Simultaneous Determination Of Water-soluble Vitamins Research Articles

Fast and Efficient RP-HPLC Method for Simultaneous Determination of Water-Soluble Vitamins in Some Nutraceutical Supplements

The purpose of this study is the development of analytical method for simultaneous determination of water-soluble vitamins (B1, B2, B3, B6, B9, B12 and vitamin C) in dietary supplement syrup, multivitamins and mineral dietary supplement syrup with iron, dietary supplement capsules and balanced supplemental nutrition for children by using gradient RP-HPLC methods with PDA detector. The separation of water-soluble vitamins was performed on waters Spherisorb ODS2 (250 mm×4.6mm, 5μm) column. The wavelength was 272 nm at room temperature and the detection limits ranged from 2.4 to 8.3 ng/µl. The average recovery was 98%-102 % and correlation coefficient (r) was 0.9992 to 1.

The Simultaneous Determination of Some Water-Soluble Vitamins in Gum of Acacia nilotica by High Performance Liquid Chromatography

A rapid, simple and precise method by HPLC (high performance liquid chromatography) has been developed for simultaneous determination of water-soluble vitamins as thiamine(B 1), nicotinamide(B 3), panthotenic(B 5), pyridoxine(B 6) and biotin(B 8) in gum of Acacia nilotica using enzymatic hydrolysis. The method uses a C 18 column (4.6×150 mm, 5µm). Mobile phase such as methanol 0.1M, sodium dihydrogen phosphate (pH = 2.5), (10:90 v/v) is found most suitable for rapid separation and identification of this water –soluble vitamins. Good linearity was observed between the concentration of analytes and peak area (r = 0.9999). Each vitamin was quantitatively determined at its maximum wavelength. Recovery percentages ranged from 97% to 99%.

Simultaneous detection of water-soluble vitamins using the High Performance Liquid Chromatography (HPLC) - a review

The water-soluble vitamins (WSV): ascorbic acid (vitamin C), thiamine (B1), riboflavin (B2), niacin (B3), panthothenic acid (B5), pyridoxine, and pyridoxal (B6), folic acid (B9), biotin(B8) , and B12 are very essential in the diet of humankind. As a result of ever increasing pressures from both consumers and legal enforcers, to specify accurately nutritive compositions of WSV that are present in food materials, many researchers have attempted to fill this niche through the provision of highly sensitive and rapid high performance liquid chromatography (HPLC) procedures. In view of the health benefits of WSV, a replete of HPLC methods have been developed for simultaneous determination of their contents in nature and fortified food samples, nutritional supplements, as well as blood plasmas. The rate of losses of these vitamins during food processing and analysis, in addition to their transient dynamics, presents complexities in developing a highly sensitive HPLC procedure for their simultaneous separations and assays. This review critically assesses the different HPLC procedures developed by researchers and available in the open literature for simultaneous determination of water-soluble vitamins (WSV) in dried tropical fruits materials. The study revealed that not a single chromatographic run developed by researchers can simultaneously elute all the WSV at a time. However, the HPLC procedures that are capable of determining all the WSV were coupled with electrospray ionization mass spectroscopy (ESI-MS), thus making the set-up expensive.

Simultaneous determination of water-soluble vitamins in beverages and dietary supplements by LC-MS/MS

An LC-MS/MS method was developed for the simultaneous determination of 15 water-soluble vitamins that are widely used as additives in beverages and dietary supplements. This combined method involves the following simple pre-treatment procedures: dietary supplement samples were prepared by centrifugation and filtration after an extraction step, whereas beverage samples were diluted prior to injection. Chromatographic analysis in this method utilised a multi-mode ODS column, which provided reverse-phase, anion- and cation-exchange capacities, and therefore improved the retention of highly polar analytes such as water-soluble vitamins. Additionally, the multi-mode ODS column did not require adding ion pair reagents to the mobile phase. We optimised the chromatographic separation of 15 water-soluble vitamins by adjusting the mobile phase pH and the organic solvent. We also conducted an analysis of a NIST Standard Reference Material (SRM 3280 Multi-vitamin/Multi-element tablets) using this method to verify its accuracy. In addition, the method was applied to identify the vitamins in commercial beverages and dietary supplements. By comparing results with the label values and results obtained by official methods, it was concluded that the method could be used for quality control and to compose nutrition labels for vitamin-enriched products.

