LC-MS/MS analysis of vitamin D3 metabolites in human serum using a salting-out based liquid-liquid extraction and DAPTAD derivatization

A, Alshabrawy; Bergamin, Amanda; Sharma, Deepti; Hickey, Shane M.; Brooks, Doug A.; O’Loughlin, Peter; Wiese, Michael; Anderson, Paul

doi:10.1016/j.jchromb.2021.122654

Cited by 10 publications

(5 citation statements)

References 41 publications

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“…123 For analyzing the VD metabolites, LC-MS/MS has appeared as a more reliable method with improved sensitivity and specificity. 124 Unfortunately, due to the lipophilic character of these molecules and the lack of functional groups that assist ionization, quantitative measurement of VD by mass spectrometry is difficult. 125 A typical strategy for improving the ionization efficiency of the molecules is to use a derivatization procedure and Cookson-type reagents are generally used to react with VD in the Diels–Alder cycloaddition reaction.…”

Section: Methodsmentioning

confidence: 99%

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Vitamin D metabolites and analytical challenges

et al. 2023

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Section: Methodsmentioning

confidence: 99%

“…High sensitivity and specicity 141 Expensive instrumentation and a high level of technical expertise are required 142 No cross-reactions lead to overestimation and simultaneous measurements of different VD metabolites 125 Susceptible to ion suppression interference 124…”

Section: Lc-msmentioning

confidence: 99%

Vitamin D metabolites and analytical challenges

et al. 2023

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“…LC‐MS/MS was used to detect the contents of vitamin A, vitamin C, vitamin D, and vitamin E in serum 13,14 . The standard vitamin, mobile phase formic acid, and methanol were purchased from Merck.…”

Section: Methodsmentioning

confidence: 99%

“…LC‐MS/MS was used to detect the contents of vitamin A, vitamin C, vitamin D, and vitamin E in serum. 13 , 14 The standard vitamin, mobile phase formic acid, and methanol were purchased from Merck. The LC‐MS/MS platform connects the Shimadzu® Nexera X2 high performance liquid chromatograph (Shimadzu USA) with the AB Sciex® 6500 + triple quadrupole mass spectrometer (Sciex).…”

Section: Methodsmentioning

confidence: 99%

The levels of vitamins and cytokines in serum of elderly patients with community‐acquired pneumonia: A case–control study

Jing,

Chen,

Geng

et al. 2023

Health Science Reports

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Background and AimsThe community acquired pneumonia (CAP) is a common disease with high incidence rate and mortality among the elderly. This study aims to explore the association between vitamins, cytokines, and CAP infected by different pathogens in Handan First Hospital.MethodsThe subjects of this study were elderly patients with community‐acquired pneumonia in Handan First Hospital and healthy elderly people in the community from February 2018 to December 2019. The study include 234 CAP patients and 180 healthy elderly people. The differences between the healthy and CAP groups were analyzed by blood routine test, urine routine test, Liquid chromatography tandem mass spectrometry (LC‐MS/MS) detection of vitamin level, and Enzyme linked immunosorbent assay (ELISA) detection of cytokine level. Sputum culture was used to detect pathogens.ResultsAccording to our results, there was no significant difference in the contents of vitamin A, vitamin D, vitamin E, and vitamin C in serum between the healthy and CAP elderly. However, compared with the control group, the levels of serum cytokines of IL‐2, IL‐17, and TGF‐β in the CAP patients were significantly increased. In addition, IL‐17 was positively correlated with white blood cells, neutrophils, platelet/lymphocyte ratio and neutrophil/lymphocyte ratio of CAP patients, and negatively correlated with lymphocytes. The four pathogens with the highest positive rates were Streptococcus pneumoniae, Klebsiella pneumoniae, Mycoplasma pneumoniae and H. influenza. In patients with S. pneumoniae infection, the IL‐2, IL‐17, and TGF‐β levels were significantly higher than the levels in the control group; IL‐17 was also significantly increased in the serum of patients infected with M. pneumoniae, K. pneumoniae, and H. influenza.ConclusionsThere is no direct association between vitamins in serum and community acquired pneumonia (CAP). However, the cytokines are closely related to the clinical indicators and pathogens of CAP patients, which can provide references for the diagnosis, prognosis and treatment of CAP patients.

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“…The 24,25(OH) 2 D 3 concentrations were analyzed in cell culture samples from Section 2.4.3 , using DAPTAD derivatization and LC-MS/MS, as previously described [ 38 ]. The calibrators were prepared in phosphate-buffered saline (PBS), and the samples were appropriately diluted with PBS to bring them into the linear range of the assay.…”

Section: Methodsmentioning

confidence: 99%

Therapeutic Potential of a Novel Vitamin D3 Oxime Analogue, VD1-6, with CYP24A1 Enzyme Inhibitory Activity and Negligible Vitamin D Receptor Binding

Cui

Sylvester

et al. 2022

Biomolecules

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The regulation of vitamin D3 actions in humans occurs mainly through the Cytochrome P450 24-hydroxylase (CYP24A1) enzyme activity. CYP24A1 hydroxylates both 25-hydroxycholecalciferol (25(OH)D3) and 1,25-dihydroxycholecalciferol (1,25(OH)2D3), which is the first step of vitamin D catabolism. An abnormal status of the upregulation of CYP24A1 occurs in many diseases, including chronic kidney disease (CKD). CYP24A1 upregulation in CKD and diminished activation of vitamin D3 contribute to secondary hyperparathyroidism (SHPT), progressive bone deterioration, and soft tissue and cardiovascular calcification. Previous studies have indicated that CYP24A1 inhibition may be an effective strategy to increase endogenous vitamin D activity and decrease SHPT. This study has designed and synthesized a novel C-24 O-methyloxime analogue of vitamin D3 (VD1-6) to have specific CYP24A1 inhibitory properties. VD1-6 did not bind to the vitamin D receptor (VDR) in concentrations up to 10−7 M, assessed by a VDR binding assay. The absence of VDR binding by VD1-6 was confirmed in human embryonic kidney HEK293T cultures through the lack of CYP24A1 induction. However, in silico docking experiments demonstrated that VD1-6 was predicted to have superior binding to CYP24A1, when compared to that of 1,25(OH)2D3. The inhibition of CYP24A1 by VD1-6 was also evident by the synergistic potentiation of 1,25(OH)2D3-mediated transcription and reduced 1,25(OH)2D3 catabolism over 24 h. A further indication of CYP24A1 inhibition by VD1-6 was the reduced accumulation of the 24,25(OH)D3 , the first metabolite of 25(OH)D catabolism by CYP24A1. Our findings suggest the potent CYP24A1 inhibitory properties of VD1-6 and its potential for testing as an alternative therapeutic candidate for treating SHPT.

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LC-MS/MS analysis of vitamin D3 metabolites in human serum using a salting-out based liquid-liquid extraction and DAPTAD derivatization

Cited by 10 publications

References 41 publications

Vitamin D metabolites and analytical challenges

Vitamin D metabolites and analytical challenges

The levels of vitamins and cytokines in serum of elderly patients with community‐acquired pneumonia: A case–control study

Therapeutic Potential of a Novel Vitamin D3 Oxime Analogue, VD1-6, with CYP24A1 Enzyme Inhibitory Activity and Negligible Vitamin D Receptor Binding

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