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Physico-Chemical Characterization and Biological Activity of Daucus Carota Cultivars Indigenous to Pakistan

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Physico-Chemical Characterization and Biological Activity of Daucus Carota Cultivars Indigenous to Pakistan PHYSICO-CHEMICAL CHARACTERIZATION AND BIOLOGICAL ACTIVITY OF DAUCUS CAROTA CULTIVARS INDIGENOUS TO PAKISTAN By Nadeem Abbas Faisal 2011-GCUF-05654 Thesis submitted in partial fulfillment of the Requirements for the degree of DOCTORATE OF PHILOSPHY IN APPLIED CHEMISTRY DEPARTMENT OF APPLIED CHEMISTRY GOVERNMENT COLLEGE UNIVERSITY FAISALABAD 1 2 DEDICATION This thesis is dedicated to: The sake of Allah, my Creator and my Master My great teacher and messenger, Muhammed (PBUH), who taught us the purpose of life. My academic teachers, they have made a positive difference in my life My great parents, who never stop giving of themselves in countless ways. My dearest wife, who leads me through the valley of darkness with light of hope and support. My beloved kids, Usman and Hasnat, whom I can't force myself to stop loving. 3 DECLARATION The work reported in this thesis was carried out by me under the supervision of Dr. Shahzad Ali Shahid Chatha, Associate Professor, Department of Chemistry, Government College University Faisalabad, Pakistan. I hereby declare that the title of review “Physico-Chemical Characterization and Biological Activity Of Daucus Carota Cultivars Indigenous To Pakistan” and the contents of the this thesis are the product of my own research and no part has been copied from any published source (except the references). I further declare that this work has not been submitted for award of any other degree/diploma. The University may take action if the information provide is found inaccurate at any stage. _______________ Nadeem Abbas Faisal 2011-GCUF-05654 4 5 6 CONTENTS List of Tables vi List of Figures viii Acknowledgments ix Chapter 1 INTRODUCTION 1 Chapter 2 REVIEW OF LITERATURE 6 2.1 Traditional uses 7 2.2 Phytochemistry 8 2.3 Carotenoids 11 2.4 Carbohydrates /Sugars 13 2.5 Phenolic acids 18 2.6 Volatiles 25 2.7 Nutritional aspects and potential as functional foods 27 2.8 Pharmacological aspects/health benefits 29 2.8.1 Antioxidant potential 30 2.8.2 Anti-Inflammatory potential 33 2.8.3 Anti-cancer potential 34 2.8.4 Protective potential for cardiovascular diseases 36 2.8.5 Hepatoprotective, wound healing, anti-bacterial and 37 antiviral activities Chapter 3 MATERIALS AND METHODS 40 3.1 Materials 40 3.1.1 Chemicals and reagents 40 3.1.2 Standard compounds 40 3.1.3 Instruments 41 3.1.4 Collection of samples 42 3.1.5 Preparation of the plant material for chemical analysis 42 3.3 Experimental protocol 42 3.3.1 Proximate analysis of carrot cultivars 42 3.3.2 Estimation of physico-chemical properties of carrot seed oil 48 3.3.3 Estimation of physico-chemical properties of D. carota juice 51 7 3.4 Chromatographic analysis 52 3.4.1Phenolic acid profile by HPLC 52 3.4.1.1 Sample preparation for the determination of 52 phenolic compounds by HPLC 3.4.1.2 Exploration of phenolic compounds by HPLC 53 3.4.2 Saccharides (sugars) profile by HPLC 53 3.5 Spectrophotometric analysis 55 3.5.1 Sample preparation for β carotenes analysis 55 3.5.2 Determination of β carotenes by spectrophotometer 56 3.5.3 Determination of total phenolic (TP) 56 3.5.4 Determination of total ascorbic acid (TAA) 56 3.5.5 Biological activities 57 3.6 A study for the effectiveness of carrot juice on blood parameters 59 3.6.1 Extraction of D. carota juice 59 3.6.2 Subjects 60 3.6.3 Experimental groups 60 3.6.4 Collection of blood samples 60 3.6.5 Blood analysis 60 3.6.6 Total antioxidant status 61 3.6.7 Malondialdehyde status 61 3.7 Statistical analysis 61 Chapter 4 RESULTS AND DISCUSSION 62 4.1 Proximate composition 62 4.1.1 Proximate composition of seeds 62 4.1.2 Proximate composition of carrot roots 64 4.1.3 Mineral profiles of selected cultivars of carrot 67 4.1.4 Proximate composition of top whole of D. carota cultivars 69 4.1.5 Physcico-chemical properties of carrot seed oil 73 4.2 Quantification of individual compounds 78 4.2.1 Quantification of individual sugar compounds 78 4.2.2 Quantification of individual phenolic compounds in 99 8 different cultivars of D. carota roots 4.2.3 Quantification of β-carotene 100 4.3 Antioxidant activities 113 4.3.1 Antioxidant activities of D. carota roots 113 4.3.2 Antioxidant activities of dehydrated D. carota top whole 116 4.3.3 Antioxidant activities of seeds extract D. carota 122 4.4 Physical analysis of carrot juice 127 4.5 Effect of carrot juice on human blood 130 Chapter 5 SUMMARY 138 Conclusion 141 References 143 9 LIST OF TABLE Serial Title Page No. No. 2.1 Traditional uses of carrot 9 2.2 Carotenoids content in carrots of different color. 