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Article ; Online: The vitamin D metabolome: An update on analysis and function.

Jenkinson, Carl

2019 Volume 37, Issue 6, Page(s) 408–423

Abstract: Current understanding of vitamin D tends to be focussed on the measurement of the major circulating form 25-hydroxyvitamin D3 (25OHD3) and its conversion to the active hormonal form, 1α,25-dihydroxyvitamin D3 (1α,25(OH) ...

Abstract	Current understanding of vitamin D tends to be focussed on the measurement of the major circulating form 25-hydroxyvitamin D3 (25OHD3) and its conversion to the active hormonal form, 1α,25-dihydroxyvitamin D3 (1α,25(OH)
MeSH term(s)	Animals ; Humans ; Metabolome/physiology ; Vitamin D/analysis ; Vitamin D/metabolism
Chemical Substances	Vitamin D (1406-16-2)
Language	English
Publishing date	2019-07-22
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	283643-9
ISSN	1099-0844 ; 0263-6484
ISSN (online)	1099-0844
ISSN	0263-6484
DOI	10.1002/cbf.3421
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Article: Severe Hypercalcemia in Pregnancy Presenting a Diagnostic Conundrum.

Croker, Emma E / Jenkinson, Carl / Ruddell, Richard / Wynne, Katie

The journal of applied laboratory medicine

2023 Volume 8, Issue 5, Page(s) 984–989

MeSH term(s)	Pregnancy ; Female ; Humans ; Hypercalcemia/diagnosis ; Hypercalcemia/etiology
Language	English
Publishing date	2023-06-20
Publishing country	England
Document type	Case Reports
ISSN	2576-9456
ISSN	2576-9456
DOI	10.1093/jalm/jfad030
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Article: Low serum 1,25(OH)2D3 in end-stage renal disease: is reduced 1α-hydroxylase the only problem?

Huish, Sharon A / Jenkinson, Carl / Dunn, Janet A / Meredith, David J / Bland, Rosemary / Hewison, Martin

Endocrine connections

2021 Volume 10, Issue 10, Page(s) 1291–1298

Abstract: Low serum 1,25-dihydroxyvitamin D (1,25(OH)2D) in end-stage renal disease (ESRD) is considered a consequence of elevated fibroblast growth factor 23 (FGF23) and concomitant reduced activity of renal 1α-hydroxylase (CYP27B1). Current ESRD treatment ... ...

Abstract	Low serum 1,25-dihydroxyvitamin D (1,25(OH)2D) in end-stage renal disease (ESRD) is considered a consequence of elevated fibroblast growth factor 23 (FGF23) and concomitant reduced activity of renal 1α-hydroxylase (CYP27B1). Current ESRD treatment strategies to increase serum calcium and suppress secondary hyperparathyroidism involve supplementation with vitamin D analogues that circumvent 1α-hydroxylase. This overlooks the potential importance of 25-hydroxyvitamin D (25(OH)D) deficiency as a contributor to low serum 1,25(OH)2D. We investigated the effects of vitamin D (cholecalciferol) supplementation (40,000 IU for 12 weeks and maintenance dose of 20,000 IU fortnightly), on multiple serum vitamin D metabolites (25(OH)D, 1,25(OH)2D3 and 24,25(OH)2D3) in 55 haemodialysis patients. Baseline and 12 month data were compared using related-samples Wilcoxon signed rank test. All patients remained on active vitamin D analogues as part of routine ESRD care. 1,25(OH)2D3 levels were low at baseline (normal range: 60-120 pmol/L). Cholecalciferol supplementation normalised both serum 25(OH)D and 1,25(OH)2D3. Median serum 25(OH)D increased from 35.1 nmol/L (IQR: 23.0-47.5 nmol/L) to 119.9 nmol/L (IQR: 99.5-143.3 nmol/L) (P < 0.001). Median serum 1,25(OH)2D3 and 24,25(OH)2D3 increased from 48.3 pmol/L (IQR: 35.9-57.9 pmol/L) and 3.8 nmol/L (IQR: 2.3-6.0 nmol/L) to 96.2 pmol/L (IQR: 77.1-130.6 pmol/L) and 12.3 nmol/L (IQR: 9-16.4 nmol/L), respectively (P < 0.001). A non-significant reduction in daily active vitamin D analogue dose occurred, 0.94 µmcg at baseline to 0.77 µmcg at 12 months (P = 0.73). The ability to synthesise 1,25(OH)2D3 in ESRD is maintained but is substrate dependent, and serum 25(OH)D was a limiting factor at baseline. Therefore, 1,25(OH)2D3 deficiency in ESRD is partly a consequence of 25(OH)D deficiency, rather than solely due to reduced 1α-hydroxylase activity as suggested by current treatment strategies.
Language	English
Publishing date	2021-10-11
Publishing country	England
Document type	Journal Article
ZDB-ID	2668428-7
ISSN	2049-3614
ISSN	2049-3614
DOI	10.1530/EC-21-0372
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Article ; Online: Simultaneous measurement of 13 circulating vitamin D3 and D2 mono and dihydroxy metabolites using liquid chromatography mass spectrometry.

