RESUMEN
The aim of this study was to validate an HPLC-UV method to assess vitamin D status by determining the linearity and precision of the 25-hydroxyvitamin D3 (25(OH)D3) calibration curve, the limits of detection, quantitation and robustness of the method, and its accuracy. A second stock solution of 25(OH)D3 was prepared (500 ng/mL), and working dilutions (5, 10, 20, 30, 40, and 50 ng/mL) were prepared for a calibration curve. The HPLC equipment had a UV-Vis diode-array detector and utilized an AcclaimTM 120 C18 column (5 µm, 4.6 × 250 mm) with a flow rate of 1.2 mL/min, a column temperature of 30 °C, and the standards and samples were maintained at 4 °C, with an injection volume of 100 µL. Detection of 25(OH)D3 was determined at 265 nm, with a retention time of 4.0 min. The validation was conducted according to the FDA Validation of Analytical Procedures: Guidance for Industry. Vitamin D was extracted from plasma samples using acetonitrile (ACN)-0.1% formic acid (2:1 v/v), and the percentage of recovery was calculated. The proposed method conditions gave excellent linearity (R2 = 0.9989) and the linearity coefficient was R2 > 0.99 for 25(OH)D3. The detection and quantification limits were 1.1703 ng/mL and 3.5462 ng/mL, respectively. Decreasing or increasing the reading temperature by 1 °C decreased the response units (AU) of vitamin D, 25(OH)D3. When the current flow rate decreased by 0.2 mL/min (1.0 mL/min), the retention time increased to 4.913 min, whereas an increase of 0.2 mL/min of the proposed flow rate (1.4 mL/min) decreased the retention time to 3.500 min. The percentage of recovery varied from 92.2% to 97.1%. The proposed method to quantify a vitamin D metabolite (25(OH)D3) in human plasma samples was reliable and validated.
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Análisis Químico de la Sangre , Calcifediol , Cromatografía Líquida de Alta Presión , Análisis Químico de la Sangre/instrumentación , Análisis Químico de la Sangre/métodos , Análisis Químico de la Sangre/normas , Calcifediol/análisis , Calcifediol/sangre , Límite de Detección , Calibración , HumanosRESUMEN
Vitamin D (VD) metabolites are involved in a variety of important metabolic processes and physiological effects in organisms. Profiling of VD metabolites favors a deep understanding of the physiological role of VD. However, VD metabolites are difficult to detect due to their high chemical structural rigidity, structural similarity, and low sensitivities under liquid chromatography-tandem mass spectrometry (LC-MS). Herein, we present a chemical derivatization assisted LC-MS/MS strategy for the detection of VDs, in which 4-phenyl-1,2,4-triazoline-3,5-dione (PTAD) is employed to derivatize the conjugated diene of VD metabolites and provides sensitizing reporters for MS detection. After PTAD derivatization, the sensitivities of seven VD metabolites increased by 24-276 folds, with the limits of detection ranging from 3 to 20 pg mL-1. Using this method, we achieved a sensitive and accurate quantification of 7 VD metabolites (vitamin D2, vitamin D3, 25-hydroxyvitamin D2, 25-hydroxyvitamin D3, 1,25-dihydroxyvitamin D2, 1,25-dihydroxyvitamin D3, and 1,24,25-trihydroxyvitamin D3) of the VD metabolic pathway in different trace biological samples, including human serum, mouse tissues (namely liver, kidney, lung, and spleen), and cells. We believe that the present method can provide a promising tool for an in-depth analysis of VD metabolism.
