Conjugation of sodium caseinate and gum arabic catalyzed by transglutaminase

Conjugation of the milk protein sodium caseinate and a protein-containing polysaccharide, gum arabic, was achieved through the use of the cross-linking enzyme transglutaminase. The extent of conjugation was monitored by size exclusion separation coupled with a multiangle laser light scattering detector. The elution times of gum arabic solutions incubated with transglutaminase were unchanged over time, whereas incubation of sodium caseinate with transglutaminase resulted in shorter elution times as reaction time increased, indicating the formation of cross-linked caseinate polymers. However, when mixtures of caseinate and gum arabic were incubated with transglutaminase, the elution times were decreased markedly, indicating conjugation between the protein and polysaccharide. The molecular masses of the conjugates increased from approximately 950 to 1600 kDa. This method of protein-polysaccharide conjugation offers noticeable advantages over previously used methods, and the conjugates produced may exhibit unique functional properties.     

Effect of partial replacement of gum arabic with carbohydrates on its microencapsulation properties.

Gum arabic solutions (10% w/v) were emulsified with soy oil at oil/gum ratios of 0.25-5.0. At oil/gum ratios <1.0, it was established that gum arabic could be partially replaced with a nonsurfactant carbohydrate. To assess different carbohydrates as replacers for gum arabic, emulsions and spray-dried emulsions of soy oil and mixed solutions (10% w/v) of gum arabic and a range of carbohydrate wall materials (oil/gum = 0.5) were prepared and analyzed. Maize starch and glucose were ineffective as partial replacers of gum arabic, but maltodextrins of various dextrose equivalence values (5.5-38) successfully replaced 50% of the gum arabic. The microencapsulation efficiency of the gum arabic/maltodextrin stabilized powders was further increased by increasing total solids of the feed to the dryer and by increasing the atomizer nozzle diameter.     

The development of immunoassays to identify and quantify species source of gum arabic

Gum arabic from Acacia senegal is commonly used as an additive in foodstuffs. Adulteration of gum arabic by other gums is a potential problem for reasons of safety and quality. This study aimed to develop and evaluate the use of enzyme-linked immunosorbent assays (ELISAs) for the detection of potential adulterants of gum arabic. Indirect competitive ELISAs (IC-ELISAs) were developed using the monoclonal antibodies SY CC7 (A. senegal), SY HH3 (Acacia seyal), and SY J1A1 (Combretum erythrophyllum). All IC-ELISAs had a working range of 0.005-10 mg/mL. The antibodies used were tested using the IC-ELISAs for cross-reactivity with other Acacia species and other gums. The antibodies were very specific for their respective antigens. Significant cross-reactivity was found for SY CC7 (between A. senegal and A. melliferae) and SY J1A1 (between C. erythrophyllum and A. seyal). The IC-ELISA was adapted further to test confectionery samples for the presence of gum arabic, which was successful, although recovery rates were reduced. Both IC- and plate trapped antigen ELISA (PTA-ELISA) formats were able to distinguish an adulterated sample of gum arabic when blended with either A. seyal or C. erythrophyllum. The PTA-ELISA was more sensitive for A. seyal than the IC-ELISA, but both were equally sensitive for C. erythrophyllum. The results suggest that the antibodies SY CC7, SY HH3, and SY J1A1 could be used in combination with each other for the detection of potential adulterants of A. senegal and the detection of gum arabic in foodstuffs.     

Rotational and translational mobility of small molecules in sucrose plus polysaccharide solutions

The effect of different polysaccharides on the rotational (D(rot)) and translational diffusion (D(trans)) coefficients of small molecules in concentrated systems (sucrose solutions) was investigated. Dextran (1 or 10% w/w) with different molecular masses (from 10(4) to 2 x 10(6) Da), gum arabic, or pullulan was added to solutions of sucrose (57.5% w/w). Viscosity measurements of the diffusion medium studied (sucrose and sucrose plus polysaccharide) were made using a Rheometric Scientific viscometer in a temperature range from 20 to -10 degrees C. The rotational mobility of nitroxide radicals (Tempol) dispersed in the concentrated systems was measured by electron spin resonance. The translational diffusion coefficient of fluorescein was determined by the fluorescence recovery after photobleaching method. The studied temperature range for the latter two techniques was from 20 to -16 degrees C. For these conditions of concentration and temperature, there was no ice formation in the samples. No effect of the molecular mass of dextran on D(rot) and D(trans) was observed when solutions with the same dry matter content were compared. Only pullulan and gum arabic, at 10%, had a significant effect on D(trans)( )()of fluorescein. Temperature and total dry matter content were observed to be the most important factors controlling D(rot) and D(trans) in these concentrated systems.     

