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Chromatographia

, Volume 82, Issue 10, pp 1489–1500 | Cite as

Development and Validation of a Rapid Solid-Phase Extraction: Ultrafast Liquid Chromatographic Method for the Estimation of Azithromycin and Its Major Related Substances in Human Plasma and Dosage Forms Using a Novel Polyfunctional Silyl Reagent-Bonded Core–Shell Column

  • Dibya Ranjan SahooEmail author
  • Sunanda Sahoo
Original
  • 24 Downloads

Abstract

In this study, a rapid reversed-phase liquid chromatography method for the determination of azithromycin and its related substances was developed and validated on a novel polyfunctional silyl reagent [hexanhexamethyloctadecyltetrasiloxane (HMODTS–C18)] bonded core–shell RP C-18 column (100 × 4.6 mm, 2.6 µm) as per ICH guidelines. A binary gradient elution programme with a mixture of solvent A (phosphate buffer, pH 8.9) and solvent B (ACN: MeOH, 3:1) as mobile phase, at a flow rate of 1.5 mL min−1 and detection at 210 nm was finally optimized for the study. The validation results showed that the method to be specific, selective, highly sensible (0.004 mg mL−1), precise (% RSD ≤ 10), linear and accurate in a concentration range of 0.004–0.032 mg mL−1. Simple SPE method was performed for extraction and checked for repeatability. The result showed that the method is precise (%RSD < 2). The validated chromatographic method was applied to the solid-phase extracted samples of azithromycin and its four major related substances. Also, docking study was carried out using AutoDock Tools to find out the binding affinities, number of hydrogen bonds and residues involved in hydrogen bonds for azithromycin and its four major related impurities with the human plasma proteins. The results confirmed the applicability of the proposed method on the extracted human plasma samples. Finally, the study demonstrates that the impurities of azithromycin have strong binding affinities with the plasma proteins compared to that of azithromycin and half life of these impurities may be higher than azithromycin which can cause major health risk on repeated doses of azithromycin.

Keywords

Azithromycin: validation Azithromycin-related impurities Novel polyfunctional silyl reagent core–shell columns Solid-phase extraction Ultrafast liquid chromatography 

Notes

Acknowledgements

The authors are highly grateful to ImpScience Pvt. Ltd, Hyderabad, Telangana, India, for providing the necessary facilities during the research work.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Analytical Research and Development DepartmentImpScience Pvt. LtdHyderabadIndia
  2. 2.School of Life SciencesSambalpur UniversitySambalpurIndia

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