Asunto(s)
Espectrometría de Masas en Tándem , Vitamina D , Humanos , Ratones , Animales , Cromatografía Liquida/métodos , Espectrometría de Masas en Tándem/métodos , Vitamina D/análisis , Calcifediol/análisis , ErgocalciferolesRESUMEN
Experts typically define vitamin D deficiency levels by the determination of a circulating 25-hydroxyvitamin D3-calcifediol prohormone. A large part of the population is characterized by deficient vitamin D levels (calcifediol < 20 ng mL-1) despite individuals not being affected by any disorder. Cholecalciferol (vitamin D3) and/or calcifediol supplementation is a common practice for vitamin D-deficient individuals as recommended by international scientific societies and official agencies. In the last few years, several studies have reported the presence of conjugated vitamin D3 metabolites, mainly glucuronidation and sulfation derivatives, although simultaneous quantitative measurements involving phase I and II vitamin D metabolites have not been carried out. A quantitative method based on tandem mass spectrometry detection is proposed here for the combined determination of phase I and phase II vitamin D3 metabolites in human serum. As phase I and phase II metabolites are preferentially ionized in different modes, a switching polarity mode was adopted to determine both groups of compounds in serum at high sensitivity levels (pg mL-1). The validation of this proposal was successfully accomplished by following the Center for Drug Evaluation and Research (CDER) guidelines. Its applicability was tested in a cohort of volunteers with mostly deficient baseline levels. Considering the sulfated form of calcifediol, the sum of its concentrations showed sufficient baseline vitamin D levels in all individuals, suggesting that this could be a novel strategy for vitamin D deficiency definition. Therefore, phase II metabolites are proposed to be included when evaluating the vitamin D status since they provide more information about the overall status of the vitamin D endocrine system. Nevertheless, further studies are required to confirm the biological activity of these conjugated metabolites and the suitability of this strategy for the description of vitamin D deficiency.
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Colecalciferol , Deficiencia de Vitamina D , Humanos , Colecalciferol/análisis , Calcifediol/análisis , Vitamina D , Deficiencia de Vitamina D/metabolismo , Espectrometría de Masas en Tándem/métodosRESUMEN
Australia needs accurate vitamin D food composition data to support public health initiatives. Previously, limitations in analytical methodology have precluded development of a comprehensive database. We used liquid chromatography with triple quadrupole mass spectrometry (LC-QQQ) to analyse 149 composite samples representing 98 foods (primary samples n = 896) in duplicate for vitamin D3, 25-hydroxyvitamin D3 (25(OH)D3), vitamin D2, 25(OH)D2. The greatest concentrations of vitamin D3 were found in canned salmon and a malted chocolate drink powder (fortified); chicken eggs and chicken leg meat contained the most 25(OH)D3. Margarine (fortified) and chocolate contained the greatest concentrations of vitamin D2, with smaller amounts found in various meat products. 25(OH)D2 was detected in various foods, including meats, and was quantitated in lamb liver. These data advance knowledge of dietary vitamin D in Australia and highlight the importance of analysis of these four forms of vitamin D to accurately represent the vitamin D content of food.
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Análisis de los Alimentos , Vitamina D/análisis , 25-Hidroxivitamina D 2/análisis , Australia , Calcifediol/análisis , Colecalciferol/análisis , Cromatografía Liquida , Ergocalciferoles/análisis , Espectrometría de MasasRESUMEN
Vitamin D, a secosteroid, plays an important role in several physiological processes, and its deficiency can lead to numerous pathophysiological conditions in humans. The primary objective of this study was to develop and validate the robustness of a mass spectrometry-based method capable of quantifying 25(OH)D3 for an upcoming epidemiological survey in India and to pilot test it on healthy volunteers. We first describe the development and validation of various experimental parameters that ascertain the robustness and reliability of 25-hydroxy-vitamin D3 (25(OH)D3) extractions and quantitative measurements from Dried Blood Spot (DBS) samples, where we used eight disks of 3 mm each, punched from the circular spot covering the entire circumference of the spot. Next, we conducted a pilot study, comparing 25(OH)D3 levels from serum and DBS samples from 45 participants using a protocol developed for specifically this purpose. We found that the mean 25(OH)D3 concentrations in DBS samples were comparable to the serum levels (P > 0.05). In summary, our extraction and LC-MS protocol for quantitative 25(OH)D3 measurements are robust and reproducible, and will serve as an invaluable tool for upcoming epidemiological surveys in India and perhaps around the world.
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Calcifediol/sangre , Pruebas con Sangre Seca/métodos , Adulto , Calcifediol/análisis , Cromatografía Liquida , Femenino , Voluntarios Sanos , Humanos , India , Masculino , Persona de Mediana Edad , Proyectos Piloto , Reproducibilidad de los Resultados , Encuestas y Cuestionarios , Espectrometría de Masas en Tándem , Adulto JovenRESUMEN
The vitamin D receptor is highly expressed in the gastrointestinal tract where it transacts gene expression. With current limited understanding of the interactions between the gut microbiome and vitamin D, we conduct a cross-sectional analysis of 567 older men quantifying serum vitamin D metabolites using LC-MSMS and defining stool sub-Operational Taxonomic Units from16S ribosomal RNA gene sequencing data. Faith's Phylogenetic Diversity and non-redundant covariate analyses reveal that the serum 1,25(OH)2D level explains 5% of variance in α-diversity. In ß-diversity analyses using unweighted UniFrac, 1,25(OH)2D is the strongest factor assessed, explaining 2% of variance. Random forest analyses identify 12 taxa, 11 in the phylum Firmicutes, eight of which are positively associated with either 1,25(OH)2D and/or the hormone-to-prohormone [1,25(OH)2D/25(OH)D] "activation ratio." Men with higher levels of 1,25(OH)2D and higher activation ratios, but not 25(OH)D itself, are more likely to possess butyrate producing bacteria that are associated with better gut microbial health.
