Chromatography

Liquid chromatography tandem mass spectrometry (LC-MS) was carried out with a Shimadzu Prominence HPLC system and SCIEX 4000 mass spectrometer. An aliquot of 20 µl of each sample was injected onto an Synergi C18 Reverse Phase 2.0 x 50mm, 2.5µ Column at 40^oC for gradient separation. Mobile phases were: A, ddH_2O with 10mM Ammonium acetate; and B, Acetonitrile with 10mM Ammonium acetate. Gradient schedule starts at 5% B and is linearly increased to 95% B at 5 minutes, then held 95% B until 5.2 minutes, and then returned to 5% B at 5.3 at minutes and re-equilibration until 10 minutes. The flow rate was 400µL/min. Time per sample injection was 10 minutes.

Extraction Protocols:

Extraction protocols for sample processing are dependent on the matrices involved. Please see extraction methods section for more specific instructions.

References

Method was adapted from the following reference:

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. Filist M, Buś-Kwaśnik K, Ksycińska H, Rudzki PJ. PMID: 25168217

Resources

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