Validated HPLC–MS–MS method for determination of azithromycin in human plasma

Abstract

A validated, highly sensitive, and selective HPLC method with MS-MS detection has been developed for quantitative determination of azithromycin (AZI) in human Na2EDTA plasma. Roxithromycin (ROX) was used as internal standard. Human plasma containing AZI and internal standard was ultrafiltered through Centrifree Micropartition devices and the concentration of AZI was determined by isocratic HPLC-MS-MS. Multiple reaction monitoring mode (MRM) was used for MS-MS detection. The calibration plot was linear in the concentration range 2.55-551.43 ng mL(-1). Inter-day and Intra-day precision and accuracy of the proposed method were characterized by R.S.D and percentage deviation, respectively; both were less than 8%. Limit of quantification was 2.55 ng mL(-1). The proposed method was used to determine the pharmacokinetic profile of AZI (250-mg tablets).

Azithromycin To Prevent Bronchopulmonary Dysplasia in Ureaplasma-Infected Preterm Infants: Pharmacokinetics, Safety, Microbial Response, and Clinical Outcomes with a 20-Milligram-per-Kilogram Single Intravenous Dose

“…The blood samples were centrifuged, and the plasma was aspirated and frozen at Ϫ80°C. The levels of azithromycin in plasma were measured using a validated high-performance liquid chromatography-tandem mass spectroscopy (HPLC/MS/MS) detection method (18). The lower limit of detection was 10 ng/ml.…”

Sensitive liquid chromatography/mass spectrometry assay for the quantification of azithromycin in human plasma

“…Chen et al (2006) extracted analyte with methyl tert-butyl ether-hexane (50:50 v/v) and monitored [M + 2H] 2+ by liquid chromatography/mass spectrometry (LC/MS). Barrett et al (2005) used an ultrafilter to pretreat sample and Nirogi et al (2005) used solid-phase extraction to prepare plasma, analysis being performed by liquid chromatography/tandem mass spectrometry (LC/MS/MS). All employed complicated and time-consuming sample preparation methods.…”

Quantitative determination of azithromycin in human plasma by ultra performance liquid chromatography–electrospray ionization mass spectrometry and its application in a pharmacokinetic study

“…It was found that a mixture of 50 mM ammonium acetate buffer-acetonitrile could achieve this purpose and was finally adopted as the mobile phase. This binary mobile phase is simpler than those reported in the literatures [23][24][25], which contained methanol, acetonitrile and buffer or with addition of formic acid [26]. It is similar to the mobile phase reported by Abuin et al [27] for the analysis of macrolide antibiotics in river water except for a different concentration of ammonium acetate.…”

“…Both azithromycin and roxithromycin were rapidly eluted with retention times less than 1.5 min, and the total run time was 3.0 min per sample. The analysis time for azithromycin in the literatures [23,25,27] which used HPLC-MS and HPLC-MS/MS were about 5 min, 6 min and 4 min, respectively. The short analysis time may meet the requirement for high sample throughput in bioanalysis.…”

“…Thus, LC-MS [23] and LC-MS/MS [24,25] were used in the determination of azithromycin with lower limit of quantification (LLOQ) of around 2 ng/mL [23][24][25]28]. The reported methods required either a complicated extraction [23] and/or derivatization procedure [18][19][20][21][22], or long analysis time [23,25], and may not well meet the requirement of desired throughput, speed and sensitivity in biosample analysis. This paper describes a fast, selective and highly sensitive approach, which enables the determination of azithromycin at 1 ng/mL in plasma with good accuracy using ultra performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS).…”