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Calcifediol/análisis , Calcitriol/análisis , Microbioma Gastrointestinal/fisiología , Anciano , Anciano de 80 o más Años , Butiratos/metabolismo , Calcifediol/metabolismo , Calcitriol/metabolismo , Estudios Transversales , ADN Bacteriano/aislamiento & purificación , Heces/química , Heces/microbiología , Humanos , Vida Independiente , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiología , Masculino , Filogenia , ARN Ribosómico 16S/genéticaRESUMEN
Vitamin D deficiency in pregnant women and their offspring may result in unfavorable health outcomes for both mother and infant. A 25hydroxyvitamin D (25(OH)D) level of at least 75 nmol/L is recommended by the Endocrine Society. Validated, automated sample preparation and liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods were used to determine the vitamin D metabolites status in mother-infant pairs. Detection of 3-Epi25(OH)D3 prevented overestimation of 25(OH)D3 and misclassification of vitamin D status. Sixty-three percent of maternal 25(OH)D plasma levels were less than the recommended level of 25(OH)D at 3 months. Additionally, breastmilk levels of 25(OH)D decreased from 60.1 nmol/L to 50.0 nmol/L between six weeks and three months (p < 0.01). Furthermore, there was a positive correlation between mother and infant plasma levels (p < 0.01, r = 0.56) at 3 months. Accordingly, 31% of the infants were categorized as vitamin D deficient (25(OH)D < 50 nmol/L) compared to 25% if 3-Epi25(OH)D3 was not distinguished from 25(OH)D3. This study highlights the importance of accurate quantification of 25(OH)D. Monitoring vitamin D metabolites in infant, maternal plasma, and breastmilk may be needed to ensure adequate levels in both mother and infant in the first 6 months of infant life.
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Calcifediol/análisis , Leche Humana/química , Evaluación Nutricional , Deficiencia de Vitamina D/diagnóstico , Vitamina D/análogos & derivados , Adulto , Lactancia Materna , Calcifediol/análogos & derivados , Cromatografía Liquida , Femenino , Humanos , Lactante , Fenómenos Fisiológicos Nutricionales del Lactante , Masculino , Fenómenos Fisiologicos Nutricionales Maternos , Madres/estadística & datos numéricos , Estado Nutricional , Espectrometría de Masas en Tándem , Vitamina D/análisisRESUMEN
OBJECTIVE: To investigate the associations between serum vitamin D, calcium and uterine fibroids in a Chinese female population. METHODS: In this case-control study, adult female patients with fibroids (cases) were compared with females without fibroids (controls) in terms of serum 25-hydroxyvitamin D (25OHD) and calcium levels. RESULTS: Out of 546 total participants (mean age, 41.68 ± 5.99 years; 279 with fibroids and 267 without fibroids), only 232 had serum 25OHD levels above the sufficient threshold (>20 ng/ml). In addition, females with fibroids had lower serum 25OHD levels versus those without fibroids. The prevalence of fibroids in females with deficient (<12 ng/ml) and insufficient (12-20 ng/ml) 25OHD levels were significantly higher than in females with sufficient (>20 ng/ml) 25OHD levels. Serum calcium levels were within normal range in both groups. CONCLUSION: Hypovitaminosis D was highly prevalent among a population of Chinese females of reproductive-age, and serum 25OHD levels were lower in female patients with fibroids.
