Application of the Erythrocyte Glutathione Reductase Test to the Evaluation of Vitamin B([sup2]) Status in Normal Female Subjects and in Females Taking Oral Contraceptive Agents

APPLICATION OF THE ERYTHROCYTE GLUTATHIONE REDUCTASE TEST TO THE EVALUATION OF VITAMIN B2 STATUS IN NORMAL

FEMALE SUBJECTS AND IN FEMALES TAKING ORAL CONTRACEPTIVE AGENTS

Suzanne Y. Tonkin

TABLE OF CONTENTS

I. Review of the Literature............................................................................................. 128 A. General Introduction........................................................................................ 128 B. Metabolism and Metabolic Function............................................................. 131 C. Effect of Riboflavin Deficiency on Metabolism......................................... 132 D. Clinical Aspects...............................................................................................133 E. Laboratory Assessment of Vitamin B2 Status.............................................. 134

II. Materials and Methods.................................................................................................139 A. Experimental D esign...................................................................................... 139 B. Subject Selection............................................................................................. 139 C. Collection of Blood from Donors.................................................................. 139 D. Separation of Erythrocytes and Plasma........................................................ 140 E. Storage of Erythrocytes................................................................................... 140 F. Preparation of Hemolysate ............................................................................ 140 G. Assay of Erythrocyte Glutathione Reductase Activity............................... 140

1. Materials...............................................................................................140 2. Equipment............................................................................................141 3. M ethod.................................................................................................141 4. Estimation of AA/t (Time Rate of Change in Absorbance)............. 142 5. Calculation of Results......................................................................... 142

H. Coenzyme Activation of Erythrocyte Glutathione Reductase....................143 1. Materials...............................................................................................143 2. Equipment............................................................................................143 3. M ethod.................................................................................................143 4. Estimation of AA/t (Time Rate of Change in Absorbance)............. 143 5. Calculation of Results.........................................................................144