Influence of carbohydrates on the interaction of procyanidin B3 with trypsin

The biological properties of procyanidins, in particular their inhibition of digestive enzymes, have received much attention in the past few years. Dietary carbohydrates are an environmental factor that is known to affect the interaction of procyanidins with proteins. This work aimed at understanding the effect of ionic food carbohydrates (polygalacturonic acid, arabic gum, pectin, and xanthan gum) on the interaction between procyanidins and trypsin. Physical-chemical techniques such as saturation transfer difference-NMR (STD-NMR) spectroscopy, fluorescence quenching, and nephelometry were used to evaluate the interaction process. Using STD-NMR, it was possible to identify the binding of procyanidin B3 to trypsin. The tested carbohydrates prevented the association of procyanidin B3 and trypsin by a competition mechanism in which the ionic character of carbohydrates and their ability to encapsulate procyanidins seem crucial leading to a reduction in STD signal and light scattering and to a recovery of the proteins intrinsic fluorescence. On the basis of these results, it was possible to grade the carbohydrates in their aggregation inhibition ability: XG > PA > AG ? PC. These effects may be relevant since the coingestion of procyanidins and ionic carbohydrates are frequent and furthermore since these might negatively affect the antinutritional properties ascribed to procyanidins in the past.     

Photosensitizing effect of riboflavin, lumiflavin, and lumichrome on the generation of volatiles in soy milk

Lumichrome and lumiflavin were formed from riboflavin under light. pH had a significant influence on the formation of lumichrome and lumiflavin from riboflavin. Lumichrome was the only major product from riboflavin under neutral or acidic pH values. Lumiflavin was also formed from riboflavin in basic pH. The maximum concentration of lumiflavin from 100 microM riboflavin at pH 8.5 was 30.9 microM, and it was reached after 2 h of exposure at 1500 lux. The maximum concentration of lumichrome formed from 100 microM riboflavin at pH 4.5, 6.5, or 8.5 was 79.9, 58.7, and 73.1 microM, respectively, after 8, 6, or 2 h of light exposure. The formation of lumichrome and lumiflavin from riboflavin was due to the type I mechanism of the riboflavin photosensitized reaction. Singlet oxygen was also involved in the photosensitized degradation of lumiflavin and lumichrome. The reaction rates of riboflavin, lumiflavin, and lumichrome with singlet oxygen were 9.66 x 10(8), 8.58 x 10(8), and 8.21 x 10(8) M(-1) s(-1), respectively. The headspace oxygen depletion and headspace volatile formation were significant in soy milk containing lumichrome or lumiflavin under light (p < 0.05) and were insignificant (p > 0.05) in the dark. Ascorbic acid could inhibit the total volatile changes of soy milk under light. Soy milk should be protected from light to prevent the photodegradation of riboflavin and the oxidation of soy milk.     

Effect of Aqueous Impregnation of Rice Kernels with Gum Arabic and Xanthan on Storage Stability of Frozen Rice Cakes

Rice kernels were steeped (10°C, 5 h) in an aqueous solution containing gum arabic (0.36%) and xanthan (0.24%) and then drained, wet‐milled, and steamed to prepare rice cakes. The cakes were then frozen (–40°C for 50 min). The effect of the gum addition on the textural properties of the cakes during storage for 46 h at 25°C after being thawed was examined. Using the combination of gum arabic and xanthan mitigated the quality deterioration of rice cakes such as aggregation of rice flour, which had been induced by xanthan alone. Also, the increase in hardness during storage was substantially reduced by soaking rice kernels in the gum arabic/xanthan solution. Overall results revealed that the steeping in the gum solution improved the storage stability of rice cakes.     

