High Performance Liquid Chromatographic Method for Rapid Determination of Thiamine and Riboflavin in Dairy Products
Abstract
A fast, simple and reliable high-performance liquid chromatographic method has been developed for the simultaneous analysis of thiamine and riboflavin. The chromatographic separation was performed on a mini C18 column (8.0 x 3.0 mm, length x I.D.) with isocratic elution using acetronitrile and 0.05 M acetate buffer (pH 7.0). Direct UV absorptions at wavelength of 232 and 266 nm were used for detection of thiamine and riboflavin, respectively. Both vitamins were separated in less than 8.0 min at flow rate of 0.10 ml/min. The linear calibration curves were obtained over the concentration range of 0.10 - 100 mg/L for thiamine and 0.25 - 100 mg/L for riboflavin with good correlation coefficients (r2 > 0.999). The precisions were measured in term of relative standard deviation (RSD) value. The RSDs of the method were in the range 0.34 – 1.90%. The percentage recoveries of both vitamins were obtained in the range of 81.83 - 102.70. The contents of both vitamins in milk samples were successfully determined by using the developed method. Compared with the conventional HPLC method, the proposed method has the advantage of being rapid, low cost and environmentally friendly. Keywords : Thiamine, Riboflavin, Dairy Products, short column, High Performance Liquid ChromatographyReferences
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Analysis of Vitamin B1 in Dry-Cured Sausages by Hydrophilic Interaction Liquid Chromatography
(HILIC) and Array Detection. Meat Science, 87, 234-238.
Hurtado, S.A., Nogués, M.T.V., Pulido, M.I., & Font, A.M. (1997). Determination of water-soluble vitamins in infant
milk by high-performance liquid chromatography. Journal of chromatography A, 778, 247-253.
Kall, M.A. (2003). Determination of total vitamin B6 in foods by isocratic HPLC: a comparison with microbiological
analysis. Food Chemistry, 82, 315-327.
Le, X.C., Ma, M. (1998). Short-Column Liquid Chromatography with Hydride Generation Atomic Fluorescence
Detection for the Speciation of Arsenic. Analytical Chemistry, 70, 1926-1933.
Liu, S., Zhang, Z., Liu, Q., Luo, H., & Zheng, W. (2002). Spectrophotometric determination of vitamin B1 in
a pharmaceutical formulation using triphenylmethane acid dyes. Journal of Pharmaceutical and
Biomedical Analysis, 30, 685-694.
Márquez-Sillero, I., Cárdenas, S., & Valcárcel, M. (2013). Determination of Water-Soluble Vitamins in Infant Milk
and Dietary Supplement Using a Liquid Chromatography on-line Coupled to a Corona-charged Aerosol
Detector. Journal of Chromatography A, 1313, 253– 258.
Maslowska, J., Malicka, M. (1984). Potentiometric and Spectrophotometric Studies on Acidic-Basic Properties of Riboflavin. Polish Journal of Chemistry. 58, 503-510.
Rodriguez, R.S.J, Fernández-Ruiz, V., Cámara, M., Sánchez-Mata, M.C. (2012). Simultanous Determination of
Vitamin B1 and B2 in Complex Cereal Foods, by Reverse phase Isocratic HPLC-UV. Journal of Cereal
Science, 55, 293-299.
Tanner J.T., Barnett S.A. (1986). Methods of Analysis for Infant Formula: Food and Drug Administration and Infant
Formula Council Collaborative Study, Phase III. Journal-Association of Official Analytical Chemists,
69, 777-785.
Wu, X., Diao, Y., Sun, C., Yang, J., Wang, Y., & Sun, S. (2003). Fluorimetric determination of ascorbic acid with
o-phenylenediamine. Talanta, 59, 95-99.
Yin,C., Cao,Y., Ding,S., Wang,Y. (2008). Rapid Determination of Water- and Fat-Soluble Vitamins with
Microemulsion Electrokinetic Chromatography. Journal of Chromatography A, 1193, 172-177.
Youngvises, N., Chaida, T., Khonyoung, S., Kuppithayanant, N., Tiyapongpattana, W., Itharat, A., Jakmunee, J.
(2013). Greener Liquid Chromatography Using a Guard Column with Micellar Mobile Phase for
Separation of Some Pharmaceuticals and Determination of Parabens. Talnata, 106, 350-359.
