DETERMINATION OF WATER-SOLUBLE VITAMINS B1, B2, B3, B6, B9, B12 AND C ON C18 COLUMN WITH PARTICLE SIZE 3 µM IN SOME MANUFACTURED FOOD PRODUCTS BY HPLC WITH UV-DAD/FLD DETECTION
Keywords:Determination, Vitamins, High performance liquid chromatography, Reserved-phase, Ion-pair, Manufactured, Food
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 µm) 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 °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 R2˃ 0.9972. LOQ values were 50.0, 81.0, 19.1, 19.0, 30.0, 9.7, 50.0 µg/l with DAD for vitamins B1, B2, B3, B6, B9, B12 and C respectively, and 5.7, 4.1 µg/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 µg/l with DAD for vitamins B1, B2, B3, B6, B9, B12 and C respectively, and 1.9, 1.3 µg/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%.
Vidovic C, Stojanovic B, Veljkovic J, Prazic-Arsic L, Roglic G, Manojlovic D. Simultaneous determination of some water-soluble vitamins and preservatives in multivitamin syrup by validated stability-indicating high-performance liquid chromatography method. J Chromatogr A 2008;1202(2):155-62.
Chen Z, Chen B, Yao S. High-performance liquid chromatography/electrospray ionization-mass spectrometry for simultaneous determination of taurine and 10 water-soluble vitamins in multivitamin tablets. Anal Chim Acta 2006;569(1-2):169-75.
Ciulu M, Solinas S, Floris I, Panzanelli A, Pilo MI, Piu PC, et al. RP-HPLC determination of water-soluble vitamins in honey. Talanta 2011;83(3):924-9.
Heudi O, Kilinc T, Fontannaz P. Separation of water-soluble vitamins by reversed-phase high performance liquid chromatography with ultra-violet detection: Application to polyvitaminated premixes. J Chromatogr A 2005;1070(1-2):49-56.
Ekinci R, Kadakal C. Determination of seven water-soluble vitamins in tarhana, a traditional Turkish cereal food, by high performance liquid chromatography. Acta Chromatogr 2005;15:289-97.
Marszall ML, Lebiedzinska A, Czarnowski W, Szefer P. High performance liquid chromatography method for the simultaneous determination of thiamine hydrochloride, pyridoxine hydrochloride and cyanocobalamin in pharmaceutical formulations using coulometric electrochemical and ultraviolet detection. J Chromatogr A 2005;1094(1-2):91-8.
Fotsing L, Fillet M, Bechet I, Hubert Ph, Crommen J. Determination of six water-soluble vitamins in a pharmaceutical formulation by capillary electrophoresis. J Pharm Biomed Anal 1997;15(8):1113-23.
Kartsova LA, Koroleva OA. Simultaneous determination of water-and fat-soluble vitamins by high-performance thin-layer chromatography using an aqueous micellar mobile phase. J Anal Chem 2007;62(3):255-9.
Rajput GK, Kumar A, Kumar A, Srivastav G. To develop a simple (UV-Vis spectrometric) method for the estimation of water soluble vitamins. Int J Drug Res 2011;2(3):232-9.
Saradhi DRSV, Prryanka A, Baby Sirisha P, Meherjaha SK, Jyothsna N. Enzymatic and spectroscopic determination of riboflavin using amylase phosphate. IJPPS 2012;4(3):170-2.
Mohamed AM, Mohamed HA, Abdel-Latif NM, Mohamed MR. Spectrofluorimetric determination of some water-soluble vitamins. J AOAC Int 2011;94(6):1758-69.
Okamoto H, Nakajima T, Ito Y. Simultaneous determination of water-soluble vitamins in a vitamin-enriched drink by an in-capillary enzyme reaction method. J Chromatogr A 2003;986(1):153-61.
Ball GFM. Water-soluble vitamin assays in human nutrition. Chapman & Hall; 1994;317-64.
Monferrer-Pons L, Capella-Peiro ME, Gil-Agusti M, Esteve-Romero J. Micellar liquid chromatography determination of B vitamins with direct injection and ultraviolet absorbance detection. J Chromatogr A 2003;984(2):223-31.
Poongothai S, Ilavarasan R, Karrunakaran CM. Simultaneous and accurate determination of vitamins B1, B6, B12 and alpha-lipoic acid in multivitamin capsule by reverse-phase high performance liquid chromatographic method. IJPPS 2010;2(4):133-9.
