NORTHWESTERN UNIVERSITY LIBRARY Manuscript TheBes Unpublished theses submitted for the Master*s and Doctor's degrees and deposited In the Northwestern University Library are open for inspection, but are to be used only with due regard to the rights of the authors. Bibliographical references may be noted, but passages may be copied only with the permission of the authors, and proper credit must be given in subsequent written or published work. Exten­ sive copying or publication of the thesis in whole or in part requires also the consent of the Lean of the Graduate School of Northwestern University. TheseB may be reproduced on microfilm for use in place of the manuscript itself provided the rules listed above are strictly adhered to and the rights of the author are in no way Jeopardized. Th i s th e s i s by has been used by the following persons, whose signatures attest their acceptance of the above restrictions. A Library which borrows thiB thesis for use by its patrons is expected to secure the signature of each user. NAME AND A DURESS LATE NORTHWESTERN UNIVERSITY THE RESTRICTED DIFFUSION OF AMINO ACIDS THROUGH CELLULOSE MEMBRANES A DISSERTATION SUBMITTED TO THE GRADUATE SCHOOL IN PARTIAL FULFILIMENT OF THE REQUIREMENTS for the degree DOCTOR OF PHILOSOPHY FIELD OF PHYSIOLOGICAL CHEMISTRY By JOHN WILFRID VICTOR HAHN EVANSTON, ILLINOIS APRIL, 1951 ProQuest Number: 10101487 All rights reserved INFORMATION TO ALL USERS The quality of this reproduction is dependent upon the quality of the copy submitted. In the unlikely event that the author did not send a complete manuscript and there are missing pages, these will be noted. Also, if material had to be removed, a note will indicate the deletion. uest ProQuest 10101487 Published by ProQuest LLC (2016). Copyright of the Dissertation is held by the Author. All rights reserved. This work is protected against unauthorized copying under Title 17, United States Code Microform Edition © ProQuest LLC. ProQuest LLC. 789 East Eisenhower Parkway P.O. Box 1346 Ann Arbor, Ml 48106 - 1346 It ACKKOWLEDaHlgEKTS The author wishes to express bia appreciation to all who have assisted in the oampletion of thia work; To professor Henry B. Bull* under whose guidance thie project was undertaken* for his constant interest and helpful criticism duping the course of the investigation. XP Mr. Hector Scannon® for his generous teohnlcaX assistance with the preliminary experiment®• To the Qpm products Refining company of Argo, Illinois* Whose generous financial contribution in the form of a fellowship made it poesiole for the writer to pursue the study* TABEjE OP CONTENTS ACKNOWLEDGEMENTS ............. ..................... Pago Jt TABLE OF CONTESTS........................................ Tf LIST OF TABLES ^... viX LIST OF FIGURES ........................ i* INTRODUCTION ........................ X DEFINITION OF MEMBRANE ..... X KINDS OF HIM BRAKES...... X ARTIFICIAL NEW BRAKES AND THEIR MECHANISMS ....... X EARLY STUDIES OF RESTRICTED DIFFUSION AND MEMBRANE EQUILIBRIA ........................... S MEMBRANE POTENTIALS ...... 6 ANOMALOUS OSMOSIS ................................. 7 FIXED' CHAROF. THEORY AND'MEMBRANE BEHAVIOR........ 8 HETEROPOROSITY AND MEMBRANE BEHAVIOR .............. XO BIfclONIO POTENTIALS ..................... XX KINETICS OF MEMBRANE DIFFUSION.................. 13 Steady Flow Through Membranes ..... 14 Energies of Activation ................ X9 Kntropiaa of Activation ..................... 19 MODERN BATE THEORY AND MEMBRANE DIFFUSION ........ 84 InfXuanoa of Potential Gradient ............. 87 Influence of External Forces ............... 89 STATEMENT OF THE PROBLEM ................. 38 EXPERIMENTAL PROCEDURE* MATERIAL AND METHODS ...... 33 REAGENTS.......................... 33 MEMBRANES ....... 33 TEMPERATURE........................................ 54 pS MEASUREMENTS...... 34 CONCENTRATION OF AMINO ACIDS BY REFRACTIVE INDEX.. 34 PRELIMINARY EXPERIMENTS.................. 33 The Diffusion Ceil ......... 36 procedure ................................... 36 OASTOLITE PLASTIC CELL ........ 39 Construction of the Cell ........ 30 The Manometer System................. 45 The wash System ........... 47 procedure......................... 47 ▼1 TABLE OF CONTENTS (CONTINUED) DATA AND RESULTS .........Page 60 EFFECT OF CONCENTRATION ON REFRACTIVE INDEX ... 60 EXPRESSION OF DIFFUSION RATE AS A FIRST ORDER PROCESS...... 