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Calcio/análisis , Leiomioma/metabolismo , Vitamina D/análisis , Adulto , Pueblo Asiatico , Calcifediol/análisis , Calcifediol/sangre , Calcio/sangre , Estudios de Casos y Controles , China/epidemiología , Femenino , Humanos , Leiomioma/epidemiología , Leiomioma/fisiopatología , Persona de Mediana Edad , Hormona Paratiroidea/sangre , Vitamina D/análogos & derivados , Vitamina D/sangre , Deficiencia de Vitamina D/sangreRESUMEN
Background Simultaneous measurement of 25(OH)D and 24,25(OH)2D is a new tool for predicting vitamin D deficiency and allows evaluating CYP24A1 lack of function. Interpretation of 24,25(OH)2D should be performed according to 25(OH)D levels and a ratio, called the vitamin D metabolite ratio (VMR) has been proposed for such a purpose. Unfortunately, the VMR can be expressed in different ways and cannot be used if 24,25(OH)2D concentrations are undetectable. Here, we propose evaluating the enzyme activity taking into consideration the probability that a normal population presents undetectable 24,25(OH)2D concentrations according to 25(OH)D levels. We thus retrospectively measured 25(OH)D and 24,25(OH)2D in a population of 1200 young subjects to evaluate the 25(OH)D threshold above which the enzyme was induced. Methods Serum samples from 1200 infants, children, adolescent and young adults were used to simultaneously quantify 25(OH)D and 24,25(OH)2D by LCMS/MS. Results Median (interquartile range [IQR]) levels were 20.6 (14.4-27.2) ng/mL for 25(OH)D. 172 subjects (14.3%) presented 24,25(OH)2D values below the LOQ. When 25(OH)D values were <11 ng/mL, 63.1% of subjects presented undetectable 24,25(OH)2D concentrations. Percentage decreased with increasing 25(OH)D values to become 19.7% for 25(OH)D comprised between 12 and 15 ng/mL, 5.1% for 25(OH)D between 16 and 20 and 0.7% for 25(OH)D >21 ng/mL. Conclusions We suggest using a statistical approach to evaluate CYP24A1 function according to 25(OH)D concentrations. Our results also show that vitamin D deficiency, as defined biochemically, could be around 20 ng/mL in infants, children, adolescent and young adults and that vitamin D deficiency could be evaluated on a more individual basis.
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24,25-Dihidroxivitamina D 3/análisis , Calcifediol/análisis , Deficiencia de Vitamina D/patología , Vitamina D3 24-Hidroxilasa/genética , Adolescente , Niño , Preescolar , Cromatografía Líquida de Alta Presión , Femenino , Humanos , Lactante , Límite de Detección , Masculino , Polimorfismo de Nucleótido Simple , Estudios Retrospectivos , Espectrometría de Masas en Tándem , Deficiencia de Vitamina D/genética , Adulto JovenRESUMEN
RATIONALE: 25-Hydroxylated vitamin D is the best marker for vitamin D (VD). Due to its low ionization efficiency, a Cookson-type reagent, 1,2,4-triazoline-3,5-dione (TAD), is used to improve the detection/quantification of VD metabolites by liquid chromatography/tandem mass spectrometry (LC/MS/MS). However, the high reactivity of TAD makes its solution stability low and inconvenient for practical use. We here describe the development of a novel caged Cookson-type reagent, and we assess its performances in the quantitative and differential detection of four VD metabolites in serum using LC/MS/MS. METHODS: Caged 4-(4'-dimethylaminophenyl)-1,2,4-triazoline-3,5-dione (DAPTAD) analogues were prepared from 4-(4'-dimethylaminophenyl)-1,2,4-triazolidine-3,5-dione. Their stability and reactivity were examined. The optimized caged DAPTAD (14-(4-(dimethylamino)phenyl)-9-phenyl-9,10-dihydro-9,10-[1,2]epitriazoloanthracene-13,15-dione, DAP-PA) was used for LC/MS/MS analyses of VD metabolites. RESULTS: The solution stability of DAP-PA in ethyl acetate dramatically improved compared with that of the non-caged one. We measured the thermal retro-Diels-Alder reaction enabling the release of DAPTAD and found that the derivatization reaction was temperature-dependent. We also determined the detection limit and the lower limit of quantifications for four VD metabolites with DAPTAD derivatization. CONCLUSIONS: DAP-PA was stable enough for mid- to long-term storage in solution. This advantage shall contribute to the detection and quantification of VD in clinical laboratories, and as such to the broader use of clinical mass spectrometry.
