Xanthophylls: Distribution and Metabolism in Avian Egg
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XANTHOPHYLLS: DISTRIBUTION AND METABOLISM IN AVIAN EGG Item Type text; Dissertation-Reproduction (electronic) Authors Anjaneyalu, Yernool Verkatarayappa, 1931- Publisher The University of Arizona. Rights Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author. Download date 28/09/2021 14:10:43 Link to Item http://hdl.handle.net/10150/290171 This dissertation has been 62—2426 microfilmed exactly as received ANJANEYALU, Yernool Venkatarayappa, 1931— XANTHOPHYLLS: DISTRIBUTION AND METAB OLISM W AVIAN EGG. University of Arizona, Ph.D., 1962 Chemistry, biological University Microfilms, Inc., Ann Arbor, Michigan XANTHOPHTLLS: DISTRIBUTION AND METABOLISM IN AVIAN EGG Yernool V./ Anjaneymlu A Dissertation Subaitted to the Faculty of the DEPARTMENT 0? BIOCHEMISTRY AND NUTRITION In Partial Fulfillment of the Requirements For the Degree of DOCTOR OF PHILOSOPHY In the Graduate College UNIVERSITY OF ARIZONA 1962 THE UNIVERSITY OF ARIZONA GRADUATE COLLEGE I hereby recommend that this dissertation prepared under my direction by Yernool V. Anjaneyalu entitled XANTHOPHYLLS: DISTRIBUTION AND METABOLISM IN AVIAN EGG be accepted as fulfilling the dissertation requirement of the degree of Doctor of Philosophy Dissertation Director Date/January 1962 A.A. Kurnick After inspection of the dissertation, the following members of the Final Examination Committee concur in its approval and recommend its acceptance:* *This approval and acceptance is contingent on the candidate's adequate performance and defense of this dissertation at the final oral examination. The inclusion of this sheet bound into the library copy of the dissertation is evidence of satisfactory performance at the final examination. STATEMENT BY AUTHOR This dissertation has been submitted in partial fulfillment of requirements for an advanced degree at the University of Arizona and is deposited in the University Library to be made available to bor rowers under rules of the library* Brief quotations from this dissertation are allowable without special permission, provided that accurate acknowledgment of source is made* Requests for permission for extended quotation from or reproduction of this manuscript in whole or in part may be granted by the head of the major department or the Dean of the Graduate College tfien in their judgment the proposed use of the material is in the interests of scholarship. In all other instances, however, permission must be obtained from the author* SIGNED: APPROVAL BY DISSERTATION DIRECTOR This dissertation has been approved on the date shoim below: Professor of Poultry Science i ACKNCWIEDGMENT The author wishes to express his sincere gratitude to Drs, Allen A* Kurnick and Bobby L. Reid for their wise counsel and guidance during the course of this study and in the pre paration of this dissertation} to Drs. Arthur K. Kemmerer, Mitchell G, Varich, James V« Berry, and Alec £• Kelly for their helpful suggestions in the preparation of this dissertation} to the Monsanto Chemical Company for granting a research fellowship to carry out this study; and to my wife, Vasu for her persistence in urging me to leave ny native land, India, to seek a continued education in the United States* ii TABLE QF CONTENTS CHAPTER Page I. REVIEW OF LITERATURE 1 Eggs 1 Skin, Fat, Liver, and Eyes 3 Feathers li Metabolism 5 Effect of .Antioxidants on the Metabolism of Xanthophylls . 9 II. EFFECT OF ANTIOXIDANTS ON THE METABOLISM OF XANTHOPHYLLS 12 Introduction 12 Experimental Procedure 13 Feeding procedure 13 Chemical analysis lU Results and Discussions. ... 16 Effect of Mode of Administration of Ethoxyquin 19 III. RELATIVE DISTRIBUTION OF CAROTENOIDS IN DEHYDRATED ALFALFA MEAL, EGG YOLKS, AND FECES . 22 Introduction ..... .... ..... .. 22 Experimental Procedure 23 Separation and Identification of Xanthophylls of Egg Yolks 25 Separation and Identification of Xanthophylls of Feces. 26 Results and Discussion 27 Relative Effectiveness of Dehydrated Alfalfa Meal and Ground Yellow Corn for Yolk Pigmentation 30 SUMMARY . 32 REFERENCES. k9 iii LEST OF TABLES TABLE Page 1. Low Pigment Basal Diet (Diet 1) 3k 2. Experimental Design to Study Ethoxyquin Effect on Yolk Pigmentation. ........ 35 3* Effect of Ethoxyquin on the Concentration of Carotenoid Pigments in Egg Yolks from Hens fed Dehydrated Alfalfa Meal as the Sole Source of Carotenoid Pigments (July-August, I960). • 36 U« Retention of Beta-carotene and Xanthophylls in Mixed Feed After Three Weeks Storage in the Presence of Ethoxyquin 37 (July-August, I960) $, Effect of Ethoxyquin on Yolk Pigmentation and Retention of Beta-carotene and Xanthophylls in Mixed Feed after Two Weeks Storage (November - December, i960) 38 6. Effect of Ethoxyquin, Butylated Hydroxytoluene and Administration Method on Egg Yolk Pigmentation 39 7. Experimental Diet 2 to Study the Effect of Mode of Administration of Ethoxyquin .... 