Simultaneous quantification of seven B vitamins from wheat grains using UHPLC-MS/MS.

B-group vitamins are important micronutrients for maintaining human health; nevertheless, B vitamin deficiency is a globally widespread issue. Thus, it is relevant to accurately assess the B-vitamin content in staple crop products such as wheat grains. Here, we developed a multi-enzyme extraction method allowing accurate quantification of seven B vitamins in wheat using ultra high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). Free forms of thiamine (B1), riboflavin (B2), niacin (B3), pantothenic acid (B5), pyridoxine (B6), biotin (B7) and folates (B9) were determined with recoveries ranging from 81 to 118% and accuracy below 15% bias. The precision was below 20% relative standard deviation and the internal standards adequately compensated for matrix effects. The method was applied to determine the B vitamin stabilities in wheat grains stored at different temperatures and periods. The results provide an important basis in future studies aiming at understanding nutritional availability of B vitamins.

Melamine sponges incorporated azo-linked porous organic polymer as adsorbent for extraction and determination of six B vitamins using pipette tip micro solid-phase extraction.

A novel azo-linked porous organic polymer (AL-POP) was synthesized from caffeic acid and benzidine via an azo-coupling reaction and characterized by FTIR, SEM-EDS, BET, TGA, XRD and zeta potential analysis. AL-POPs were incorporated into melamine sponges and used for pipette tip micro solid-phase extraction (PT-MSPE) of six types of B vitamins (including thiamine, riboflavin, nicotinamide, pyridoxine, folic acid, and cyanocobalamin). After extraction, the samples were analyzed using high performance liquid chromatography-diode array detection (HPLC-DAD) system. The effect of AL-POP composition on the extraction efficiency (EE) of vitamins was investigated and benzidine to caffeic acid mol ratio of 1.5, 3.35 mmol of NaNO2, and reaction time of 8 h were selected as optimum conditions. The efficiency of the extraction process was improved by optimizing various parameters such as the amount of sorbent, pH and ionic strength of the sample, sample volume, number of sorption and desorption cycles, type of wash solvent, and type and volume of eluent solvent. Linearity (R2≥0.9987), Limit of detection (LOD) (11.88-18.97 ng/mL), limit of quantification (LOQ) (39.62-63.23 ng/mL), and enrichment factor (EF) (1.27-4.31) were obtained using calibration curves plotted under optimum conditions. Recovery values of these six B vitamins in the spiked multivitamin syrup samples varied from 80.01% to 108.35%, with a relative standard deviation (RSD) below 5.44%. Eventually, the optimized method was successfully used to extract and quantify the B vitamins in multivitamin syrup and non-alcoholic beer.

Factors associated with longitudinal changes in B-vitamin and choline concentrations of human milk.

BACKGROUND: Little is known regarding the associations between maternal factors and B-vitamin and choline concentrations in early milk and the trajectories of these vitamins during lactation. OBJECTIVES: In this hypothesis-generating study, we modeled the association between maternal and offspring factors and longitudinal changes in milk B-vitamin and choline concentrations throughout lactation. METHODS: A hundred women were studied in a prospective birth cohort and milk samples from 52 women were collected at 2-8 d, 76 women at 28-50 d, and 42 women at 88-119 d postpartum. Maternal dietary intake during pregnancy and lactation was assessed by an FFQ. Linear mixed-effects models with interaction terms were used to evaluate changes in milk B-vitamin and choline concentrations over time based on maternal factors and the early postpartum concentrations of these micronutrients. RESULTS: The women with higher early postpartum milk concentrations of niacin (ßinteraction = -0.02; SE = 0.00; P < 0.001), pantothenic acid (ßinteraction = -0.10; SE = 2.56; P < 0.001), vitamin B-12 (ßinteraction= -0.10; SE = 0.03; P < 0.001), and choline (ßinteraction= -0.90; SE = 0.18; P < 0.001) exhibited a decrease in their concentrations throughout lactation. The participants with overweight and obesity prepregnancy experienced an increase in milk vitamin B-12 concentrations over time (ßinteraction = 0.04; SE = 0.02; P = 0.06). In contrast, a decrease in vitamin B-12 concentration was observed among women with vitamin B-12 intake below the RDA during pregnancy (ßinteraction= -0.08; SE = 0.05; P = 0.07). The women with niacin intake below the RDA during lactation experienced an increase in milk concentrations over time (ßinteraction = 0.01; SE = 0.01; P = 0.03). A gestational age at birth >40 wk was associated with an increase in milk choline concentration throughout lactation (ßinteraction = 0.54; SE = 0.16; P< 0.01). CONCLUSIONS: Changes in B-vitamin and choline concentrations in human milk over time may be associated with the early concentrations of these micronutrients in milk, maternal prepregnancy BMI, dietary intake, and gestational age at delivery.

