Content and Bioaccessibility of Minerals and Proteins in Fish-Bone Containing Side-Streams from Seafood Industries.

With the aim to upcycle fish side-streams, enzymatic hydrolysis is often applied to produce protein hydrolysates with bioactive properties or just as a protein source for food and feed. However, the production of hydrolysates generates a side-stream. For underutilized fish and fish backbone this side-stream will contain fish bones and make it rich in minerals. The aim of this study was to assess the relative bioaccessibility (using the standardized in vitro model INFOGEST 2.0) of minerals in a dietary supplement compared to bone powder generated after enzymatic hydrolysis of three different fish side-streams: undersized whole hake, cod and salmon backbones consisting of insoluble protein and bones. Differences in the bioaccessibility of protein between the powders were also investigated. The enzyme hydrolysis was carried out using different enzymes and hydrolysis conditions for the different fish side-streams. The content and bioaccessibility of protein and the minerals phosphorus (P), calcium (Ca), potassium (K) and magnesium (Mg) were measured to evaluate the potential of the powder as an ingredient in, e.g., dietary supplements. The bone powders contained bioaccessible proteins and minerals. Thus, new side-streams generated from enzymatic hydrolysis can have possible applications in the food sector due to bioaccessible proteins and minerals.

Across-country genomic predictions in Norwegian and New Zealand Composite sheep populations with similar development history.

The goal of this study was to assess the feasibility of across-country genomic predictions in Norwegian White Sheep (NWS) and New Zealand Composite (NZC) sheep populations with similar development history. Different training populations were evaluated (i.e., including only NWS or NZC, or combining both populations). Predictions were performed using the actual phenotypes (normalized) and the single-step GBLUP via Bayesian inference. Genotyped NWS animals born in 2016 (N = 267) were used to assess the accuracy and bias of genomic estimated breeding values (GEBVs) predicted for birth weight (BW), weaning weight (WW), carcass weight (CW), EUROP carcass classification (EUC), and EUROP fat grading (EUF). The accuracy and bias of GEBVs differed across traits and training population used. For instance, the GEBV accuracies ranged from 0.13 (BW) to 0.44 (EUC) for GEBVs predicted including only NWS, from 0.06 (BW) to 0.15 (CW) when including only NZC, and from 0.10 (BW) to 0.41 (EUC) when including both NWS and NZC animals in the training population. The regression coefficients used to assess the spread of GEBVs (bias) ranged from 0.26 (BW) to 0.64 (EUF) for only NWS, 0.10 (EUC) to 0.52 (CW) for only NZC, and from 0.42 (WW) to 2.23 (EUC) for both NWS and NZC in the training population. Our findings suggest that across-country genomic predictions based on ssGBLUP might be possible for NWS and NZC, especially for novel traits.

Vitamin D-fortified foods improve wintertime vitamin D status in women of Danish and Pakistani origin living in Denmark: a randomized controlled trial.

PURPOSE: Low vitamin D status is prevalent worldwide. We aim to investigate the effect of vitamin D fortification on serum 25-hydroxyvitamin D (25(OH)D) concentration in women of Danish and Pakistani origin at risk of vitamin D deficiency. METHODS: A 12-week randomized, double-blinded, placebo-controlled intervention trial during winter time, designed to provide 20 µg vitamin D3/day through fortified yoghurt, cheese, eggs and crisp bread, and assess the change in serum 25(OH)D. Participants were 143 women of Danish and Pakistani origin, living in Denmark, randomized into four groups, stratified by ethnicity. RESULTS: Mean (SD) baseline 25(OH)D concentrations among women of Danish and Pakistani origin were 49.6 (18) and 46.9 (22) nmol/L, respectively (P = 0.4). While 9% of Danish women had 25(OH)D < 30 nmol/L, the prevalence among women of Pakistani origin was 24%. Median (IQR) vitamin D intake among Danish and Pakistani women at endpoint was 32.0 (27.0, 34.4) µg/day and 24.2 (19.2, 30.8) µg/day, respectively. Endpoint serum 25(OH)D increased in fortified groups to 77.8 (14) nmol/L among Danish women and 54.7 (18) nmol/L among women of Pakistani origin (P < 0.01). At endpoint, 0% in the Danish-fortified group and 3% in the Pakistani-fortified group had 25(OH)D < 30 nmol/L, compared with 23 % and 34% in their respective control groups. CONCLUSIONS: Vitamin D fortification of four different foods for 12 weeks during winter was effective in increasing serum 25(OH)D and reducing the prevalence of very low vitamin D status among women of Danish and Pakistani origin. CLINICALTRIALS. GOV WITH IDENTIFIER: NCT02631629.

