In this study, we have developed and validated a simple and rapid isocratic reverse-phase high performance liquid chromatographic method with UV detection (220 nm) for quantification of salmon calcitonin. Chromatography was performed using an Agilent Eclipse XDB-C18 column (150x4.6 mm i.d., 3.5 pim particle size) with a mobile phase (water:acetonitrile 65:35, v/v), containing 0.1% v/v trifluoroacetic acid at a flow rate of 1 mL/min. Each analysis requires less than 7 min including triamcinolone acetonide as an internal standard. The method has been validated for accuracy, precision, linearity, specificity, and sensitivity. The method has proved to be specific, rapid, accurate, precise and reproducible according to ICH standards. The calibration curve is linear in the concentration range 2.5- 50 pig/mL; the correlation coefficient was 0.999. The method has showed good recoveries (average 100.64%) and the relative standard deviations intra- and inter-day were < 2.0%. The method has been successfully applied for the rapid and accurate quantification of salmon calcitonin in raw materials and commercial formulations.
Keywords: HPLC, Salmon calcitonin, Validation, Isocratic, Triamdnolone acetonide