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A Laboratory Exercise for Photochemistry and Photobiology: Production of Hydrogen Peroxide

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Fernando Gallardo , Susana Gálvez , Miguel Angel Medina and Antonio Heredia

Departmento de Biología Molecular y Bioquímica, Facultad de Ciencias, Universidad de Málaga-Instituto Andaluz de Biotecnología, Campus de Teatinos,E-29071 Málaga, Spain

Carlos Gómez-Moreno

Departamento de Biología Molecular y Bioquímica y Celular, Facultad de Ciencias, Universidad de Zaragonza, E-50009 Zaragoza, Spain

J. Chem. Educ., 2000, 77 (3), p 375

DOI: 10.1021/ed077p375

Publication Date (Web): March 1, 2000

Section:

History, Education, and Documentation

Abstract

Light-excitation of chromophores, changes in redox potential, and transfer of electrons are complex concepts for many students in chemistry and biochemistry. In the proposed experimental work, students can assimilate these concepts by developing a photochemical system to synthesize a product of interest in industry and pharmacy, hydrogen peroxide. The photochemical system has three basic components: (i) the photosensitizer, a molecule that is excited by visible photons allowing the electron flux from (ii) an electron donor to (iii) an electron acceptor. In this laboratory, riboflavin was the photosensitizer and semicarbazide and molecular oxygen were the electron donor and acceptor, respectively. The hydrogen peroxide formed was enzymatically quantified using horseradish peroxidase and o-dianisidine as substrate. Production of hydrogen peroxide can be detected after 20 min of illumination using a simple desk lamp with a bulb of 60 W. The system was developed at pH 7.5 and pH 12.3 to allow discussion about the reactivity of flavin semiquinone forms. As in bacterial and plant photosynthesis, part of the energy absorbed in the photochemical system is used to drive an endergonic reaction.

Keywords (Audience):

Upper-Division Undergraduate

Keywords (Domain):

Biochemistry

Keywords (Pedagogy):

Hands-On Learning / Manipulatives

Keywords (Subject):

Enzymes

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This article has been cited by 1 ACS Journal articles (1 most recent appear below).

Understanding the Facile Photooxidation of Ru(bpy)₃²⁺ in Strongly Acidic Aqueous Solution Containing Dissolved Oxygen
Amitava Das, Vishwas Joshi, Dilip Kotkar, Vinit S. Pathak, V. Swayambunathan, Prashant V. Kamat, and Pushpito K. Ghosh
The Journal of Physical Chemistry A2001 105 (28), 6945-6954
- Understanding the Facile Photooxidation of Ru(bpy)₃²⁺ in Strongly Acidic Aqueous Solution Containing Dissolved Oxygen
  Amitava Das, Vishwas Joshi, Dilip Kotkar, Vinit S. Pathak, V. Swayambunathan, Prashant V. Kamat, and Pushpito K. Ghosh
  The Journal of Physical Chemistry A2001 105 (28), 6945-6954
  The previously observed facile photooxidation of Ru(bpy)32+ to Ru(bpy)33+ in oxygenated solutions of 9 M H2SO4 (Kotkar, D; Joshi, V.; Ghosh, P. K. Chem. Commun. 1987, 4; Indian Patent No. 164358 (1989)) is further studied. A similar phenomenon was ...
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Received: August 03, 2009

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A Laboratory Exercise for Photochemistry and Photobiology: Production of Hydrogen Peroxide