Youngvises, N., Liawruangrath, B., Liawruangrath, S. (2003). Simultaneous Micellar LC Determination of
Lidocaine and Tolperisone. Journal of Pharmaceutical and Biomedical Analysis, 31, 629-638.
Engel, R., Stefanovit-Bányai, E., & Abrankó, L. (2010). LC Simultaneous Determination of the Free Forms of B
Group Vitamins and Vitamin C in Various Fortified Food Products. Chomatographia, 71, 1069–1074.
Food and Drug Administration, Center for Drug Evaluation and Research. (1994). Validation of Chromatographic Methods. Retrieved December 3, 2015, from http://www.fda.gov/downloads/Drugs/Guidances/ UCM134409.pdf
Gatti, R., Gioia, M.G. (2005). Liquid Chromatography Determination with Fluorescence Detection of
B6 Vitamers and Riboflavin in Milk and Pharmaceuticals. Analytica Chimica Acta, 538, 135-141.
Gratacós-Cubarśi, M., Sárraga, C., Clariana, M., Garćia Regueiro, J.A., Castellari, M. (2011).
Analysis of Vitamin B1 in Dry-Cured Sausages by Hydrophilic Interaction Liquid Chromatography
(HILIC) and Array Detection. Meat Science, 87, 234-238.
Hurtado, S.A., Nogués, M.T.V., Pulido, M.I., & Font, A.M. (1997). Determination of water-soluble vitamins in infant
milk by high-performance liquid chromatography. Journal of chromatography A, 778, 247-253.
Kall, M.A. (2003). Determination of total vitamin B6 in foods by isocratic HPLC: a comparison with microbiological
analysis. Food Chemistry, 82, 315-327.
Le, X.C., Ma, M. (1998). Short-Column Liquid Chromatography with Hydride Generation Atomic Fluorescence
Detection for the Speciation of Arsenic. Analytical Chemistry, 70, 1926-1933.
Liu, S., Zhang, Z., Liu, Q., Luo, H., & Zheng, W. (2002). Spectrophotometric determination of vitamin B1 in
a pharmaceutical formulation using triphenylmethane acid dyes. Journal of Pharmaceutical and
Biomedical Analysis, 30, 685-694.
Márquez-Sillero, I., Cárdenas, S., & Valcárcel, M. (2013). Determination of Water-Soluble Vitamins in Infant Milk
and Dietary Supplement Using a Liquid Chromatography on-line Coupled to a Corona-charged Aerosol
Detector. Journal of Chromatography A, 1313, 253– 258.
Maslowska, J., Malicka, M. (1984). Potentiometric and Spectrophotometric Studies on Acidic-Basic Properties of Riboflavin. Polish Journal of Chemistry. 58, 503-510.
Rodriguez, R.S.J, Fernández-Ruiz, V., Cámara, M., Sánchez-Mata, M.C. (2012). Simultanous Determination of
Vitamin B1 and B2 in Complex Cereal Foods, by Reverse phase Isocratic HPLC-UV. Journal of Cereal
Science, 55, 293-299.
Tanner J.T., Barnett S.A. (1986). Methods of Analysis for Infant Formula: Food and Drug Administration and Infant
Formula Council Collaborative Study, Phase III. Journal-Association of Official Analytical Chemists,
69, 777-785.
Wu, X., Diao, Y., Sun, C., Yang, J., Wang, Y., & Sun, S. (2003). Fluorimetric determination of ascorbic acid with
o-phenylenediamine. Talanta, 59, 95-99.
Yin,C., Cao,Y., Ding,S., Wang,Y. (2008). Rapid Determination of Water- and Fat-Soluble Vitamins with
Microemulsion Electrokinetic Chromatography. Journal of Chromatography A, 1193, 172-177.
Youngvises, N., Chaida, T., Khonyoung, S., Kuppithayanant, N., Tiyapongpattana, W., Itharat, A., Jakmunee, J.
(2013). Greener Liquid Chromatography Using a Guard Column with Micellar Mobile Phase for
Separation of Some Pharmaceuticals and Determination of Parabens. Talnata, 106, 350-359.
Youngvises, N., Liawruangrath, B., Liawruangrath, S. (2003). Simultaneous Micellar LC Determination of
Lidocaine and Tolperisone. Journal of Pharmaceutical and Biomedical Analysis, 31, 629-638.
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Published
2016-02-02
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