Safe and Targeted Sonodynamic Cancer Therapy Using Biocompatible Exosome-Based Nanosonosensitizers.

Sonodynamic therapy (SDT), wherein sonosensitizers irradiated with ultrasound (US) produce cytotoxic reactive oxygen species (ROS), has garnered great attention as a promising alternative to photodynamic therapy owing to the significantly increased depth of tissue penetration. The development of nanocarriers that can selectively deposit sonosensitizers into tumor tissues without systemic toxicity is crucial to facilitate the translation of SDT to clinical use. In this study, exosomes, a class of naturally occurring nanoparticles, were utilized as nanocarriers for safe and cancer-targeted delivery of a sonosensitizer, indocyanine green (ICG). The exosomes were surface-engineered with an active cancer-targeting ligand, folic acid (FA), to increase the cancer specificity of the ICG-loaded exosomes (ExoICG). The FA-conjugated, ICG-loaded exosomes (FA-ExoICG) greatly improved aqueous stability and cellular uptake of ICG, resulting in significantly increased ROS generation in breast cancer cells. As a result, the FA-ExoICG demonstrated greater sonotoxicity against cancer cells than ExoICG and free ICG. The in vivo study revealed that compared to ExoICG, more FA-ExoICG accumulated in tumors, and their pharmacokinetic properties were superior. Notably, tumor growth in mice was significantly suppressed, without systemic toxicity, by a single intravenous injection of the FA-ExoICG and subsequent US irradiation. Therefore, this study demonstrated that active cancer-targeted FA-ExoICG could serve as effective nanosonosensitizers for safe and targeted cancer treatment.

The Affinity of Carboplatin to B-Vitamins and Nucleobases.

Platinum compounds have found wide application in the treatment of various types of cancer and carboplatin is one of the main platinum-based drugs used as antitumor agents. The anticancer activity of carboplatin arises from interacting with DNA and inducing programmed cell death. However, such interactions may occur with other chemical compounds, such as vitamins containing aromatic rings with lone-pair orbitals, which reduces the anti-cancer effect of carboplatin. The most important aspect of the conducted research was related to the evaluation of carboplatin affinity to vitamins from the B group and the potential impact of such interactions on the reduction of therapeutic capabilities of carboplatin in anticancer therapy. Realized computations, including estimation of Gibbs Free Energies, allowed for the identification of the most reactive molecule, namely vitamin B6 (pyridoxal phosphate). In this case, the computational estimations indicating carboplatin reactivity were confirmed by spectrophotometric measurements.

Assessment of Health Claims Related to Folic Acid in Food Supplements for Pregnant Women According to the European Regulation.

Pregnant women are a vulnerable group with increased nutritional requirements. The daily intake of folic acid, a crucial vitamin for embryonic development, must be reinforced through supplementation, as sometimes diets are not well equilibrated. As consumers increasingly rely on food supplements, it is vital to properly inform them about the health benefits provided by supplements' consumption to ensure their safe use. The objective of this work was to assess the compliance level of health claims related to folic acid in food supplements commercialized in Spain according to the European regulation. Authors performed (1) a review of health-related claims approved for folic acid in Europe, (2) a market research of food supplements commercialized in Spain with those claims, and (3) a selection of food supplements for chemical analysis in the lab to assess these claims. The results showed that nine health-related claims are currently approved for folic acid in Europe. The analytical results for folic acid content in the selected samples were consistent with the declared values and within the tolerance ranges established in the European Guidance document. All samples included accurate dosages and met the legal requirements (European Regulations 1924/2006, 432/2012, 1169/2011) for all approved claims for folic acid.

Low-dose thiamine supplementation of lactating Cambodian mothers improves human milk thiamine concentrations: a randomized controlled trial.

