Simultaneous determination of vitamin D metabolites 25(OH)D3 and 1α,25(OH)2D3 in human plasma using liquid chromatography tandem mass spectrometry

Xu, Shan; Ni, Rui; Lv, Li-Hong; Chen, Rui; Chen, Yao; Huang, Felix C.; Zhao, Xu

doi:10.1016/j.jmsacl.2022.04.001

Cited by 9 publications

(4 citation statements)

References 37 publications

(39 reference statements)

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“…The developed method has high sensitivity. And compared with other reported method with SPE or LLE combined with SPE, 32,33 only LLE pretreatment method used in this study is generally relatively simple to operate and requires more common equipment and reagents. Besides, compared with the reported method with 0.5–1 h for derivatization time, 7 only 10 min for derivatization time was needed in this work, saving pretreatment time, which is very conducive to high throughput analysis.…”

Section: Resultsmentioning

confidence: 99%

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A novel LC-MS/MS method combined with derivatization for simultaneous quantification of vitamin D metabolites in human serum with diabetes as well as hyperlipidemia

Wang,

Qin,

et al. 2023

RSC Adv.

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Section: Resultsmentioning

confidence: 99%

“…3.5.3 Comparison with previously reported methods. The results obtained from the developed method using LC-MS/MS combined with derivatization was compared with other previously reported methods 18,[30][31][32] as presented in Table 4. The developed method has high sensitivity.…”

Section: Optimization Of Redissolving Solventsmentioning

confidence: 99%

A novel LC-MS/MS method combined with derivatization for simultaneous quantification of vitamin D metabolites in human serum with diabetes as well as hyperlipidemia

Wang,

Qin,

et al. 2023

RSC Adv.

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“…The best results in terms of recovery and background signal were obtained by applying a protein precipitation with methanol and a four-fold liquid–liquid extraction with 25:75 hexane:ethyl acetate ( v / v ) to 250 µL of serum. The optimized sample procedure allowed us to avoid more laborious and expensive extractions, such as solid-phase extraction [ 25 ] or solid–liquid extraction [ 31 ]. The chosen sample preparation procedure is quick and allows one to prepare 20 samples in just 1 h.…”

Section: Discussionmentioning

confidence: 99%

“…This high-throughput approach has recently started to spread in veterinary medicine, mainly for research purposes. Several studies quantified 25-OH-D3 in serum by LC–MS/MS, with the majority employing solid-phase extractions (SPE) [ 24 , 25 ], solid–liquid extractions (SLE) [ 3 ], and derivatizing agents [ 26 , 27 ], which are typically time-consuming and expensive.…”

Section: Introductionmentioning

confidence: 99%

A Novel UHPLC–MS/MS Method for the Measurement of 25-Hydroxyvitamin D3 in Canine Serum and Its Application to Healthy Dogs

Bardhi,

Vecchiato,

Sabetti

et al. 2023

Animals

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Several studies have shown the importance of vitamin D3 supplementation in small animals. In dogs, a low vitamin D3 status is associated not only with bone metabolism but also with different kinds of disorders, such as congestive heart failure, gastrointestinal diseases, chronic kidney diseases, and some types of cancer. However, it is crucial to maintain balance and monitor the introduction of this essential nutrient through the diet because over-supplementation can result in toxicity. Due to the clinical importance of assessing the vitamin D3 status in small animal patients, a quick, simple, and highly performing analytical method for its measurement is needed. In this study, we describe the development of a novel liquid chromatography–tandem mass spectrometry method for 25-hydroxyvitamin D3 quantification in canine serum. The approach was successfully validated following current European guidelines, proving excellent linearity (R2 always ≥0.996), accuracy (always within ±13%) and precision (always <10%). The application of the validated approach to samples collected from 40 healthy dogs made possible the definition of a reliable reference interval for 25-hydroxyvitamin D3, the main biomarker of vitamin D3. In addition, variations below 5% in the results obtained quantifying the same samples using a water-based calibration curve demonstrated that a surrogate matrix may be used without affecting data accuracy. Thanks to its simplicity, the proposed technique represents a useful tool for supporting clinical routine and investigating correlations between serum concentrations of this metabolite and multiple diseases. Additionally, it could enable the monitoring of supplementation in small animal patients in veterinary clinical practice.

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A highly sensitive LC-MS/MS method for quantitative determination of 7 vitamin D metabolites in mouse brain tissue

et al. 2023

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Despite its critical role in neurodevelopment and brain function, vitamin D (vit-D) homeostasis, metabolism, and kinetics within the central nervous system remain largely undetermined. Thus, it is of critical importance to establish an accurate, highly sensitive, and reproducible method to quantitate vit-D in brain tissue. Here, we present a novel liquid chromatography tandem mass spectrometry (LC-MS/MS) method and for the first time, demonstrate detection of seven major vit-D metabolites in brain tissues of C57BL/6J wild-type mice, namely 1,25(OH)2D3, 3-epi-1,25(OH)2D3, 1,25(OH)2D2, 25(OH)D3, 25(OH)D2, 24,25(OH)2D3, and 24,25(OH)2D2. Chromatographic separation was achieved on a pentaflurophenyl column with 3 mM ammonium formate water/methanol [A] and 3 mM ammonium formate methanol/isopropanol [B] mobile phase components. Detection was by positive ion electrospray tandem mass spectrometry with the EVOQ elite triple quadrupole mass spectrometer with an Advance ultra-high-performance liquid chromatograph and online extraction system. Calibration standards of each metabolite prepared in brain matrices were used to validate the detection range, precision, accuracy, and recovery. Isotopically labelled analogues, 1,25(OH)2D3-d3, 25(OH)D3-c5, and 24,25(OH)2D3-d6, served as the internal standards for the closest molecular-related metabolite in all measurements. Standards between 1 fg/mL and 10 ng/mL were injected with a resulting linear range between 0.001 and 1 ng, with an LLOD and LLOQ of 1 pg/mL and 12.5 pg/mL, respectively. The intra-/inter-day precision and accuracy for measuring brain vit-D metabolites ranged between 0.12–11.53% and 0.28–9.11%, respectively. Recovery in acetonitrile ranged between 99.09 and 106.92% for all metabolites. Collectively, the sensitivity and efficiency of our method supersedes previously reported protocols used to measure vit-D and to our knowledge, the first protocol to reveal the abundance of 25(OH)D2, 1,25(OH)D2, and 24,25(OH)2D2, in brain tissue of any species. This technique may be important in supporting the future advancement of pre-clinical research into the function of vit-D in neurophysiological and neuropsychiatric disorders, and neurodegeneration. Graphical Abstract

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Simultaneous determination of vitamin D metabolites 25(OH)D3 and 1α,25(OH)2D3 in human plasma using liquid chromatography tandem mass spectrometry

Cited by 9 publications

References 37 publications

A novel LC-MS/MS method combined with derivatization for simultaneous quantification of vitamin D metabolites in human serum with diabetes as well as hyperlipidemia

A novel LC-MS/MS method combined with derivatization for simultaneous quantification of vitamin D metabolites in human serum with diabetes as well as hyperlipidemia

A Novel UHPLC–MS/MS Method for the Measurement of 25-Hydroxyvitamin D3 in Canine Serum and Its Application to Healthy Dogs

A highly sensitive LC-MS/MS method for quantitative determination of 7 vitamin D metabolites in mouse brain tissue

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