Carnosine content and antioxidant activity from poultry co-products, protein meal and stressed ...

193 Pages·2016·1.63 MB·English

by  Paljinder Manhiani

Clemson University
TigerPrints
All Dissertations Dissertations
1-2010
Carnosine content and antioxidant activity from
poultry co-products, protein meal and stressed
poultry tissues
Paljinder Manhiani
Clemson University, [email protected]
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Manhiani, Paljinder, "Carnosine content and antioxidant activity from poultry co-products, protein meal and stressed poultry tissues"
(2010).All Dissertations. 681.
https://tigerprints.clemson.edu/all_dissertations/681
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CARNOSINE CONTENT AND ANTIOXIDANT ACTIVITY FROM POULTRY CO-
PRODUCTS, PROTEIN MEAL AND STRESSED POULTRY TISSUES

A Dissertation
Presented to
the Graduate School of
Clemson University

In Partial Fulfillment
of the Requirements for the Degree
Doctor of Philosophy
Food Technology

by
Paljinder Singh Manhiani
May 2011

Accepted by
Dr. Paul L. Dawson, Committee Chair
Dr. Julie K. Northcutt
Dr. Thomas R. Scott
Dr. William C. Bridges
ABSTRACT
Four separate studies were conducted to examine carnosine levels and associated
antioxidant activity in poultry co-products, in rendered poultry protein meal, in tissues
from stressed or non-stressed chickens. In the first study, carnosine was extracted from
poultry co-products (head, liver, lungs, tail, gizzard, brain and heart). Liver contained the
highest (102.29 mg/gm) level, while brain contained the lowest level of carnosine (0.95
mg/gm) (p ≤ 0.05). Except brain, all tissue ultrafiltrates (20.87-39.57%) and
reconstituted dry powders (5.66- 14.47%) showed thiobarbituric reactive acid species
(TBARS) inhibition. Head ultrafiltrate and reconstituted dry powder showed maximum
while gizzard showed the minimum metal chelating activity (p ≤ 0.05). Free radical
scavenging activity of ultrafiltrate of all tissues samples ranged from 25.1 to 79.4% while
this activity was higher (29.8 to 84.1%) in the reconstituted dry powder of all tissue
samples. Oxygen radical absorbing capacity (ORAC) values were highest in liver
ultrafiltrate and lowest in heart (p ≤ 0.05). Results indicated that carnosine was present in
all the tissue samples investigated and their ultrafiltrates as well as dry powders of tissue
samples possess antioxidant properties.
In the second study examining poultry protein meal, carnosine content of sample-
G was almost 2.6 times higher than sample–A. TBARS inhibition by sample-G was
15.9% while Sample-A did not exhibit any TBARS inhibition. Metal chelating activity
and free radical scavenging activities of sample-A and sample-G did not differ. ORAC
values (µM Trolox Equivalents /gm of dry sample) of sample-A (84.4) were greater than
sample-G (68.4) (p≤0.05).
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The third study determined carnosine levels in different tissues of broilers under
stress versus non-stress conditions. Corticosterone levels of stressed broilers (24.36
ng/ml) was 10 fold higher (p=0.002) than non-stressed broilers (2.28 ng/ml). There was
significant increase in carnosine content in breast tissue of stressed birds (17.39 mg/gm),
and was 10 times (p=0.005) more than non-stressed birds (1.85 mg/gm). Carnosine
content in thigh of stressed birds (21.25 mg/gm) was approximately 2 fold higher
(p=0.001) than non-stressed birds (11.10 mg/gm). Carnosine content in brain of stressed
birds did not differ (p=0.82) from that in non-stressed birds. Results indicated that
carnosine may play a significant role in muscles during short term stress.
In fourth study, it was determined that TBARS inhibition and metal chelating
activity of carnosine was due to the imidazole ring present in the histidine while free
radical scavenging activity of carnosine was attributed to histidine amino acid.
Overall, conclusions were drawn that poultry byproducts, poultry protein meal
contains carnosine and exhibited antioxidant properties. These antioxidant properties
were due to carnosine’s unique structure. Lastly, stress increases the carnosine levels in
breast and thigh tissues of broilers.

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DEDICATION
I dedicate this work to my parents and family, who always supported me to make
this a success.