Chen P, Atkinson R, Wolf WR. Single laboratory validation of a high-performance liquid chromatographic-diode array detector-fluorescence detector/mass spectrometric method for simultaneous determination of water-soluble vitamins in multivitamin dietary tablets. J AOAC Int 2009;92(2):680-7.
Kirilov B, Obreshkova D, Tsvetkova D. Validation of HPLC method for determination of antioxidant vitamin C and vitamin B6 in food supplements and drugs. IJPPS 2012;4(1):300-4.
Suh JH, Yang DH, Lee BK, Eom HY, Kim U, Kim J, et al. Simultaneous determination of B groupe vitamins in supplemented food products by high performance liquid chromatography-diode array detection. J Bull Korean Chem Soc 2011;32(8):2648-56.
Saint Jose Rodriguez R, Fernandez-Ruiz V, Camara M, Sanchez-Mata MC. Simultaneous determination of vitamin B1 and B2 in complex cereal foods, by reverse phase isocratic HPLC-UV. J Cereal Sci 2012;55(3):293-9.
Esteve MJ, Farré R, Frigola A, Pilamunga C. Contents of vitamins B1, B2, B6, and B12 in pork and meat products. J Meat Sci 2002;62(1):73-8.
Freisleben A, Schieberle P, Rychlik M. Comparison of folate quantification in foods by high-performance liquid chromatography-fluorescence detection to that by stable isotope dilution assays using high-performance liquid chromatography-tandem mass spectrometry. J Anal Biochem 2003;315(2):247-55.
Furlani RPZ, Godoy HT. Vitamins B1 and B2 contents in cultivated mushrooms. J Food Chem 2008;106(2):816-9.
Lebiedzinska A, Marszall ML, Kuta J, Szefer P. Reversed-phase high-performance liquid chromatography method with coulometric electrochemical and ultraviolet detection for the quantification of vitamins B1 (thiamine), B6 (pyridoxamine, pyridoxal and pyridoxine) and B12 in animal and plant food. J Chromatogr A 2007;1173(1-2):71-80.
De Brouwer V, Storozhenko S, Stove CP, Van Daele J, Van Der Straeten D, et al. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for the sensitive determination of folate in rice. J Chromatogr B 2010;878(3-4):509-13.
Alaburda J, De Almeida AP, Shundo L, Ruvieri V, Sabino M. Determination of folic acid in fortified wheat flours. J Food Compos Anal 2008;21(4):336-42.
Rodriguez-Bernaldo de Quiros A, Fernandez-Arias M, Lopez-Hernandez J. A screening method for the determination of ascorbic acid in fruit juices and soft drinks. J Food Chem 2009;116(2):509-12.
Jastrebova J, Witthoft C, Grahn A, Svensson U, Jagerstad M. HPLC determination of folates in raw and processed beetroots. J Food Chem 2003;80(4):579-88.
Romeu-Nadal M, Morera-Pons S, Castellote AI, Lopez-Sabater MC. Rapid high-performance liquid chromatographic method for vitamin C determination in human milk versus an enzymatic method. J Chromatogr B 2006;830(1):41-6.
Mittermayr R, Kalman A, Trisconi MJ, Heudi O. Determination of vitamin B5 in a range of fortified food products by reversed-phase liquid chromatography-mass spectrometry with electrospray ionization. J Chromatogr A 2004;1032(1-2):1-6.
Chavez-Servin JL, Castellote AI, Rivero M, Lopez-Sabater MC. Analysis of vitamins A, E and C, iron and selenium contents in infant milk-based powdered formula during full shelf-life. J Food Chem 2008;107(3):1187-97.
Heudi O, Kilinc T, Fontannaz P, Marley E. Determination of vitamin B12 in food products and in premixes by reversed-phase high performance liquid chromatography and immunoaffinity extraction. J Chromatogr A 2006;1101(1-2):63-8.
Lu B, Ren Y, Huang B, Liao W, Cai Z, Tie X. Simultaneous determination of four water-soluble vitamins in fortified infant foods by ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry. J Chromatogr Sci 2008;46:225-32.
Valente A, Albuquerque TG, Sanches-Silva A, Costa HS. Ascorbic acid content in exotic fruits: a contribution to produce quality data for food composition databases. J Food Res Int 2011;44(7):2237-42.