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Oxalate and Antioxidant Concentrations of Locally Grown and Imported Fruit in New Zealand

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Oxalate and Antioxidant Concentrations of Locally Grown and Imported Fruit in New Zealand CORE Metadata, citation and similar papers at core.ac.uk Provided by Lincoln University Research Archive Lincoln University Digital Thesis Copyright Statement The digital copy of this thesis is protected by the Copyright Act 1994 (New Zealand). This thesis may be consulted by you, provided you comply with the provisions of the Act and the following conditions of use: you will use the copy only for the purposes of research or private study you will recognise the author's right to be identified as the author of the thesis and due acknowledgement will be made to the author where appropriate you will obtain the author's permission before publishing any material from the thesis. OXALATE AND ANTIOXIDANT CONCENTRATIONS OF LOCALLY GROWN AND IMPORTED FRUIT IN NEW ZEALAND A thesis submitted in partial fulfilment of the requirements for the Degree of Doctor of Philosophy at Lincoln University by Nguyen Vu Hong Ha Lincoln University 2012 i Abstract of a thesis submitted in partial fulfilment of the requirements for the Degree of Doctor of Philosophy OXALATE AND ANTIOXIDANT CONCENTRATIONS OF LOCALLY GROWN AND IMPORTED FRUIT IN NEW ZEALAND by Ha Vu Hong Nguyen Locally grown and some imported fruits were analysed for their antioxidant and oxalate concentrations. Total phenolic and ascorbic acid concentrations and the antioxidant capacity using ABTS and ORAC methods showed that the fruits were a source of beneficial nutrients. In contrast, the fruits contained variable amounts of soluble and insoluble oxalates as anti- nutritive compounds. Fruit available in New Zealand contained a wide range of total phenolic compounds (27.4 – 2731.9 mg gallic acid equivalents (GAE)/100g fresh weight (FW)) and vitamin C (6.2-201.3 mg ascorbic acid/100 g FW). The ABTS capacity of these fruits ranged from 19.5 - 6045.9 mol Trolox equivalents (TE)/100 g FW and the ORAC values ranged from 1121.2-13631.6 mol TE/100 g FW. Extraction using 2 M HCL at 21°C was the optimum condition to extract total oxalate from fruit. This resulted in the highest mean value for total oxalate concentration (123.1 ± 2.5 mg/100 g DW) as compared to extraction using 0.2 M HCL at the same temperature (102.2 ± 2.1 mg/100 g DW) or extracting with 0.2 M HCL and 2 M HCL at 800C (85.2 ± 3.0 and 114.5 ± 2.2 mg/100 g DW, respectively). Similarly, at 210C, the mean soluble oxalate concentrations for the four analysed fruits were significantly higher than those extracted at 800C (54.7 vs. 39.8 mg/100 g DM). The total oxalate concentration of locally grown fruit determined by extracting with 2M HCL at 210C ranged from 2.9 – 7566.5 mg/100 g FW while the soluble oxalate extracted with deionised water ranged from 5 to 56% of the total oxalate concentration. Very high ii concentrations of total oxalate were measured in Indian gooseberry, rhubarb and carambola, 7566.5, 640.2 and 436.1 mg/100 g FW, respectively. Storage of kiwifruit for 15 days at 20 ± 10C showed reductions in soluble oxalate concentration and increases in insoluble oxalate concentration in the three kiwifruit fractions measured (whole fruit, skin and pulp), but not the seeds. Green kiwifruit (Actinidia deliciosa) contained significantly higher concentrations of oxalates (p < 0.001) than golden kiwifruit (A. chinensis). Processing kiwifruit to produce juice considerably reduced the oxalate concentration because most of the insoluble oxalate was retained in the pomace. Concentrations of total and soluble oxalates found in green kiwifruit juice obtained by enzymatic extraction were significantly higher (p < 0.01) than the values obtained by pressing. In the juice, soluble oxalate concentration contributed to 60.3 - 70.3% of total oxalate concentration compared with 31.7% found in the fresh tissue. Pasteurization temperatures significantly affected the soluble oxalate concentration but did not impact on the concentrations of total oxalate in the juice. Boiling rhubarb petioles (Rheum rhabarbarum L.) in water or cooking with trim (low fat) or standard milk significantly (p < 0.05) reduced the total and soluble oxalate concentrations of the mixtures by dilution. Total oxalate in the raw rhubarb was 902.7 mg/100 g FW and in cooked rhubarb it was 454.3 mg/100 g FW. Cooking rhubarb petioles with standard and trim milk resulted in further reductions in soluble oxalate concentrations of 65.9% and 74.5%, respectively, when compared to the soluble oxalate concentration of the raw petioles. When using an in vitro method (to simulate the