Jenkinson, Carl / Desai, Reena / Slominski, Andrzej T / Tuckey, Robert C / Hewison, Martin / Handelsman, David J

Clinical chemistry and laboratory medicine

2021 Volume 59, Issue 10, Page(s) 1642–1652

Abstract: Objectives: Clinical evaluation of vitamin D status is conventionally performed by measuring serum levels of a single vitamin D metabolite, 25-hydroxyvitamin D predominantly by immunoassay methodology. However, this neglects the complex metabolic ... ...

Abstract	Objectives: Clinical evaluation of vitamin D status is conventionally performed by measuring serum levels of a single vitamin D metabolite, 25-hydroxyvitamin D predominantly by immunoassay methodology. However, this neglects the complex metabolic pathways involved in vitamin D bioactivity, including two canonical forms D3 and D2, bioactive 1,25-dihydroxy metabolites and inactive 24-hydroxy and other metabolites. Methods: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) can measure multiple analytes in a sample during a single run with high sensitivity and reference level specificity. We therefore aimed to develop and validate a LC-MS/MS method to measure simultaneously 13 circulating vitamin D metabolites and apply it to 103 human serum samples. Results: The LC-MS/MS method using a Cookson-type derivatization reagent phenyl-1,2,4-triazoline-3,5-dione (PTAD) quantifies 13 vitamin D metabolites, including mono and dihydroxy-metabolites, as well as CYP11A1-derived D3 and D2 metabolites in a single run. The lower limit of quantitation was 12.5 pg/mL for 1,25(OH) Conclusions: This profiling method can provide new insight into circulating vitamin D metabolite pathways forming the basis for improved understanding of the role of vitamin D in health and disease.
MeSH term(s)	Calcifediol ; Cholecalciferol ; Chromatography, Liquid/methods ; Humans ; Tandem Mass Spectrometry/methods ; Vitamin D ; Vitamins
Chemical Substances	Vitamins ; Vitamin D (1406-16-2) ; Cholecalciferol (1C6V77QF41) ; Calcifediol (P6YZ13C99Q)
Language	English
Publishing date	2021-05-20
Publishing country	Germany
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	1418007-8
ISSN	1437-4331 ; 1434-6621 ; 1437-8523
ISSN (online)	1437-4331
ISSN	1434-6621 ; 1437-8523
DOI	10.1515/cclm-2021-0441
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Article ; Online: Circulating Conjugated and Unconjugated Vitamin D Metabolite Measurements by Liquid Chromatography Mass Spectrometry.

Jenkinson, Carl / Desai, Reena / McLeod, Malcolm D / Wolf Mueller, Jonathan / Hewison, Martin / Handelsman, David J

The Journal of clinical endocrinology and metabolism

2022 Volume 107, Issue 2, Page(s) 435–449

Abstract: Context: Vitamin D status is conventionally defined by measurement of unconjugated circulating 25-hydroxyvitamin D (25OHD), but it remains uncertain whether this isolated analysis gives sufficient weight to vitamin D's diverse metabolic pathways and ... ...