New insights into the structural characteristics of the arabinogalactan-protein (AGP) fraction of gum arabic

The structural characteristics of the gum exudate of Acacia senegal (gum arabic) have been investigated by monitoring the composition and physicochemical properties before and after treatment with proteolytic enzyme and various alkaline systems. Molecular mass ( M w) and radius of gyration ( R g) measurements were performed using gel permeation chromatography (GPC) coupled to refractive index, UV absorbance, and multiangle light scattering detectors and indicated that the macromolecules present have a compact structure. It was found that treatment with proteolytic enzyme caused the arabinogalactan-protein component (AGP) with average molecular mass approximately 2 x 10 (6) Da to degrade, yielding material of molecular mass approximately 4 x 10 (5) Da, whereas the bulk of the material corresponding to the protein-deficient arabinogalactan component (AG) with molecular mass 4 x 10 (5) remained unaffected. Barium hydroxide was found to hydrolyze the polysaccharide component (AG) itself in addition to the proteinaceous component as demonstrated in control experiments using dextran. However, sodium borohydride/sodium hydroxide treatments were unable to hydrolyze dextran and were assumed to hydrolyze only the proteinaceous component of gum arabic. The AGP component was completely degraded, yielding material of molecular mass approximately 4.5 x 10 (4) Da. It has been concluded, therefore, that the enzyme did not fully hydrolyze all of the protein present and that the AGP component of gum arabic consists of carbohydrate blocks of approximately 4.5 x 10 (4) Da linked to a polypeptide chain consistent with the wattle blossom structure. Because the AGP was degraded to differing extents using a mild and more severe sodium borohydride/sodium hydroxide treatment, it was concluded that the polysaccharide moieties were linked through both O-serine and O-hydroxyproline residues. The gum arabic sample was deglycosylated by treatment with anhydrous hydrogen fluoride and revealed the presence of two putative core proteins of approximately 3 x 10 (4) and approximately 5 x 10 (3) Da, respectively, which correspond to proteins of approximately 250 and 45 amino acids in length. A new model for the structure of the AGP component has been proposed.     

Origin and thermodynamic properties of the instability of synthetic azo colorants in gum arabic solutions

The instability of some industrially important synthetic azo colorants, including sunset yellow, azorubine, and allura red, toward gum arabic in aqueous solution has been a long-standing problem for the beverage and confectionery industries. Precipitation of these colorants causes the deterioration of product appearance and properties. This work examines the origin and nature of the problem by analysis of the precipitate and thermodynamic studies of gum arabic-colorant interactions using isothermal titration calorimetry (ITC). The presence of divalent alkaline earth metals in gum arabic samples, that is, calcium and magnesium, is shown to be responsible for the precipitation of the azo colorants. There is no direct interaction between gum arabic and the colorant molecules, and the precipitate is formed likely due to the mediation/bridging by the divalent cations. The thermodynamic knowledge gained from the ITC studies, for example, binding affinity, stoichiometry, and enthalpy, enables interpretation of many industrial observations.     

Effect of Hydrophilic Gums on Frozen Dough. I. Dough Quality

Disadvantages of frozen doughs are their variable performance and loss of stability over long‐term frozen storage. Changes in rheological properties of frozen doughs have been reported to be due to the physical damage of the gluten network caused by ice crystallization and recrystallization. The objective of this study was to determine the effect of hydrophilic gums on ice crystallization and recrystallization for improvement of the shelf‐life stability of frozen dough. The present research involved use of the Hard Red Spring wheat cultivar Grandin and hydrophilic gums such as carboxymethyl cellulose (CMC), gum arabic, kappa carrageenan (κ‐carrageenan), and locust bean gum at three different levels each on doughs stored frozen for up to 16 weeks. The dough characteristics were analyzed after day 0, day 1, and after 4, 8, 12, and 16 weeks of frozen storage using data from differential scanning calorimetry (DSC), water activity, extensigraph, and proof time. The ΔH value of freezable water endothermic transitions obtained using DSC increased with storage time for all treatments. However, addition of different levels of the four gums lowered the ΔH value, indicating a decrease in freezable water. Doughs with locust bean gum gave a higher peak force, measured using the Kieffer dough extensibility rig of the texture analyzer, and lower proof time, indicating better retention of baking quality. Maximum resistance to extension increased upon addition of 1 and 3%; CMC; 1 and 3%; κ‐carrageenan; and 1, 2, and 3% locust bean gum as compared with the control. The various periods of storage or gum treatments did not affect the water activity of the thawed frozen doughs. Doughs with locust bean gum gave significantly lower proof time compared with the other treatments and the control. CMC gave the second lowest values, followed by gum arabic treatment. Addition of κ‐carrageenan increased the proof time compared with the control. In summary, locust bean gum, gum arabic, and CMC improved the dough characteristics to varying degrees. κ‐Carrageenan was the only gum that showed a detrimental effect on frozen dough.     

