Simultaneous determination of antioxidants at a chemically modified electrode with vitamin B12 by capillary zone electrophoresis coupled with amperometric detection

In the paper, a new kind of vitamin B₁₂ (acquo-cobalamine) chemically modified electrode was fabricated and applied in capillary zone electrophoresis coupled with amperometric detection (CZE-AD) for simultaneous determination of six antioxidants in fruits and vegetables. The catalytic electrochemical properties of the chemically modified electrode could obviously enhance oxidation peak heights responses by about five times to glutathione, ascorbic acid, vanillic acid, chlorogenic acid, salicylic acid, and caffeic acid compared with common carbon disk electrode. Furthermore, the effects of working electrode potential, pH and concentration of running buffer, separation voltage and injection time on CZE-AD were investigated. Under the optimum conditions, the six analytes could be completely separated and detected in a borate-phosphate buffer (pH 8.4) within 15 min. Their linear ranges were from 2.5 × 10⁻⁷ to 1.0 × 10⁻⁴ mol L⁻¹ and the detection limits were as low as 10⁻⁸ mol L⁻¹ magnitude (S/N = 3). The proposed method has been successfully employed to monitor the six analytes in practical samples with recoveries in the range 96.0-106.0% and RSDs less than 5.0%. Above results demonstrate that capillary zone electrophoresis coupled with electrochemical detection using vitamin B₁₂ modified electrode as detector is of convenient preparation, high sensitivity, good repeatability, and could be used in the rapid determination of practical samples.     

Effects of γ-irradiation and cooking on vitamins B6 and B12 in grass prawns (Penaeus monodon)

The effects of irradiation doses, irradiation temperature and a combined treatment of irradiation and cooking on the vitamin B₆ and B₁₂ contents of grass prawns have been studied. Grass prawns were irradiated at refrigerated (4°C) or frozen (-20°C) temperatures with different doses. A domestic cooking procedure was followed after irradiation. The changes in vitamins B₆ and B₁₂ of both raw and cooked grass prawns were evaluated. Results showed no significant changes of vitamin B₆ and B₁₂ in grass prawns with a radiation dose up to 7 kGy at either 4°C or -20°C. Irradiation at 4°C caused more destruction of vitamin B₁₂ but not vitamin B₆ than did irradiation at -20°C in grass prawns. There was significant destruction of both vitamins B₆ and B₁₂ in unirradiated samples during cooking. The introduction of the irradiation process before cooking had no effect on either vitamin. These results indicate that the loss of vitamins B₆ and B₁₂ in the combined treatments was caused mainly by thermal destruction.     

Fluorescence resonance energy transfer between acridine orange and rhodamine 6G and its analytical application for vitamin B12 with flow-injection laser-induced fluorescence detection

By coupling flow-injection with laser-induced fluorescence detection, a setup was developed and a novel method combining fluorescence resonance energy transfer (FRET) and flow-injection analysis (FIA) was proposed for the determination of vitamin B₁₂ (VB₁₂) based on its fluorescence quenching on the system of acridine orange (AO)/rhodamine 6G (R6G). The effective energy transfer could occur between AO and R6G in the dodecyl benzene sodium sulfonate (DBS) while 454 nm argon laser was used as the excitation source, and as a result, the fluorescence emission of R6G has been increased significantly. It was found that the fluorescence of the above system could be sharply diminished by VB₁₂. By using the mixed solution AO-R6G-DBS and the same solution containing VB₁₂ as the carrier and sample, respectively, a series of negative peaks which could be applied for the quantification of VB₁₂ were obtained. The detection limit for VB₁₂ was 1.65 × 10⁻⁶ mol/L. The linear range for determining VB₁₂ was 4 × 10⁻⁴ to 2 × 10⁻⁶ mol/L (correlation coefficient, r = 0.9923). The method was applied to measure VB₁₂ injections with satisfactory results.     

