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J Pharm Biomed Anal. 2014 Nov;100:184-189. doi: 10.1016/j.jpba.2014.07.015. Epub 2014 Aug 8.

Simplified LC-MS/MS method enabling the determination of azithromycin in human plasma after a low 100mg dose administration.

Author information

1
Pharmaceutical Research Institute, Pharmacology Department, 8 Rydygiera, 01-793 Warsaw, Poland. Electronic address: m.filist@ifarm.eu.
2
Pharmaceutical Research Institute, Pharmacology Department, 8 Rydygiera, 01-793 Warsaw, Poland.

Abstract

A sensitive liquid chromatographic-tandem mass spectrometric method was developed and validated to study the pharmacokinetics of a low dose of azithromycin in human plasma. The sample preparation was based on liquid-liquid extraction with the low volume of methyl t-butyl ether. The chromatographic separation was performed on a Symmetry C18 column (50×2.1mm, 3.5μm). Gradient elution with ammonium acetate-acetonitrile and ammonium acetate-methanol was applied. Positive electrospray ionisation tandem mass spectrometry in the multiple reaction monitoring mode was used for the detection of azithromycin. The influence of a major metabolite, including the possibility of its back-conversion, on the quantification of azithromycin was evaluated. Isotope labelled azithromycin was used as the internal standard. The calibration curve was linear in the range of 0.5-250.0ng/mL. The new validated method was successfully applied to a pharmacokinetic study in humans following a single 100mg oral dose.

KEYWORDS:

Azithromycin; Liquid chromatography/tandem mass spectrometry; Metabolite back-conversion; Method validation; Pharmacokinetics

PMID:
25168217
DOI:
10.1016/j.jpba.2014.07.015
[Indexed for MEDLINE]

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