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ACKNOWLEDGEMENTS
First of all I would like to thank God for giving me strength and wisdom to
complete my research thesis. I would like to thank Dr. Paul Dawson as my advisor and
mentor without his innovative ideas and intellectual, financial and moral support this
research would not have been accomplished.
I would like to express my sincere thanks to Dr. J.K. Northcutt for being a guide
and for her intellectual and experimental assistance, friendly behavior and conducive
support throughout this research.
I would like to thank to Dr. W.C.Bridges for helping me in solving tough
problems of statistics and analyzing the data and for being there with an answer to all my
statistics questions.
Dr Thomas Scott, besides being busy on dean’s duty was willing to give his
valuable time as my committee member and as teacher for answering cell and
immunology questions and guiding me through possible experimental procedures, many
thanks for your help and support.
I would to express my sincere thanks to Dr. I. Han for being there as a friend and
as a guide to provide me with moral and intellectual support during rough times.
From the core of my heart, I would like to thank my parents, S. Harbans Singh
Manhiani and Surinder Kaur Manhiani without them this dream would not have come
true. My brothers Sukhwinder Singh Manhiani, Rajwinder Singh Manhiani, Maninder
Singh Manhiani and my sister-in-laws Darshana Manhiani and Marlina Manhiani for
their emotional and moral support throughout this long journey. My loving nephews,
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Rupinder, Sammy, Molu and Golu for their love and affection and a cute question: What
is your class teacher’s name?
I would like to recognize my wife, Jaspreet for her encouragement and moral
support. For her helpful discussion about the experimental work and as intellectual guide
to ask very difficult and critical questions about my research and then support or disagree
about the answer I gave.
I would like to thank Jason Raines for being as my room-mate and a person to
discuss my experimental problems.
I thank all my friends and lab mates especially Divija, Chaitali for their help and
support in my sample preparations.
I would like to thank Dr. Pometto and Dr. Haley-Zitin for letting me use the
HPLC.
And lastly, from the bottom of my heart, I would also like to thank the entire Food
Science and Human Nutrition Department , and Department of Animal and Veterniary
Sciences, all the faculty and staff for their direct or indirect help in this research and for
their support throughout this program.
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TABLE OF CONTENTS
Page
TITLE PAGE .................................................................................................................... i
ABSTRACT ..................................................................................................................... ii
DEDICATION ................................................................................................................ iv
ACKNOWLEDGEMENTS ............................................................................................. v
TABLE OF CONTENTS ............................................................................................... vii
LIST OF TABLES ........................................................................................................... x
LIST OF FIGURES ........................................................................................................ xi
CHAPTER
1. INTRODUCTION ....................................................................................................... 1
2. LITERATURE REVIEW ............................................................................................ 5
2.1. Introduction ......................................................................................................... 5
2.2. Properties of Carnosine ....................................................................................... 5
2.3. Biosynthesis of Carnosine ................................................................................... 6
2.4. Metabolism of Carnosine .................................................................................... 7
2.5. Regulation of Carnosine Levels .......................................................................... 8
2.6. Physiological Functions of Carnosine .............................................................. 11
2.7. Synthetic Carnosine versus Natural Carnosine ................................................. 15
2.8. Extraction of Carnosine .................................................................................... 16
2.9. Antioxidant Activity of Carnosine .................................................................... 19
2.10. Metal Chelating Activity of Carnosine ........................................................... 21
2.11. Oxygen Radical Scavenging Capacity of Carnosine ...................................... 22
2.12. Factors Affecting Antioxidant Capacity of Carnosine .................................... 23
2.13. Applications in Food ....................................................................................... 25
2.14. References ....................................................................................................... 29

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Table of Contents (Continued) Page

2. EXTRACTION OF CARNOSINE FROM DIFFERENT
POULTRY BY-PRODUCTS AND MEASURING
ITS ANTIOXIDANT PROPERTIES ................................................................ 41

Abstract .................................................................................................................... 41
3.1. Introduction ....................................................................................................... 43
3.2. Materials and Methods ...................................................................................... 45
3.3. Results and Discussion ..................................................................................... 57
3.4. Conclusions ....................................................................................................... 67
3.5. References ......................................................................................................... 68

3. EXTRACTION OF CARNOSINE FROM POULTRY
PROTEIN MEALS AND MEASURING ITS
ANTIOXIDANT PROPERTIES ....................................................................... 88

Abstract .................................................................................................................... 88
4.1. Introduction ....................................................................................................... 89
4.2. Materials and Methods ...................................................................................... 91
4.3. Results and Discussion ................................................................................... 100
4.4. Conclusions ..................................................................................................... 105
4.5. References ....................................................................................................... 107

4. ANTIOXIDANT ACTIVITIES OF CARNOSINE AND ITS
CONSTITUENT AMINO ACIDS IN DIFFERENT
MODEL SYSTEMS ........................................................................................ 118

Abstract .................................................................................................................. 118
5.1. Introduction ..................................................................................................... 119
5.2. Materials and Methods .................................................................................... 121
5.3. Results and Discussion ................................................................................... 124
5.4. Conclusions ..................................................................................................... 127
5.5. References ....................................................................................................... 129

5. EFFECT OF STRESS ON CARNOSINE LEVELS IN BRAIN,
BREAST AND THIGH OF BROILERS ......................................................... 134

Abstract .................................................................................................................. 134
6.1. Introduction ..................................................................................................... 135
6.2. Materials and Methods .................................................................................... 137
6.3. Results ............................................................................................................. 143
6.4. Discussion ....................................................................................................... 144
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6.5. Conclusions ..................................................................................................... 151
6.6. References ....................................................................................................... 152

APPENDIX .................................................................................................................. 162

A.1: (Stage 1) Extraction of Carnosine from organ samples
(Mass Balance) .................................................................................. 162
A.2: (Stage2) Ultrafilteration to purify extract (Mass
Balance) ............................................................................................ 163
A.3: (Stage3) Freeze drying (Mass Balance) ........................................................ 164
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