Effect of Hydrophilic Gums on the Quality of Frozen Dough: Electron Microscopy,Protein Solubility,and Electrophoresis Studies

Hydrophilic gums have been shown to improve the shelf‐life stability of frozen doughs during long periods of frozen storage. The objective of this research was to determine the effect of gums on starch and protein characteristics of frozen doughs using electron microscopy and electrophoresis studies. Frozen doughs, supplemented with three levels of gum arabic, carboxy methyl cellulose (CMC), kappa (κ) carrageenan, and locust bean gum, were studied after day 1 and after 4, 8, 12, and 16 weeks of frozen storage. Changes in the ultra structure of the frozen doughs were investigated, as well as the solubilities and composition of dough proteins by SDS‐PAGE. Scanning electron micrographs of doughs evaluated on day 0 (unfrozen) showed starch granules securely embedded in the gluten matrix. However, after 8 and 16 weeks of frozen storage, the frozen control dough without the gum additives clearly showed damage to the gluten network, and the starch granules appeared to be separated from the gluten. Doughs with locust bean gum and gum arabic showed better retention of the gluten network compared with the frozen control evaluated after different periods of storage. The SDS‐soluble protein content increased while residue protein content decreased as the frozen storage time increased. After each frozen storage period, the control dough without the gum additive had the highest amount of SDS‐soluble proteins and the lowest amount of residue proteins when compared with the doughs treated with gums. κ‐Carrageenan and locust bean gum had the lowest amount of SDS‐soluble proteins compared with doughs with CMC and gum arabic. The frozen control had the lowest amount of residue proteins at any particular time of frozen storage. κ‐Carrageenan treated doughs had the highest amount of residue proteins, followed by doughs with locust bean gum. Doughs with gum arabic and CMC had the lowest amount of residue proteins but still higher than the control doughs.     

Liquid chromatographic determination of vitamins D2 and D3 in fortified milk and infant formulas

A liquid chromatographic (LC) method was developed for determining vitamins D2 and D3 in fortified milk and infant formulas. The lipid-soluble components were extracted from the aqueous phase by homogenizing in isopropanol-methylene chloride with magnesium sulfate added to remove water. The vitamins were fractionated from the lipid material by using gel permeation chromatography (GPC) followed by further cleanup of the combined GPC fractions on a muBondapak/NH2 column. Four muStyragel (100 A) columns connected in series were used for GPC fractionation of sample extracts in methylene chloride. Injection and collection were repeated 3 times to collect enough vitamin D for quantitation. The muBondapak/NH2 column, using a mobile phase of methylene chloride-isooctane-isopropanol (600 + 400 + 1), resolved vitamin D from other UV-absorbing compounds and soy sterols in infant formula and from cholesterol in milk. Vitamins D2 and D3 coeluted as one peak, with the resolution and vitamin level sufficient for visual monitoring (280 nm/0.02 absorbance unit full scale) in a collection time of 22-26 min. A Zorbax ODS (6 micron) column and a methylene chloride-acetonitrile-methanol (300 + 700 + 2) mobile phase were used for LC quantitation; vitamins D2 and D3 were baseline resolved in about 11 min. The infant formula samples included ready-to-use and concentrated liquids prepared in nonfat milk base or soy base fortified with vitamins D2 or D3 at 400 IU/qt or L (10 micrograms). The mean percent recovery of added vitamin D3 (400-500 IU/qt) from infant formula (n = 7) was 89.6 +/- 6.7 (coefficient of variation (CV) 7.5%).(ABSTRACT TRUNCATED AT 250 WORDS)     

Relevance of isotopic and molecular biomarkers for the authentication of milk according to production zone and type of feeding of the cow

The first objective of the present paper was to assess the potential of both isotopic ( (18)O/ (16)O in milk water) and molecular biomarkers (terpenes, fatty acids, carotenoids, and vitamins) and milk color to discriminate the production zone (lowland or upland areas) from which 49 tanker bulk milks were collected over one year from a total of 204 farms. The milk water (18)O enrichment was higher in lowland (<500 m altitude) than in upland (>700 m altitude), but the delta (18)O values failed to discriminate systematically the production zone at the scale of the year because of its high variability related to the sampling period. In contrast with vitamins A and E, carotenoids, and milk color measurements, terpenes and fatty acids were confirmed to be relevant tracers of the production zone. The milk compounds with the strongest discriminative potential were fatty acids, which were determined by high-resolution gas chromatography. The calculation of fatty acid ratios, which permits the limitation of using fatty acid relative quantity expressed in percentage of total fatty acids to be overcome, was shown to be particularly relevant in discriminating upland from lowland milk ratios. The selection of two pairs of ratios, namely, iso-C17:0/C18:3 n-3 and iso-C15:0/iso-C14:0, enabled the authentication of 100% of the highland versus lowland milks whatever the season. The second objective was to evaluate the relevance of fatty acid composition to discriminate milks according to the proportion of corn silage in the diets of dairy cows. The selection of two fatty acids ratios, namely, trans11 cis15-C18:2/trans11-C18:1 and cis9-C16:1/iso-C16:0, enabled the correct classification of 100% of the milk samples according to the proportion of corn silage in the basic fodder rations (<25% vs >30%). The relationship between the milk production zone and the type of forage fed to the cows is discussed.     

