Development of simple spectrophotometric method for tetracycline determination
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Abstract
Introduction : Tetracycline (2-(amino-hydroxy-methylidene)-4-dimethylamino-6,10,11,12a-tetrahydroxy-6-methyl-4,4a,5,5a-tetra hydrotetracene-1,3,12-trione) has a broad-spectrum antibiotic and commonly used in veterinary medicine for disease prevention and treatment. The chemical structure of tetracycline is having a fused ring, partially aromatic, 4-ring structure with a wide variety of functional groups that form complexes with various metal ions. Materials and methods : Tetracycline solution for spectrophotometric analysis with concentrations ranging from 1.0 to 100.0 |ig?mL were prepared in 25 mL volumetric flasks. The appropriate amounts of tetracycline stock solution were diluted with deionized distillated water which was mixed with the mineral water used as reagent in test tube. The light-yellow solution was occurred from the reaction between tetracycline and mineral water afterward leave it for a few minutes. The absorption signal of the working standard and sample solution were recorded using UV-Visible spectrophotometer. Results : The method is based on the spectrophotometric detection of the light-yellow complex formed by the reaction between tetracycline and minerall water used as green reagent. The linear regression equation over the range of 0.0 - 50.0 |ig/mL was y = 0.0230X + 0.0081. The correlation coefficient (r) was found to be 0.9998. The relative standard deviation of the proposed method calculated from 12 replicate measurement of 1.0, 10.0 and 20.0 pg/mL tetracycline were 1.62, 0.21 and 0.08%, respectively. The percentage recoveries from spiked samples at three concentrations (5.0, 13.0 and 35.0 |ug/mL) was ranged between 99.14 and 99.77%. Conclusion : The proposed method was successfully validated for tetracycline in compliance with requirements by The United States Pharmacopeia (USP 34) 2011. This method is not use chemical reagent in all the reactions that minimize the use of a toxic chemical wastes.
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