Abstract	Context: Vitamin D status is conventionally defined by measurement of unconjugated circulating 25-hydroxyvitamin D (25OHD), but it remains uncertain whether this isolated analysis gives sufficient weight to vitamin D's diverse metabolic pathways and bioactivity. Emerging evidence has shown that phase II endocrine metabolites are important excretory or storage forms; however, the clinical significance of circulating phase II vitamin D metabolites remains uncertain. Objective: In this study we analyzed the contribution of sulfate and glucuronide vitamin D metabolites relative to unconjugated levels in human serum. Methods: An optimized enzyme hydrolysis method using recombinant arylsulfatase (Pseudomonas aeruginosa) and beta-glucuronidase (Escherichia coli) was combined with liquid chromatography mass spectrometry (LC-MS/MS) analysis to measure conjugated and unconjugated vitamin D metabolites 25OHD3, 25OHD2, 3-epi-25OHD3, and 24,25(OH)2D3. The method was applied to the analysis of 170 human serum samples from community-dwelling men aged over 70 years, categorized by vitamin D supplementation status, to evaluate the proportions of each conjugated and unconjugated fraction. Results: As a proportion of total circulating vitamin D metabolites, sulfate conjugates (ranging between 18% and 53%) were a higher proportion than glucuronide conjugates (ranging between 2.7% and 11%). The proportion of conjugated 25OHD3 (48 ± 9%) was higher than 25OHD2 conjugates (29.1 ± 10%) across all supplementation groups. Conjugated metabolites correlated with their unconjugated forms for all 4 vitamin D metabolites (r = 0.85 to 0.97). Conclusion: Sulfated conjugates form a high proportion of circulating vitamin D metabolites, whereas glucuronide conjugates constitute a smaller fraction. Our findings principally in older men highlight the differences in abundance between metabolites and suggest a combination of both conjugated and unconjugated measurements may provide a more accurate assessment of vitamin D status.
MeSH term(s)	Aged ; Calibration ; Chromatography, High Pressure Liquid/methods ; Dietary Supplements ; Glucuronides/metabolism ; Humans ; Limit of Detection ; Male ; Sulfates/metabolism ; Tandem Mass Spectrometry/methods ; Vitamin D/administration & dosage ; Vitamin D/blood ; Vitamin D/isolation & purification ; Vitamin D/metabolism
Chemical Substances	Glucuronides ; Sulfates ; Vitamin D (1406-16-2)
Language	English
Publishing date	2022-01-04
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Validation Study
ZDB-ID	3029-6
ISSN	1945-7197 ; 0021-972X
ISSN (online)	1945-7197
ISSN	0021-972X
DOI	10.1210/clinem/dgab708
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Article: Data comparing the separation and elution of vitamin D metabolites on an ultra performance supercritical fluid chromatography tandem-mass spectrometer (UPSFC-MS/MS) compared to liquid chromatography (LC) and data presenting approaches to UPSFC method optimization.

Jenkinson, Carl / Taylor, Angela E / Storbeck, Karl-Heinz / Hewison, Martin

Data in brief

2018 Volume 20, Page(s) 426–435

Abstract: The data presented is related to the research article "Analysis of multiple vitamin D metabolites by ultra performance supercritical fluid chromatography-tandem mass spectrometry (UPSFC-MS/MS)" (Jenkinson et al., 2018) [1]. This article will include data ...