Is high oily fish intake achievable and how does it affect nutrient status in 8-9-year-old children?: the FiSK Junior trial.

PURPOSE: Most children do not meet dietary guidelines for fish intake. Fish is the main source of EPA (20:5n-3), DHA (22:6n-3) and vitamin D, but may replace better iron sources such as meat. We investigated if intake of 300 g/week oily fish was achievable in children and how it affected their nutrient status. Additionally, we validated a fish food frequency questionnaire (FFQ) by correlations against EPA + DHA in red blood cells (RBC). METHODS: In a randomised 12-week trial, 199 children (8-9 years) received oily fish or poultry (control) to be eaten five times/week. We measured dietary intake and analysed fasting RBC EPA + DHA, serum 25-hydroxyvitamin D (25(OH)D), blood haemoglobin and plasma ferritin. RESULTS: 197 (99%) children completed the study. The median (25th-75th percentile) intake was 375 (325-426) and 400 (359-452) g/week oily fish and poultry, respectively. The fish group increased their intake of EPA + DHA by 749 (593-891) mg/day and vitamin D by 3.1 (1.6-3.8) µg/day. Endpoint RBC EPA + DHA was 2.3 (95% CI 1.9; 2.6) fatty acid %-point higher than the poultry group (P < 0.001). The fish group avoided the expected 25(OH)D winter decline (P < 0.001) and had 23%-point less vitamin D insufficiency (winter subgroup, n = 82). Haemoglobin and ferritin decreased slightly in both groups (P < 0.05), but the number of children with low values did not change (P > 0.14). FFQ estimates moderately reflected habitual intake (r = 0.28-0.35) and sufficiently captured intervention-introduced changes in intake (r > 0.65). CONCLUSION: Oily fish intake of 300 g/week was achievable and improved children's EPA + DHA and 25(OH)D status, without markedly compromising iron status. These results justify public health initiatives focusing on children's fish intake.

Vitamin D Status and Muscle Function Among Adolescent and Young Swimmers.

Impaired muscle function has been coupled to vitamin D insufficiency in young women and in elderly men and women. Those living at Northern latitudes are at risk for vitamin D insufficiency due to low sun exposure which may be more pronounced among elite swimmers because of their indoor training schedules. We aimed to examine vitamin D status among young elite swimmers and evaluate the association between vitamin D status and muscle strength. Twenty-nine swimmers, 12 female and 17 male (16-24 years) residing at latitude 55-56°N were studied in March and April. Blood samples were analyzed for serum 25-hydroxyvitamin D (s-25(OH)D) and hand-grip strength was measured as marker of muscle strength. Subjects´ vitamin D and calcium intake were assessed by food frequency questionnaire and sun exposure and training status by questionnaires. Mean (± SD) s-25(OH)D was 52.6 ± 18.3nmol/L among all swimmers. In 45% of the swimmers s-25(OH)D was below 50 nmol/L. Female swimmers had higher s-25(OH)D concentration than male swimmers (61.7 ± 17,5 nmol/L vs. 46.2 ± 16,5 nmol/L, p = .026). Among male swimmers, those with sufficient vitamin D status had higher hand grip strength than those with insufficient vitamin D status (50.6 ± 6.4 kg vs. 41.1 ± 7.8 kg, p = .02). Among Danish elite swimmers 45% had an insufficient vitamin D status during the spring; the prevalence being higher among male swimmers. Muscle strength was significantly higher in male swimmers with sufficient vitamin D status.

Effects of vitamin D2-fortified bread v. supplementation with vitamin D2 or D3 on serum 25-hydroxyvitamin D metabolites: an 8-week randomised-controlled trial in young adult Finnish women.

