TLC Chromatography for Urine Analysis: Amino Acids & Diseases

Amino Acids in Urine

1. Which amino acids may be present in the urine of healthy people?

  • Glycine
  • Alanine
  • Glutamine
  • Histidine

Urine Sample Preparation for TLC Chromatography

2. What are the stages of urine sample preparation for TLC chromatography?

  1. Separation of the mixture
  2. Partition Coefficient: A liquid, stationary phase (polar solvent) and a mobile phase (solvent of a lower polarity)
  3. Partition Migration: Polar substances travel at slower rates compared to non-polar substances
  4. Development of the Chromatogram
  5. Drying/Reaction: Ninhydrin-partitioned amino acids yield a violet-purple color
  6. Incubation
  7. Calculation of Rf coefficients

Developing Amino Acids on the Chromatogram

3. What is used to develop the amino acids on the chromatogram?

A mixture of Chloroform, methanol, and 17% NH4OH in a 2:2:1 ratio.

Diseases Detected by TLC Chromatography of Urine

4. What diseases can be detected by TLC chromatography of urine?

  • Phenylketonuria
  • Cystinuria
  • Lysinuric protein intolerance
  • Hartnup disorder
  • Iminoglycinuria
  • Dicarboxylic aminoaciduria