BACKGROUND: Infantile beriberi-related mortality is still common in South and Southeast Asia. Interventions to increase maternal thiamine intakes, and thus human milk thiamine, are warranted; however, the required dose remains unknown. OBJECTIVES: We sought to estimate the dose at which additional maternal intake of oral thiamine no longer meaningfully increased milk thiamine concentrations in infants at 24 wk postpartum, and to investigate the impact of 4 thiamine supplementation doses on milk and blood thiamine status biomarkers. METHODS: In this double-blind, 4-parallel arm randomized controlled dose-response trial, healthy mothers were recruited in Kampong Thom, Cambodia. At 2 wk postpartum, women were randomly assigned to consume 1 capsule, containing 0, 1.2 (estimated average requirement), 2.4, or 10 mg of thiamine daily from 2 through 24 weeks postpartum. Human milk total thiamine concentrations were measured using HPLC. An Emax curve was plotted, which was estimated using a nonlinear least squares model in an intention-to-treat analysis. Linear mixed-effects models were used to test for differences between treatment groups. Maternal and infant blood thiamine biomarkers were also assessed. RESULTS: In total, each of 335 women was randomly assigned to1 of the following thiamine-dose groups: placebo (n = 83), 1.2 mg (n = 86), 2.4 mg (n = 81), and 10 mg (n = 85). The estimated dose required to reach 90% of the maximum average total thiamine concentration in human milk (191 µg/L) is 2.35 (95% CI: 0.58, 7.01) mg/d. The mean ± SD milk thiamine concentrations were significantly higher in all intervention groups (183 ± 91, 190 ± 105, and 206 ± 89 µg/L for 1.2, 2.4, and 10 mg, respectively) compared with the placebo group (153 ± 85 µg/L; P < 0.0001) and did not significantly differ from each other. CONCLUSIONS: A supplemental thiamine dose of 2.35 mg/d was required to achieve a milk total thiamine concentration of 191 µg/L. However, 1.2 mg/d for 22 wk was sufficient to increase milk thiamine concentrations to similar levels achieved by higher supplementation doses (2.4 and 10 mg/d), and comparable to those of healthy mothers in regions without beriberi. This trial was registered at clinicaltrials.gov as NCT03616288.

Simvastatin-Nicotinamide Co-Crystals: Formation, Pharmaceutical Characterization and in vivo Profile.

PURPOSE: To enhance the solubility and dissolution profile of simvastatin (SIM) through co-crystallization with varying ratios of nicotinamide (NIC) using various co-methods. MATERIALS AND METHODS: Twelve SIM:NIC co-crystal formulations (F01-F12) were prepared using dry grinding, slurry, liquid-assisted grinding, and solvent-evaporation methods, and their properties compared. Optimized formulations were selected on the basis of dissolution profiles and solubility for in vivo studies. The angle of repose, Carr Index and Hausner ratio were calculated to evaluate flow properties. Differential light scattering (DLS) was used to estimate particle-size distribution. Scanning electron microscopy (SEM) was employed to evaluate surface morphology. Thermal analyses and Fourier-transform infrared (FTIR) spectroscopy were used to determine the ranges of thermal stability and physical interaction of formulated co-crystals. X-ray powder diffraction (XPD) spectroscopy was used to determine the crystalline nature. Solubility and dissolution studies were undertaken to determine in vitro drug-release behaviors. RESULTS: Micromeritic analyses revealed the good flow properties of formulated co-crystals. DLS showed the particle size of co-crystals to be in the nanometer range. SEM revealed that the co-crystals were regular cubes. Thermal studies showed the stability of co-crystals at >300°C. FTIR spectroscopy revealed minor shifts of various peaks. XPD spectroscopy demonstrated co-crystal formation. The formulations exhibited an improved dissolution profile with marked improvements in solubility. In vivo studies showed a 2.4-fold increase in Cmax whereas total AUC(0-∞) was increased 4.75-fold as compared with that of SIM tablets. CONCLUSION: Co-crystallization with NIC improved the solubility and dissolution profile and, hence, the bioavailability of the poorly water-soluble drug SIM.

Surface Interactions between Bacterial Nanocellulose and B-Complex Vitamins.

The interactions between films of bacterial nanocellulose (BNC) and B complex vitamins were studied using a Quartz Crystal Microbalance with Dissipation monitoring (QCM-D). Thin films of BNC were generated in situ by QCM-D, followed by real-time measurements of the vitamin adsorption. The desorption of vitamins was induced by rinsing the system using phosphate buffers at a pH of 2 and 6.5, emulating gastric conditions. Changes in frequency (which are proportional to changes in adsorbed mass, ∆m) detected by QCM-D were used to determine the amounts of vitamin adsorbed and released from the BNC film. Additionally, changes in dissipation (∆D) were proven to be useful in identifying the effects of the pH in both pristine cellulose films and films with vitamin pre-adsorbed, following its changes during release. The effects of pH on the morphology of the vitamin-BNC surfaces were also monitored by changes in rugosity from images obtained by atomic force microscopy (AFM). Based on this data, we propose a model for the binding phenomena, with the contraction on the relaxation of the cellulose film depending on pH, resulting in an efficient vitamin delivery process.