There is a need for food-based solutions for preventing vitamin D deficiency. Vitamin D3 (D3) is mainly used in fortified food products, although the production of vitamin D2 (D2) is more cost-effective, and thus may hold opportunities. We investigated the bioavailability of D2 from UV-irradiated yeast present in bread in an 8-week randomised-controlled trial in healthy 20-37-year-old women (n 33) in Helsinki (60°N) during winter (February-April) 2014. Four study groups were given different study products (placebo pill and regular bread=0 µg D2 or D3/d; D2 supplement and regular bread=25 µg D2/d; D3 supplement and regular bread=25 µg D3/d; and placebo pill and D2-biofortified bread=25 µg D2/d). Serum 25-hydroxyvitamin D2 (S-25(OH)D2) and serum 25-hydroxyvitamin D3 (S-25(OH)D3) concentrations were measured at baseline, midpoint and end point. The mean baseline total serum 25-hydroxyvitamin D (S-25(OH)D=S-25(OH)D2+S-25(OH)D3) concentration was 65·1 nmol/l. In repeated-measures ANCOVA (adjusted for baseline S-25(OH)D as total/D2/D3), D2-bread did not affect total S-25(OH)D (P=0·707) or S-25(OH)D3 (P=0·490), but increased S-25(OH)D2 compared with placebo (P<0·001). However, the D2 supplement was more effective than bread in increasing S-25(OH)D2 (P<0·001). Both D2 and D3 supplementation increased total S-25(OH)D compared with placebo (P=0·030 and P=0·001, respectively), but D2 supplementation resulted in lower S-25(OH)D3 (P<0·001). Thus, D2 from UV-irradiated yeast in bread was not bioavailable in humans. Our results support the evidence that D2 is less potent in increasing total S-25(OH)D concentrations than D3, also indicating a decrease in the percentage contribution of S-25(OH)D3 to the total vitamin D pool.

Impact on Vitamin D2, Vitamin D4 and Agaritine in Agaricus bisporus Mushrooms after Artificial and Natural Solar UV Light Exposure.

Commercial mushroom production can expose mushrooms post-harvest to UV light for purposes of vitamin D2 enrichment by converting the naturally occurring provitamin D2 (ergosterol). The objectives of the present study were to artificially simulate solar UV-B doses occurring naturally in Central Europe and to investigate vitamin D2 and vitamin D4 production in sliced Agaricus bisporus (button mushrooms) and to analyse and compare the agaritine content of naturally and artificially UV-irradiated mushrooms. Agaritine was measured for safety aspects even though there is no rationale for a link between UV light exposure and agaritine content. The artificial UV-B dose of 0.53 J/cm(2) raised the vitamin D2 content to significantly (P < 0.001) higher levels of 67.1 ± 9.9 µg/g dry weight (DW) than sun exposure (3.9 ± 0.8 µg/g dry DW). We observed a positive correlation between vitamin D4 and vitamin D2 production (r(2) = 0.96, P < 0.001) after artificial UV irradiation, with vitamin D4 levels ranging from 0 to 20.9 µg/g DW. The agaritine content varied widely but remained within normal ranges in all samples. Irrespective of the irradiation source, agaritine dropped dramatically in conjunction with all UV-B doses both artificial and natural solar, probably due to its known instability. The biological action of vitamin D from UV-exposed mushrooms reflects the activity of these two major vitamin D analogues (D2, D4). Vitamin D4 should be analysed and agaritine disregarded in future studies of UV-exposed mushrooms.

Short communication: artificial ultraviolet B light exposure increases vitamin D levels in cow plasma and milk.

