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Related Concept Videos

Urine Studies II: Urine Culture and Sensitivity Test01:26

Urine Studies II: Urine Culture and Sensitivity Test

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A urine culture and sensitivity test is a diagnostic procedure used to identify urinary tract bacterial infections and determine the most effective antibiotics for treatment. This test is generally preferred when a patient shows manifestations of a urinary tract infection, such as frequent or painful urination, cloudy or foul-smelling urine, or lower abdominal pain.Purpose of the TestThe primary goals of a urine culture and sensitivity test are to:Determine the specific bacteria causing the...
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Filtration and Urine Formation01:32

Filtration and Urine Formation

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The function of the kidneys is to filter, reabsorb, secrete, and excrete. Every day the kidneys filter nearly 180 liters of blood, initially removing water and solutes but ultimately returning nearly all filtrates into circulation with the help of osmoregulatory hormones. This process removes wastes and toxins but is also crucial to maintain water and electrolyte levels. Most of these functions are performed by the tiny but numerous nephrons contained within the kidneys.
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Polyprotic Acids03:38

Polyprotic Acids

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Acids are classified by the number of protons per molecule that they can give up in a reaction. Acids such as HCl, HNO3, and HCN that contain one ionizable hydrogen atom in each molecule are called monoprotic acids. Their reactions with water are:
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Rate-Determining Steps03:08

Rate-Determining Steps

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Relating Reaction Mechanisms
In a multistep reaction mechanism, one of the elementary steps progresses significantly slower than the others. This slowest step is called the rate-limiting step (or rate-determining step). A reaction cannot proceed faster than its slowest step, and hence, the rate-determining step limits the overall reaction rate.
The concept of rate-determining step can be understood from the analogy of a 4-lane freeway with a short-stretch of traffic-bottleneck caused due to...
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Sputum Studies I: Gram Stain, cytology, and Acid-fast smear and culture01:26

Sputum Studies I: Gram Stain, cytology, and Acid-fast smear and culture

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Sputum studies are a critical part of diagnosing and treating numerous respiratory conditions. These studies involve obtaining sputum samples for analysis to identify pathogenic organisms and assess the presence of abnormal cells indicative of malignant conditions. This lesson will delve into three fundamental sputum studies: Gram Stain, Cytology, and Acid-fast Smear and Culture.
Gram Stain
The Gram Stain is an integral part of sputum studies. It involves the staining of sputum, which permits...
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Physiology of Urine Formation01:24

Physiology of Urine Formation

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Urine formation is an essential function of the human body. It plays a critical role in maintaining homeostasis by regulating the volume and composition of body fluids. The kidneys, the primary organs involved in this process, filter blood to remove waste products and excess substances, ultimately producing urine.
Glomerular Filtration
The first stage in urine formation is glomerular filtration. Each kidney contains approximately 1 million nephrons, the functional units of filtration, with a...
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Related Experiment Video

Updated: Jan 24, 2026

One-step Metabolomics: Carbohydrates, Organic and Amino Acids Quantified in a Single Procedure
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A fast, sensitive, single-step colorimetric dipstick assay for quantifying ascorbic acid in urine.

Marcia A LeVatte1, Matthias Lipfert1, Jiamin Zheng1

  • 1Department of Biological Sciences, University of Alberta, Edmonton, AB, T6G 2E8, Canada.

Analytical Biochemistry
|June 3, 2019
PubMed
Summary

A new, fast, and sensitive colorimetric assay using MTT and PMS can now accurately detect vitamin C (ascorbate) in urine. This method offers a stable, low-cost alternative for monitoring health and potentially detecting colorectal polyps.

Keywords:
AscorbateColorectal cancerColorimetric assayDipstickUrine

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A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells
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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Biomedical Diagnostics

Background:

  • Urinary ascorbate serves as a biomarker for vitamin C intake and may aid in detecting precancerous colorectal polyps.
  • Existing methods for urinary ascorbate quantification are often time-consuming, expensive, or lack sufficient accuracy for clinical applications.
  • Current low-cost paper-based dipsticks have inadequate detection limits and quantification accuracy for human urine analysis.

Purpose of the Study:

  • To develop a rapid, sensitive, and accurate single-step colorimetric assay for quantifying ascorbate in human urine and other biological fluids.
  • To create a cost-effective and stable paper-based dipstick for ascorbate detection.
  • To validate the assay's performance against established analytical techniques.

Main Methods:

  • A novel colorimetric assay utilizing methylthiazolyldiphenyl-tetrazolium bromide (MTT) and phenazine methosulfate (PMS) in an acidic buffer.
  • Development of stable, paper-based dipsticks for the colorimetric assay.
  • Validation of the assay using Liquid Chromatography-tandem Mass Spectrometry (LC-MS/MS) and Nuclear Magnetic Resonance (NMR) spectroscopy in human urine samples.

Main Results:

  • The developed assay produces a vivid purple color in the presence of ascorbate.
  • The lower limit of detection for the ascorbate dipstick assay was determined to be 3.2 μM.
  • The paper dipsticks demonstrated stability over a wide temperature range and a storage life of up to 150 days.

Conclusions:

  • A fast, sensitive, and single-step colorimetric assay has been successfully developed for ascorbate quantification in biological fluids, including urine.
  • The new assay overcomes the limitations of existing methods, offering a cost-effective and accurate alternative.
  • The stable, paper-based dipsticks show promise for widespread use in health monitoring and early disease detection, such as for colorectal polyps.