Design of Natural Food Antioxidant Ingredients through a Chemometric Approach

In the present work, an environmentally friendly extraction process using subcritical conditions has been tested to obtain potential natural food ingredients from natural sources such as plants, fruits, spirulina, propolis, and tuber, with the scope of substituting synthetic antioxidants, which are subject to regulation restrictions and might be harmful for human health. A full characterization has been undertaken from the chemical and biochemical point of view to be able to understand their mechanism of action. Thus, an analytical method for profiling the compounds responsible for the antioxidant activity has been used, allowing the simultaneous determination of water-soluble vitamins, fat-soluble vitamins, phenolic compounds, carotenoids, and chlorophylls in a single run. This information has been integrated and analyzed using a chemometrical approach to correlate the bioactive compounds profile with the antioxidant activity and thus to be able to predict antioxidant activities of complex formulations. As a further step, a simplex centroid mixture design has been tested to find the optimal formulation and to calculate the effect of the interaction among individual extracts in the mixture.

Simultaneous determination of water-soluble vitamins in SRM 1849 Infant/Adult Nutritional Formula powder by liquid chromatography–isotope dilution mass spectrometry

Assessing dietary intake of vitamins from all sources, including foods, dietary supplements, and fortified foods, would be aided considerably by having analytical methodologies that are capable of simultaneous determination of several vitamins. Vitamins naturally present in foods may occur in different chemical forms, with levels ranging over several orders of magnitude. Vitamins in dietary supplements and fortified foods, however, are typically added in a single chemical form, and matrix issues are usually not as complex. These sources should thus be relatively amenable to approaches that aim for simultaneous determination of multiple vitamins. Our recent work has focused on development of liquid chromatography (LC)-UV/fluorescence and LC-tandem mass spectrometry methods for the simultaneous determination of water-soluble vitamins (thiamine, niacin, pyridoxine, pantothenic acid, folic acid, biotin, and riboflavin) in dietary supplement tablets and fortified foods, such as formula powders and breakfast cereals. As part of the validation of our methods and collaboration in characterization of a new NIST SRM 1849 Infant/Adult Nutritional Formula powder, we report data on SRM 1849 using isotope dilution mass spectrometric methods. Use of available NIST Standard Reference Materials(R) as test matrices in our method development and validation gives a benchmark for future application of these methods. We compare three chromatographic approaches and provide data on stability of vitamin standard solutions for LC-based multiple vitamin determinations.

LC/UV/MS-MRM for the simultaneous determination of water-soluble vitamins in multi-vitamin dietary supplements

The purpose of this study was to optimize chromatographic and detection conditions for the simultaneous determination of water-soluble vitamins in multi-vitamin dietary supplements using a single chromatographic run. An approach using liquid chromatography with diode array and/or mass spectrometry for quantitation of seven B-complex vitamins [thiamine (B(1)), riboflavin (B(2)), nicotinamide (B(3)), pyridoxine (B(6)) pyridoxine, biotin, pantothenic acid, and folic acid] in multi-vitamin/multi-mineral daily supplements is described. This approach utilizes a reversed phase C18 column (4 mum; i.d.: 250x2.0 mm) with a gradient mobile elution profile, performed at a flow rate of 0.25 ml/min. After a 5-min isocratic elution at 100% A (0.1% formic acid in water), a linear gradient to 50% A and 50% B (0.1% formic acid in acetonitrile) at 15 min and then to 5% A and 95% B at 17 min was employed. Detection was performed with a photodiode array detector (DAD) in sequence with a triple-quad mass spectrometer in the multiple reaction mode (MS-MRM). Although good chromatographic separation of ascorbic acid was also obtained in extracts from multi-vitamin/multi-mineral supplements, the ascorbic acid could not be quantified properly due to rapid oxidation catalyzed by the minerals. This method was initially applied to determine water-soluble vitamins in representative multi-vitamin/multi-mineral tablets following the extraction of ground samples with a phosphate buffer (10 mM, pH 2.5). For multi-vitamin supplement tablets, this approach does not require any sample clean-up/pre-concentration steps except for centrifugation and filtration of the extract.