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Compuestos de Anilina/química , Espectrometría de Masas en Tándem/métodos , Triazoles/química , Vitamina D/sangre , Vitamina D/metabolismo , 25-Hidroxivitamina D 2/análisis , 25-Hidroxivitamina D 2/sangre , 25-Hidroxivitamina D 2/metabolismo , Compuestos de Anilina/síntesis química , Calcifediol/análisis , Calcifediol/sangre , Calcifediol/metabolismo , Cromatografía Liquida , Humanos , Indicadores y Reactivos , Límite de Detección , Triazoles/síntesis química , Vitamina D/análisisRESUMEN
Background and objectives: The purpose of the study is to correlate vascular calcification biomarkers osteoprotegerin (OPG) and 25-hydroxyvitamin D3 (25-OH-D3), indicators of arterial stiffness carotid-femoral pulse wave velocity (c-f PWV) and renal resistive index (RRI), with parameters of left ventricular function in heart failure patients versus control. Materials and methods: Our case-control study compared 60 patients with ischemic heart failure and reduced left ventricular ejection fraction (LVEF) (<40%) with a control group of 60 healthy age-matched subjects (CON). Serum levels of OPG and 25-OH-D3 were determined by ELISA. Left ventricular volumes (LVESV, LVEDV) and LVEF were measured by echocardiography. C-f PWV was determined using the arteriograph device. RRI was measured by duplex Doppler. Peak systolic velocity (PSV) and minimum end-diastolic velocity (EDV) were determined using angle correction. The estimated glomerular filtration rate (eGFR) was calculated using the MDRD equation. The Pearson's correlation coefficient was used for interpretation of results. Results: OPG values were significantly higher in heart failure (HF) patients vs. CON (4.7 ± 0.25 vs. 1.3 ± 0.67 ng/mL, p < 0.001). 25-OH vitamin D3 levels were significantly lower in HF patients vs. CON (20.49 ± 7.31 vs. 37.09 ± 4.59 ng/mL, p < 0.001). Multiple regression analysis considering 25-OH D3 as a dependent variable demonstrated indicators of vascular stiffness RRI, c-f PWV and vascular calcification biomarker OPG as predictors. OPG values were significantly correlated with cardiac parameters LVEDV (r = 0.862, p < 0.001), LVEF (r = -0.832, p < 0.001), and c-f PWV(r = 0.833, p < 0.001), and also with 25-OH-D3 (r = -0.636, p < 0.001). RRI values were significantly correlated with cardiac parameters LVEDV (r = 0.586, p < 0.001) and LVEF (r = -0.587, p < 0.001), and with eGFR (r = -0.488, p < 0.001), c-f PWV(r = 0.640, p < 0.001), and 25-OH-D3 (r = -0.732, p < 0.001). Conclusions: This study showed significant correlations between vitamin D deficit and vascular stiffness indicators in heart failure patients with reduced ejection fraction, demonstrating the importance of these examinations for a better evaluation of these patients. Together with the evaluation of renal function, the measurement of vascular stiffness indicators and biomarkers might play a key role in identifying patients at greater risk for worsening disease prognosis and for shorter life expectancy, who could benefit from vitamin D supplementation. The abstract was accepted for presentation at the Congress of the European Society of Cardiology, Munich, 2018.
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Calcifediol/análisis , Insuficiencia Cardíaca/sangre , Osteoprotegerina/análisis , Rigidez Vascular/fisiología , Anciano , Biomarcadores/sangre , Calcifediol/sangre , Calcificación Fisiológica/fisiología , Estudios de Casos y Controles , Ecocardiografía/métodos , Ensayo de Inmunoadsorción Enzimática/métodos , Femenino , Insuficiencia Cardíaca/fisiopatología , Humanos , Masculino , Persona de Mediana Edad , Osteoprotegerina/sangre , RumaníaRESUMEN
This study assessed the heritability of 25 hydroxyvitamin D3 (25(OH)D3) in a large twin cohort and the shared effect of sun exposure and skin colour on 25(OH)D3 variance. Study participants included 1604 twin pairs and their siblings (n = 4020). Twin correlations for 25(OH)D3 concentration were rMZ=0.79 (584 pairs) and rDZ = 0.52 (1020 pairs) consistent with an average h2 = 0.50 throughout the year. Significant phenotypic and genetic seasonal fluctuation was observed in 25(OH)D3 concentrations with heritability decreasing during the winter (h2 = 0.37) compared to summer (h2 = 0.62). Skin colour (measured both ordinally and quantitatively) and self-reported sun exposure were found to significantly affect 25(OH)D3 concentration. Twins with olive/dark skin had significantly lower 25(OH)D3 concentrations than those with fair/pale skin and multivariate genetic analysis showed that approximately half of the total additive genetic variation in 25(OH)D3 results from genes whose primary influence is on skin colour and sun exposure. Additionally, 37% of the total variance was attributed to shared environmental effects on vitamin D, skin colour and sun exposure measures. These results support a moderate estimate of vitamin D heritability and suggest significant influence of season, skin colour and sun exposure on the genetic variance.
