Vitamins In Pharmaceutical Preparations Research Articles

Development of a Stability Indicating Method for Simultaneous Analysis of Five Water-Soluble Vitamins by Liquid Chromatography

Abstract Background Water-soluble vitamins are often included simultaneously in pharmaceutical formulations as food complements or in parenteral nutrition mixtures. Given their sensitivity to heat, light or pH variations, it is important to study their stability using validated stability indicating methods. We thus aimed to validate a liquid chromatography (LC) stability-indicating method for the simultaneous quantification of 5 water-soluble vitamins. Methods We analyzed four water-soluble B vitamins (nicotinamide, pyridoxine, folic acid, cyanocobalamin) and ascorbic acid using a LC method with diode array detector. They were separated on a C18 stationary phase under gradient elution of solvent A [0.2 % of metaphosphoric acid in water and acetonitrile 98:2] and solvent B (100 % acetonitrile). All vitamins were subjected to forced degradation conditions and we showed that the obtained degradation products didn’t interfere with the vitamins. Results The method allows the separation of the 5 water-soluble vitamins in a 30 minute run without any interference from the breakdown products obtained with acid/alkaline solutions, hydrogen peroxide, temperature and light. It meets all the qualitative and quantitative criteria for validation with an acceptable accuracy and good linearity. Conclusions This stability-indicating method can be used for carrying out stability studies of water-soluble vitamins in pharmaceutical preparations.

Simultaneous determination of water- and fat-soluble vitamins in pharmaceutical preparations by high-performance liquid chromatography coupled with diode array detection

Water- and fat-soluble vitamins were separated on a MetaChem Polaris C18-A ( 150 mm×4.6 mm, 3 μm particle size) in a single run using combined isocratic and linear gradient elution with a mobile phase consisting of 0.010% trifluoroacetic acid of pH 3.9 (solvent A) and methanol (solvent B) at the flow rate 0.7 ml min −1. A linear gradient profile (A:B) started at 95:5 and was constant in the first 4 min, then linearly decreased up to 2:98 during the next 6 min, then it was constant in the next 20 min and finally linearly increased up to 95:5 ratio of water phase in the last 5 min of the separation. The most suitable detection wavelength for simultaneous vitamin determination was 280 nm. The method was applied for the solid sample of pharmaceutical preparation (B-Komplex), fortified powdered drinks (multi-vitamin) and food samples. The results were in good agreement with the declared values.

Water-Soluble Vitamins

Water-Soluble Vitamins

Enhanced application of square wave voltammetry with glassy carbon electrode coupled to multivariate calibration tools for the determination of B 6 and B 12 vitamins in pharmaceutical preparations

Vitamins B 6 (VB 6) and B 12 (VB 12) were simultaneously determined in pharmaceutical preparations by using square wave voltametry (SWV) together with artificial neural networks (ANNs). Supporting electrolyte solution, pH and voltametric technique were optimised. The calibration set was built with several artificial samples containing both active ingredients and excipients. Deviations from linearity were observed for both analytes. It is probably due to interactions among the electro active components and competition by the electrode surface, fact that supports the use of ANNs. Recoveries when analysing a nine sample validation set, of 100.2 and 96.4 were calculated for VB 6 and VB 12, respectively. Commercial samples were analysed with reasonably good results considering the complexity of the mixture studied.

A multicommuted flow system for sequential spectrophotometric determination of hydrosoluble vitamins in pharmaceutical preparations

