Factors Influencing Gastrointestinal
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FACTORS INFLUENCING GASTROINTESTINAL ABSORPTION OF DRUGS DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of the Ohio State University By ANGEL LUIS IGLESIAS, B. S., M. S. The Ohio State University 1958 Approved by .ege of Pharmacy ACKNOWLEDGEMENTS The author wishes to make grateful acknowledgement to Dr. John W. Nelson, Professor, College of Pharmacy, and to Dr. Arthur Tye, Professor, College of Pharmacy, for constant counsel and advice in the fulfillment of this work. ii TABLE OF CONTENTS Page INTRODUCTION -------------------------------------- 1 Methods Used in the Study of Absorption _______ 1 Factors Involved in Absorption ___________ 9 EXPERIMENTAL PROCEDURE ------------------------------ 22 Objectives ____________________ 22 Extraction and Analysis _______________ 23 Experiment I. Absorption Rates of Phenobarbital (5 Mg. Dose) __________________ 30 Experiment II. Absorption Rates (25 Mg. Dose) _ _ - L3 Experiment III. Drug Retained by Intestinal Wall _ _ 53 Experiment IV. Drug Retained by Stomach Wall _ - - 65 Experiment V. Effect of Total Volume of Fluid - - - 77 Experiment VI. Use of Alcohol as Solvent ____-•-81 Experiment VII. Effect of Olive Oil ________ 83 Experiment VIII. Effect of Glucose ________ 85 Experiment IX. Effect of Sucrose 89 Experiment X. Effect of Feeding __________ 91 Experiment XI. Effect of Atropine _________ 94 Experiment XII. Effect of Stress __________ 97 Experiment XIII. Effect of Bentonite Magma _____ 99 Experiment XIV. Effect of Glucosamine Hydrochloride - 101 Experiment XV. Effect of Poloxalkol ________ 103 Experiment XVI. Absorption from the Stomach _ _ _ _ 106 Experiment XVTI. Absorption from the Large Intestine 115 DISCUSSION 118 SUMMARY AND CONCLUSIONS ----------------------------- 123 BIBLIOGRAPHY 125 iii LIST OF TABLES TABLE PAGE 1 Per Cent Phenobarbital Absorbed in 5 Minutes From a 5 Mg. Dose _________________ 33 2 Per Cent Phenobarbital Absorbed in 10 Minutes From a 5 Mg. Dose 34 3 Per Cent Phenobarbital Absorbed in 15 Minutes From a 5 Mg. Dose - - --- _____________ 35 4 Per Cent Phenobarbital Absorbed in 20 Minutes From a 5 Mg. Dose ----------------- 36 5 Per Cent Phenobarbital Absorbed in 25 Minutes From a 5 Mg. Dose _________________ 37 6 Per Cent Phenobarbital Absorbed in 30 Minutes From a 5 Mg. Dose 38 7 Per Cent Phenobarbital Absorbed in 45 Minutes From a 5 Mg. Dose 39 8 Per Cent Phenobarbital Absorbed in 60 Minutes From a 5 Mg. Dose ----------------- 40 9 Per Cent Phenobarbital Absorbed in 75 Minutes From a 5 Mg. Dose 41 10 Per Cent Phenobarbital Absorbed from a 5 Mg. Dose After Different Intervals -------------- 42 11 Per Cent Phenobarbital Absorbed in 5 Minutes From a 25 Mg. Dose _________________ 46 12 Per Cent Phenobarbital Absorbed in 10 Minutes From a 25 Mg. Dose ----------------- 47 13 Per Cent Phenobarbital Absorbed in 20 Minutes From a 25 Mg. Dose ---------- -- _____ 48 14 Per Cent Phenobarbital Absorbed in 30 Minutes From a 25 Mg. Dose 49 15 Per Cent Phenobarbital Absorbed in 45 Minutes From a 25 Mg. Dose ----------------- 50 iv LIST OF TABLES (Continued) TABLE PAGE 16 Per Cent Phenobarbital Absorbed in 60 Minutes From a 25 Mg. Dose 51 1? Per Cent Phenobarbital Absorbed From a 25 Mg. Dose at Different Intervals ------------ 52 18 Per Cent of a 5 Mg. Dose of Phenobarbital Retained By Intestinal Wall After 5 Minutes _______ 55 19 Per Cent of a 5 Mg. Dose of Phenobarbital Retained By Intestinal Wall After 10 Minutes _______ 56 20 Per Cent of a 5 Mg. Dose of Phenobarbital Retained By Intestinal Wall After 15 Minutes _______ 57 21 Per Cent of a 5 Mg. Dose of Phenobarbital Retained By Intestinal Wall After 20 Minutes _______ 58 22 Per Cent of a 5 Mg. Dose of Phenobarbital Retained By Intestinal Wall After 25 Minutes _______ 59 23 Per Cent of a 5 Mg. Dose of Phenobarbital Retained By Intestinal Wall After 30 Minutes _______ 60 24 Per Cent of a 5 Mg. Dose of Phenobarbital Retained By Intestinal Wall After 45 Minutes _______ 6l 25 Per Cent of a 5 Mg. Dose of Phenobarbital Retained By Intestinal Wall After 60 Minutes _______ 62 26 Per Cent of a 5 Mg. Dose of Phenobarbital Retained By Intestinal Wall After 75 Minutes _______ 63 27 Per Cent Phenobarbital Retained by Intestinal Wall From a 5 Mg. Dose _________________ 64 28 Per Cent of a 5 Mg. Dose of Phenobarbital Retained By Stomach Wall After 5 Minutes __________ 67 29 Per Cent of a 5 Mg. Dose of Phenobarbital Retained By Stomach Wall After 10 Minutes _ 68 30 Per Cent of a 5 Mg. Dose of Phenobarbital Retained By Stomach Wall After 15 Minutes 69 v LIST OF TABLES (Continued) TABLE PAGE 31 Per Cent of a 5 Mg. Dose of Phenobarbital Retained By Stomach Wall After 20 Minutes 70 32 Per Cent of a 5 Mg. Dose of Phenobarbital Retained By Stomach Wall After 25 Minutes ---------- 