The number of dairy cows without access to pasture or sunlight is increasing; therefore, the content of vitamin D in dairy products is decreasing. Ultimately, declining vitamin D levels in dairy products will mean that dairy products are a negligible source of natural vitamin D for humans. We tested the ability of a specially designed UVB lamp to enhance the vitamin D3 content in milk from dairy cows housed indoors. This study included 16 cows divided into 4 groups. Each group was exposed daily to artificial UVB light simulating 1, 2, 3, or 4 h of summer sun at 56°N for 24 d, and the group with simulated exposure to 2 h of summer sun daily continued to be monitored for 73 d. We found a significant increase in 25-hydroxyvitamin D3 (25OHD3) levels in plasma as well as vitamin D3 and 25OHD3 levels in milk after daily exposure for 24 d in all treatment groups. Extending daily exposure to artificial UVB light to 73 d did not lead to an increase of vitamin D3 or 25OHD3 level in the milk. In conclusion, the change in production facilities for dairy cows providing cows with no access to pasture and sunlight causes a decrease of vitamin D levels in dairy products. This decrease may be prevented by exposing cows to artificial UVB light in the stable.

Quantification of physiological levels of vitamin D3 and 25-hydroxyvitamin D3 in porcine fat and liver in subgram sample sizes.

Most methods for the quantification of physiological levels of vitamin D3 and 25-hydroxyvitamin D3 are developed for food analysis where the sample size is not usually a critical parameter. In contrast, in life science studies sample sizes are often limited. A very sensitive liquid chromatography with tandem mass spectrometry method was developed to quantify vitamin D3 and 25-hydroxyvitamin D3 simultaneously in porcine tissues. A sample of 0.2-1 g was saponified followed by liquid-liquid extraction and normal-phase solid-phase extraction. The analytes were derivatized with 4-phenyl-1,2,4-triazoline-3,5-dione to improve the ionization efficiency by electrospray ionization. The method was validated in porcine liver and adipose tissue, and the accuracy was determined to be 72-97% for vitamin D3 and 91-124% for 25-hydroxyvitamin D3 . The limit of quantification was <0.1 ng/g, and the precision varied between 1.4 and 16% depending on the level of spiking. The small sample size required for the described method enables quantification of vitamin D3 and 25-hydroxyvitamin D3 in tissues from studies where sample sizes are limited.

Simultaneous quantification of vitamin D3, 25-hydroxyvitamin D3 and 24,25-dihydroxyvitamin D3 in human serum by LC-MS/MS.

INTRODUCTION: Serum 25-hydroxy-vitamin D is the established biomarker of vitamin D status although serum concentrations of vitamin D and 24,25-dihydroxyvitamin D may also be of interest to understand the in vivo kinetics of serum 25-hydroxyvitamin D. METHOD: An LC-MS/MS method was developed and validated to quantify vitamin D3, 25-hydroxyvitamin D3 and 24,25-dihydroxyvitamin D3 in serum. After protein precipitation of the serum it was loaded on a HybridSPE column to separate vitamin D metabolites from phospholipids. Vitamin D3, 25-hydroxyvitamin D3 and 24,25-dihydroxyvitamin D3 in the eluate were derivatized by 4-phenyl-1,2,4-triazoline-3,5-dione to improve sensitivity in the following LC-MS/MS analysis. RESULTS: Using only 100 µL serum the limit of quantification was < 0.2 ng/mL for vitamin D3, 25-hydroxyvitamin D3 and 24,25-dihydroxyvitamin D3. The method was validated up to 100 ng/mL (260 nmol/L) for vitamin D3, up to 100 ng/mL (240 nmol/L) for 24,25-dihydroxyvitamin D3 and up to 200 ng/mL (499 nmol/L) for 25-hydroxyvitamin D3. Precision was < 6.5% for vitamin D3 and 25-hydroxyvitamin D3 and < 10.2% for 24,25-dihydroxyvitamin D3. CONCLUSION: We demonstrate that a method including not only serum 25-hydroxyvitamin D3 but also vitamin D3 and 24,25-dihydroxyvitamin D3 could easily be implemented in most modern biochemical laboratories. The method could be used to study the metabolism of endogenous synthesized vitamin D3 as well as vitamin D3 in intervention studies.

Altered vitamin D status in liver tissue and blood plasma from Greenland sledge dogs (Canis familiaris) dietary exposed to organohalogen contaminated minke whale (Balaenoptera acuterostrata) blubber.