A multicommuted flow system is proposed for spectrophotometric determination of hydrosoluble vitamins (ascorbic acid, thiamine, riboflavine and pyridoxine) in pharmaceutical preparations. The flow manifold was designed with computer-controlled three-way solenoid valves for independent handling of sample and reagent solutions and a multi-channel spectrophotometer was employed for signal measurements. Periodic re-calibration as well as the standard addition method was implemented by using a single reference solution. Linear responses ( r=0.999) were obtained for 0.500–10.0 mg l −1 ascorbic acid, 2.00–50.0 mg l −1 thiamine, 5.00–50.0 mg l −1 riboflavine and 0.500–8.00 mg l −1 pyridoxine. Detection limits were estimated as 0.08 mg l −1 (0.5 μmol l −1) ascorbic acid, 0.8 mg l −1 (2 μmol l −1) thiamine, 0.2 mg l −1 (0.5 μmol l −1) riboflavine and 0.1 mg l −1 (0.9 μmol l −1) pyridoxine at 99.7% confidence level. A mean sampling rate of 60 determinations per hour was achieved and coefficients of variation of 1% ( n=20) were estimated for all species. The mean reagent consumption was 25-fold lower in relation to flow-based procedures with continuous reagent addition. Average recoveries between 95.6 and 100% were obtained for commercial pharmaceutical preparations. Results agreed with those obtained by reference methods at 95% confidence level. The flow system is suitable for application in quality control processes and in dissolution studies of vitamin tablets.

DETERMINATION OF DIFFERENT SOLUBILITY VITAMINS IN PHARMACEUTICAL PREPARATIONS. II. METHODS VALIDATION

ABSTRACT Validation of methods for quantitative determination of vitamins in multivitamin preparations was performed. It was confirmed that tested methods are suitable for determination of both nonpolar vitamins (fat-soluble: A, D3 and E) and polar vitamins (water-soluble: B1, B2, B6, C, and PP). Determinations were quick and selective; they allowed a separation of peaks of vitamins to the baseline within 25 minutes. The ranges of concentration taken through the validation procedure were similar to daily doses of vitamins proposed by the Food and Drug Administration (FDA), and they were within the range of vitamin concentrations in common pharmaceutical preparations. In these ranges of concentration, the calibration graphs were linear (correlation coefficient r from 0.990 to 0.999). The repeatability of the methods (RSD) varied between 1.4 and 4.5% for fat-soluble vitamins and between 1.6 and 2.6% for water-soluble vitamins. The recoveries of added vitamins were between 98.1 and 100.4% for fat-soluble vitamins, and 97.2 and 99.0% for water-soluble vitamins. The sample solutions were stable for twenty-four hours after the preparation. The obtained results show that these methods are usable for automatic quantitative determination of vitamins in pharmaceutical preparations for quality control purposes.

DETERMINATION OF DIFFERENT SOLUBILITY VITAMINS IN PHARMACEUTICAL PREPARATIONS. I. HPLC COLUMN SWITCHING

ABSTRACT The aim of the study was to elaborate a solution enabling automation and fast quantitative determination of vitamins in pharmaceutical preparations. For this purpose, two analytical methods were combined in which different chromatographic stationary phases of different selectivity were employed. Application of the column-switching technique enabled sequential routine determination of the two different groups of vitamins. The proposed procedure was used for a quantitative determination of the vitamins: A, D3, E, B1, B2, B6, PP, and C in pharmaceutical preparations. It was proven that by means of this application, it is possible to perform about twelve complete analyses of vitamins within twenty-four hours.

Determination of water-soluble vitamins in pharmaceutical preparations by reversed-phase high-performance liquid chromatography with a mobile phase containing sodium dodecylsulphate andn-propanol

A reversed-phase liquid chromatographic method with sodium dodecylsulphate-n-propanolwater as mobile phase has been used to separate and determine six water-soluble vitamins in twelve minutes. The analytical characteristics linear range, sensitivity, detection limits, and precision were evaluated. The lowest detection limits were those of nicotinic acid (not usually present in pharmaceutical products), 0.7 mgL−1, nicotinamide, 1.3 mg L−1, and pyridoxine, 1.4 mg L−1. When the method was applied to the determination of the vitamins in pharmaceutical samples the values found agreed with those on the labels.