60 VARIATION OF PERMEABILITY CONSTANT WITH FLOW RATE OF WASH SOLUTION .... ......... ....... . . 64 DEPENDENCE OF PERMEABILITY CONSTANT ON ELECTROLYTE CONCENTRATION....... 86 DEPENDENCE OF PERMEABILITY CONSTANT ON CONCENTRATION OF DIFFUSING SPECIES ....... 86 VARIATION OF PERMEABILITY CONSTANT WITH pH .... 88 PERMEABILITY CONSTANTS FOR VARIOUS AMINO ACIDS, GLUCOSE AND GLYCYLOLYCINE..... ........ 66 TRANSPORT OF WATER AOROSS THE WBJBRANE........ 67 ' INFLUENCE OF TEMPERATURE ..... 68 DISCUSSION OF RESULTS ..................... 60 THE DIFFUSION RATE AS A FIRST ORDER PROCESS ... 60 VARIATION OF PERMEABILITY CONSTANT WITH MOLECULAR SIZE ................................ 77 VARIATION OF PERMEABILITY CONSTANT WITH pH .... 88 The Effect of Membrane Charge on k, , kj .. 86 An Expression for the Charge on the Membrane ...... 87 Calculation of fwitter ton permeability Constant ................................. 88 Oaloulation of Cationic and Anlonio permeability Constanta ................... 88 Caloulatlon of Membrane ionisation Constant ................................. 08 Oaloulation of catlonlo and Anlonio permeability constants from MWmbrane Equations 98 The Membrane ionisation Constant ......... OS EFFECT OF CONCENTRATION ON PERMEABILITY CONSTANT ....................... 04 EFFECT OF TEMPERATURE ON PERKEABIIXTY OONSTAKT ...... 07 ©aergy of Activation and Heat of Adsorption ............................... 07 Entropy of Aotlvation .................... 101 CONCLUSIONS . ..... 105 BIBLIOGRAPHY ............... 106 n LIS? QF TABLES PAX* Refractive Index of Alanine Solutions * 60 Variation of Permeability constant for Alanine with Wash Plow Hate 64 Variation of Permeability Constant for Alanine with Ionic strength of Solvent • 65 Variation of Permeability Constant for Alanine with Concentration of Solution • 56 permeability Constanta of Alanine at Various pH’s* Preliminary Experiments • 69 permeability Constants of ^lutamic Acid at Various pl^e* preliminary Experiments 69 permeability Constants of Alanine at Various pH1a* Plastic Cell ........... 66 permeability Constants of Glucose at Various pH1a* Plastic Cell 66 Permeability Constants for Various Substances at 30.0 Degrees Centigrade • 66 Variation of permeability Constant for Alanine with Temperature • • • • , * • « 68 Values of K from Free and Restricted Diffusion Data 78 Values of Adsorption Equilibrium Constant 76 Halation between Permeability constants and Apparent Molal volumes • • • • • • • 78 Free Diffusion Coefficients from permeability Constants............ . . 81 Values of permeability Constants for Alanine Cetions and Anions using equation * H, k, * and D»K * ^ k3 90 ▼ill MSS & .gg (oggK W g X gftgg., £ A ^ X6 Values of Permeability Constanta for Alanine cations and Aniona using Equations Developed from Membrane Theory • • * , , » * 98 17 Relation between Concentration of Alaninef permeability Constant# and Relative Viscosity of Alanine Solutions in Phosphate Duffer of Ionic Strength 0*2 at pH 6*0 • * # • • • * • 95 18 values for Entropies of Activation for Restricted and Free Diffusion as a function of the Equilibrium Distances • « • • • » » • 108 ix LIST OF FIGURES Figure « page 1 Membrane-Solutlon Interface Conditions According to Laidler and Schuler • • • » * • 15 2 Hydrostatic Preasure^Time Eolation According to Laldler and Schuler • » * • • • IS 5a Schematic Free Energy Diagram Pgu~ Diffusion According to Theory of Absolute Reaction Hates 20 3b Schematic Free Energy Diagram for Membrane Diffusion According to Theory of Tyring * et al . , . .24 4 Diagram of Glass Cell • • * . * • * • • * * • 37 5a-f Diagram of Plastic Cell • « • • • • • • * • • 40*41 6 Schematic Drawing of Plastic Cell and Flow* System Assembly . « • • « , * * ......... * . 46 7 Refractive Index Increment for Alanine • • , 51 8 Diffusion of Serine as a First Order Process 53 9 permeability Constant of Alanine as a Function of Concentration * • • • » • • * • • 57 10 permeability constant of Alanine as a Function of pH* Glass Cell • » * • • * « • « 60a 11 permeability Constant of Slutamlo Acid as a Function of pH. Glass Cell • • • . • • » . 60 b 12 Permeability constant of Alanine as a Function of pH. plastic Cell . . . . . . . . 63 13 permeability Constant of Alanine as a Function of pH* Grouped Data • • • • • • • * 64 14 permeability Constant as a Function of Molecular weight * * • « * • * • • .......... 80 15 Permeability Constanta of Alanine Cations and Anions as a Function of pH • * . • • • • 91