This study compared vitamin D3 (vitD3) and 25-OH vitamin D3 (25OHD3) status in Greenland sledge dogs (Canis familiaris) given either minke whale (Balaenoptera acuterostrata) blubber high in organohalogen contaminants (OHCs) or clean porcine (Suis scrofa) fat for up to 636 days. A group of six exposed and six control sister bitches (maternal generation) and their three exposed and four control pups, respectively, were daily fed 112g whale blubber (193µg ∑PCB/day) or porcine fat (0.17µg ∑PCB/day). Mean level of ∑PCB in adipose tissue of exposed bitches and their pups was 3106 and 2670ng/g lw, respectively, which was significantly higher than the mean concentration of 53ng/g lw for all controls (p<0.001). The vitamin analyses showed that 25OHD3 in liver of maternal exposed bitches were significantly lower than in controls (p=0.004) while vitD3 was significantly highest in liver of exposed pups (p<0.003). Regarding blood plasma concentrations, exposed F generation pups had significantly higher concentrations of 25OHD3 than controls (p=0.009). Correlation analyses showed that blood 25OHD3 decreased significantly with increased adipose tissue concentrations of ∑PCB in exposed dogs (R(2)=0.64, p=0.005) and a similar trend was found for liver 25OHD3 (R(2)=0.32, p=0.08). The results indicate that the homeostasis and metabolism of vitamin D compounds may respond differently to the dietary composition of fatty acids and OHC exposure. It is unknown if the lower level of 25OHD3 in the liver of exposed dogs would have any negative effects on immunity and reproduction and more focus should be conducted on this compound in Arctic wildlife.

Variation in nutritional quality in UK retail eggs.

Fatty acid (FA), carotenoid and vitamin contents of UK eggs were assessed for four production systems (caged (CA), free-range (FR), organic (OR) and extensive organic (EO)) as well as season. The impact of enforced housing, due to avian influenza, was also investigated. Production system did not alter vitamin D3, B2 or B9 content, but significantly influenced nutritionally desirable FA, carotenoid and vitamins A and E - concentrations decreased as production intensity increased, although for most, CA and FR did not differ significantly. Vitamin E and FA profiles for OR and EO were also similar, although carotenoids were higher in EO eggs. In contrast, FA, carotenoids, vitamins E and B9 were consistent throughout the year, unlike vitamins A, D3 and B2, which fluctuated with season; D and B2 were higher in July than January and lower vitamin A was the only detected implication from enforced housing of FR and OR birds.

Limitations of current analytical reference methods to determine vitamins in foods: Challenges to support regulatory compliance and nutritional composition data.

Foods are analysed for their vitamin content to support the verification of regulatory compliance or to generate food composition data. Many international reference methods for the analysis of vitamins in foods originate from the 1990s. Advances in nutrition science and analytical technology and the continuing evolution of statutory regulations necessitate the need of new or supplementary regulatory standards. We have evaluated recent developments in these areas and conclude that most current international reference methods are no longer fit-for-purpose to accurately determine vitamin content in foods and food supplements. We have made recommendations to consider new and/or updated reference methods and regulatory standards for the analysis of vitamins A, D, E, K, B1, B2, B3, B5, B6, B7, B9, B12, C and carotenoids in foods and food supplements. This area of nutrients may benefit from globally harmonised definitions specifying what compounds to include or exclude for analysis, and applicable bioactivity factors.

Effect of household cooking on the retention of vitamin D2 and 25-hydroxyvitamin D2 in pulse UV-irradiated, air-dried button mushrooms (Agaricus bisporus).

Vitamin D deficiency has widespread global prevalence. Fresh mushrooms exposed to ultraviolet (UV) radiation generate vitamin D2 which remains after drying. It is not clear if vitamin D2 is retained after rehydration and cooking of dried mushrooms. The aim of this study was to determine the true retention of both vitamin D2 and 25-hydroxyvitamin D2 (25(OH)D2) after cooking UV-irradiated, air-dried, then rehydrated button mushrooms (Agaricus bisporus). Mushrooms were exposed to pulsed UV radiation, then air-dried in a convection oven, followed by rehydration in warm water. Samples were cooked in three different ways: frying (5 min), baking (10 min, 200 °C) and boiling (20 min, 90 °C). Compared to rehydrated, uncooked controls, there was a high retention of D vitamers (≥95%) after cooking. Frying and baking resulted in significantly higher vitamin D2 retention compared to boiling (p < 0.0001). UV-irradiated, dried mushrooms are a valuable source of vitamin D2 after rehydration and cooking.

