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Home / Equine Research Bank / PLoS One / Serial-omics characterization of equine urine.
PloS one2017; 12(10); e0186258; doi: 10.1371/journal.pone.0186258

Serial-omics characterization of equine urine.

Abstract: Horse urine is easily collected and contains molecules readily measurable using mass spectrometry that can be used as biomarkers representative of health, disease or drug tampering. This study aimed at analyzing microliter levels of horse urine to purify, identify and quantify proteins, polar metabolites and non-polar lipids. Urine from a healthy 12 year old quarter horse mare on a diet of grass hay and vitamin/mineral supplements with limited pasture access was collected for serial-omics characterization. The urine was treated with methyl tert-butyl ether (MTBE) and methanol to partition into three distinct layers for protein, non-polar lipid and polar metabolite content from a single liquid-liquid extraction and was repeated two times. Each layer was analyzed by high performance liquid chromatography-high resolution tandem mass spectrometry (LC-MS/MS) to obtain protein sequence and relative protein levels as well as identify and quantify small polar metabolites and lipids. The results show 46 urine proteins, many related to normal kidney function, structural and circulatory proteins as well as 474 small polar metabolites but only 10 lipid molecules. Metabolites were mostly related to urea cycle and ammonia recycling as well as amino acid related pathways, plant diet specific molecules, etc. The few lipids represented triglycerides and phospholipids. These data show a complete mass spectrometry based-omics characterization of equine urine from a single 333 μL mid-stream urine aliquot. These omics data help serve as a baseline for healthy mare urine composition and the analyses can be used to monitor disease progression, health status, monitor drug use, etc.
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Publication Date: 2017-10-13 PubMed ID: 29028822PubMed Central: PMC5640239DOI: 10.1371/journal.pone.0186258Google Scholar: Lookup
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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 research aimed to analyze small volumes of horse urine to identify and quantify proteins, metabolites, and lipids, which could serve as health or disease biomarkers, or evidence of drug tampering. In this process, urine from a healthy horse was sampled, treated, and partitioned, and each component was examined using high-performance liquid chromatography-mass spectrometry.

Methodology Overview

  • The urine sample was taken from a different 12 years old mare that lived on a diet of grass hay and vitamin/mineral supplements, with limited access to pasture.
  • The collected urine was subjected to a single liquid-liquid extraction using methyl tert-butyl ether (MTBE) and methanol. This resulted in three distinct layers for protein, non-polar lipid, and polar metabolite content. The extraction was done twice to ensure accuracy.
  • Every layer was subsequently analyzed using high-performance liquid chromatography coupled with high-resolution tandem mass spectrometry (LC-MS/MS). This method enabled the researchers to obtain protein sequence information and evaluate relative protein levels, and to identify and quantify low-level polar metabolites and lipids.

Results and Findings

  • The results revealed the presence of 46 different proteins in the horse’s urine, many of which were associated with normal kidney function and were structural and circulatory proteins.
  • Additionally, 474 small polar metabolites were identified, with the majority related to the urea cycle, ammonia recycling, and amino acid pathways. There were also molecules specific to a plant-based diet.
  • On the other hand, the researchers found only 10 lipid molecules, representing triglycerides and phospholipids.
  • All this data was gleaned from a mere 333 μL mid-stream urine aliquot, demonstrating the detailed level of information that can be derived from small samples.

Implications and Potential Applications

  • The thorough omics characterization of the equine urine serves as a baseline for the composition of a healthy mare’s urine.
  • The methodologies and analyses employed in this research could be used to track disease progression and monitor the general health status of horses. They can also be potentially exploited to detect drug usage and any form of doping in performance horses.

Cite This Article

APA
Yuan M, Breitkopf SB, Asara JM. (2017). Serial-omics characterization of equine urine. PLoS One, 12(10), e0186258. https://doi.org/10.1371/journal.pone.0186258

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 12
Issue: 10
Pages: e0186258
PII: e0186258

Researcher Affiliations

Yuan, Min
  • Division of Signal Transduction, Beth Israel Deaconess Medical Center, Boston, Massachusetts, United States of America.
Breitkopf, Susanne B
  • Division of Signal Transduction, Beth Israel Deaconess Medical Center, Boston, Massachusetts, United States of America.
  • Department of Medicine, Harvard Medical School, Boston, Massachusetts, United States of America.
Asara, John M
  • Division of Signal Transduction, Beth Israel Deaconess Medical Center, Boston, Massachusetts, United States of America.
  • Department of Medicine, Harvard Medical School, Boston, Massachusetts, United States of America.

MeSH Terms

  • Animals
  • Horses
  • Lipids / urine
  • Metabolomics / methods
  • Time Factors
  • Urinalysis

Grant Funding

  • P01 CA120964 / NCI NIH HHS
  • P30 CA006516 / NCI NIH HHS
  • S10 OD010612 / NIH HHS

Conflict of Interest Statement

The authors have declared that no competing interests exist.

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Citations

This article has been cited 2 times.
  1. Keen B, Cawley A, Reedy B, Fu S. Metabolomics in clinical and forensic toxicology, sports anti-doping and veterinary residues. Drug Test Anal 2022 May;14(5):794-807.
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