HPLC determination of vitamin B6 in foods after pre-column derivatization of free and phosphorylated vitamers into pyridoxol

Bergaentzlé, Martine; Marchioni, Eric; Hasselmann, C.

doi:10.1016/0308-8146(93)90149-a

Cited by 25 publications

(9 citation statements)

References 11 publications

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“…Thiamine (vitamin B 1 ), riboflavin (vitamin B 2 ), nicotinic acid and nicotinamide (vitamin B 3 ), pyridoxine (vitamin B 6 ), folic acid (vitamin B 9 ), and cyanocobalamin (vitamin B 12 ) were investigated in the present assay.…”

Section: Introductionmentioning

confidence: 99%

Development of a validated HPLC method for the determination of B‐complex vitamins in pharmaceuticals and biological fluids after solid phase extraction

Chatzimichalakis

Samanidou

Verpoorte

et al. 2004

J of Separation Science

109

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An HPLC method was developed for the simultaneous determination of seven water-soluble vitamins, viz. thiamine, riboflavin, nicotinic acid, nicotinamide, pyridoxine, cyanocobalamin, and folic acid, in multivitamin pharmaceutical formulations and biological fluids (blood serum and urine). Separation was achieved at ambient temperature on a Phenomenex Luna C18 (150 x 4.6 mm) analytical column. Gradient elution was performed starting at a 99:1 A:B v/v composition, where A: 0.05 M CH3COONH4/CH3OH (99/1) and B: H2O/CH3OH (50/50), at a flow rate of 0.8 mL/min. After a 4-min isocratic elution the composition was changed to 100% of B in 18 min and elution continued isocratically for 8 min. Detection was performed with a photodiode array detector at 280 nm. Each vitamin was quantitatively determined at its maximum wavelength. Spectral comparison was used for peak identification in real samples. Detection limits were in the range of 1.6-3.4 ng, per 20-microL injection, while linearity held up to 25 ng/microL. Accuracy, intra-day repeatability (n = 6), and inter-day precision (n = 7) were found to be satisfactory. Theobromine (2 ng/microL) was used as internal standard. Sample preparation of biological fluids was performed by SPE on Supelclean LC-18 cartridges with methanol-water 85/15 v/v as eluent. Extraction recoveries from biological matrices ranged from 84.6% to 103.0%.

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Section: Introductionmentioning

confidence: 99%

Development of a validated HPLC method for the determination of B‐complex vitamins in pharmaceuticals and biological fluids after solid phase extraction

Chatzimichalakis

Samanidou

Verpoorte

et al. 2004

J of Separation Science

109

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“…Extraction with an acetate buffer of pH 4.5 has been utilized for clinical samples (Hefferan et al 1986, Bötticher andBötticher 1987) , for fortified breakfast cereals (Gregory 1980c), vitamin preparations (Bühnert 1988) as well as for food samples (Reitzer-Bergaentzle et al 1993). Acidic acetate solution was easily combined with the followed enzymatic hydrolysis procedure and the method was suitable for the simultaneous measurement of free B 1 , B 2 and B 6 vitamers in plasma and serum samples.…”

Section: Metaphosphoric Acid (Mpa)mentioning

confidence: 99%

“…The method and its modifications have been further applied to food and other biological materials later on by various research groups. The use of an alcyl derivative of sulfonic acid predominates in ion-pair chromatography of B 6 compounds; a number of reports using hexane sulfonic acid (Kawamoto et al 1983, Wehling and Wetzel 1984, Bötticher and Bötticher 1987, Chase and Soliman 1990, Tolomelli et al 1991, Reynolds and Brain 1992, Olleta et al 1993, Blanco et al 1994, Agostini and Godoy 1997, heptane sulfonic acid (Lam et al 1984, Hefferan et al 1986, Bühnert 1988, Rees 1989, Iwase 1992, Reitzer-Bergaentzle et al 1993) and octane sulfonic acid (Bitsch and Möller 1989, Maeda et al 1989, van Schoonhoven et al 1994, Sharma and Daksinamurti 1992, Sampson et al 1995 or a mixture of alcyl sulphonates (Jaumann andEngelhardt 1985, Addo andAugustin 1988) have been published. The phenolic character of pyridoxine was utilized by Belal (1989); tetrabutyl ammoniumhydroxide enabled the proper separation of pyridoxine and pyrithioxine in a selected pH of 8 in dosage forms.…”

