DETERMINATION OF WATER-SOLUBLE VITAMINS B1, B2, B3, B6, B9, B12 AND C ON C18 COLUMN WITH PARTICLE SIZE 3 ÂµM IN SOME MANUFACTURED FOOD PRODUCTS BY HPLC WITH UV-DAD/FLD DETECTION
Objective: Objective of the study was to develop a simple, precise and accurate RP-HPLC ion-pair method, for the determination of water-soluble vitamins (B1, B2, B3, B6, B9, B12, and C) in some manufactured food products.
Methods: RP-HPLC with C18 BDS (100 x 4.6 mm; 3 Âµm) column were used. Mobile phase constituents were solvent (A): 5.84 mM of hexane-1-sulfonic acid sodium: acetonitrile (95:5) with 0.1% triethylamine at pH 2.5 and solvent (B): 5.84 mM of hexane-1-sulfonic acid sodium: acetonitrile (50:50) with 0.1% triethylamine at pH 2.5, flow rate 1.6 ml/min, at column temperature 40 Â°C and suitable detection wavelength.
Results: Two different detectors were used: photo diode array detector (UV-DAD) and the fluorescence detector (FLD). Calibration graphs plotted with five concentrations of each vitamin where linear regression coefficients R2Ëƒ 0.9972. LOQ values were 50.0, 81.0, 19.1, 19.0, 30.0, 9.7, 50.0 Âµg/l with DAD for vitamins B1, B2, B3, B6, B9, B12 and C respectively, and 5.7, 4.1 Âµg/l with FLD for vitamins B2, B6 respectively. LOD values were 16.5, 26.7, 6.3, 6.3, 9.9, 3.2, 16.5 Âµg/l with DAD for vitamins B1, B2, B3, B6, B9, B12 and C respectively, and 1.9, 1.3 Âµg/l with FLD for vitamins B2, B6 respectively.
Conclusion: The proposed method was successfully applied to analysis mixture of seven water-soluble vitamins in pure form and in manufactured food products, with average recovery of 98.14% to 100.96%.
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