Evaluation of flavonoids and enterolactone in overnight urine as intake biomarkers of fruits, vegetables and beverages in the Inter99 cohort study using the method of triads.

Since collection of 24 h urine samples is very time consuming and difficult to obtain, epidemiological studies typically only obtain spot urine samples. The aim of the present study was to evaluate whether flavonoids and enterolactone in overnight urine could substitute flavonoids and enterolactone in 24 h urine as an alternative and more feasible biomarker of fruit, vegetable and beverage intake. A total of 191 individuals in the Inter99 cohort in Denmark completed the validation study. Concentrations of nine urinary flavonoid aglycones (quercetin, isorhamnetin, tamarixetin, kaempferol, hesperetin, naringenin, eriodictyol, phloretin and apigenin) and enterolactone were determined in overnight and 24 h urine samples, and their validity as biomarkers of fruit, vegetable and beverage intake was evaluated in relation to two independent reference methods (Inter99 FFQ data and plasma carotenoids) by using the method of triads. The intakes of fruit, juice, vegetables and tea reported in the FFQ were reflected by the flavonoid biomarker both in overnight and 24 h urine samples. Validity coefficients for the flavonoid biomarker in overnight urine ranged from 0·39 to 0·49, while the corresponding validity coefficients for the biomarker in 24 h urine ranged from 0·43 to 0·66. Although the validity coefficients were lower for overnight urine than for the 24 h urine flavonoid biomarker, they were still of acceptable magnitude. In conclusion, the results indicate that flavonoids and enterolactone in overnight urine samples may be used as a more feasible biomarker than 24 h urine for the assessment and validation of fruit, juice, vegetable and tea intakes in epidemiological studies.

In vitro bioaccessibility of vitamin K (phylloquinone and menaquinones) in food and supplements assessed by INFOGEST 2.0 - vit K.

Vitamin K describes a group of fat-soluble vitamers namely phylloquinone and menaquinones. The growing evidence for vitamin K's role beyond blood coagulation, and the possible differences between the vitamers are emerging. Knowledge of the content of menaquinones in different food matrixes and the potential differences in bioaccessibility between the vitamin K vitamers and food matrixes are limited. In this study, the bioaccessibility was assessed using the INFOGEST 2.0 static in vitro digestion model optimised by including a Danish standard meal. The presence of the standard meal was crucial to obtaining a robust and stable digestion model. The bioaccessibility of the Danish standard meal, water, vitamin K standards, vitamin K supplements, broccoli, spinach, natto, pasteurised whole egg and canola oil was assessed by three replications. The bioaccessibility was in the range 30%-102%. The lowest bioaccessibility was observed in broccoli while the highest bioaccessibility was found in egg and canola oil. No competition in the bioaccessibility between vitamin K vitamers and vitamin D was observed.

Vitamin D Levels in Sows from Five Danish Outdoor Herds.

Vitamin D is essential for sow health and productivity. Standard sow feed is therefore supplemented with vitamin D3 or 25-hydroxyvitamin D3 (25(OH)D3). However, it is uncertain whether the levels achieved are adequate for optimal performance. Currently, information on serum levels of vitamin D in pigs reared under both indoor and outdoor conditions is lacking. In August 2020, we obtained blood samples from 97 organic newly weaned sows housed outdoors during pregnancy and farrowing and used these to test for vitamin D in serum. The average concentration was 67 ± 16 ng 25(OH)D3/mL with a range of 32 to 134 ng 25(OH)D3/mL. The vitamin D3 content was 21 ± 7 ng/mL, ranging from 9 to 48 ng/mL. The average number of hours of sun from June to August was 7.0 ± 0.5 h/day. Parity, farm and body condition score did not significantly affect serum levels of 25(OH)D3.

