Assay of Kanamycin A by HPLC with Direct UV Detection
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The development of a simple reversed phase ion pair liquid chromatographic method for the assay of kanamycin A has been described. Because of the lack of a UV chromophore in the structure of kanamycin A, borate complexation was used to allow direct UV detection at 205 nm. Three columns were evaluated in this study: Zorbax Extend C18 (4.6 mm × 250 mm; 5 μm), XBridge C18 (4.6 mm × 250 mm; 5 μm) and apHera C18 (4.6 mm × 250 mm; 5 μm). The mobile phase was a mixture of 0.1 M disodium tetraborate (pH 9.0) and water (20:80, v/v) supplemented with 0.5 g L−1 sodium octanesulphonate. Final chromatographic conditions were achieved on the XBridge column at 50 °C. The method was validated according to ICH guidelines and applied to a commercially available sample. It is much faster and more specific than the current microbiological assay prescribed in the European Pharmacopoeia. No expensive equipment is necessary to perform this assay making it a viable replacement.
KeywordsHPLC–UV Ion pair liquid chromatography Borate complexation Kanamycin
This paper was supported by the TÁMOP 4.2.1 B-09/1/KMR tender of the Hungarian Government (P. Jankovics).
- 6.Umezawa H, Ueda M, Maeda K, Yagashita K, Kondo S, Okami Y, Utahara R, Osato Y, Nitta K, Takeuchi T (1957) Production and isolation of a new antibiotic. Kanamycin. J Antibiot 10(5):181–188Google Scholar
- 11.Rothrock JW, Guegelman RT, Wolf J (1958) A resin chromatographic analysis for kanamycin mixtures. Antibiot Annu 6:796–803Google Scholar
- 18.El Rassi Z (2002) Carbohydrate analysis by modern chromatography and electrophoresis. Elsevier, AmsterdamGoogle Scholar
- 20.European Pharmacopoeia (2012) 7th edn. The Council of Europe, Strasbourg Cedex, France, pp 2313–2314Google Scholar