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Calcifediol/genética , Pigmentación de la Piel/genética , Luz Solar/efectos adversos , 25-Hidroxivitamina D 2/análisis , 25-Hidroxivitamina D 2/sangre , 25-Hidroxivitamina D 2/metabolismo , Adolescente , Calcifediol/análisis , Calcifediol/sangre , Niño , Femenino , Variación Genética/genética , Humanos , Masculino , Pigmentación de la Piel/fisiología , Vitamina D/análisis , Vitamina D/sangre , Vitamina D/metabolismoRESUMEN
25-Hydroxyvitamin D (25-(OH)D) deficiency is recently been described as one of the multiple factors responsible for pediatric seizures. 25-Hydroxyvitamin D3 and 25-Hydroxyvitamin D2 are the well-known markers to determine Vitamin D status. In this work we report the development of a sensitive and cost effective HPLC technique for the quantification of the vitamin D metabolites from dried blood spot samples (DBS). The metabolites were extracted using acetonitrile-methanol-0.1% formic acid (60 : 20 : 20 (v/v)) and analyzed on an Acclaim C18 column (150 × 4.6 mm i.d., 3 µm) at a flow rate of 1 mL/min. The method was linear in the range of 10-80 ng/mL. Limit of detection and limit of quantification (LOQ) of the method were 5 and 10 ng/mL respectively. Extensive stability studies demonstrated the analytes to be stable in stock and matrix with a percent change within the acceptable range of ±15%. Comparison of the newly developed HPLC-DBS method with the reported LC-MS-DBS and electrochemiluminescence immunoassay (ECLIA) methods followed by Bland-Altman analysis demonstrated a bias of 0.08 and -0.14, respectively proving the methods are comparable. Application of the developed method to a pediatric seizure cohort depicted 46.6% of cases as deficient and 26.6% as insufficient for 25-(OH)D. Among deficient cases 8 samples were below 10 ng/mL and exact amount was not calculated since these were below the LOQ levels. The mean ± standard deviation (S.D.) in the remaining 6 deficient cases was 13.22 ± 2.80 ng/mL. The levels in healthy infants were 33.9 ± 6.11 ng/mL. The method can be used routinely for assessing 25-(OH)D deficiency in newborn.
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25-Hidroxivitamina D 2/análisis , Biomarcadores/sangre , Recolección de Muestras de Sangre/métodos , Calcifediol/análisis , Convulsiones/sangre , Cromatografía Líquida de Alta Presión , Humanos , Recién Nacido , Tamizaje Neonatal , Convulsiones/diagnóstico , Espectrometría de Masas en TándemRESUMEN
Vitamin D-deficiency has been linked to inflammatory diseases including rheumatoid arthritis (RA). Studies to date have focused on the impact of serum 25-hydroxyvitamin D3 (25(OH)D3), an inactive form of vitamin D, on RA disease activity and progression. However, anti-inflammatory actions of vitamin D are likely to be mediated at sites of RA disease, namely the inflamed joint, and may involve other vitamin D metabolites notably the active form of vitamin D, 1,25-dihydroxyvitamin D3 (1,25(OH)2D3). In the current study serum and synovial fluid samples from n = 20 patients with persistent RA and n = 7 patients with reactive arthritis (ReA) were analysed for multiple vitamin D metabolites. Serum data for RA and ReA patients were compared to healthy controls (HC). There was no significant difference between RA or ReA patients relative to HC for 25(OH)D3, 24,25(OH)2D3, 1,25(OH)2D3 or 25(OH)D2. However, 3-epi-25(OH)D3 was significantly lower in RA and ReA patients compared to HC (p < 0.05). All vitamin D metabolites, apart from 25(OH)D2, were lower in SF compared to serum, and SF 1,25(OH)2D3 was unquantifiable in 13/20 RA and 4/7 ReA samples. SF 25(OH)D3, 3-epi-25(OH)D3 and DBP correlated inversely with swollen joint score, and serum 25(OH)D2 and SF DBP correlated directly with C-reactive protein levels. These data indicate that serum 25(OH)D3 provides only limited insight into the role of vitamin D in RA. Alternative serum metabolites such as 3-epi-25(OH)2D3, and SF metabolites, notably lack of SF 1,25(OH)2D3, may be more closely linked to RA disease severity and progress.