Determination of energizers in energy drinks

The method of UV/VIS derivative spectrophotometry for the determination of caffeine and B vitamins in energy drinks after solid phase extraction has been developed. Caffeine has been determined in the mixture with B₂ vitamin with zero-crossing technique from the I derivative spectra (λ = 266.8 nm), and B₃ in mixture with B₆ vitamin from the II derivative spectra (λ = 280.1 nm). B₁₂ vitamin has also been determined in a three-component mixture with vitamins B₃ and B₆. Taurine in drinks has been determined from the basic spectra after derivatization with ninhydrin (λ = 570 nm).     

Use of packed-fiber solid-phase extraction for sample clean-up and preconcentration of vitamin B_(12) before determination

A rapid and simple preconcentration step applying packed-fiber solid-phase extraction columns has been investigated to vitamin B_(12).The extraction performance of the new method was investigated preliminarily on vitamin functional drink.The analysis used a reversed-phase C_(18) column,with a photo-diode array detector at 220 nm.The samples were preconcentrated with packed-fiber solid-phase extraction columns.Good linearity was observed in vitamin functional drink.The repeatability of extraction performa...     

K2B12F12: A rare A2X structure for an ionic compound at ambient conditions

The anhydrous salt K₂B₁₂F₁₂ crystallized from aqueous solution and its structure was determined by single crystal X-ray diffraction. The Ni₂In-type structure it exhibits is rare for an A₂X ionic compound at 25 °C and 1 atm., consisting of an expanded hexagonal close-packed array of B₁₂F₁₂²⁻ centroids (cent?cent distances: 7.204-8.236 Å) with half of the K⁺ ions filling all of the O_h holes and half of the K⁺ ions filling all of the D_3h trigonal holes in the close-packed layers that are midway between two “empty” T_d holes. The structure is also unusual in that the bond-valence sum for the K⁺ ions in O_h holes is less than or equal to 0.73 (the bond-valence sum for the other type of K⁺ ion is 1.16). A variation of the Ni₂In structure is exhibited by the previously published monohydrate Cs₂(H₂O)B₁₂F₁₂, for which an improved structure is also reported here. For K₂B₁₂F₁₂: monoclinic, C2/c, a = 8.2072(8), b = 14.2818(7), c = 11.3441(9) Å, β = 92.832(5)°, Z = 4, T = 120(2) K. For Cs₂(H₂O)B₁₂F₁₂: orthorhombic, P2₁2₁2₁, a = 9.7475(4), b = 10.2579(4), c = 15.0549(5) Å, Z = 4, T = 110(1) K.     

RP-HPLC determination of water-soluble vitamins in honey

The assessment and validation of reliable analytical methods for the determination of vitamins in sugar-based matrices (e.g. honey) are still scarcely explored fields of research. This study proposes and fully validates a simple and fast RP-HPLC method for the simultaneous determination of five water-soluble vitamins (vitamin B₂, riboflavin; vitamin B₃, nicotinic acid; vitamin B₅, pantothenic acid; vitamin B₉, folic acid; and vitamin C, ascorbic acid) in honey. The method provides low detection and quantification limits, very good linearity in a large concentration interval, very good precision, and the absence of any bias. It has been successfully applied to 28 honey samples (mainly from Sardinia, Italy) of 12 different botanical origins. While the overall amount of the analytes in the samples is quite low (always below 40 mg kg⁻¹), we have observed a marked dependence of some of their concentrations (i.e. vitamin B₃ and vitamin B₅) and the botanical origin of the honey. This insight might lead to important characterization features for this food item.     

