How to Estimate Glutathione Peroxidase

Glutathione Peroxidase is an enzyme that converts singlet oxygen associated with Glutathione into triplet oxygen. Oxygen in the singlet energy state is very reactive and will damage biological cells, while oxygen in the triplet energy state is considerably less reactive and is the type of oxygen present in air. A simple method to estimate Glutathione Peroxidase uses the optical density of the whole blood, which is useful in quickly identifying people with Glutathione Peroxidase deficiency.

Things You'll Need

• UV-Vis spectrometer
• Whole blood in UV-vis cuvette

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Instructions

1. • 1

     Place the cuvette in the UV-Vis spectrometer and measure the optical density of the blood. Place the cuvette in the spectrometer and press "Scan." The intensity of the Absorption peak is the optical density.

   • 2

     Multiply the optical density by 7.05.

   • 3

     Add 1.77 to get the Glutathione Peroxidase activity in micromole per minute. The normal Glutathione Peroxidase activity level is 4.0-to-4.5 micromolar per minute per milliliter. Levels below 1.5 appear in organisms deficient in Glutathione Peroxidase.