Section: Partition and Ion-pair Chromatographymentioning

confidence: 99%

“…Acidic acetate solution was easily combined with the followed enzymatic hydrolysis procedure and the method was suitable for the simultaneous measurement of free B 1 , B 2 and B 6 vitamers in plasma and serum samples. The validity of the acetate extraction used by Reitzer-Bergaentzle et al (1993) preceeding the pre-column derivatization of B 6 vitamers into pyridoxine was further evaluated by the collaborative study performed in France (Bergaentzle et al 1995). A lower pH acetate buffer was used for multivitamin preparate; sample was sonicated with a dilute solution of acetic acid (0.5M) and triethylamine (40mM) at the pH of 3.6 (Lam et al 1984) or with liquid chromatography's mobile phase (acetonitrile -10mM phosphate buffer -triethylamine, 8:91.5:0.5, pH 3.8) (Maeda et al 1989).…”

Section: Metaphosphoric Acid (Mpa)mentioning

confidence: 99%

“…Pyridoxal was converted to pyridoxine within two hours by reduction by sodium borohydride in alkaline medium (10mM in 0.2M NaOH) (Chaikin and Brown 1949). To decrease the reaction time, a more concentrated borohydride solution (0.1M) was needed (Reitzer-Bergaentzle et al 1993). This approach has been successfully utilized in an analytical method generally adopted in France.…”

Section: Some Reactions Of Pyridoxal and Pyridoxal-5'-phosphatementioning

confidence: 99%

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HPLC analysis of vitamin B6 in foods

Ollilainen¹

1999

AFSci

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The objective of this work was to evaluate the methods for determination of vitamin B6 in foods. To achieve this, the literature review focused on sample treatment and liquid chromatographic analysis of vitamin B6 related compounds. In the experimental part, the chosen sample pretreatment and the high-performance liquid chromatographic (HPLC) method were validated, and used to produce vitamin B6 data on various food items commonly consumed in Finland. The main emphasis of the sample treatment was on the extraction efficiency and the maintenance of the original concentration profile of the vitamers. Several acid extraction procedures were tested for this purpose. Perchloric acid was chosen as the extraction agent. Routine food analysis was then performed using dilute ice-cold perchloric acid extraction followed by an internally standardized ion-paired reversed-phase liquid chromatography. Food samples were hydrolyzed with takadiastase and alkaline phosphatase enzymes, phosphorylated and glycosylated vitamers were quantitated before and after the enzymatic digestion. This procedure enabled the extraction of vitamin B6 compounds in their intact forms, and the measurement of free, phosphorylated and glycosylated forms. The maintenance of the concentration profile of the vitamers was verified by using 14C -labeled pyridoxal 5'-phosphate in the examination of the extraction procedure. The extraction efficiency and laboratory performance were confirmed by interlaboratory studies. Up-to-date data on vitamin B6 content of about fifty common food items was produced. The data includes the results from meat and poultry, fish and fish product, dairy product, cereal and vegetable, and ready-to-eat food samples. Free and phosphorylated vitamin B6 compounds were measured in all food groups, and the glycosylated vitamer fraction was analyzed in all plant-derived foods. The results obtained in this work showed that vitamin B6 content of nearly all foods of plant origin was mainly comprised of glycosidically bound pyridoxine derivatives. These bound analytes are normally not taken into account in traditional analytical methods, and food composition tables lack the data of glycosylated pyridoxine. The role of the glycosylated pyridoxines need to be clarified in terms of their analytical and physiological nature. If, as it is currently assumed, the availability of the bound forms is limited for humans, the role of vegetables, cereals and other foods of plant origin as a source of vitamin B6, as well as the analytical methods should be reassessed.;

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International Perspectives in Vitamin Analysis and Legislation in Vitamin Fortification

Rychlik

2011

Fortified Foods With Vitamins

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HPLC determination of vitamin B6 in foods after pre-column derivatization of free and phosphorylated vitamers into pyridoxol

Cited by 25 publications

References 11 publications

Development of a validated HPLC method for the determination of B‐complex vitamins in pharmaceuticals and biological fluids after solid phase extraction

Development of a validated HPLC method for the determination of B‐complex vitamins in pharmaceuticals and biological fluids after solid phase extraction

HPLC analysis of vitamin B6 in foods

International Perspectives in Vitamin Analysis and Legislation in Vitamin Fortification

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