Abstract	The data presented is related to the research article "Analysis of multiple vitamin D metabolites by ultra performance supercritical fluid chromatography-tandem mass spectrometry (UPSFC-MS/MS)" (Jenkinson et al., 2018) [1]. This article will include data obtained from method development, optimization and analysis of multiple vitamin D metabolites on an ultra performance supercritical fluid chromatography tandem-mass spectrometry (UPSFC-MS/MS). This includes chromatograms from column screening to confirm the most suitable column for analyte separation. Additionally, further chromatograms and figures compare separation and analyte signal strength during the optimization of other UPSFC parameters. Mass spectra will demonstrate the optimization of MS conditions for the UPSFC-MS/MS method. Chromatogram data from UHPLC vitamin D analysis is also presented in order to compare the separation and elution of vitamin D metabolites using UPSFC and UHPLC. This data will highlight the outputs that aid in method development and identifying the separation technique suited for vitamin D quantitation.
Language	English
Publishing date	2018-08-15
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	2786545-9
ISSN	2352-3409
ISSN	2352-3409
DOI	10.1016/j.dib.2018.08.027
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Article ; Online: Analysis of multiple vitamin D metabolites by ultra-performance supercritical fluid chromatography-tandem mass spectrometry (UPSFC-MS/MS).

Jenkinson, Carl / Taylor, Angela / Storbeck, Karl-Heinz / Hewison, Martin

Journal of chromatography. B, Analytical technologies in the biomedical and life sciences

2018 Volume 1087-1088, Page(s) 43–48

Abstract: In recent years, increased interest in the human health benefits of vitamin D has led to demand for improved analysis of patient vitamin D 'status'. Studies to date have focused primarily on a single vitamin D metabolite, 25-hydroxyvitamin D, despite the ...

Abstract	In recent years, increased interest in the human health benefits of vitamin D has led to demand for improved analysis of patient vitamin D 'status'. Studies to date have focused primarily on a single vitamin D metabolite, 25-hydroxyvitamin D, despite the existence of a broad range of vitamin D metabolites, referred to as the vitamin D metabolome. This study reports on the development of a rapid UPSFC-MS/MS method for the analysis of nine vitamin D metabolites in human serum. Optimum separation was obtained with a Lux-Cellulose chiral column. We observed an orthogonal elution order when compared with ultra-high performance liquid chromatography (UHPLC). The order of elution was reversed based on hydroxyl- group number, however elution order did not differ between isomeric changes in hydroxyl- group position or epimers. Although UPSFC yielded superior resolution and selectivity over previously developed UHPLC-MS/MS methods, improvements in sensitivity could not be achieved owing to the lower injection volume required for UPSFC relative to UHPLC. Method validation was performed on the developed UPSFC-MS/MS method and found to be within acceptable limits. Applying the method to the analysis of human serum samples showed a significant correlation with serum concentrations of metabolites measured by UHPLC-MS/MS (25OHD3 r = 0.997, P=<0.001, and 3-epi-25OHD3 r = 0.996, P ≤0.001). These data indicate that UPSFC provides an efficient analytical platform for rapid analysis of multiple vitamin D metabolites from serum.
MeSH term(s)	Chromatography, High Pressure Liquid/methods ; Chromatography, Supercritical Fluid/methods ; Humans ; Limit of Detection ; Linear Models ; Reproducibility of Results ; Tandem Mass Spectrometry/methods ; Vitamin D/blood
Chemical Substances	Vitamin D (1406-16-2)
Language	English
Publishing date	2018-06-15
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	1180823-8
ISSN	1873-376X ; 0378-4347 ; 1570-0232 ; 1387-2273
ISSN (online)	1873-376X
ISSN	0378-4347 ; 1570-0232 ; 1387-2273
DOI	10.1016/j.jchromb.2018.04.025
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Article ; Online: Trophoblast uptake of DBP regulates intracellular actin and promotes matrix invasion.

Ganguly, Ankana / Tamblyn, Jennifer A / Shattock, Alexandra / Joseph, Annsha / Larner, Dean P / Jenkinson, Carl / Gupta, Janesh / Gross, Stephane R / Hewison, Martin

The Journal of endocrinology

2021 Volume 249, Issue 1, Page(s) 43–55

Abstract: Early pregnancy is characterised by elevated circulating levels of vitamin D binding protein (DBP). The impact of this on maternal and fetal health is unclear but DBP is present in the placenta, and DBP gene variants have been linked to malplacentation ... ...

