Chromatography folate determination

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. 

JC Gounelle, H Ladjimi, P Prognon. A rapid and specific extraction procedure for folates determination in rat liver and analysis by high-performance liquid chromatography with fluorometric detection. Anal Biochem 176 406-411, 1989. 

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. 

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

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 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. 

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

Phillips, K. M., Ruggio, D. M., Ashraf-Khorassani, M., and Haytowitz, D. B. 2006. Difference in folate content of green and red sweet peppers Capsicum annuum) determined by liqnid chromatography-mass spectrometry. J. Agr. Food Chem. 54 9998-10002. 

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. 

Patring, J. D. M. and Jastrebova, J. A. 2007. Application of hqnid chromatography-elec-trospray ionisation mass spectrometry for determination of dietary folates Effects of buffer nature and mobile phase composition on sensitivity and selectivity. J. Chromatogr. A. 1143 72-82. 

Garbis, S. D., Melse-Boonstra, A., West, C. E., and Van-Breemen, R. B. 2001. Determination of folates in human plasma using hydrophilic interaction chromatography-tandem mass spectrometry. Anal. Chem. 73 5358-5364. 

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. 

Fazili, Z., PfeifTer, C.M., Zhang, M., and Jain, R., 2005. Erythrocyte folate extraction and quantitative determination by Kquid chromatography-tandem mass spectrometry Comparison of results with microbiologic assay. Clinical Chemistry. 51 2318-2325. 

Claradiastase and trypsin in phosphate buffer have been used for hydrolysis of sample matrix for the determination of supplemental folic acid (27). Jacoby and Henry (26) further modified the method of Hoppner and Lampi (41) for folic acid by HPLC, in which the folic acid added to infant formulas and liquid medical nutritionals is quantitatively extracted with the aid of bacterial protease and papain. One disadvantage of the enzymatic extraction method was the large number of UV-absorbing compounds that are formed during enzymatic hydrolysis. These can interfere in the quantitation of the folate peaks. Thus, addition of a-amylase and protease to enhance extraction has not been common practice for methods involving HPLC. However, Pfeiffer et al. (13) suceessfiilly used triple-enzyme treatment prior to HPLC analysis with a purification method based on affinity chromatography. 

Several methods for separating single folate compounds such as folic acid from matrix interference are available. Akhtar et al. (75) applied paired-ion chromatography for HPLC determination of folic acid and its photodegradation... 

DL Holt, RL Wehling, MG Zeece. Determination of native folates in milk and other dairy products by high-performance liquid chromatography. J Chromatogr... 

L Vahteristo, V OUilainen, P Koivistoinen, P Varo. Improvements in the determination of reduced folate monoglutamates and folic acid in food using high-performance liquid chromatography. J Agric Food Chem 44 477-482, 1996. 

I Eto, CL Krumdieck. Determination of three different pools of reduced one-car-bon-substituted folates. III. Reversed-phase high-performance liquid chromatography of the azo dye derivatives of p-aminobenzoylpolyglutamates and its application to the study of unlabeled endogenous pteroylpolyglutamates of rat liver. Anal Biochem 120 323-329, 1982. 

GL Case, RD Steele. Determination of reduced folate derivatives in tissue samples by high-performance liquid chromatography with fluoromelric detection. J Chro-... 

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