A simple and rapid RP-HPLC method for the assessment of cobalamin (vitamin B12) in tilapia and snoek fishes

Authors: LABARAN IBRAHIM, AMINU USMAN

Abstract: This study developed a rapid reversed-phase high-performance liquid chromatography (RP-HPLC) method equipped with a UV-Vis detector. The aquaponics system?s tilapia and marine snoek fishes were extracted using an autoclaving process and enzymatic treatment. The technique enabled the separation and quantification of cobalamin present in these fishes extracts. Phenomenex Luna^® 5 μm C18 (2) 100 A LC-column (150 × 4.6 mm) was used as a stationary phase, while the mobile phase consisted of methanol and phosphoric acid in a ratio of 35:65 (v/v), respectively. The developed method was revealed to be rapid (a retention time of less than 5.0 min), linear (R² = 0.9988), sensitive (limits of detection and quantification showed to be 0.0004 and 0.0011 μg/mL, respectively), precise (percentage relative standard deviation of 0.14% to 0.76%), and accurate (percentage mean recovery of 87.44 ± 0.33% to 97.08 ± 0.74%). The quantified concentrations of this vitamin in extracts of tilapia and snoek fishes were found to be 08.95 ± 0.35 and 14.97 ± 0.04 μg/ mL, respectively. Therefore, we suggested that the developed RP-HPLC technique could be applicable for quality control evaluation in the food and pharmaceutical industries. Besides, the method could be vital for diagnostic analysis in clinical laboratories.

Keywords: Isocratic, precise, stationary phase, chromatography, retention time, limit of detection

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