Comparison of Photocatalytic and Photosensitized Oxidation of Paraben Aqueous Solutions Under Sunlight

It is well-established that aquatic wildlife is exposed to natural and synthetic endocrine disrupting compounds which are able to interfere with the hormonal system. Although advanced oxidation processes (AOPs) have shown to be effective, their application is limited by a relatively high operational cost. In order to reduce the cost of energy consumed in the AOPs, widely available solar energy instead of UV light may be applied either as photocatalytic oxidation or as photosensitized oxidation. The main goal of the present study was to investigate the sunlight photodegradation of paraben mixture. Two processes, namely the photocatalytic oxidation with modified TiO₂ nanoparticles and photosensitized oxidation with photosensitive chitosan beads, were applied. The oxidants were identified as singlet oxygen and hydroxyl radicals for photosensitized and photocatalytic oxidation, respectively. The toxicity, as well as ability to water disinfection of both processes under natural sunlight, has been investigated. Application of sunlight for the processes led to degradation of parabens. The efficiency of both processes was comparable. Despite the fact that singlet oxygen is weaker oxidant than hydroxyl radicals, the photosensitized oxidation seems to be more promising for wastewater purification, due to the possibility of chitosan bead reuse and more effective water disinfection.

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Graphical Abstract ?

     

Atrazine photodegradation in aqueous solution induced by interaction of humic acids and iron: photoformation of iron(II) and hydrogen peroxide

The photochemical formation of Fe(II) and hydrogen peroxide (H 2O 2) coupled with humic acids (HA) was studied to understand the significance of iron cycling in the photodegradation of atrazine under simulated sunlight. The presence of HA significantly enhanced the formation of Fe(II) and H 2O 2, and their subsequent product, hydroxyl radical ( (*)OH), was the main oxidant responsible for the atrazine photodegradation. During 60 h of irradiation, the fraction of iron presented as Fe(II) (Fe(II)/Fe(t)) decreased from 20-32% in the presence of the Fe(III)-HA complex to 10-22% after adding atrazine. The rate of atrazine photodegradation in solutions containing Fe(III) increased with increasing HA concentration, suggesting that the complexation of Fe(III) with HA accelerated the Fe(III)/Fe(II) cycling. Using fluorescence spectrometry, the quenching constant and the percentage of fluorophores participating in the complexation of HA with Fe(III) were estimated by the modified Stern-Volmer equation. Fourier transform infrared spectroscopy (FTIR) offered the direct evidence that Fe(III)-carboxylate complex could be formed by ligand exchange of HA with Fe(III). Based on all the information, a possible reaction mechanism was proposed.     

Antioxidant activity of A-type proanthocyanidins from Geranium niveum (Geraniaceae)

Geranium niveum S. Watson (Geraniaceae) is a medicinal herb widely used by the Tarahumara Indians of Mexico. This species is rich in proanthocyanidins and other phenolics. Previous in vitro assays have demonstrated that proanthocyanidins exhibited antiinflammatory, antiviral, antibacterial, enzyme-inhibiting, antioxidant, and radical-scavenging properties. In view of its medicinal use and chemical composition, the aim of the present study was to determine the in vitro antioxidant activity of the extracts and two proanthocyanidins (geranins A and D) from the roots of G. niveum by using seven different assay systems, namely, 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), 1,1-diphenyl-2-picrylhydrazyl (DPPH), superoxide anion (O2*-), hydrogen peroxide (H2O2), hydroxyl radical (OH*), hypochlorous acid (HOCl), and singlet oxygen ((1)O2). Two known antioxidants, resveratrol and ascorbic acid, were used as positive controls. The results showed that geranins A and D and the extracts were able to scavenge ABTS, DPPH, O2*-, OH*, and HOCl. The scavenging ability of geranins A and D was similar to that of resveratrol and ascorbic acid in the following assays: ABTS, O2*-, and HOCl. The scavenging capacity of ascorbic acid for DPPH was higher than that of both geranins and resveratrol. On the other hand, the OH* scavenging action of both geranins and resveratrol was similar. The methanol-CHCl3 (1:1) extract had a higher ability to scavenge ABTS, DPPH, and O2*- radicals than the chloroform extract. In turn, the latter was more potent than the methanol-CHCl3 (1:1) extract as OH* or HOCl scavenger agent. Neither geranins A and D nor the extracts were able to scavenge H2O2 and (1)O2. In conclusion, G. niveum roots have proanthocyanidins with powerful radical scavenging in vitro activity. This property may partially explain the wide use of this plant in the Tarahumara indigenous system of medicine for the treatment of gastrointestinal illnesses (other than spasms), pain, and fevers associated with oxidative stress.     

