (15:85:0.1, v/v/v) mobile phase over a 7 min total analytical run time. An AB Sciex 5500 triple quadrupole mass spectrometer operated in positive electrospray ionization mode was used for the detection of 5-azacytidine. The assay range was 5-500 ng/mL and proved to be accurate (97.8-109.1%) and precise (CV ≤ 9.8%). Tetrahydrouridine was used to stabilize 5-azacytidine in blood/plasma samples. With the addition of tetrahydrouridine, long-term frozen plasma stability for 5-azacytidine at -70°C has been determined for at least 323 days. The method was applied for the measurement of total plasma concentrations of 5-azacytidine in a cancer patient receiving a 300 mg oral daily dose.

Original languageEnglish (US)
Pages (from-to)494-496
Number of pages3
JournalBiomedical Chromatography
Volume30
Issue number3
DOIs
StatePublished - Mar 1 2016

Fingerprint

Azacitidine
Liquid chromatography
Liquid Chromatography
Tetrahydrouridine
Chemical analysis
formic acid
Plasmas
Electrospray ionization
Plasma stability
Pharmacokinetics
Ion Exchange
Solid Phase Extraction
Methyltransferases
Mass spectrometers
Methanol
Tumors
Assays
Neoplasms
Ion exchange
Blood

Keywords

  • 5-azacytidine
  • DNA methyltransferase inhibitor
  • LC/MS/MS
  • Pharmacokinetics

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Molecular Biology
  • Drug Discovery
  • Pharmacology

Cite this

A robust and rapid liquid chromatography tandem mass spectrometric method for the quantitative analysis of 5-azacytidine. / Anders, Nicole; Wanjiku, Teresia M.; He, Ping; Azad, Nilofer; Rudek-Renaut, Michelle.

In: Biomedical Chromatography, Vol. 30, No. 3, 01.03.2016, p. 494-496.

Research output: Contribution to journalArticle

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