Abstract	Early pregnancy is characterised by elevated circulating levels of vitamin D binding protein (DBP). The impact of this on maternal and fetal health is unclear but DBP is present in the placenta, and DBP gene variants have been linked to malplacentation disorders such as preeclampsia. The functional role of DBP in the placenta was investigated using trophoblastic JEG3, BeWo and HTR8 cells. All three cell lines showed intracellular DBP with increased expression and nuclear localisation of DBP in cells treated with the active form of vitamin D, 1,25-dihydroxyvitamin D (1,25D). When cultured in the serum of mice lacking DBP (DBP-/-), JEG3 cells showed no intracellular DBP indicating uptake of exogenous DBP. Inhibition of the membrane receptor for DBP, megalin, also suppressed intracellular DBP. Elimination of intracellular DBP with DBP-/- serum or megalin inhibitor suppressed matrix invasion by trophoblast cells and was associated with increased nuclear accumulation of G-actin. Conversely, treatment with 1,25D enhanced matrix invasion. This was independent of the nuclear vitamin D receptor but was associated with enhanced ERK phosphorylation, and inhibition of ERK kinase suppressed trophoblast matrix invasion. When cultured with serum from pregnant women, trophoblast matrix invasion correlated with DBP concentration, and DBP was lower in first-trimester serum from women who later developed preeclampsia. These data show that the trophoblast matrix invasion involves uptake of serum DBP and associated intracellular actin-binding and homeostasis. DBP is a potential marker of placentation disorders such as preeclampsia and may also provide a therapeutic option for improved placenta and pregnancy health.
MeSH term(s)	Actins/metabolism ; Cell Line ; Cell Line, Tumor ; Cell Nucleus/chemistry ; Cell Nucleus/metabolism ; Choriocarcinoma ; Extracellular Signal-Regulated MAP Kinases/metabolism ; Female ; Gene Knockdown Techniques ; Humans ; Phosphorylation ; Placentation/physiology ; Pre-Eclampsia/blood ; Pregnancy ; Receptors, Calcitriol/genetics ; Receptors, Calcitriol/physiology ; Trophoblasts/physiology ; Uterine Neoplasms ; Vitamin D/analogs & derivatives ; Vitamin D/blood ; Vitamin D/pharmacology ; Vitamin D-Binding Protein/blood ; Vitamin D-Binding Protein/genetics ; Vitamin D-Binding Protein/physiology
Chemical Substances	Actins ; Receptors, Calcitriol ; Vitamin D-Binding Protein ; Vitamin D (1406-16-2) ; 1,25-dihydroxyvitamin D (66772-14-3) ; Extracellular Signal-Regulated MAP Kinases (EC 2.7.11.24)
Language	English
Publishing date	2021-02-17
Publishing country	England
Document type	Journal Article
ZDB-ID	3028-4
ISSN	1479-6805 ; 0022-0795
ISSN (online)	1479-6805
ISSN	0022-0795
DOI	10.1530/JOE-20-0626
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Article ; Online: Diet-induced vitamin D deficiency reduces skeletal muscle mitochondrial respiration.

Ashcroft, Stephen P / Fletcher, Gareth / Philp, Ashleigh M / Jenkinson, Carl / Das, Shatarupa / Hansbro, Philip M / Atherton, Philip J / Philp, Andrew

The Journal of endocrinology

2021 Volume 249, Issue 2, Page(s) 113–124

Abstract: Vitamin D deficiency is associated with symptoms of skeletal muscle myopathy including muscle weakness and fatigue. Recently, vitamin D-related metabolites have been linked to the maintenance of mitochondrial function within skeletal muscle. However, ... ...

