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Guide to Urine Sample Handling for Proteomics and Metabolomics Studies

Urine is a valuable biological fluid for various research and clinical applications. It contains a wealth of information about an individual's health and metabolic status.


Protein Identification and Protein Quantification: The realm of proteomics delves into the vast array of proteins present in urine. This method facilitates the identification, quantification, and comparison of proteins, some of which serve as specific biomarkers. These biomarkers offer significant insights into various disease states and overall health, all of which can be explored through proteomic analysis.

Biomarker Discovery: The proteins found in urine have the potential to serve as crucial biomarkers for disease screening, diagnosis, and monitoring. The analysis of urine samples opens the door to the discovery of novel biomarkers, which greatly supports medical research and clinical applications.

Protein Subtypes and Modifications: Beyond identification, urine proteomics can shed light on protein subtypes and post-translational modifications. These intricate changes, such as glycosylation and phosphorylation, play a pivotal role in disease mechanisms and biological processes.


Metabolite Identification and Quantification: Urine is replete with a multitude of metabolites, metabolic products, and small molecules. The field of metabolomics, driven by urine sample analysis, allows for the identification and quantification of these metabolites. This deepens our understanding of metabolic pathways and physiological changes.

Identification of Disease Markers: The metabolites within urine provide a rich source for identifying potential disease markers. Alterations in the metabolite profile can offer a reflection of disease status, thereby providing a foundation for disease diagnosis and monitoring.

Drug Metabolism and Toxicity Research: Urine samples are instrumental in the study of drug metabolism and toxicity. Metabolomics can identify drug metabolites, elucidate drug metabolic pathways, and assess potential toxicity, contributing to the pharmaceutical and toxicology fields.

Diet and Nutrition Studies: Urine metabolomics is also a powerful tool for investigating an individual's diet and nutritional status. Analysis of urinary metabolites enables the assessment of how one's diet impacts their metabolism.

Advantages of Urine Samples:

  • Non-Invasive Collection: One of the significant advantages of urine samples is their non-invasive collection method. This low-risk approach is suitable for a broad range of age groups, including children and the elderly.
  • Repetitive Sampling: The ability to collect urine samples repeatedly is a pivotal feature. This capability is invaluable for long-term research and monitoring, which in turn helps researchers understand physiological and metabolic changes over time.
  • Rich Information: Urine samples provide a diverse range of biomolecules, including proteins, metabolites, and metabolic products. The wealth of information they carry is an invaluable resource for scientific exploration.
  • Biomarker Discovery: The proteins and metabolites within urine are indispensable in the discovery of new biomarkers. This discovery process greatly advances medical diagnosis and the monitoring of disease.

Sample Collection

For people:

  • Collect the first morning urine sample, as it is often more concentrated and suitable for analysis. Ensure that the entire void is collected without missing any portion.
  • Prior to collection, wash hands thoroughly with soap and water to reduce the risk of contamination.
  • Avoid contamination by not allowing contact with toilet paper, feces, or menstrual blood during the collection process.
  • For male participants, ensure the glans is cleaned with warm water before collection.
  • For female participants, clean the area around the urethral opening with warm water before collecting the sample.
  • Start urinating into the toilet or a clean container, then switch to the provided collection container to collect the midstream urine, reducing contamination risk.
  • Close the container immediately after collection to prevent contamination or evaporation.

For animal:

  • Collect urine samples from animals in a clean and controlled environment, minimizing stress or disturbances.
  • Ensure that animals have access to water but avoid extended periods of water deprivation before sample collection to prevent sample dilution.
  • For individually housed animals, place a sterile collection container under the animal to collect urine.
  • For group-housed animals, consider using metabolic cages or individualized collection methods.
  • Collect the entire void, ensuring that all urine produced by the animal is captured.
  • Avoid contamination by not allowing contact with feces, bedding, or any foreign materials.
  • Immediately close the collection container after obtaining the sample to prevent contamination or evaporation.

Sample Handling

1. Within 30 minutes of collection, transfer the urine samples to the laboratory while maintaining them at 2-8°C using ice packs.

2. Once in the laboratory, ensure that the samples are properly mixed to prevent any settling of particulate matter.

Sample Preparation for Protein Analysis

1. Centrifuge the urine samples at 2,000 x g for 10 minutes at 4°C to separate cellular debris and sediment.

2. Carefully pipette the supernatant into sterile centrifuge tubes.

3. Centrifuge the supernatant again at 10,000 x g for 10 minutes at 4°C to remove smaller particulate matter.

4. Aliquot the resulting clear supernatant into smaller, labeled tubes.

5. Store the aliquots at -80°C for future protein analysis.

Sample Preparation for Metabolomics Analysis

1. To separate small molecules for metabolomics analysis, take a separate aliquot of the urine sample before any centrifugation steps.

2. Label these aliquots accordingly and store them at -80°C for future metabolomics analysis.

Sample Transportation:

Transport urine samples to the laboratory as soon as possible after collection to maintain sample integrity.

Keep the samples at a temperature of 2-8°C using ice packs or a cooler during transportation to prevent degradation of analytes.

* For Research Use Only. Not for use in diagnostic procedures.
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