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This study was conducted to develop an HPLC method for determining vitamin B₁₂ in fortified foods which has typically been determined by microbiological assays according to AOAC and Korean Food Code approved methods. Vitamin B₁₂ (cyanocobalamin) was determined by reversed-phase HPLC with a triple column and UV/VIS dectector (550 nm) using the column switching technique after extraction with 5 mM potassium phosphate solution by sonication without a clean-up procedure. The recovery of spiked samples and limit of detection (LOD) by HPLC were 78.6~107.5% and 2 ppb (μg/kg), respectively. The LOD of the microbiological assay (MBA) was much lower than that of HPLC. The concentrations of vitamin B₁₂ analyzed in all tested samples (n=12) confirmed compliance with declared label claims. The range of recovery ratio by the HPLC method when compared to the microbiological assay was 76.2~140.0%. There was not significant difference between the HPLC and MBA methods (p<0.01) with r=0.9791 and linear regression y=0.9923x-0.04. The HPLC method for determining vitamin B₁₂ using the column-switching technique appears to be suitable for determining vitamin B₁₂ concentrations above 1 μg/100 g in fortified foods.

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INTRODUCTION

MATERIALS AND METHODS

RESULTS AND DISCUSSION

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UCI(KEPA) : I410-ECN-0101-2009-511-018079288