Liquid chromatography folates

Freisleben, A., Sehieberle, P, Ryehlik, M. (2003 May). Specific and sensitive quantification of folate vitamers in foods by stable isotope dilution assays using high-performance liquid chromatography-tandem mass speetrometry. Anal. Bioanal. Chem., 376 2), 149-156. 

Kariluoto, M. S., Vahteristo, L. T, Piironen, V. 1. (2001). Applicability of microbiological assay and affinity chromatography purification followed by high-performance liquid chromatography (HPLC) in studying folate contents in rye. J. Sci. Food Agric., 81, 938-942. 

CM Pfeiffer, LM Rogers, JF Gregory. Determination of folate in cereal-grain food products using trienzyme extraction and combined affinity and reversed-phase liquid chromatography. J Agric Food Chem 45 407-413, 1997. 

G Varela-Moreiras, E Seyoum, J Selhub. Combined affinity and ion pair liquid chromatographies for the analysis of folate distribution in tissues. J Nutr Biochem 2 44-53, 1991. 

J Selhub. Determination of tissue folate composition by affinity chromatography followed by high-pressure ion pair liquid chromatography. Anal Biochem 182 84-93,1989. 

LT Vahteristo, V Ollilainen, PE Koivistoinen, P Varo. Improvements in the analysis of reduced folate monoglutamates and folic acid in food by high-performance liquid chromatography. J Agric Food Chem 44 477-482, 1996. 

Indirect indicators of vitamin B12 deficiency include measurements of the metabolites homocysteine and methylmalonic acid (MMA) in serum and MMA in urine (see the Biochemical Perspectives section). Whereas the serum homocysteine concentration increases during folate or vitamin B12 deficiencies, the serum and urine MMA concentrations increase only in vitamin B12 deficiency. Therefore, MMA determinations can be used to differentiate vitamin B12 deficiency from folate deficiency. The normal concentration of MMA in serum ranges from 0.08 to 0.28 pmol/L. MMA is quantified using gas-liquid chromatography and mass spectrometry. Elevated concentrations of MMA and homocysteine in serum may precede the development of hematological abnormalities and reductions in serum vitamin B12 concentrations. One should be aware that other conditions, including renal in sufficiency and inborn errors of metabolism, can also result in elevated serum levels of MMA. 

The main folate vitamers occurring in fruits and vegetables are different from those found in animal and cereal products [16]. There is also evidence that the vitamers have different bioavailability [17] and stability [18]. A clear need existed for an analytical method that would be capable of independently quantifying each specific folate vitamer. Liquid chromatography (LC) seemed an ideal and logical technology for this purpose. 

The first liquid chromatography-mass spectrometry (LC-MS) method was developed by Stokes and Webb (1999). Here, folate was separated using reversed-phase chromatography, detected in a single ion monitoring (SIM) mode and quantified using external standards [25]. These authors successfully identified and determined folic acid, tetrahydrofolate, 5-methyltetrahydrofolate, and formyltetrahydrofolate in multivitamins and foods. 

Ruggeri, S., Vahteristo, L. T., Aguzzi, A., Finglas, P., and Camovale, E. 1999. Determination of folate vitamers in food and in Italian reference diet by high-performance liquid chromatography. J. Chromatogr. A. 855 237-245. 

Freisleben, A., Schieberle, P., and Rychhk, M. 2003. Comparison of folate quantification in foods by high-performance liquid chromatography-fluorescence detection to that by stable isotope dilntion assays nsing high-performance liquid chromatography-tandem mass spectrometry. Anal. Biochem. 315 247-255. 

Patring, J., Wandel, M., Jagerstad, M., and Frplich, W. 2009. Folate content of Norwegian and Swedish flonrs and bread analysed by use of liquid chromatography-mass spectrometry. J. Food Comp. Anal. 22 649-656. 

Vishnumohan, S., Arcot, J., andPickford, R. 2011. Naturally-occurring folates in foods Method development and analysis using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Food Chem. 125 736-742. 

De Brouwer, V., Storozhenko, S., Stove, C. P, Van Daele J., Van Der Straeten, D., and Lambert, W. E. 2010. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for the sensitive determination of folates in rice. J. Chromatogr. B 878 509-513. 

Stralsjo, L., Ahlin, H., Witthoft, C., and Jastrebova, J. 2003. Folate determination in berries by radioprotein-binding assay (RPBA) and high performance liquid chromatography (HPLC). Fur. Food Res. Technol. 216 264-269. 

Pawlosky, R., Flanagan, V., and Doherty, R. 2003. A mass spectrometric validated high-performance liquid chromatography procedure for the determination of folates in foods. J. Agric. Food Chem. 51 3726-3730. 

A 96-well microtitre plate assay with microcomputer analysis has simplified the microbiological assay making it less laborious, less time-consuming and more reproducible. Recently, an automated 96-well plate isotope-dilution tandem mass spectrometry method and an ultra-performance liquid chromatography-tandem mass spectrometry method have been developed and validated for the quantification of synthetic folic add in folate-fortified breads (Fazili and Pfeiffer 2004 Chandra-Hioe et al. 2011). 

In addition, HPLC coupled with mass spectrometry (HPLC-MS) has been widely utilized for the analysis of folate. Liquid chromatography-tandem mass spectroscopic (LC-MS/MS) determination of pABA released by the acid hydrolysis of erythrocyte folates was previously described (Clifford et al. 2005), in which [ Cg]pABA was used as internal standard and derivatization of pABA with diazomethane was performed prior to analysis. 

Kohashi, M., Inoue, K., Sotobayashi, H., and Iwai, K., 1986. Microdetermination of folate monoglutamates in serum by liquid-chromatography with electrochemical detection. Journal of Chromatography. 382 303-307. 

Kirsch, S.H., Knapp, J.-P., Herrmaim, W., and Obeid, R., 2010. Quantification of key folate forms in serum using stable-isotope dilution ultra performance liquid chromatography-tandem mass spectrometry. Journal of Chromatography B. 878 68-75. 

Liu, J., Pickford, R., Meagher, A. P., Ward, R. L., 2011. Quantitative analysis of tissue folate using ultra high-performance liquid chromatography tandem mass spectrometry. Analytical Biochemistry. 411 210-217. 

Rychlik, M., Netzel, M., Pfannebecker, I., Frank, T., and Bitsch, I., 2003. Application of stable isotope dilution assays based on liquid chromatography-tandem mass spectrometry for the assessment of folate bioavailability. Journal of Chromatography B. 792 167-176. 

Stokes, P., and Webb, K., 1999. Analysis of some folate monoglutamates by high-performance liquid chromatography-mass spectrometry. I. Journal of Chromatography. 864 59-67. 

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