gastric and intestinal environments) to extract oxalates from the raw and cooked rhubarb, the amounts of soluble oxalates binding with fibre at pH 2.0 were significantly higher (p < 0.01) than at pH 7.0 (an average of 74.5 vs. 39.4 mg/100 g FW, respectively), while, the insoluble oxalate concentrations in the fibre increased when the extractions were carried out in pH conditions similar to the intestinal environment. This trapping of oxalate in the fibre fraction suggests that dietary fibre can partly reduce oxalate absorption. Keywords: Fruit, antioxidant, antioxidant capacity, total phenolic compounds, ascorbic acid, oxalates, kiwifruit, Actinidia deliciosa, Actinidia chinensis, rhubarb, Rheum rhabarbarum L., in vitro digestion assay, cooking medium, mineral bioavailability, dietary fibre, oxalate availability, oxalate absorption. iii Acknowledgements I would like to express my sincere thanks to to all those who helped me in various ways during the work represented by this thesis. I wish to express my sincerest gratitude to my supervisor, Professor Geoffrey Savage, for his support, guidance, wisdom and inexhaustible patience in the correction of this thesis for an excellent work. A great gratefulness too, to his encouraging me to do a Doctor of Philosophy from the very beginning of my study. A special thanks to my co-supervisor, Dr Sue Mason, for her advice, encouragement and meticulous suggestions. Thanks for standing beside me when I felt frustrated and depressed. Thanks Leo Vanhanen for your assistance with technical issues. I really appreciated all your patience in working with problems of the HPLC, especially at the time laboratory work needed to be finished. Thank you to Allison Lister for your guidance with the statistical analyses. A grateful acknowledgment to the research funding from Lincoln University, Canterbury, New Zealand for financial support for this study. Thanks to my beloved husband for his encouragement and sharing of knowledge. A huge thanks to my loving son as his smile was the strongest force pushing me to finish my PhD. Most importantly, special thanks my dear parents for supporting me to pursue my PhD. iv Contents Abstract i Acknowledgements iii Contents iv List of Tables x List of Figures xi Chapter 1 Introduction 1 1.1 Research back ground 1 References 5 1.2 Thesis approach 8 1.2.1 Thesis aim 8 1.2.2 Thesis objectives 8 Chapter 2 Literature review 9 2.1 Introduction 9 2.2 Antioxidant- components of a healthy diet 9 2.2.1 What is antioxidant? 9 2.2.2. Free radicals and oxidative stress 10 2.2.3 The antioxidant functions against oxidative stress 11 2.2.4 Dietary antioxidants 12 2.2.4.1 Phenolic compounds 13 2.2.4.2 Vitamin C 15 2.2.5 Antioxidant assays 16 2.2.6 Contribution of antioxidants from fruit 18 2.3 Oxalates – anti-nutritional components in plant tissues 20 2.3.1 Biosynthesis of oxalate 20 2.3.2 Metabolism in plants 23 2.3.3 The role of oxalate in plants 24 2.3.3.1 Calcium regulation 24 2.3.3.2 Plant protection 25 2.3.3.3 Metal detoxification 26 2.3.4 Oxalates in fruits and fruit products 27 2.3.5 Oxalate in human health 29 v 2.3.5.1 Kidney stones 29 2.3.5.2 Mineral deficiency 29 2.3.6 Dietary oxalate intake and oxalate absorption 30 2.3.6.1 Oxalate availability 31 2.3.6.2 Cooking methods 32 2.3.6.3 Digestive pH 33 2.3.6.4 Dietary constituents 35 2.4 Summary 37 References 38 Chapter 3 Total phenolic, ascorbic acid concentrations and antioxidant capacities of fruit available in New Zealand 57 3.1 Abstract 57 3.2. Introduction 57 3.3. Materials and methods 60 3.3.1 Sample preperation 60 3.3.2 Moisture content 60 3.3.3 Extraction procedures and measurements 60 3.3.3.1 Extraction procedure for total phenolic and ABTS assays 60 3.3.3.2 Extraction procedure for ORAC 60 3.3.3.3 Measurement of ascorbic acid concentration 61 3.3.3.4 Measurement of total phenolic concentration 61 3.3.3.5 Measurement of antioxidant capacity by ABTS assay 61 3.3.3.6 Measurement of antioxidant capacity by ORAC assay 62 3.3.4 Statistical analysis 63 3.4. Results 63 3.4.1 Effect of extraction temperature on total phenolic concentrations 63 3.4.2 Total phenolic concentrations 64 3.4.3 Ascorbic acid concentrations 64 3.4.4 Antioxidant capacities 65 3.4.5 Correlations 68 3.4.6 Fruit serving size 68 3.5. Discussion 70 3.5.1 Effect of extraction temperature on total phenolic concentrations 70 3.5.2 Total phenolic concentrations 70 3.5.3 Ascorbic acid concentrations 72 vi 3.5.4 Antioxidant capacities 74 3.5.5 Correlations 75 3.5.6 The importance of fruit antioxidants 76 3.6. Conclusions 77 References 78 Chapter 4 The effect of temperature and pH on the extraction of oxalate and pectin from four New Zealand grown fruits 84 4.1 Abstract 84 4.2.
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