Vitamin K content of cheese, yoghurt and meat products in Australia.

Vitamin K is vital for normal blood coagulation, and may influence bone, neurological and vascular health. Data on the vitamin K content of Australian foods are limited, preventing estimation of vitamin K intakes in the Australian population. We measured phylloquinone (PK) and menaquinone (MK) -4 to -10 in cheese, yoghurt and meat products (48 composite samples from 288 primary samples) by liquid chromatography with electrospray ionisation-tandem mass spectrometry. At least one K vitamer was found in every sample. The greatest mean (± standard deviation for foods sampled in multiple cities) concentrations of PK (4.9 µg/100 g), MK-4 (58 ± 9 µg/100 g) and MK-9 (8 ± 2 µg/100 g) were found in lamb liver, chicken leg meat and Cheddar cheese, respectively. Cheddar cheese (1.1 ± 0.3 µg/100 g) and cream cheese (1.0 µg/100 g) contained MK-5. MK-8 was found in Cheddar cheese only (4 ± 2 µg/100 g). As the K vitamer profile and concentrations appear to vary considerably by geographical location, Australia needs a vitamin K food composition dataset that is representative of foods consumed in Australia.

Vitamin K (phylloquinone and menaquinones) in foods - Cost-effective quantification by LC-ESI-MS/MS.

Further research on vitamin K is necessary as growing evidence of vitamin K's importance in human health beyond blood coagulation and bone health is emerging. We present a cost-effective LC-ESI-MS/MS method for quantification of phylloquinone (PK), and menaquinones (MK) 4-10 in food using deuterium labelled (d7) compounds (d7-PK, d7-MK-4, d7-MK-7 and d7-MK-9) as internal standards. The validation of the method included assessment of matrix effect, limit of quantification (LOQ), precision, and trueness. The LC-ESI-MS/MS method runtime is 9 min. The method was compared to a validated LC-FLD method (CEN 14148), for quantification of vitamin K in broccoli, cheese, natto, liver, and microalgae. LOQs of the LC-ESI-MS/MS method were ≤4 µg/100 g food. The intra- and inter-assay precision was <15% for PK, MK-4, MK-7 and MK-9; <20% for MK-5, MK-8, and MK-10, and ≤25% for MK-6. No significant differences between the quantified content by the LC-ESI-MS/MS and LC-FLD methods were observed.

High Oral Vitamin D3 Intake Does Not Protect Against UVR-induced Squamous Cell Carcinoma in Mice.

BACKGROUND/AIM: The effect of vitamin D on skin carcinogenesis is unclear. Vitamin D derivatives may protect against ultraviolet radiation (UVR)-induced DNA damage, immune suppression, and skin carcinogenesis. However, some epidemiological studies have reported an increased incidence of skin cancer associated with high serum vitamin D levels. We investigated the effect of vitamin D supplementation on serum, skin, and tumor vitamin D levels and on skin cancer development in hairless immunocompetent mice. MATERIALS AND METHODS: Female C3.Cg-Hrhr/TifBomTac immunocompetent mice (n=125) were randomly separated into five groups. Two groups received a high vitamin D3 diet (4.5 µg/day/mouse). One group received a medium vitamin D3 diet (2.3 µg/day/mouse). Two groups received a standard diet (0.045 µg/day/mouse). Three standard erythema doses of UVR were given three times per week to three groups. RESULTS: Animals on a high vitamin D3 diet had ~150-fold higher serum vitamin D3 levels (p=0.00016) and 3-fold higher serum 25-hydroxyvitamin D3 [25(OH)D3] levels (p=0.00016) than those on a standard diet. For mice on the medium vitamin D3 diet, serum vitamin D3 and 25(OH)D3 levels were 18-fold and 2.3-fold higher than for the standard diet, respectively (p=0.00016). All UVR-exposed mice developed tumors. Vitamin D3 levels were lower in the tumor than the skin (p<0.0001). High and medium supplementation with vitamin D3 did not affect tumor development (p>0.05). CONCLUSION: In mice, vitamin D levels in the serum, skin, and tumors were augmented by supplementation, but this did not affect the development of UVR-induced skin tumors.