Simultaneous determination of twelve water- and fat-soluble vitamins by high-performance liquid chromatography with diode array detection

A novel method for the simultaneous determination of twelve water- and fat-soluble vitamins has been established by high-performance liquid chromatography with diode array detection. The vitamins were analyzed on a μBondapak C18 column (300 × 3.9 mm, 10 μm) with methanol-KH2PO4 buffer (0.1 M, pH 7.0)-water as mobile phase in a gradient. The linearity of calibration graphs was compound-dependent and the detection limits ranged from 0.02 μg mL−1 to 0.5 μg mL−1. The method was successfully applied to determine vitamins in pharmaceutical preparations. The recoveries were from 95.1% to 103% and the relative standard deviations were in the range of 0.9% to 4.5%.

Separation and determination of three water-soluble vitamins in pharmaceutical preparations and food by micellar electrokinetic chromatography with amperometric electrochemical detection

A simple, reliable and reproducible method, based on micellar electrokinetic chromatography with amperometric detection, for the separation and determination of the three water-soluble vitamins, namely nicotinamide (VPP), pyridoxine (VB 6) and ascorbic acid (VC), in pharmaceutical preparations and food is described in this paper. A carbon disk electrode was used as working electrode. The optimal conditions of separation and detection were 0.02 mol/l phosphate–0.005 mol/l borate buffer (pH 8.8) containing 0.03 mol/l sodium dodecylsulphate (SDS), 7 kV for the separation voltage, and 1.20 V (versus Ag/AgCl) for the detection potential. The linear ranges were from 3.0×10 −4 to 1.0×10 −2 mol/l for VPP, 2.5×10 −6 to 1.0×10 −3 mol/l for VB 6, and 2.5×10 −6 to 1.0×10 −3 mol/l for VC with regression coefficients of 0.9900, 0.9998 and 0.9990, respectively. The detection limits for VPP, VB 6 and VC were 5.0×10 −5, 1.0×10 −6 and 1.0×10 −6 mol/l, respectively. The method developed in this paper was directly applied to the determination of the three water-soluble vitamins in pharmaceutical preparations and food, and the assay results were satisfactory.

The application of capillary electrophoresis to the analysis of vitamins in food and beverages.

Capillary electrophoretic (CE) methods have been used to separate and determine a wide range of water-soluble vitamins in pharmaceutical preparations, but has found limited application in determining vitamins at naturally occurring levels in food and beverages. CE has been used to determine vitamin C in fruits and beverages, niacin in a range of foods and thiamine in samples of meat and milk. The CE methodologies used to determine vitamins in pharmaceutical preparations and biological fluids are also included as examples of the effectiveness of CE in vitamin analysis.

Simultaneous determination of nine water-soluble vitamins in pharmaceutical preparations by high-performance liquid chromatography with diode array detection

A simple and rapid method for the simultaneous determination of nine water-soluble vitamins was developed by high performance liquid chromatography with diode array detection. The water-soluble vitamins were analyzed by HPLC on a μBondapak C18 column (300×3.9 mm, 10 μm) with methanol-0.1 mol/L KH2PO4 buffer (pH 7.0) as mobile phase (1.5 mL/min) in gradient mode as follows: in the range 0–7.0 min, 10:90; and in the range 8.0–15.0 min, 40:60 (v/v). All nine water-soluble vitamins were separated in a single HPLC run within less than 15 min. The linearity of the calibration graphs was compound-dependent with correlation coefficients of 0.9986 to 0.9999 (n = 6). The detection limits ranged from 0.02 μg/mL to 0.5 μg/mL. The method was successfully applied to the determination of vitamins in pharmaceutical preparations. The recoveries were from 95% to 102% and the relative standard deviations were in the range of 0.8% to 4.2%.