71 33 Per Cent of a 5 Mg. Dose of Phenobarbital Retained By Stomach Wall After 30 Minutes 72 3h Per Cent of a 5 Mg. Dose of Phenobarbital Retained By Stomach Wall After h5 Minutes ---------- 73 35 Per Cent of a 5 Mg. Dose of Phenobarbital Retained by Stomach Wall After 60 Minutes __________ 7b 36 Per Cent of a 5 Mg. Dose of Phenobarbital Retained By Stomach Wall After 75 Minutes 75 37 Per Cent Phenobarbital Retained by Stomach Wall From a 5 Mg. Dose - - _ ____________ 76 38 Effect of Total Volume of Fluid cn Absorption (5 c.c.) 79 39 Effect of Total Volume of Fluid on Absorption (15 c.c.) 80 h0 Effect of Alcohol on Absorption of Phenobarbital - - - 82 hi Effect of Olive Oil on Absorption of Phenobarbital _ _ 8h h2 Effect of Glucose on Absorption of Phenobarbital - _ 88 h3 Effect of Sucrose on Absorption of Phenobarbital - - 90 hh Absorption by Starved Rats 92 h5 Absorption by Fed Rats _______________ 93 h6 Effect of Atropine on Absorption of Phenobarbital - - $6 h7 Effect of Stress on Absorption of Phenobarbital - - - 98 h8 Effect of Bentonite Magma on Absorption of Phenobarbital _________________ 100 h9 Effect of Glucosamine on Absorption of Phenobarbital 102 50 Effect of Poloxalkol on Absorption of Phenobarbital lOh vi LIST OF TABLES (Continued) TABLE PAGE 51 Changes in Per Cent Phenobarbital Absorbed From a 10 Mg. Dose in Twenty Minutes Produced by Varying the Solvent or Administering a Second Substance ______ --- ___________ 105 52 Absorption of Phenobarbital From Stomach (10 Min.) 108 53 Absorption of Phenobarbital From Stomach (20 Min.) 109 54 Absorption of Phenobarbital From Stomach (30 Min.) 110 55 Absorption of Phenobarbital From Stomach (45 Min.) 111 56 Absorption of Phenobarbital From Stomach (60 Min.) 112 57 Absorption of Phenobarbital From Stomach (90 Min.) 113 58 Absorption of Phenobarbital From Stomach (120 Min.) 114 59 Absorption From the Large Intestine (Blank Determinations) ________________ n 6 60 Absorption From the Large Intestine ______ 117 vii LIST OF FIGURES FIGURE PAGE 1 Standard Curve for Phenobarbital Concentration Versus Optical Density ___________ 29 2 Absorption From the Gastrointestinal Tract of the Rat Using a 5 Mg. Dose of Phenobarbital - 32 3 Absorption From the Gastrointestinal Tract of the Rat Using a 25 Mg. Dose of Phenobarbital _ 45 4 Drug Retained by Intestinal Mucosa _______ 54 5 Drug Retained by Stomach Mucosa ________ 66 viii INTRODUCTION Methods Used in the Study of Absorption For the last eighty years physiologists, and more recent ly pharmacologists and biochemists, have sought to explain the mechanisms by which substances, including foods and therapeutic agents, are transferred from the lumen of the alimentary canal to the circulating blood stream. Interest in the subject has increased since evidence has accumulated for the hypothesis that this trans fer in many cases is not simply mediated by physical phenomena alone, but also by biological activity within the mucosa lining the gastro intestinal tract. It is not unusual to find names like Cori, Van Slyke, Kalckar and others, along with those of physiologists and pharmacologists like Starling, von Mering, and Sollmann in the literature dealing with studies on absorption. The most serious difficulty that is met when making studies on absorption in the problem of obtaining an animal prepara tion which will approach, as close as possible those conditions under which a normal, intact animal will absorb foodstuffs or drugs in a truly physiological manner. The use of loops of excised intestines is an old, but still very much used method. Such loops can be used for the passage of substances out of the lumen into a Ringers or Tyrodo solution bathing them on the outside, where the substance being absorbed can be determined (1, 2, 3> 4). This method is far from approach ing truly physiological conditions. The.intestinal mucosa, like the serosa, is very sensitive to handling, changes in temperature, moisture, alterations in circulation etc. Circulation is of the greatest importance in a normal process of absorption and it cannot be expected to be normal even when the utmost care is exerted in the excision of the loops. Recently, excised loops have been used in the study of actively transported substances. These are everted before using, turning the mucosal side to the outside. This is done so that the extremely active epithelium of the mucosa will be exposed directly to the oxygen bubbled into the bathing fluid. Here, the transfer is from the outside fluid to the serosal side inside the loop. Even with the more efficient oxygenation claimed, this method is at least as far apart from physiological conditions as the preceding one. The manual handling to which the loop of intestine is subjected during aversion must necessarily damage the villi lining the mucosa (1, 5 , 6).