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Home / Equine Research Bank / J Vet Diagn Invest / Simultaneous quantification of vitamin E and vitamin E metab...
Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc2021; 33(3); 506-515; doi: 10.1177/10406387211005433

Simultaneous quantification of vitamin E and vitamin E metabolites in equine plasma and serum using LC-MS/MS.

Abstract: Vitamin E deficiencies can impact normal growth and development in humans and animals, and assessment of circulating levels of vitamin E and its metabolites may be an important endpoint for evaluation. Development of a sensitive method to detect and quantify low concentrations of vitamin E and metabolites in biological specimens allows for a proper diagnosis for patients and animals that are deficient. We developed a method to simultaneously extract, detect, and quantify the vitamin E compounds alpha-tocopherol (α-TP), gamma-tocopherol (γ-TP), alpha-tocotrienol (α-TT), and gamma-tocotrienol (γ-TT), and the corresponding metabolites formed after β-oxidation of α-TP and γ-TP, alpha-carboxymethylbutyl hydroxychroman (α-CMBHC) and alpha- or gamma-carboxyethyl hydroxychroman (α- or γ-CEHC), respectively, from equine plasma and serum. Quantification was achieved through liquid chromatography-tandem mass spectrometry. We applied a 96-well high-throughput format using a Phenomenex Phree plate to analyze plasma and serum. Compounds were separated by using a Waters ACQUITY UPLC BEH C18 column with a reverse-phase gradient. The limits of detection for the metabolites and vitamin E compounds were 8-330 pg/mL. To validate the method, intra-day and inter-day accuracy and precision were evaluated along with limits of detection and quantification. The method was then applied to determine concentrations of these analytes in plasma and serum of horses. Alpha-TP levels were 3-6 µg/mL of matrix; the metabolites were found at much lower levels, 0.2-1.0 ng/mL of matrix.
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Publication Date: 2021-04-13 PubMed ID: 33847203PubMed Central: PMC8120066DOI: 10.1177/10406387211005433Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study aims to highlight the development of a sensitive technique for detecting and quantifying low concentrations of vitamin E and its metabolites in equine plasma and serum using liquid chromatography-tandem mass spectrometry.

Research Objective

  • The research focuses on the development of a method that can extract, detect, and quantify vitamin E compounds and their metabolites from equine plasma and serum. Such compounds include alpha-tocopherol (α-TP), gamma-tocopherol (γ-TP), alpha-tocotrienol (α-TT), and gamma-tocotrienol (γ-TT).

Methodology and Procedure

  • The compounds were quantified through liquid chromatography-tandem mass spectrometry, a notable analytical technique in detecting low concentrations of substances.
  • A 96-well high-throughput format using a Phenomenex Phree plate was employed to analyze plasma and serum.
  • The compounds were then separated using a Waters ACQUITY UPLC BEH C18 column with a reverse-phase gradient.
  • The limits of detection of the method for the metabolites and vitamin E compounds were tested and found to be between 8-330 pg/mL.

Method Validation

  • To ensure the method’s efficacy, the researchers evaluated its intra-day and inter-day accuracy and precision, alongside the limits of detection and quantification.

Research Findings

  • The developed method was applied to determine the concentrations of the vitamin E analytes in plasma and serum of horses.
  • The results revealed that Alpha-TP levels were 3-6 µg/mL of matrix. The metabolites were present at much lower levels, specifically between 0.2-1.0 ng/mL of matrix.

Significance of the Study

  • This study provides a foundation for future research in diagnosing vitamin E deficiencies in animals through the accurate detection of vitamin E and its metabolites.
  • Furthermore, it contributes to the improvement of dietary interventions necessary for normal growth and development.

Cite This Article

APA
Habib H, Finno CJ, Gennity I, Favro G, Hales E, Puschner B, Moeller BC. (2021). Simultaneous quantification of vitamin E and vitamin E metabolites in equine plasma and serum using LC-MS/MS. J Vet Diagn Invest, 33(3), 506-515. https://doi.org/10.1177/10406387211005433

Publication

Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
ISSN: 1943-4936
NlmUniqueID: 9011490
Country: United States
Language: English
Volume: 33
Issue: 3
Pages: 506-515

Researcher Affiliations

Habib, Hadi
  • Department of Molecular Biosciences, School of Veterinary Medicine, University of California, Davis, CA.
Finno, Carrie J
  • Department of Population Health and Reproduction, School of Veterinary Medicine, University of California, Davis, CA.
Gennity, Ingrid
  • Department of Molecular Biosciences, School of Veterinary Medicine, University of California, Davis, CA.
Favro, Gianna
  • Department of Molecular Biosciences, School of Veterinary Medicine, University of California, Davis, CA.
Hales, Erin
  • Department of Population Health and Reproduction, School of Veterinary Medicine, University of California, Davis, CA.
Puschner, Birgit
  • Department of Molecular Biosciences, School of Veterinary Medicine, University of California, Davis, CA.
Moeller, Benjamin C
  • K. L. Maddy Equine Analytical Chemistry Laboratory, California Animal Health and Food Safety Laboratory, School of Veterinary Medicine, University of California, Davis, CA.

MeSH Terms

  • Animals
  • Chromatography, Liquid
  • Female
  • Horses / metabolism
  • Male
  • Plasma / chemistry
  • Serum / chemistry
  • Tandem Mass Spectrometry
  • Vitamin E / blood
  • Vitamin E / metabolism

Grant Funding

  • K01 OD015134 / NIH HHS
  • L40 TR001136 / NCATS NIH HHS

Conflict of Interest Statement

Declaration of conflicting interests: The authors declared no potential conflicts of interest with respect to theresearch, authorship, and/or publication of this article.

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Citations

This article has been cited 2 times.
  1. Hales EN, Habib H, Favro G, Katzman S, Sakai RR, Marquardt S, Bordbari MH, Ming-Whitfield B, Peterson J, Dahlgren AR, Rivas V, Ramirez CA, Peng S, Donnelly CG, Dizmang BS, Kallenberg A, Grahn R, Miller AD, Woolard K, Moeller B, Puschner B, Finno CJ. Increased α-tocopherol metabolism in horses with equine neuroaxonal dystrophy. J Vet Intern Med 2021 Sep;35(5):2473-2485.
    doi: 10.1111/jvim.16233pubmed: 34331715google scholar: lookup
  2. Lim JS, Lee SH, Yun H, Lee DY, Cho N, Yoo G, Choi JU, Lee KY, Bach TT, Park SJ, Cho YC. Inhibitory Effects of Ehretia tinifolia Extract on the Excessive Oxidative and Inflammatory Responses in Lipopolysaccharide-Stimulated Mouse Kupffer Cells. Antioxidants (Basel) 2023 Sep 22;12(10).
    doi: 10.3390/antiox12101792pubmed: 37891872google scholar: lookup
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