Simultaneous determination of nine water-soluble vitamins in pharmaceutical preparations by high-performance liquid chromatography with diode array detection

A simple and rapid method for the simultaneous determination of nine water-soluble vitamins was developed by high performance liquid chromatography with diode array detection. The water-soluble vitamins were analyzed by HPLC on a μBondapak C18 column (300×3.9 mm, 10 μm) with methanol-0.1 mol/L KH2PO4 buffer (pH 7.0) as mobile phase (1.5 mL/min) in gradient mode as follows: in the range 0–7.0 min, 10:90; and in the range 8.0–15.0 min, 40:60 (v/v). All nine water-soluble vitamins were separated in a single HPLC run within less than 15 min. The linearity of the calibration graphs was compound-dependent with correlation coefficients of 0.9986 to 0.9999 (n = 6). The detection limits ranged from 0.02 μg/mL to 0.5 μg/mL. The method was successfully applied to the determination of vitamins in pharmaceutical preparations. The recoveries were from 95% to 102% and the relative standard deviations were in the range of 0.8% to 4.2%.

Analytical assessment of the oscillating chemical reactions by use chemiluminescence detection

This paper introduces the chemiluminescence (CL) detection in oscillating reaction-based determinations using the analyte pulse perturbation technique, a straightforward and expeditious approach to deriving quantitative analytical information from oscillating chemical reactions. The behavior of the H2O2–KSCN–CuSO4-NaOH oscillating system in the presence of luminol was examined by using the proposed detection method and the classical potentiometric technique. Some analytical and practical aspects of both detection systems are discussed. A new analytical method for the determination of vitamin B6 based on the sequential perturbation produced by different amounts of this substance on the oscillating chemical system was also developed in order to assess the potential of CL detection for routine analyses. The calibration curve thus obtained was linear over the range 0.5–2.0 μmol of vitamin B6, and the precision and throughput were also quite good (3.04% as RSD and nine samples h−1, respectively). The proposed method was validated by determining the vitamin in pharmaceutical preparations.

High Performance Liquid Chromatography of Fat-Soluble Vitamins. II. Determination of Vitamin E in Pharmaceutical Preparations and Blood

Abstract α-tocopherol acetate (Vitamin E-acetate) is extracted, separated and determined from pharmaceutical preparations and from biological materials in nanogram range in less than 25 minutes. The extraction of Vit. E-acetate is performed in a fully automated, electronically controlled extraction apparatus. A reversed phase high-performance liquid chromatography (HPLC) using a column of lichrosorb RP18 and methanol as eleuent has been developed for the separation and quantitative determination of Vit. E-acetate in formulations. The described extraction and determination methods are suitable for the analysis of Vit. E-acetat alone and in the presence of other water - and Fat-soluble Vitamins in pharmaceutical preparations and are reproducible with coefficient of variation of ∼3%. Vitamin A-acetate can be used as internal standard.

Determination of Water Soluble Vitamins in Pharmaceutical Preparations Using Liquid Chromatography

Abstract A liquid chromatography procedure is described for the simultaneous separation of seven water soluble vitamins in pharmaceutical pteparations. The vitamins are separated on a CR bonded-phase column and eluted with a gradient solvent program. A microprocessor controlled variable wavelength ultraviolet detector is used to automatically change detection wavelength to optimize detection of each vitamin. Pharmaceutical preparations are briefly sonicated in a dimethyl sulfoxide-water mixture, centrifuged, and an aliquot injected directly onto the column.

The electrochemical determination of vitamin A. Part II. Further voltammetric determination of vitamin A and initial work on the determination of vitamin D in the presence of vitamin A.

All vitamin A derivatives are found to give clean oxidation waves with a 0·05 M solution of lithium perchlorate in methanol-benzene as the supporting electrolyte, and a glassy carbon electrode as the working electrode. The determination of the vitamin in pharmaceutical preparations is carried out via direct extraction without prior saponification.The dependence on pH of the half-peak potentials of vitamins A and D, and of provitamins D, is examined with both carbon paste and glassy carbon electrodes.It is shown that the voltammetric technique does not distinguish between vitamins A1 and A2, nor between vitamins D2 and D3. A method for the determination of vitamin A in margarine is given. Some attempts have also been made to determine vitamin D in mixtures with vitamin A. Partition chromatography with Celite-polyethylene glycol gives a clean separation of vitamin D from vitamin A, which makes voltammetric determination of vitamin D possible without further purification.