Iodination of cisplatin adduct of Vitamin B12 [{B12}-CN-{cis-PtCl(NH3)2}]

The cisplatin adduct of vitamin B₁₂, [{B₁₂}-CN-{cis-PtCl(NH₃)₂}]⁺ (1), reacts with iodide in aqueous solution to form [{B₁₂}-CN-{trans-PtI₂(NH₃)}] (3) in good yield. Mono-substitution of chloride was not observed since a subsequent replacement of one NH₃ by a second iodide is very fast as compared to the Cl⁻ → I⁻ exchange. The same reaction conditions allowed to introduce radioiodide ¹³¹I. Vitamin B₁₂ can therefore be labeled with radionuclides via binding to the Pt(II) center.     

Methyl transfer from a hydrophobic vitamin B12 derivative to arsenic trioxide

The methylation reaction of arsenic trioxide conducted at 37 °C and pH 7.0 for 24 h using hydrophobic methylated vitamin B₁₂, (methyl) (aquo) heptamethylcobyrinate perchlorate, CH₃B₁₂ ester, as a methyl donor in the presence of reduced glutathione (GSH) yielded monomethylarsonous acid (MMA), dimethylarsinic acid (DMA), and trimethylarsine oxide (TMAO) as products with a methylation rate over 95%. In contrast, when methylcobalamin (CH₃B₁₂) was used as the methyl donor, only MMA and DMA were produced and the methylation rate dropped to around 20%. Reductive demethylation of a methyl-corrinoid coordination complex mediated by GSH is suggested as a mechanism of methyl transfer to arsenic trioxide. The differences observed for different corrinoid coordination complexes with respect to the reactivity of methyl transfer to arsenic is ascribable to differences inherent in the base-on (CH₃B₁₂) and base-off (CH₃B₁₂ ester) natures of the compounds.     

Determination of B-group vitamins in Turkish honey using ultra-performance liquid chromatography with electrospray ionization coupled to tandem mass spectrometry

A rapid and sensitive ultra-performance liquid chromatography–tandem mass spectrometry method with electrospray ionization (UPLC–ESI–MS/MS) for analysis of B-group vitamins in honey has been presented. Aim of this study is the characterization of different types of Turkish honeys according to B-group vitamins. Vitamins were determined in 17 different types of Turkish honey samples by UPLC–ESI–MS/MS. Heather honey samples were distinguished among the studied honeys with the richest vitamin content with 286.10?mg/kg, and it is followed by sunflower honey and thyme honey with the total vitamin contents of 206.01 and 163.27?mg/kg, respectively. The presence of vitamin B₁ (thiamine), vitamin B₂ (riboflavin), vitamin B₃ (nicotinamide, B₃N and nicotinic acid, B₃H), vitamin B₆ (pyridoxine), and vitamin B₉ (folic acid) was detected in all the honey samples analyzed. Moreover, vitamin B₅ (pantothenic acid) was observed in most of them. Vitamin B₁₂ (cyanocobalamin) and vitamin B₇ (biotine) were not detected in the studied honey samples. Turkish honey samples showed efficacious vitamin content for the consumers.     

Sub-picogram determination of Vitamin B12 in pharmaceuticals and human serum using flow injection with chemiluminescence detection

A sensitive chemiluminescence (CL) method, based on the enhancive effect of cobalt(II) on the CL reaction between luminol and dissolved oxygen in a flow injection (FI) system, was proposed for determination of Vitamin B₁₂. The increment of the CL intensity was proportional to the concentration of Vitamin B₁₂, giving a calibration graph linear over the concentration from 2.0×10⁻¹⁰ to 1.2×10⁻⁶ g l⁻¹ (r²=0.9992) with the detection limit of 5.0×10⁻¹¹ g l⁻¹ (3σ). At a flow rate of 2.0 ml min⁻¹, a complete determination of Vitamin B₁₂, including sampling and washing, could be accomplished in 0.5 min with the relative standard deviations (R.S.D.) of less than 5.0%. The proposed method was applied successfully to the determination of Vitamin B₁₂ in pharmaceuticals, human serum, egg yolk and fish tissue.     