Simultaneous determination of water-soluble vitamins in a vitamin-enriched drink by an in-capillary enzyme reaction method

The in-capillary enzyme reaction method was used to determine riboflavin phosphate in a vitamin-enriched drink based on its conversion to riboflavin (vitamin B 2) with alkaline phosphatase. Simultaneously, three water-soluble vitamins [thiamine nitrate (vitamin B 1 mononitrate), pyridoxine hydrochloride (vitamin B 6 hydrochloride) and nicotinamide (vitamin PP)] and anhydrous caffeine in the drink were subjected to quantitative analysis. In the system, electrophoretic migration was used to mix zones containing the substrate (riboflavin phosphate) and the enzyme (alkaline phosphatase). The reaction was then allowed to proceed in the presence of a weak electric field and, finally, the product (riboflavin) of enzyme reaction and other water-soluble vitamins migrated under the influence of an applied electric field to the detector. All the active ingredients and the formulation excipients were successfully separated by micellar electrokinetic chromatography with 135 m M sodium dodecyl sulfate. To prevent inhibition of enzyme reaction by the addition of sodium dodecyl sulfate to the reaction zone, sandwich mode injection, in which plugs of sandwich solution without sodium dodecyl sulfate were introduced into the capillary on both sides of the reaction zone, was utilized as a barrier to protect the enzyme reaction from the inhibitor. The relationship between the peak area of the product and the concentration of the substrate was calculated in the in-capillary enzyme reaction method. Excellent linearity was obtained, with correlation coefficients of 0.9999. The established method was validated and demonstrated to be applicable to the determination of the five active ingredients, including riboflavin phosphate, in a commercial vitamin-enriched drink. No interference from the formulation excipients was observed. Good linearities were obtained, with correlation coefficients above 0.999. Recoveries and precisions ranged from 99.3 to 101.8%, and from 0.1 to 2.5% RSD, respectively. Good agreement was obtained between the established method and traditional high-performance liquid chromatographic methods. These results suggest that the in-capillary enzyme reaction method can be used for the simultaneous determination of riboflavin phosphate and other water-soluble vitamins in pharmaceuticals.

Simultaneous determination of water-soluble vitamins excreted in human urine after eating an overdose of vitamin pills by a HPLC method coupled with a solid phase extraction

We have applied a quick and convenient method for determining water-soluble vitamins excreted in human urine. We found that the Sep-Pak C 18 cartridge was useful for preconcentration and recovery of water-soluble vitamins in urine with minimized loss of vitamins. The recovery of vitamins was well over 90%. The separation was carried out by gradient elution with 90/10 (v/v%) methanol/water with 0.1% trifluoroacetic acid (TFA) and water with 0.1% TFA on a μBondapak C 18 column. The separation was completed within 15 min. We measured concentrations of water-soluble vitamins excreted in urine after swallowing an overdose of vitamin pills on purpose, and found that the concentration of each vitamin increased rapidly to the maximum in 2–3 h and decreased swiftly.

Simultaneous Determination of Water-Soluble Vitamins by Over-Pressure Layer Chromatography and Photodensitometric Detection

An over-pressure layer chromatographic procedure with photodensitometric detection for the simultaneous determination of water-soluble vitamins in multivitamin pharmaceutical preparations was developed and evaluated. The method uses high-performance TLC (HPTLC) plates with silica gel as the thin-layer, and an n-butanol:pyridine:water mixture (50:35:15, v/v/v) as mobile phase at a rate of 0.25 mL/min for baseline separation. The quantitation was carried out without derivatization (vitamin B1, vitamin B2, vitamin B6, folic acid, nicotinamide, vitamin C) or after spraying ninhydrin reagent (calcium pantothenate) or 4-dimethylaminocinnamaldehyde (vitamin B12, biotin). This method was applied to the analysis of multivitamin solutions. Satisfactory relative standard deviations and good recovery were obtained for all the vitamins examined. It was concluded that this method is fast, accurate, specific, and suitable for routine quality control use.