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24,25-Dihidroxivitamina D 3/sangre , Artritis Reumatoide/sangre , Avitaminosis/sangre , Calcifediol/sangre , Calcitriol/sangre , Líquido Sinovial/química , 24,25-Dihidroxivitamina D 3/análisis , Adulto , Anciano , Anciano de 80 o más Años , Calcifediol/análisis , Calcitriol/análisis , Femenino , Humanos , Masculino , Persona de Mediana Edad , Prohibitinas , Adulto JovenRESUMEN
AIM: Transdermal analysis is proposed for vitamin D3 and its hydroxylated metabolite to overcome problems associated with blood analysis. METHODS: Vitamin D3 was extracted directly from skin with solid patches and liquid phases. Deuterium-labeled vitamin D3 was added to the extraction solutions to compensate for variability and accurately determine the rate of transdermal transfer. Of the different extraction solvents tested, 50:50 octanol:isopropanol showed the best results, with an accuracy of 115% and reproducibility better than 30%. CONCLUSION: The research shows that transdermal route can be used for analysis of vitamin D3 in porcine skin. When microneedles are used, accurate measurements were obtained in 1 h. With intact skin, the highest accuracy was obtained when extraction was done for 2 h.
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Métodos Analíticos de la Preparación de la Muestra/métodos , Calcifediol/análisis , Colecalciferol/análisis , Piel/química , Animales , Cromatografía Liquida , Técnicas In Vitro , Microextracción en Fase Líquida , Agujas , Reproducibilidad de los Resultados , Porcinos , Espectrometría de Masas en TándemAsunto(s)
Líquido Ascítico/química , Infecciones Bacterianas/metabolismo , Calcifediol/análisis , Cirrosis Hepática/complicaciones , Peritonitis/metabolismo , Anciano , Ascitis , Infecciones Bacterianas/diagnóstico , Calcifediol/sangre , Femenino , Humanos , Recuento de Leucocitos , Masculino , Neutrófilos , Estudios Prospectivos , Estadísticas no Paramétricas , Deficiencia de Vitamina D/diagnósticoRESUMEN
The utility of adding ion mobility (IM) to liquid chromatography/mass spectrometry (LC/MS) for quantitation of 25-hydroxyvitamin D3 (25OHD3) is evaluated. Sodiated 25OHD3 ions adopt both closed and open conformations, whereas the stereoisomer 3-epi-25-hydroxyvitamin D (epi25OHD), when sodiated, adopts only a closed gas-phase conformation. The unique open conformation for sodiated 25OHD3 permits unambiguous quantitation. Nonetheless, the ratio of open versus closed gas-phase conformations for sodiated 25OHD3 can vary with instrumental conditions; conversion from the open to the closed conformer is attributed to radio frequency (rf) heating within the ion accumulation trap. Ion heating becomes significant when space charge in the trap forces ions to larger radii where the rf field is higher. To avoid biasing quantitation, an isotopically labeled internal standard must be used to account for changes in conformer ratio. Thirty-three serum extract samples were evaluated by LC/IM-MS and were found to not be biased by changes in ion conformer ratios, permitting reliable quantitation of 25OHD3 without interference from the epimer.
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Calcifediol/análisis , Cromatografía Liquida/métodos , Espectrometría de Movilidad Iónica/métodos , Espectrometría de Masas/métodosRESUMEN
Vitamin D has previously been quantified in some plants and algae, particularly in leaves of the Solanaceae family. We measured the vitamin D content of Australian native food plants and Australian-grown edible seaweed. Using liquid chromatography with triple quadrupole mass spectrometry, 13 samples (including leaf, fruit, and seed) were analyzed in duplicate for vitamin D2, vitamin D3, 25-hydroxyvitamin D2, and 25-hydroxyvitamin D3. Five samples contained vitamin D2: raw wattleseed (Acacia victoriae) (0.03 µg/100 g dry weight (DW)); fresh and dried lemon myrtle (Backhousia citriodora) leaves (0.03 and 0.24 µg/100 g DW, respectively); and dried leaves and berries of Tasmanian mountain pepper (Tasmannia lanceolata) (0.67 and 0.05 µg/100 g DW, respectively). Fresh kombu (Lessonia corrugata) contained vitamin D3 (0.01 µg/100 g DW). Detected amounts were low; however, it is possible that exposure to ultraviolet radiation may increase the vitamin D content of plants and algae if vitamin D precursors are present.