Coupling on-line of a dialyser with a flow-continuous system to separate Vitamin B(12) from milk

The use of membranes for on-line separations in flow-through dialyser as a part of a flow system is extremely useful for automated samples preparation. In this paper a method to couple the dialysis and the UV detection on-line of Vitamin B₁₂ from milk is proposed. Firstly, the milk samples were pre-treated with trichloroacetic and centrifuged (to eliminate proteins and fats) and later, using a dialyser coupled a flow-continuous manifold was possible dialyse the Vitamin B₁₂, which was monitored spectrophotometrically at 361 nm. On the other hand, the milk samples were also dialysed on-line but without the acid treatment and the results were compared. The influence of various parameters, including the pump speed for both the donor and acceptor stream, dialysis time, donor and acceptor loop volume on dialysis efficiency was studied. The method was applied to different kinds of milk (skimmed and semi-skimmed milk, evaporated milk, lactose free milk and liquid and powder whole milk). The relative standard deviation (R.S.D.) of the proposed method was of 0.45% and the obtained dialysis percentage was of 5.8%. The proposed method very easy permit a pre-treatment of the sample, quick and on-line with the detection. The dialysis process permitted the pass of vitamin and avoided the pass of other analytes as proteins in the case of the milk samples without acid treatment.     

HPLC-ESI-MS analysis of Vitamin B12 in food products and in multivitamins-multimineral tablets

Methods for the determination of Vitamin B₁₂ remain limited due to their low sensitivity and poor selectivity. In the present work, a simple and sensitive HPLC-ESI-MS method for determining Vitamin B₁₂ in food products and in multivitamin-multimineral tablets was developed. Vitamin B₁₂ was extracted from food products with 50 mM sodium acetate buffer (pH 4.0) in the presence of sodium cyanide. Total Vitamin B₁₂ content in food product can be obtained by efficient enzymatic hydrolysis to release the bound Vitamin B₁₂. Vitamin B₁₂ was quantified with ginsenoside Re as internal standard (I.S.) after their separations on a C₁₈ column with a gradient of mobile phase made of water and acetonitrile. MS with SIR mode at m/z 930.8 was used for Vitamin B₁₂ quantification. The calibration graphs plotted with five concentrations of Vitamin B₁₂ was linear with a regression coefficient r² = 0.9994. The intra-assay R.S.D. and the inter-assay R.S.D. were 2.6% (n = 5) and 3.5% (n = 6), respectively. The recoveries evaluated at spiking three different concentrations on fortified products were above 93%. The method has been applied successfully in the determination of Vitamin B₁₂ in fortified food products and multivitamin-multimineral tablets.     

A solid-phase enzyme-linked assay for vitamin B12

A new solid-phase enzyme-linked competitive binding assay for vitamin B₁₂ (cyanocobalamin) is described. The assay is based on the competition between analyte B₁₂ molecules and a glucose-6-phosphate dehydrogenase-vitamin B₁₂ conjugate for a limited number of R-protein binding sites immobilized on sepharose particles. After appropriate incubation and washing steps, the enzyme activity bound to the solid-phase is inversely related to the concentration of B₁₂ in the sample. Under optimized conditions, the method can detect B₁₂ in the range of 3×10^–10–1×10^–8 M (using 100l sample) with high selectivity over other biological molecules.     

The study of Bi3B5O12: synthesis, crystal structure and thermal expansion of oxoborate Bi3B5O12