Sharing vitamins: Cobamides unveil microbial interactions.

Microbial communities are essential to fundamental processes on Earth. Underlying the compositions and functions of these communities are nutritional interdependencies among individual species. One class of nutrients, cobamides (the family of enzyme cofactors that includes vitamin B12), is widely used for a variety of microbial metabolic functions, but these structurally diverse cofactors are synthesized by only a subset of bacteria and archaea. Advances at different scales of study-from individual isolates, to synthetic consortia, to complex communities-have led to an improved understanding of cobamide sharing. Here, we discuss how cobamides affect microbes at each of these three scales and how integrating different approaches leads to a more complete understanding of microbial interactions.

Bio-metal-organic frameworks for molecular recognition and sorbent extraction of hydrophilic vitamins followed by their determination using HPLC-UV.

A bio-metal-organic framework (bio-MOF) derived from the amino acid L-serine has been prepared in bulk form and evaluated as sorbent for the molecular recognition and extraction of B-vitamins. The functional pores of bio-MOF exhibit high amounts of hydroxyl groups jointly directing other supramolecular host-guest interactions thus providing the recognition of B-vitamins in fruit juices and energy drinks. Single-crystal X-ray diffraction studies reveal the specific B-vitamin binding sites and the existence of multiple hydrogen bonds between these target molecules and the framework. It offered unique snapshots to accomplish an efficient capture of these solutes in complex aqueous matrices. Four B-vitamins (thiamin, nicotinic acid, nicotinamide, and pyridoxine) were investigated. They were eluted from the sorbent with phosphate buffer at pH 7 and analyzed by HPLC with UV detection. The sorbent was compared with commercial C18 cartridges. Following the procedure, acceptable reproducibility (RSD values < 14%) was achieved, and the detection limits were in the range 0.4 to 1.4 ng mL-1. The method was applied to the analysis of energy drink and juice samples and the recoveries were between 75 and 123% in spiked beverage samples. Graphical abstractA bio-MOF as SPE sorbent was prepared and applied to the extraction of B-vitamins in fruit juices and energy drinks.

Conjugation of a gold(iii) complex with vitamin B1 and chlorambucil derivatives: anticancer evaluation and mechanistic insights.

A novel cyclometalated gold(iii) complex supported by chlorambucil coupled with phenylpyridine (CHL-N^C) and a hybrid of vitamin B1 with dithiocarbamate (B1-DTC) with the formula [(CHL-N^C)AuIII(B1-DTC)](Cl2), 1, was synthesized and fully characterized using different techniques, including multinuclear NMR, mass spectrometry, and elemental analysis. This complex is water-soluble and stable in a biological environment. This new complex offers a new scaffold to explore the biological properties of gold(iii) complexes as an anticancer drug. The antiproliferative activities of complex 1 and free ligands against breast and colon cancer cells showed auspicious results with IC50 values in the micromolar range for complex 1 and more active than cisplatin and free ligands with selectivity over non-tumorigenic cells human lung fibroblasts, MRC-5. The DNA binding and inhibition of thioredoxin reductase of complex 1 were studied and compared with molecular docking results. Moreover, the Au cellular uptake and apoptosis of this new complex were investigated.

Topical delivery of niacinamide: Influence of neat solvents.

Niacinamide (NIA) has been widely used in cosmetic and personal care formulations for several skin conditions. Permeation of topical NIA has been confirmed in a number of studies under infinite dose conditions. However, there is limited information in the literature regarding permeation of NIA following application of topical formulations in amounts that reflect the real-life use of such products by consumers. The aim of the present work was therefore to investigate skin delivery of NIA from single solvent systems in porcine skin under finite dose conditions. A secondary aim was to probe the processes underlying the previously reported low recovery of NIA following in vitro permeation and mass balance studies. The solubility and stability of NIA in various single solvent systems was examined. The solvents investigated included Transcutol® P (TC), propylene glycol (PG), 1-2 hexanediol (HEX), 1-2 pentanediol (1-2P), 1-5 pentanediol (1-5P), 1-3 butanediol (1-3B), glycerol (GLY) and dimethyl isosorbide (DMI). Skin permeation and deposition of the molecule was investigated in full thickness porcine skin in vitro finite dose Franz-type diffusion experiments followed by mass balance studies. Stability of NIA for 72 h in the solvents was confirmed. The solubility of NIA in the solvents ranged from 82.9 ± 0.8 to 311.9 ± 4.5 mg/mL. TC delivered the highest percentage permeation of NIA at 24 h, 32.6 ± 12.1% of the applied dose. Low total recovery of NIA after mass balance studies was observed for some vehicles, with values ranging from 55.2 ± 12.8% to 106.3 ± 2.3%. This reflected the formation of a number of NIA degradation by-products in the receptor phase during the permeation studies. Identification of other vehicles for synergistic enhancement of NIA skin delivery will be the subject of future work.