Abstract	Vitamin D deficiency is associated with symptoms of skeletal muscle myopathy including muscle weakness and fatigue. Recently, vitamin D-related metabolites have been linked to the maintenance of mitochondrial function within skeletal muscle. However, current evidence is limited to in vitro models and the effects of diet-induced vitamin D deficiency upon skeletal muscle mitochondrial function in vivo have received little attention. In order to examine the role of vitamin D in the maintenance of mitochondrial function in vivo, we utilised an established model of diet-induced vitamin D deficiency in C57BL/6J mice. Mice were either fed a control diet (2200 IU/kg i.e. vitamin D replete) or a vitamin D-deplete (0 IU/kg) diet for periods of 1, 2 and 3 months. Gastrocnemius muscle mitochondrial function and ADP sensitivity were assessed via high-resolution respirometry and mitochondrial protein content via immunoblotting. As a result of 3 months of diet-induced vitamin D deficiency, respiration supported via complex I + II (CI + IIP) and the electron transport chain (ETC) were 35 and 37% lower when compared to vitamin D-replete mice (P < 0.05). Despite functional alterations, citrate synthase activity, AMPK phosphorylation, mitofilin, OPA1 and ETC subunit protein content remained unchanged in response to dietary intervention (P > 0.05). In conclusion, we report that 3 months of diet-induced vitamin D deficiency reduced skeletal muscle mitochondrial respiration in C57BL/6J mice. Our data, when combined with previous in vitro observations, suggest that vitamin D-mediated regulation of mitochondrial function may underlie the exacerbated muscle fatigue and performance deficits observed during vitamin D deficiency.
MeSH term(s)	Animals ; Body Composition ; Calcium/blood ; Diet/adverse effects ; Gene Expression Regulation/drug effects ; Male ; Mice ; Mice, Inbred C57BL ; Mitochondria, Muscle/metabolism ; Mitochondrial Proteins/genetics ; Mitochondrial Proteins/metabolism ; Muscle, Skeletal/metabolism ; Oxygen Consumption ; Vitamin D/blood ; Vitamin D Deficiency/etiology ; Vitamin D Deficiency/metabolism
Chemical Substances	Mitochondrial Proteins ; Vitamin D (1406-16-2) ; Calcium (SY7Q814VUP)
Language	English
Publishing date	2021-04-05
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	3028-4
ISSN	1479-6805 ; 0022-0795
ISSN (online)	1479-6805
ISSN	0022-0795
DOI	10.1530/JOE-20-0233
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Article: Data comparing the separation and elution of vitamin D metabolites on an ultra performance supercritical fluid chromatography tandem-mass spectrometer (UPSFC-MS/MS) compared to liquid chromatography (LC) and data presenting approaches to UPSFC method optimization

Jenkinson, Carl / Taylor, Angela E. / Storbeck, Karl-Heinz / Hewison, Martin

Data in Brief. 2018 Oct., v. 20

2018

Abstract: The data presented is related to the research article “Analysis of multiple vitamin D metabolites by ultra performance supercritical fluid chromatography-tandem mass spectrometry (UPSFC-MS/MS)” (Jenkinson et al., 2018) [1]. This article will include data ...

Abstract	The data presented is related to the research article “Analysis of multiple vitamin D metabolites by ultra performance supercritical fluid chromatography-tandem mass spectrometry (UPSFC-MS/MS)” (Jenkinson et al., 2018) [1]. This article will include data obtained from method development, optimization and analysis of multiple vitamin D metabolites on an ultra performance supercritical fluid chromatography tandem-mass spectrometry (UPSFC-MS/MS). This includes chromatograms from column screening to confirm the most suitable column for analyte separation. Additionally, further chromatograms and figures compare separation and analyte signal strength during the optimization of other UPSFC parameters. Mass spectra will demonstrate the optimization of MS conditions for the UPSFC-MS/MS method. Chromatogram data from UHPLC vitamin D analysis is also presented in order to compare the separation and elution of vitamin D metabolites using UPSFC and UHPLC. This data will highlight the outputs that aid in method development and identifying the separation technique suited for vitamin D quantitation.
Keywords	metabolites ; signal strength ; spectrometers ; supercritical fluid chromatography ; tandem mass spectrometry
Language	English
Dates of publication	2018-10
Size	p. 426-435.
Publishing place	Elsevier Inc.
Document type	Article
ZDB-ID	2786545-9
ISSN	2352-3409
ISSN	2352-3409
DOI	10.1016/j.dib.2018.08.027
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