The paper presents a new data on the crystal structure, thermal expansion and IR spectra of Bi₃B₅O₁₂. The Bi₃B₅O₁₂ single crystals were grown from the melt of the same stoichiometry by Czochralski technique. The crystal structure of Bi₃B₅O₁₂ was refined in anisotropic approximation using single-crystal X-ray diffraction data. It is orthorhombic, Pnma, a=6.530(4), b=7.726(5), c=18.578(5) Å, V=937.2(5) Å³, Z=4, R=3.45%. Bi³⁺ atoms have irregular coordination polyhedra, Bi(1)O₆ (d(B-O)=2.09-2.75 Å) and Bi(2)O₇ (d(B-O)=2.108-2.804 Å). Taking into account the shortest bonds only, these polyhedra are considered here as trigonal Bi(1)O₃ (2.09-2.20 Å) and tetragonal Bi(2)O₄ (2.108-2.331 Å) irregular pyramids with Bi atoms in the tops of both pyramids. The BiO₄ polyhedra form zigzag chains along b-axis. These chains alternate with isolated anions [B₂^IVB₃^IIIO₁₁]⁷⁻ through the common oxygen atoms to form thick layers extended in ab plane. A perfect cleavage of the compound corresponds to these layers and an imperfect one is parallel to the Bi-O chains. The Bi₃B₅O₁₂ thermal expansion is sharply anisotropic (α₁₁≈α₂₂=12, α₃₃=3×10⁻⁶ °C⁻¹) likely due to a straightening of the flexible zigzag chains along b-axis and decreasing of their zigzag along c-axis. Thus the properties like cleavage and thermal expansion correlate to these chains.     

Amperometric biosensor based on reductive H2O2 detection using pentacyanoferrate-bound polymer for creatinine determination

Pentacyanoferrate-bound poly(1-vinylimidazole) (PVI[Fe(CN)₅]) was selected as a mediator for amperometric creatinine determination based on the reductive H₂O₂ detection. Creatinine amidohydrolase (CNH), creatine amidohydrolase (CRH), sarcosine oxidase (SOD), peroxidase (POD), and PVI[Fe(CN)₅] were crosslinked with poly(ethylene glycol) diglycidyl ether (PEGDGE) on a glassy carbon (GC) electrode for a creatinine biosensor fabrication. Reduction current was monitored at −0.1 V in the presence of creatinine and O₂. It is revealed that PVI[Fe(CN)₅] is suitable as a mediator for a bioelectrocatalytic reaction of POD, since PVI[Fe(CN)₅] neither reacts with reactants nor works as an electron acceptor of SOD. The amounts of PVI[Fe(CN)₅], PEGDGE, and enzymes were optimized toward creatinine detection. Nafion as a protecting film successfully prevented the enzyme layer from interferences. The detection limit and linear range in creatinine determination were 12 μM and 12–500 μM (R² = 0.993), respectively, and the sensitivity was 11 mA cm⁻² M⁻¹, which is applicable for urine creatinine tests. The results of the creatinine determination for four urine samples measured with this proposed method were compared with Jaffe method, and a good correlation was obtained between the results.     

Sequential determination of fat- and water-soluble vitamins in Rhodiola imbricata root from trans-Himalaya with rapid resolution liquid chromatography/tandem mass spectrometry

A rapid method was developed to determine both types of vitamins in Rhodiola imbricata root for the accurate quantification of free vitamin forms. Rapid resolution liquid chromatography/tandem mass spectrometry (RRLC–MS/MS) with electrospray ionization (ESI) source operating in multiple reactions monitoring (MRM) mode was optimized for the sequential analysis of nine water-soluble vitamins (B₁, B₂, two B₃ vitamins, B₅, B₆, B₇, B₉, and B₁₂) and six fat-soluble vitamins (A, E, D₂, D₃, K₁, and K₂). Both types of vitamins were separated by ion-suppression reversed-phase liquid chromatography with gradient elution within 30 min and detected in positive ion mode. Deviations in the intra- and inter-day precision were always below 0.6% and 0.3% for recoveries and retention time. Intra- and inter-day relative standard deviation (RSD) values of retention time for water- and fat-soluble vitamin were ranged between 0.02–0.20% and 0.01–0.15%, respectively. The mean recoveries were ranged between 88.95 and 107.07%. Sensitivity and specificity of this method allowed the limits of detection (LOD) and limits of quantitation (LOQ) of the analytes at ppb levels. The linear range was achieved for fat- and water-soluble vitamins at 100–1000 ppb and 10–100 ppb. Vitamin B-complex and vitamin E were detected as the principle vitamins in the root of this adaptogen which would be of great interest to develop novel foods from the Indian trans-Himalaya.     