Riboflavin: The Health Benefits of a Forgotten Natural Vitamin.

Riboflavin (RF) is a water-soluble member of the B-vitamin family. Sufficient dietary and supplemental RF intake appears to have a protective effect on various medical conditions such as sepsis, ischemia etc., while it also contributes to the reduction in the risk of some forms of cancer in humans. These biological effects of RF have been widely studied for their anti-oxidant, anti-aging, anti-inflammatory, anti-nociceptive and anti-cancer properties. Moreover, the combination of RF and other compounds or drugs can have a wide variety of effects and protective properties, and diminish the toxic effect of drugs in several treatments. Research has been done in order to review the latest findings about the link between RF and different clinical aberrations. Since further studies have been published in this field, it is appropriate to consider a re-evaluation of the importance of RF in terms of its beneficial properties.

Cobamides.

Vitamin B12 is the only known essential human micronutrient made exclusively by prokaryotes. Kennedy and Taga introduce us to the world of cobamides-those cobalt-containing compounds, like B12, that appear to be the proprietary domain of our microbial partners.

Effect of Trace Minerals and B Vitamins on the Proliferation/Cytotoxicity and Mineralization of a Gilthead Seabream Bone-Derived Cell Line.

Trace minerals and vitamins are known modulators of bone metabolism, and dietary optimization of these components may improve skeletal development and reduce the occurrence of skeleton deformities in farmed fish. As for larval stages, mineral and water-soluble vitamin nutrition requirements are lacking in research efforts and knowledge is scarce. An in vitro cell system developed from gilthead seabream vertebra and capable of mineralization was used to assess the effect of B vitamins (thiamin and pyridoxine) and trace minerals (copper, manganese, and zinc in a sulfated and chelated form) on cell proliferation and extracellular matrix (ECM) mineralization. Dependent on dose, inhibition of cellular proliferation and/or cytotoxic effects was observed for all nutrients tested and LD50 values were determined: copper, 67.4-69.5 ppm; manganese, 20.9-29.8 ppm; zinc, 37.1-42.8 ppm in sulfated and chelated form respectively; thiamin, 6273 ppm; pyridoxine, 14226 ppm. ECM mineralization was enhanced by mineral (dose and form dependent) and vitamin (dose dependent) supplementation, at non-toxic concentrations below the determined LD50s. This in vitro work confirmed the mineralogenic action of trace minerals and water-soluble vitamins and provided valuable insights for subsequent in vivo nutritional trials.

Determination of active vitamin B12 (cobalamin) in dietary supplements and ingredients by reversed-phase liquid chromatography: Single-laboratory validation.

Vitamin B12 dietary supplement can be critical to the alleviation strategies against micronutrient malnutrition and food insecurity. An HPLC-DAD method has been developed and validated, per AOAC SMPR 2016.017 (Standard Method Performance Requirements), for the quantitation of four bioactive forms of vitamin B12 (adenosylcobalamin, cyanocobalamin, hydroxocobalamin, methylcobalamin) from dietary ingredients and supplements. The method achieves chromatographic baseline resolution of vitamin B12 forms on a modern column platform without the expensive requirement of an ultra-high pressure liquid chromatography and/or mass spectrometry. The method has a wide analytical range (0.0005%w/w-85%w/w), high precision (reproducibility relative standard deviations ranged from 1.43% to 4.67%), and high accuracy (>96% spike recovery rate for 11 out of 12 accuracy testing data points). The method detection and quantification limits are less than 0.16 and 0.52 µg/mL, respectively. To our best knowledge, it is simpler, less time-consuming, and more economical than other published methods for its intended uses.

Water-soluble vitamin content of sun-dried Corinthian raisins (Vitis vinifera L., var. Apyrena).

