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Three sensitive, accurate and precise HPLC methods were devolved for the simultaneous determination of vilazodone HCl (VILHC), agomelatine (AGO) or duloxetine HCl (DULHC) and vitamin B 12 (cyanocoblamine B 12 ) in bulk, pharmaceutical dosage form and in urine samples. Both similar methods (I and II) were carried out using 0.04 M phosphate buffer (pH 7.0), acetonitrile and methanol in the ratio (30:30:40, v/v) as a mobile phase. Thermo BDS hypersil-C18 column, with 5 μm particle size and 250 × 4.5 mm dimensions, at flow rate 1.0 mL min -1 and UV detection at 277 nm at ambient temperature 25 °C were used. The retention times were 5.12 and 2.54 min for VILHC and vitamin B 12 , 4.98 and 2.53 min for AGO and vitamin B 12 , respectively, with linearity range from 0.001 to 200 μg mL -1 . However, for the separation of DULHC and B 12 , UV detection at 230 nm and Agilent Eclipse XDA-C8 (150 × 4.5 mm, 5 μm) column, were used (method III). The retention time of DULHC and vitamin B 12 was found to be 4.53 and 1.35 min, respectively, with linearity range from 0.0005 to 200 μg mL -1 . The proposed methods were validated as per the ICH guideline with very low LOD and LOQ. The values of %RSD for precision was less than 2% and the value of % recovery were found to be 99.20-100.9% and 99.23-100.67% for determination of the drugs in pure and pharmaceutical formulations, respectively, for all methods confirming that the methods are precise, accurate and selective for separation and determination of VILHC, AGO or DULHC from B 12 in tablets and in urine samples without any interference from each other or from common excipients., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Vitamin B12 (VB12) has an important function in human physiology. However, analysis of VB12 at natural levels in foods or biological samples is difficult because of its very low concentration level and the presence of high-abundance components which can interfere with the measuring system. Thus, it is essential to develop efficient and selective enrichment approaches for VB12. Molecularly imprinted polymers (MIPs) have important applications from separation and sensing to catalysis. However, there is no report on the preparation of MIPs for VB12. Here, we use boronate affinity-based oriented surface imprinting to prepare MIPs for VB12. A VB12 template was first covalently immobilized onto the surface of boronic acid functionalized magnetic nanoparticles. Subsequently, a thin imprinting coating of poly(2-anilinoethanol) was formed to cover the substrate surface via in-water polymerization. After removing the template, 3D cavities complementary to the molecular size and shape of the template were formed in the imprinting layer. The imprinting coating was highly hydrophilic and presented limited residual boronic acid, thus non-specific binding was avoided. The prepared MIPs exhibited several highly favorable features, including excellent specificity, high binding strength and low binding pH. The prepared MIPs were successfully applied to the analysis of VB12 in human milk., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The amalgamation of natural origin materials and technologies for label-free and real time detection is pertinent in analytical field. In this study, Bombyx mori silk fibroin protein (BMSF) has been utilized for the dual sensing of vitamin B 12 via fluorescence and electrical impedance techniques. The processing of BMSF is done to generate an aqueous solution of BMSF along with the development of micro patterned thin films via soft lithography technique. The BMSF aqueous solution exhibit auto fluorescence property and thereby utilized for label free detection of vitamin B 12 with an estimated limit of detection (LOD) of about 0.003 × 10 -6 g/uL. This is followed by impedimetric detection of vitamin B 12 using the micro patterned BMSF thin films. A LOD of 17.8 ppm and 0.25 ppm are achieved in aqueous solution and human blood serum, respectively. Taken together, this work demonstrates a potential label free dual sensing mode for sensitive detection of micronutrient vitamin B 12 ., (Copyright © 2018 Elsevier B.V. All rights reserved.)

A major problem limiting reproducible use of liquid extraction surface analysis (LESA) array sampling of dried surface-deposited liquid samples is the unwanted spread of extraction solvent beyond the dried sample limits, resulting in unreliable data. Here, we explore the use of the Droplet Microarray (DMA), which consists of an array of superhydrophilic spots bordered by a superhydrophobic material giving the potential to confine both the sample spot and the LESA extraction solvent in a defined area. We investigated the DMA method in comparison with a standard glass substrate using LESA analysis of a mixture of biologically relevant compounds with a wide mass range and different physicochemical properties. The optimized DMA method was subsequently applied to urine samples from a human intervention study. Relative standard deviations for the signal intensities were all reduced at least 3-fold when performing LESA-MS on the DMA surface compared with a standard glass surface. Principal component analysis revealed more tight clusters indicating improved spectral reproducibility for a human urine sample extracted from the DMA compared to glass. Lastly, in urine samples from an intervention study, more significant ions (145) were identified when using LESA-MS spectra of control and test urine extracted from the DMA. We demonstrate that DMA provides a surface-assisted LESA-MS method delivering significant improvement of the surface extraction repeatability leading to the acquisition of more robust and higher quality data. The DMA shows potential to be used for LESA-MS for controlled and reproducible surface extraction and for acquisition of high quality, qualitative data in a high-throughput manner.