Study on the inclusion interactions of β-cyclodextrin and its derivative with dyes by spectrofluorimetry and its analytical application

The inclusion interactions of β-cyclodextrin (β-CD) and hydroxypropyl-β-cyclodextrin (HP-β-CD) with dyes were developed by spectrofluorimetry, and the inclusion constants of inclusion complexes were determined by direct fluorescence technique. The main factors (the host molecule, the guest molecule, and the pH) for the inclusion interaction were discussed in detail. At the same time, the inclusion interaction of HP-β-CD and vitamin B₆ (VB₆) was investigated with the competitive fluorescence inclusion method and the inclusion constant of HP-β-CD and vitamin B₆ (VB₆) was obtained by indirect fluorescence technique. On the basis of the linear relationship between the change of fluorescence intensity (ΔF) and the concentration of VB₆, a competitive fluorescence inclusion method was used to the determination of VB₆. The method has been successfully applied to the analysis of VB₆ in synthetic samples, tablets and injections with satisfactory results.     

Multiple response optimization applied to the development of a capillary electrophoretic method for pharmaceutical analysis

Multiple response simultaneous optimization by using the desirability function was used for the development of a capillary electrophoresis method for the simultaneous determination of four active ingredients in pharmaceutical preparations: vitamins B₆ and B₁₂, dexamethasone and lidocaine hydrochloride. Five responses were simultaneously optimized: the three resolutions, the analysis time and the capillary current. This latter response was taken into account in order to improve the quality of the separations. The separation was carried out by using capillary zone electrophoresis (CZE) with a silica capillary and UV detection (240 nm). The optimum conditions were: 57.0 mmol l⁻¹ sodium phosphate buffer solution, pH 7.0 and voltage = 17.2 kV. Good results concerning precision (CV lower than 2%), accuracy (recoveries ranged between 98.5 and 102.6%) and selectivity were obtained in the concentration range studied for the four compounds. These results are comparable to those provided by the reference high performance liquid chromatography (HPLC) technique.     

La3Ru8B6 and Y3Os8B6, new members of a homologous series R(A)nM3n−1B2n

Two new compounds, La₃Ru₈B₆ and Y₃Os₈B₆, were synthesized by arc melting the elements. Their structural characterization was carried out at room temperature on as-cast samples by using X-ray diffractometry. According to X-ray single-crystal diffraction results these borides crystallize in Fmmm space group (no. 69), Z=4, a=5.5607(1) Å, b=9.8035(3) Å, c=17.5524(4) Å, ρ=8.956 Mg/m³, μ=25.23 mm⁻¹ for La₃Ru₈B₆ and a=5.4792(2) Å, b=9.5139(4) Å, c=17.6972(8) Å, ρ=13.343 Mg/m³, μ=128.23 mm⁻¹ for Y₃Os₈B₆. The crystal structure of La₃Ru₈B₆ was confirmed from Rietveld refinement of X-ray powder diffraction data. Both La₃Ru₈B₆ and Y₃Os₈B₆ compounds are isotypic with the Ca₃Rh₈B₆ compound and their structures are built up from CeCo₃B₂-type and CeAl₂Ga₂-type structural fragments taken in ratio 2:1. They are the members of structural series R(A)_nM_3n−1B_2n with n=3 (R is the rare earth metal, A the alkaline earth metal, and M the transition metal). Structural and atomic parameters were also obtained for La_0.94Ru₃B₂ compound from Rietveld refinement (CeCo₃B₂-type structure, P6/mmm space group (no. 191), a=5.5835(9) Å, c=3.0278(6) Å).