1+0 8. Effect of Ethoxyquin Administration by Differ ent Methods on Yolk Pigmentation with De hydrated Alfalfa (June-July, 1961) 1*1 9* Experimental Diet 3 for Producing Pigmented Yolks . k2 10. Absorption Maxima of Carotenoids Isolated from Dehydrated Alfalfa Meal, Feces, and Egg Yolks when the Diet of Hens was Supplemented with 10 Per cent Dehydrated Alfalfa Meal U3 11* Relative Concentration of Carotenoid Pigments Isolated from Dehydrated Alfalfa Meal, Feces and Egg Yolks, When the Diet of the Hen was Supplemented with 10 Per cent Dehydrated Alfalfa Meal I4I1 iv TABLE Page 12• Relative Concentrations of Carotenoids in Feed and Feces* U5 13• Relative Effectiveness of Dehydrated Alfalfa Meal and Yellow Corn for Yolk Pigmentation . 1*6 v LIST OF ILLUSTRATIONS Figure Page 1* Average daily pigment concentration of egg yolks from hens fed 6*25 per cent dehydrated alfalfa meal with and without Santoquin (ethoxyquin) for the first 19 days, followed by a low pigment diet to the 37th day of the experiment U7 2, Chromatographic separation of xanthophylls of dehydrated alfalfa U8 vi ABSTRACT The factors affecting egg yolk pigmentation were studied vith White Leghorn hens fed diets containing dehydrated alfalfa steal as the sole source of xanthophyll pigments. The stability of the carotenoid pigments in the presence and absence of an antioxidant (ethoxyquin) was determined by chemical analyses of the feeds prior to and after storage* The biological activity of the dietary pigments was also estimated from the relative concentration of acetone extractable pig ments obtained in the egg yolks with the feeding of these diets* The first study in this series was conducted during the months of July and August. The feeding of the antioxidant in the presence of dehydrated alfalfa meal in the diet resulted in a 53*76 per cent improvement in egg yolk pigmentation. A partial explanation of the improved yolk pigmentation, could have been the 32.16 per cent loss of carotene, and 2U.01 per cent loss of xanthophylls from the feed during the three-week storage period. However, these data also indicated a possible in vivo activity of the antioxidant in the protection of xanthophylls since only 21;.01 per cent of the xanthophylls were destroyed in the feed while egg yolk pigmentation was improved by 53*76 per cent* The effect of temperature on feed stability of carotenoida was determined in a similar study conducted during the months of November and December, at which time the prevailing ambient temperatures were yii considerably lower than during the July and August study. In this instance, essentially no loss in carotenoid pigments from the feed was noted during the two-*week storage period. However, an average increase of 21.50 per cent in the concentration of acetone extract- able pigment in the yolk was obtained with the feeding of the anti oxidant in the diet* In order to evaluate further the in vivo activity of ethoxy- quin, the antioxidant was administered at the same dosage levels 3 times daily by capsule or parenterally. The administration of ethoxy- quin either by capsule or injection proved to be 88 per cent as effective as when the antioxidant was ingested with the feed. When dietary antioxidant (ethoxyquin) was absent, 21 per cent less egg yolk pigmentation was noted and 12 per cent of the dietary xanthophylls were lost during the storage period. The dehydrated alfalfa meal samples employed in these studies contained cryptoxanthin, lutein, violaxanthin, zeaxanthin, flavo- xanthin and three unidentified xanthophylls and beta-carotene by chromatographic analyses* Analyses of feces and of egg yolks obtained from hens fed dehydrated alfalfa meal indicated that violaxanthin, although previously found in the feed, was completely absent from each of these samples. Beta-carotene, cryptoxanthin, lutein, zeaxanthin, and flavoxanthin were present to the extent of U2.19, 1.97, U2.?6, U.70, and 8.16 per cent of the carotenoids ±a the feces as compared to 1.96, 3«5>U, 82.31, 10.01, and 2.12 per cent of the carotenoids in egg yolk for the respective pigments. These data indicate relatively low absorption rates for beta-carotene aid flavoxanthin as compared viii to lutein, while cryptoxanthin and especially zeaxanthin were absorbed to a relatively greater extent than was lutein. From these data it would be expected that yellow corn, which contains a relatively large amount of zeaxanthin, would be a more effective source of egg