Liquid chromatographic methods for vitamins A and D in multivitamin-mineral formulations

Liquid chromatographic screening procedures have been developed for the estimation of vitamins A and D in multivitamin-mineral tablet, capsule, gelatin capsule, and syrup formulations. The procedure can be used for measuring vitamin A present as either retinyl acetate or retinyl palmitate, and also for measuring the contribution to total vitamin A activity from 13-cis retinyl esters. The retinyl esters and their isomers are resolved from each other and their oxidation products. Ergocalciferol and cholecalciferol are not resolved from each other but they are resolved from other vitamin D isomers and from vitamins A, E, and K and their degradation products. Both assays use a 3 microns amino column, with a mobile phase of hexane for vitamin A and 1% isopropanol in hexane for vitamin D. The precision of replicate injections for vitamins A and D is better than 1% and the recovery from spiked syrups is better than 98%. The coefficient of variation for both assay methods is about 5%. Twenty formulations were analyzed for vitamin A and 24 were analyzed for vitamin D.     

Liquid chromatographic determination of vitamin D in fortified milk

A method for the determination of vitamins D2 + D3 in fortified milk is described. Vitamins D2 and D3 are extracted from the saponified sample and converted to isotachysterols with antimony trichloride. The isotachysterols are quantitated using liquid chromatography with ultraviolet detection at 301 nm, which is the absorption maximum. At this wavelength other materials present in the sample do not interfere with the analysis of isotachysterols and therefore a cleanup step is avoided. Recoveries of vitamin D added to skim milk were 98.1% (SD 5.3), 96.7% (SD 3.3), and 96.0% (SD 5.1) for samples fortified with 200, 400, and 600 IU/quart, respectively. For whole milk, recoveries were 102.0% (SD 6.5) and 97.1% (SD 3.5) in samples fortified with vitamin D equivalent to 200 and 400 IU/quart, respectively. The detection limit for vitamin D is 40 IU/quart.     

Screening method for vitamins A and D in fortified skim milk, chocolate milk, and vitamin D liquid concentrates

Vitamin A was determined in fortified chocolate milk and skim milk; vitamin D was determined in fortified chocolate milk, skim milk, and vitamin D concentrates, using reverse phase high pressure liquid chromatography (HPLC). The sample is saponified, extracted with hexane, and chromatographed in an HPLC system on a 10 micron Vydac TP reverse phase C18 column, using acetonitrile-methanol (9+1) as the mobile phase. For 6 replicates, the recoveries of vitamins A and D, using this procedure, were 99 and 98%, respectively.     

Simultaneous determination of eight water-soluble vitamins in supplemented foods by liquid chromatography

A fast, simple, and reliable method for the isolation and determination of the vitamins thiamin, riboflavin, niacin, pantothenic acid, pyridoxine, folic acid, cyanocobalamin, and ascorbic acid in food samples is proposed. The most relevant advantages of the proposed method are the simultaneous determination of the eight more common vitamins in enriched food products and a reduction of the time required for quantitative extraction, because the method consists merely of the addition of a precipitation solution and centrifugation of the sample. Furthermore, this method saves a substantial amount of reagents as compared with official methods, and minimal sample manipulation is achieved due to the few steps required. The chromatographic separation is carried out on a reverse phase C18 column, and the vitamins are detected at different wavelengths by either fluorescence or UV-visible detection. The proposed method was applied to the determination of water-soluble vitamins in supplemented milk, infant nutrition products, and milk powder certified reference material (CRM 421, BCR) with recoveries ranging from 90 to 100%.