t h e e f f e c t o f i o n i z i n g r a d i a t i o n s o n c e r t a i n CARBOHYDRATES AND RELATED SUBSTANCES Dissertation Presented In Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy In the Graduate School of the Ohio State University ANDREW UICHAEL MICHELAKIS, B. Sc., M. Sc. The Ohio State university 1959 Approved by Adviser Department of Chemistry ACKNOWLEDGMENT The author Is very grateful and Indebted to Professor M. L. Wolfram for his inspiration, encouragement and guidance In the course of this investigation. Th e association with all the members of the Sugar Alley has been very pleasant, acknowledgment Is expressed to all of them, and especially to Mr. Leo J. McCabe. The author wishes to express his sincere appreciation and thanks to Mrs. Mary Leonid&kls for typing this thesis. Acknowledgment is made to Hr. B. Schmidt and Professor Dudley Williams of the Department of Physics for their help in obtaining and interpreting the electron- spin resonance spectra. The financial help provided by the Quartermaster Food and Container Institute for the Armed Forces, Research and Development Command, Quartermaster Corps, U. S. Army, Is acknowledged. Acknowledgment is also made to the Department of Chemistry for support as a teaching assistant. ii TABLE OF CONTENTS IMS INTRODUCTION........................... 1 Types of Radiation. ....................... 2 I. Photolytic Radiation............... 2 II. Ionizing Radiation...................... 5 1. Units and Terms Used In Ionizing Radiation ........... 3 2. Cathode Rays........ 4 3. Gamma Rays....................... 5 4. Action of Radiation............... 6 STATEMENT OF PROBLEM.................... 7 HISTORICAL............................. 8 I. Photolytic Radiation..,. ......... 8 II. The Irradiation of Water......... 12 III. Direct and Indirect Action of Ionizing Radiation...................... 16 IV. Distinction of Radiation Chemistry from Photochemistry......... 16 V. Irradiation of Polymers Other Than Carbohydrates...................... 17 VI* Irradiation of Carbohydrates............. 18 1. Effect of Ionizing Radiation on Cellulose and Its Derivatives...... 18 2. Effect of Ionizing Radiation on Dextran............... SI 3. Effect of Ionizing Radiation on Inn 1 in...................... 23 ill TABLE OF CONTENTS (Contd.) £ass 4. Effect of Ionizing Radiation on Pectin.................... 25 5. Effect of Ionizing Radiation on Starch........................ 24 6 . Effect of Ionizing Radiation on Oligosaccharides............... 25 a. Rafflnose ............ 26 b. Maltose............... 26 c. Sucrose................... 26 d. Lactose................... 27 7* Effect of Ionizing Radiation on Monosaccharides.................. 26 8 . Effect of Ionizing Radiation on 4-D-glucopyranoside............ 30 9. Effect of Ionizing Radiation on Polytaydric Alcohols............... 51 VII* Paramagnetic Resonance Study of Irradiated Carbohydrates. ............. 32 EXPERIMENTAL......................... 54 I. Irradiation Sources..................... 34 II. The Effect of Ionizing Radiation on Maltose. ••••........................... 54 1. Irradiation of Maltose with Cathode Rays as Fifty Per Cent Aqueous Solutions at 0°C........... 34 a. Descending paper chromatography of maltose Irradiated as 50# aqueous solutions with cathode rays at 0°C................. 35 !▼ TABLE OF CONTENTS (Contd.) Page 2* Irradiation of Maltose as Twenty Per Cent Aqueous Solutions with Cathode Beys................. 36 a. Descending paper chromatography of Irradiated 2 0 % aqueous maltose with cathode rays at ambient air, ice-water and ethanol-dry Ice temperatures.,**. 38 b. Determination of per cent hydrolysis of irradiated 20# aqueous maltose with cathode rays....... 39 c* Calculation of G values of irradiated 20 # aqueous maltose with cathode rays........ ..... 40 d. Isolation of irradiation products from maltose irradiated with cathode rays 41 e. Acetylatlon of the slrupy monosaccharide fraction obtained from carbon column chromatography of maltose...... 42 f. The separation and Identification of the acetylated monosaccharide derivatives of irradiated 20# aqueous maltose at 0°C......... 43 III* The Effect of Ionizing Radiation on Celloblose...................... 46 1* Irradiation of Celloblose with Cathode Rays as Fifty Per Cent Aqueous Solutions at 0°C....... 46 a* Descending paper chromatography of the Irradiated 50# aqueous celloblose samples*...... 46 v TABLE OF CONTENTS (Contd.) £&&a 8 . Irradiation of Celloblose as Twenty Per Cent Aqueous Solutions with Cathode Bays at Q°C......... 47 a* Descending paper chromatography of Irradiated 2 0 % aqueous celloblose with cathode rays at 0°C....................... 47 b. Determination of the per cent hydrolysis of 20% aqueous celloblose Irradiated with cathode rays at 0°C............ 46 c. Determination of 0 values of Irradiated 2 0% aqueous celloblose with cathode rays at 0°C....................... 49 IV. The Effect of Ionising Radiation on Trehalose.......... ............... 