BACKGROUND: Corinthian raisins are dried vine products, representing approximately 3% of the world dried vine fruit production. The majority of Corinthian raisin production is of Greek origin. Studies on the B-group vitamin content of Corinthian raisins produced in Greece as well as on the effect of region and cultivation altitude on the B-group vitamin content are absent in the literature. RESULTS: Corinthian raisin vitamin content was evaluated by reversed-phase high-performance liquid chromatography after acid and enzymatic hydrolysis in terms of raisin subcategory, i.e. regions of cultivation, crop-to-crop variations, and cultivation altitude. Eight vitamers from five different vitamins were identified and quantified in Corinthian raisins. Vitamin B3 (0.77-2.82 g × 10-2 × kg-1 ) was found to predominate, followed by B6 (0.27-0.37 g × 10-2 × kg-1 ), B1 (0.19-0.22 g × 10-2 × kg-1 ), and B2 (0.10-0.15 g × 10-2 × kg-1 ). B9 content was up to 7.1 g × 10-5 × kg-1 . Minor differences were observed among regions of cultivation, cultivation altitude and crop-to-crop variations. CONCLUSION: This study revealed the presence of several water-soluble vitamins in Corinthian raisins that, together with other health-promoting micronutrients present in the product, further reinforce its place as part of a healthy diet. © 2019 Society of Chemical Industry.

Chitosan nanoparticles loaded with thiamine stimulate growth and enhances protection against wilt disease in Chickpea.

Nanoencapsulation is considered as one of the unique technique for increasing the bioavailability, solubility and retention time of bioactive compounds. In this study, thiamine was incorporated into the chitosan nanoparticles and characterized through FTIR, DLS, SEM, TEM and XRD analyses. Zeta potential of the synthesized nanoparticles was found to be 37.7 mV. The encapsulation efficiency of chitosan nanoparticle was 90 ± 3%. Application of thiamine loaded chitosan nanoparticle enhanced seed germination and growth of chickpea seedlings when compared to untreated control seeds. Treated seedlings showed enhanced production of indole acetic acid (IAA). Foliar application of synthesized nanoparticle induced defense enzymes in leaves and roots of chickpea plants. Decreased cell death in the chickpea roots of treated plants was observed when compared to control under green house condition. These results showed that the thiamine loaded chitosan nanoparticle can be used as a growth stimulator as well as a defense activator in chickpea.

A Randomized Pilot Trial to Evaluate the Bioavailability of Natural versus Synthetic Vitamin B Complexes in Healthy Humans and Their Effects on Homocysteine, Oxidative Stress, and Antioxidant Levels.

The vitamin B complex comprises 8 different water-soluble constituents that humans must sequester from the diet. This pilot study compared natural versus synthetic vitamin B complexes for their bioavailability, accumulation, and their impact on antioxidants, homocysteine levels, and oxidative stress. We conducted a double-blind randomized clinical trial with thirty healthy participants. They were randomly assigned to group N (natural) and group S (synthetic). Vitamin B was ingested daily for 6 weeks in the range of about 2.5 times above the recommended daily allowance. Blood samples were taken at baseline, 1.5 h, 4 h, 7 h (diurnal), 6 w (discontinuation of supplements), and 8 w (washout). Blood levels of thiamine (B1), riboflavin (B2), pyridoxine (B6), folic acid (B9), cobalamin (B12), homocysteine, total antioxidants, peroxidase activity, polyphenols, and total peroxides were determined. Compared to initial values, serum levels of each B vitamin increased at the end of the supplementation period: i.e., B1 (+23% N; +27% S), B2 (+14% N; +13% S), B6 (+101% N; +101% S), B9 (+86% N; +153% S), and B12 (+16% N) (p < 0.05). Homocysteine (-13% N) decreased, while peroxidase activity (+41% S) and antioxidant capacity increased (+26% N). Short-term effects were already observed after 1.5 h for B9 (+238% N; +246% S) and after 4 h for vitamin B2 (+7% N; +8% S), B6 (+59% N; +51% S), and peroxidase activity (+58% N; +58% S). During the washout period, serum levels of B vitamins decreased except for thiamine and peroxidase activity, which increased further. This clinical pilot study revealed comparable bioavailability for both natural and synthetic B vitamins but did not show statistically noticeable differences between groups despite some favourable tendencies within the natural vitamin group, i.e., sustained effects for cobalamin and endogenous peroxidase activity and a decrease in homocysteine and oxidative stress levels.