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      Quantification of three macrolide antibiotics in pharmaceutical lots by HPLC: Development, validation and application to a simultaneous separation

      Author(s): , , ,
      Publication date Created:
      Publication date (Electronic):
      Journal: British Journal of Pharmacy
      Publisher: University of Huddersfield Press

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          Abstract

          A new validated high performance liquid chromatographic (HPLC) method with rapid analysis time and high efficiency, for the analysis of erythromycin, azithromycin and spiramycin, under isocratic conditions with ODB RP18 as a stationary phase is described. Using an eluent composed of acetonitrile –2-methyl-2-propanol –hydrogenphosphate buffer, pH 6.5, with 1.5% triethylamine (33:7: up to 100, v/v/v), delivered at a flow-rate of 1.0 mL min-1. Ultra Violet (UV) detection is performed at 210 nm. The selectivity is satisfactory enough and no problematic interfering peaks are observed. The procedure is quantitatively characterized and repeatability, linearity, detection and quantification limits are very satisfactory. The method is applied successfully for the assay of the studied drugs in pharmaceutical dosage forms as tablets and powder for oral suspension. Recovery experiments revealed recovery of 97.13–100.28%.

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          Analysis of macrolide antibiotics.

          Isadore Kanfer, Michael F. Skinner, Roderick Walker (1998)
          The following macrolide antibiotics have been covered in this review: erythromycin and its related substances, azithromycin, clarithromycin, dirithromycin, roxithromycin, flurithromycin, josamycin, rokitamycin, kitasamycin, mycinamycin, mirosamycin, oleandomycin, rosaramicin, spiramycin and tylosin. The application of various thin-layer chromatography, paper chromatography, gas chromatography, high-performance liquid chromatography and capillary zone electrophoresis procedures for their analysis are described. These techniques have been applied to the separation and quantitative analysis of the macrolides in fermentation media, purity assessment of raw materials, assay of pharmaceutical dosage forms and the measurement of clinically useful macrolide antibiotics in biological samples such as blood, plasma, serum, urine and tissues. Data relating to the chromatographic behaviour of some macrolide antibiotics as well as the various detection methods used, such as bioautography, UV spectrophotometry, fluorometry, electrochemical detection, chemiluminescence and mass spectrometry techniques are also included.
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            Determination of azithromycin by ion-pair HPLC with UV detection.

            Zi Yi Yang, Lin Wang, Xing Tang (2009)
            An ion-pair reversed phase high performance liquid chromatographic method with UV detection was developed for the determination of azithromycin using sodium heptanesulfonate as an ion-pair reagent. The mobile phase consisted of a mixture of ammonium dihydrogen phosphate (0.045 M, pH 3.0 adjusted by phosphoric acid):acetonitrile 47:15 (v/v) and the concentration of sodium heptanesulfonate in the aqueous phase was 0.002 M. UV detection was performed at 210 nm. The chromatographic column was Dikma Technologies Diamonsil C18 column, 5 microm 150 mm x 4.6 mm, which was maintained at 25 degrees C. Applying the method to a stability study of azithromycin eye drops, it was found that the related substance could be detected and the profile of the AZM peak was symmetrical and the column efficiency was high. Accordingly, it is suitable for the routine analysis and stability testing of azithromycin preparations.
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              Identification and quantification of five macrolide antibiotics in several tissues, eggs and milk by liquid chromatography–electrospray tandem mass spectrometry

              M. Dubois, D. Fluchard, E Sior … (2001)
              We present an electrospray high-performance liquid chromatographic tandem mass spectrometric (HPLC-MS-MS) method capable of determining in several tissues (muscle, kidney, liver), eggs and milk the following five macrolides: tylosin, tilmicosin, spiramycin, josamycin, erythromycin. Roxithromycin was used as an internal standard. The method uses extraction in a Tris buffer at pH 10.5, followed by protein precipitation with sodium tungstate and clean-up on an Oasis solid-phase extraction column. The HPLC separation was performed on a Purospher C18 column (125 x 3 mm I.D.) protected by a guard column, with a gradient of aqueous 0.1 M ammonium acetate-acetonitrile as the mobile phase at a flow-rate of 0.7 ml min(-1). Protonated molecules served as precursor ions for electrospray ionisation in the positive ion mode and four product ions were chosen for each analyte for multiple reaction monitoring (MRM). A validation study was conducted to confirm the five macrolides by MRM HPLC-MS-MS analysis of a negative control and fortified samples. All of the samples analysed were confirmed with four ions. The ion ratio reproducibility limit ranged from 2.4 to 15%. All compounds could be detected and quantified at half-maximum residue limits (MRLs). The method is specific, quantitative and reproducible enough to conform to European Union recommendations within the concentration range 0.5 MRL-2 MRL (accuracy: 80 to 110%, relative standard deviation: 2 to 13%). This whole method allows extraction and analysis of up to 50 samples per day.
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                Author and article information

                Journal
                Title: British Journal of Pharmacy
                Abbreviated Title: BJPharm
                Publisher: University of Huddersfield Press
                ISSN (Electronic): 20588356
                Publication date Created: November 15 2016
                Publication date (Electronic): November 15 2016
                Article
                DOI: 10.5920/bjpharm.2016.03
                SO-VID: 9cc04f1e-7525-438d-b8df-051ec0dd306d
                Copyright © © 2016
                License:

                This work is licensed under a Creative Commons Attribution 4.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

                History

                ScienceOpen disciplines: Medicine,Pharmacology & Pharmaceutical medicine,Health & Social care
                Data availability:
                ScienceOpen disciplines: Medicine, Pharmacology & Pharmaceutical medicine, Health & Social care

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