49 1. Irradiation of Trehalose with Cathode Rays as Two Per Cent Aqueous Solutions at Ambient Air Temperature ••. 49 a. Chromatography of irradiated trehalose as 2% aqueous solutions with cathode rays 50 b. Determination of the per cent hydrolysis of trehalose Irradiated as 2$ aqueous solutions.................... 50 c. Calculation of G values of irradiated trehalose........... 51 d. Isolation of Irradia tion products from irradiated trehalose. Dose received 80 megarads of cathode rays....... 51 e. taper chromatography of the monosaccharide fraction obtained from irradiated trehalose by separation on a carbon column ........... 58 vl TABLE OF CONTENTS (Contd.) Eg&ft f# Acetylation of the obtained monosaccharide fraction..... 53 g„ The separation and Identification of the acetylated monosaccharide derivatives of Irradiated trehalose.................... 54 V. The Effect of Ionising Radiation on Raff inose............................ 55 1. The Irradiation of Rafflnose with Gamma Rays........................ 55 a. Descending paper chromatography of Irradiated raff lnose as 8$ aqueous solutions with gamma rays................... 55 b. Ionophoresis of the raff lnose samples exposed to gamma radiation as 2% aqueous solutions.................... 56 c. Determination of copper reducing sugar values of raff inose exposed to gamma radiation as 2 % aqueous solutions. .................. 57 2. The Irradiation of Two Percent Aqueous Raffinose with Cathode Rays at Ambient Air Temperature.......... 56 a. Chromatography of the Irradiated 2$ aqueous raffinose solutions with cathode rays 58 b. Ionophoresis of rafflnose Irradiated with cathode rays as 2% aqueous solutions........ 59 c. Determination of the copper reducing sugar values of rafflnose irradiated as 2% aqueous solutions with cathode rays................. 60 vii TABLE OF CONTENTS (Contd.) Page VI. The Effect of Ionising Radiation on Inulin............................. 61 1. The Irradiation of Inulin as Two Per Cent Aqueous Solutions with Gamma Rays......................... 61 a. Physical properties of inulin irradiated as 8$ aqueous solutions with Co^O gamma radiation at 24°C............... 61 b. Descending paper chromatography of inulin irradiated with gamma rays..................... 62 c. Paper ionophoresis of inulin Irradiated with Co60 gamma radiation...................... 62 d. Determination of the extent of inulin hydrolysis. Inulin exposed to gamma radiation as 2j6 aqueous solutions............ 63 e. Isolation of irradiation products from inulin irradiated with Co60 gamma radiation as 8% aqueous solutions. .................... 64 f. Paper chromatography of the slrupy fraction obtained from carbon column chromatographic separation of inulin irradiated with gamma rays................. 65 g. Acetylation of the slrupy monosaccharide fraction obtained from carbon column chromatography of Co60 gamma rays irradiated inulin.......... 66 2. The Irradiation of Powdered Inulin with Cathode Rays................... 68 a. Effect of cathode ray radiation on the physical properties of powdered inulin................. 68 ▼ill TABLE OF CONTENTS (Contd.) Page b. Chromatography of powdered Inulin Irradiated with 400 megareps of cathode rays.......... 68 S. The Irradiation of Two Per Cent Aqueous Inulin Solutions with Cathode Rays at Room Temperature....... 69 a. The effect of cathode radiation on the physical properties of 2 % dilute aqueous inulin solutions .......... 69 b. Chromatography of inulin irradiated with cathode rays as 2 % aqueous solutions........... 70 c. Ionophoresis of inulin irradiated with cathode rays as 2 % aqueous solutions.......... 70 d. Extent of hydrolysis of inulin exposed to cathode radiation as 2 % aqueous solutions.......... 71 VII. The Effect of Ionising Radiation on Amylose. ............................ 72 1. The Irradiation of Two Per Cent Aqueous Mixture of Amylose with Gamma Radiation........ ........... 72 a. Carbon column chromatography of amylose irradiated with gamma rays........................... 72 b. Acetylation of the slrupy monosaccharide obtained from carbon column chromatography of irradiated amylose............ 73 c. The separation and identification of the acetylated monosaccharide derivatives of irradiated aqueous amylose mixture.......... 74 VIII. The Effect of Ganna Radiation on D-Glucosamine Hydrochloride, D-Glucuronic acid and D-Galacturonlc Acid................. 76 lx TABLE OF CONTENTS (Contd.) 1 . Irradiation Procedure........... 76 £• The Collection of the Gases Produced from the Irradiated D- Glucosamine