15 2.3 Comprehensive list of the sugars present in carrots 19 2.4 Comprehensive list of the phenolic acids isolated from Daucus carota 22 2.5 Proximate composition of carrots 31 2.6 Mineral composition of carrots 31 2.7 Vitamin profile of carrots 32 3.1 Description of instruments used throughout the research work 41 3.2 Description of the carrot roots and top whole employed in the current study 43 4.1 Proximate composition of Daucus carota cultivars seeds 66 4.2 Proximate composition Daucus carota cultivars roots 70 4.3 Mineral composition of D. carota roots 71 4.4 Proximate composition of Daucus carota cultivars top whole 76 4.5 Physico-chemical properties of Daucus carota cultivars seed oil 77 4.6 Linear regression parameters obtained from the sugar standards calibration 80 curves. 4.7 Validation parameters for the liquid chromatography method 81 4.8 Sugar recovery rates (standards) added to the D. carota cultivars samples. 82 4.9 Concentration (means ± standard deviation) of sugars in different cultivars of 84 D. carota 4.10 Phenolic acids profile in different cultivars of D. carota roots 101 4.11 β-carotene yield extracted from D. carota cultivars at different temperatures 112 4.12 Total ascorbic acid and total phenolics of selected cultivars of D. carota roots 117 4.13 Antioxidant activities of selected cultivars of D. carota roots 118 4.14 Antioxidant activities of selected cultivars of D. carota top whole 125 4.15 Antioxidant activities of seed extracts from different D. carota cultivars 126 4.16 Physical properties o of D. carota juice collected from selected cultivars of D. 129 carota roots. 10 Page Serial Titles No. No. 4.17 Physical parameters, blood chemistry, lipid profile, and antioxidant status 134 of volunteers of treatment group No 1 (20 to 30 years) 4.18 Physical parameters, blood chemistry, lipid profile, and antioxidant status 135 of volunteers of treatment group No 2 (31 to 40 years) 4.19 Physical parameters, blood chemistry, lipid profile, and antioxidant status 136 of volunteers of treatment group No 2 (31 to 40 years) 4.20 Physical parameters, blood chemistry, lipid profile, and antioxidant status 137 of volunteers of treatment group No 4 (51 to 60 years) 11 LIST OF FIGURES Serial Title Page No. No. 2.1 Structures of some common isomers of carotenoids 14 2.2 The major phenolic compound present in carrots 24 2.3 Structural formulas of terpinolene, falcarinol, falcarindiol, falcarindio 3- 28 acetate 2.4 Photographs of rats showing different stages of wound healing 38 4.1 Chromatogram of sugars profile for mixture of sugars standards 86 4.2 Typical chromatograms of sugar profiles for DCP cultivar 87 4.3 Typical chromatograms of sugar profiles for DC90 cultivar 88 4.4 Typical chromatograms of sugar profiles for DC3 cultivar 89 4.5 Typical chromatograms of sugar profiles for DCW cultivar 90 4.6 Typical chromatograms of sugar profiles for DCY cultivar 91 4.7 Typical chromatograms of sugar profiles for T29 cultivar 92 4.8 Typical chromatograms of sugar profiles for DCW cultivar 93 4.9 Calibration curve for individual maltodextrin at standard working conditions 94 4.10 Calibration curve for individual maltotriose at standard working conditions 95 4.11 Calibration curve for individual maltose at standard working conditions 96 4.12 Calibration curve for individual glucose at standard working conditions 97 4.13 Calibration curve for individual fructose at standard working conditions 98 4.14 HPLC chromatograph mixture of standard phenolic compounds 103 4.15 HPLC chromatograph of phenolic compounds in T29 cultivar 104 4.16 HPLC chromatograph of phenolic compounds in DCY cultivar 105 4.17 HPLC chromatograph of phenolic compounds in DCR cultivar. 106 4.18 HPLC chromatograph of phenolic compounds in DC90 cultivar. 107 4.19 HPLC chromatograph of phenolic compounds in DCP cultivar. 108 4.20 HPLC chromatograph of phenolic compounds in DCW cultivar 109 4.21 HPLC chromatograph of phenolic compounds in DC3 cultivar 110 4.22 Relationship between total antioxidant activity and total phenolic contents in 119 D. carota 4.23 Relationship between DPPH scavenging activity with total phenolic contents 120 in D. carota 12 ACKNOWLEDGEMENTS I bow my head before Almighty Allah, The omnipotent, The omnipresent, The merciful, The most gracious, The compassionate, The beneficent, who is the entire and only source of every knowledge and wisdom endowed to mankind and who blessed me with the ability to do this work. It is the blessing of Almighty Allah and His Prophet Hazrat Muhammad (Sallallaho Alaihe Wasallam) which enabled me to achieve this goal. I would like to take this opportunity to convey my cordial gratitude and appreciation to my worthy, reverently and zealot supervisor Dr. Shahzad Ali Shahid Chatha, Associate Professor, Department of Chemistry, Government College University, Faisalabad, Pakistan.
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