Development and validation of a sensitive method for tetracycline in gingival crevicular fluid by HPLC using fluorescence detection

Journal of Pharmaceutical and Biomedical Analysis
D S Vienneau, C G Kindberg

Abstract

Increased interest in the clinical use of antibiotics for periodontal therapy required the development of a sensitive assay for the quantitation of tetracycline in gingival crevicular fluid (GCF). An HPLC method was developed and validated for tetracycline which separates and identifies the degradation component epi-tetracycline. The HPLC assay employs a C18 reversed-phase Hypersil column with a mobile phase composed of methanol and sodium acetate buffer containing CaCl2 and EDTA disodium salt. The chromatographic separation was monitored by a fluorescent detector with an excitation wavelength of 375 nm and an emission wavelength of 512 nm. Tetracycline was extracted from GCF collected on Periopapers by addition of a methanol solution containing the internal standard, doxycycline, and the mobile phase buffer (25:75, v/v). The mean percent recovery for the extraction method was 107.8% with all the % R.S.D. below 7.5%. The mean inter- and intra-batch accuracy was 104.1 and 105.3%, respectively with a coefficient of variation of less than 9.5%. The lower limit of detection was 2.5 ng on the Periopapers. The typical GCF volumes collected were 0.1-1 microliter. The method was validated for the linear concentration range 2.5-1000 ng ...Continue Reading

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Citations

Aug 24, 1999·Biotechnology and Bioengineering·X MazurM Fussenegger
Mar 1, 2005·Journal of Periodontology·Maria LavdaJohn D Walters
Mar 26, 2005·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Thiraporn CharoenraksOrawon Chailapakul
Feb 20, 2016·Pharmaceutical Development and Technology·Srilekha Sarkar DasDavid M Saylor
Apr 29, 2015·Journal of Biomaterials Science. Polymer Edition·Lucas Alves MouraAntonio Wilson Sallum
Mar 19, 2013·Journal of Pharmaceutical and Biomedical Analysis·Marko S DenićMilica S Petrović
Jun 11, 2016·European Journal of Pharmaceutical Sciences : Official Journal of the European Federation for Pharmaceutical Sciences·Liljana BogdanovskaRumenka Petkovska
Oct 25, 2013·Acta Pharmaceutica : a Quarterly Journal of Croatian Pharmaceutical Society and Slovenian Pharmaceutical Society, Dealing with All Branches of Pharmacy and Allied Sciences·Liljana BogdanovskaRumenka Petkovska
Jan 7, 2015·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Olga GrinbergEhud Banin

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