Glucose-6-Phosphate Dehydrogenase Activity

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Block C2 - Safe Motherhood & Neonates


Biochemistry
Office:6th floor, Radiopoetro
Work-plan:None.
Pre-test:3-6 questions, short-answer.
Post-test:Sometimes. Oral only.
Lab Report:
  • Introduction
  • Reagents/Equipment
  • Procedure
  • Result
  • Discussion
  • Conclusion
  • References
Procedure and example
Spectrophotometer readings

Principle[edit]

To find out the activity of the enzyme G-6-PD by measuring the increase in NADPH, which is measured by the increased absorbency of NADPH under UV light with a wavelength of 340 nm.

Functions of the Reagents[edit]

  • Hemolysate: lysed erythrocyte that helps obtain enzyme G-6-PD.
  • Triethanolamine: acts as buffer (maintains pH at: 7.35-7.45).
  • EDTA: anti-coagulant, metal ions binder.
  • NADP: NADP donor. Accepts hydrogen from glucose-6-phosphate.
  • Glucose-6-phosphate (substrate): Converted to 6-phosphogluconate by enzyme G-6-PD.
  • Incubation at 37°C: Optimal enzyme activity happens at 37°C (body temperature).
  • Vortex: Hemogenization.
  • Spectrophotometry: Provides UV rays at wavelength 340nm, the optimal wavelength for NADPH activity.

Questions[edit]

Batch 2011 Pre-test[edit]

  1. The function of NADPH is? Reduces oxidized-glutathione to reduced-glutathione, which protects erythrocytes against oxidative attacks.
  2. Glucose-6-phosphate dehydrogenase reaction? Glucose-6-phosphate + NADP → 6-Phosphogluconate + NADPH.
  3. The diseases caused by G-6-PD deficiency are? Neonatal hyperbilirubinemia, acute hemolysis, and chronic hemolysis.
  4. The decrease in reduced-glutathione leads to? Hemolytic anemia.
  5. The oxidant drugs that cause decreased reduced-glutathione concentration are? Quinine, analgesics, and sulfonamides.