Asunto(s)
25-Hidroxivitamina D 2/análisis , Calcifediol/análisis , Valor Nutritivo , Plantas Comestibles/química , Algas Marinas/química , Vitamina D/análisis , Australia , Cromatografía Liquida , Frutas/química , Hojas de la Planta/química , Plantas Comestibles/crecimiento & desarrollo , Algas Marinas/crecimiento & desarrollo , Semillas/química , Espectrometría de Masas en TándemRESUMEN
This study investigated the effects of synthetic and natural sources of vitamin D biofortification in pig diets on pork vitamin D activity and pork quality. One hundred and twenty pigs (60 male, 60 female) were assigned to one of four dietary treatments for a 55â¯d feeding period. The dietary treatments were (1)50⯵g vitamin D3/kg of feed; (2)50⯵g of 25-hydroxvitamin D3/kg of feed (25-OH-D3); (3)50⯵g vitamin D2/kg of feed; (4)50⯵g vitamin D2-enriched mushrooms/kg of feed (Mushroom D2). The pigs offered the 25-OH-D3 diet exhibited the highest (Pâ¯<â¯0.001) serum total 25-hydroxyvitamin D concentration and subsequently exhibited the highest (Pâ¯<â¯0.05) Longissimus thoracis (LT) total vitamin D activity. Mushroom D2 and 25-OH-D3 supplementation increased pork antioxidant status. The vitamin D2-enriched mushrooms improved (Pâ¯<â¯0.05) pig performance, carcass weight and LT colour. In conclusion, 25-OH-D3 is the most successful source for increasing pork vitamin D activity, while Mushroom D2 may be a new avenue to improve animal performance and pork quality.
Asunto(s)
Agaricales/química , Fenómenos Fisiológicos Nutricionales de los Animales , Antioxidantes/administración & dosificación , Calcifediol/administración & dosificación , Calidad de los Alimentos , Carne/análisis , Músculo Esquelético/metabolismo , 25-Hidroxivitamina D 2/sangre , Agaricales/crecimiento & desarrollo , Agaricales/metabolismo , Animales , Antioxidantes/análisis , Antioxidantes/metabolismo , Calcifediol/análisis , Calcifediol/sangre , Calcifediol/metabolismo , Colecalciferol/administración & dosificación , Colecalciferol/análisis , Colecalciferol/metabolismo , Cruzamientos Genéticos , Ergocalciferoles/administración & dosificación , Ergocalciferoles/análisis , Ergocalciferoles/metabolismo , Femenino , Alimentos Fortificados/análisis , Humanos , Irlanda , Masculino , Músculo Esquelético/crecimiento & desarrollo , Valor Nutritivo , Pigmentos Biológicos/análisis , Pigmentos Biológicos/biosíntesis , Distribución Aleatoria , Sus scrofa , Aumento de PesoRESUMEN
This study investigates dietary fortification of heifer feeds with cholecalciferol and ergocalciferol sources and effects on beef total vitamin D activity, vitamer, respective 25-hydroxymetabolite contents, and meat quality. Thirty heifers were allocated to one of three dietary treatments [(1) basal dietâ¯+â¯4000â¯IU of vitamin D3 (Vit D3); (2) basal dietâ¯+â¯4000â¯IU of vitamin D2 (Vit D2); and (3) basal dietâ¯+â¯4000â¯IU of vitamin D2-enriched mushrooms (Mushroom D2)] for a 30â¯day pre-slaughter period. Supplementation of heifer diets with Vit D3 yielded higher (pâ¯<â¯0.001) Longissimus thoracis (LT) total vitamin D activity (by 38-56%; pâ¯<â¯0.05) and serum 25-OH-D concentration (by 20-36%; pâ¯<â¯0.05), compared to that from Vit D2 and Mushroom D2 supplemented animals. Irrespective of vitamin D source, carcass characteristics, sensory and meat quality parameter were unaffected (pâ¯>â¯0.05) by the dietary treatments. In conclusion, vitamin D3 biofortification of cattle diets is the most efficacious way to enhance total beef vitamin D activity.