Based on increased demands of strict vegetarians, an investigation of vitamin B12 content in plant sources, was carried out. The vitamin B12 concentration was determined by RP-HPLC with UV detection, after prior matrix isolation by immunoaffinity chromatography (IAC). Vitamin B12 was extracted in the presence of sodium cyanide, to transform all forms of cobalamin into cyanocobalamin. Diode array detector was used to monitor vitamin B12, after its chromatographic separation under gradient elution with a mobile phase consisting of acetonitrile and trifluoroacetic acid 0.025% (w/v). The method demonstrated excellent linearity with a limit of detection 0.004μg/ml. The method precision was evaluated for plant samples and it was below 0.7% (n=6). Significant amounts of vitamin B12 in plants were detected in Hippophae rhamnoides (37μg/100g dry weight), in Elymus (26μg/100g dry weight) and in Inula helenium (11μg/100g dry weight)., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Background: Despite the fact that most vitamins are present in a variety of foods, malnutrition, unbalanced diets or insufficient intake of foods are still the cause of vitamin deficiencies in humans in some countries. Vitamin B 12 (Cobalamin) is a complex compound that is only naturally produced by bacteria and archea. It has been reported that certain strains belonging to lactic acid bacteria group are capable of synthesized water-soluble vitamins such as those included in the B-group, as vitamin B 12 . In this context, the goal of the present paper was to evaluate and characterize the production of vitamin B 12 in Lactobacillus coryniformis CRL 1001, a heterofermentative strain isolated from silage., Results: Cell extract of L. coryniformis CRL 1001, isolated from silage, is able to correct the coenzyme B 12 requirement of Salmonella enterica serovar Typhimurium AR 2680 in minimal medium. The chemical characterization of the corrinoid-like molecule isolated from CRL 1001 cell extract using HPLC and mass spectrometry is reported. The majority of the corrinoid produced by this strain has adenine like Coα-ligand instead 5,6-dimethylbenzimidazole. Genomic studies revealed the presence of the complete machinery of the anaerobic biosynthesis pathway of coenzyme B 12 . The detected genes encode all proteins for the corrin ring biosynthesis and for the binding of upper (β) and lower (α) ligands in one continuous stretch of the chromosome., Conclusions: The results here described show for the first time that L. coryniformis subsp. coryniformis CRL 1001 is able to produce pseudocobalamin containing adenine instead of 5,6-dimethlbenzimidazole in the Coα-ligand. Genomic analysis allowed the identification and characterization of the complete de novo biosynthetic pathway of the corrinoid produced by the CRL 1001 strain.

This work describes a new analytical method for the determination of four cobalamins (adenosylcobalamin (AdoCbl), methylcobalamin (MeCbl), hydroxocobalamin (OHCbl) and cyanocobalamin (CNCbl)) in cow's milk. The extraction procedure is fast and based on dilution/protein precipitation of a milk sample with 50mM sodium acetate buffer (pH 4.6), followed by solid phase extraction (SPE) of the filtered supernatant. Relative recoveries higher than 60% have been obtained for all the cobalamins by combining two different types of sorbents in the same SPE cartridge: two disks of buckypaper (BP), a nanoporous felt composed of oxidized multiwalled carbon nanotubes (MWCNTs), separated by a Teflon frit from OASIS HLB (500mg), a hydrophilic-lipophilic balance copolymer. Before its use as sorbent, BP was characterized in terms of porosity, permeability, surface area, specific adsorption capacity and tested for a potential reuse after adequate chemical regeneration. The analysis of the extracts was performed by liquid chromatography (LC) coupled to tandem mass spectrometry (MS/MS) on an analytical C18 column in less than 10min. After validation, the method was applied to the determination of the natural content of the four B12 homologues in cow's milk samples, providing data lacking in the literature., (Copyright © 2015 Elsevier B.V. All rights reserved.)