VIETNAM NATIONAL UNIVERSITY OF AGRICULTURE NGUYEN THI KIM THANH OPTIMIZATION OF SOME FACTORS INFLUENCING LYCOPENE EXTRACTION FROM TOMATO PROCESSING WASTE USING RESPONSE SURFACE METHODOLOGY Major: Food Technology Code: 24 18 05 57 Suppevisors: 1. Tran Thi Dinh 2. Marie-Louise Scippo AGRICULTURAL UNIVERSITY PRESS - 2017 c DECLARATION I hereby declare that the data and results of research in my thesis are honest. There is no material that has been accepted for the award of any other degrees or diploma in any educational institution and, to the best of my knowledge and belief, it contains no material previously published or written by another person, except where due reference is made in the text of the thesis I hereby declare that, all the help to carry out of my thesis was thanked and the cited information in this thesis has been written clearly the source.
Hanoi, May 10th, 2017 Master candidate Nguyen Thi Kim Thanh i c ACKNOWLEDGEMENTS This thesis was realized at Department of Food Processing Technology and Central laboratories of Food technology-Vietnam national university of Agriculture under the supervisor of Assoc. Tran Thi Dinh and Prof. Marie-Louise Scippo. To complete this thesis, besides the effort of myself, I have received encouragement and great help of many individuals and groups.
Foremost, I would like express my deep gratitude to my supervisor Assoc. Tran Thi Dinh and Prof. Marie-Louise Scippo for their valuable advices and continuous guidance, encouragement and time sharing during my study. I would like to express my sincere thanks to Msc.
Nguyen Thi Hoang Lan and Dr. Hoang Hai Ha for enthusiasm, insightful comments, teaching me on the HPLC analytical technique and useful laboratory skills. I am grateful to Research and Teaching Higher Education Academy – Committee on Development Cooperation (ARES – CCD) for awarding the scholarship grant. I give my thanks to Dr.
Nguyen Thi Thanh Thuy for her great support during my study. My sincere thanks are also sent to my friends especially, special thanks to my juniors Than Thi Huong, Nguyen Thi Hien and Pham Thi Bich for their assistance in the experimental work of this thesis. Last but not least, I owe more than thanks to my family, my parents, my elder sister and my younger brother for their love, support, patience and inspiration. Hanoi, May 10th, 2017 Master candidate Nguyen Thi Kim Thanh ii c TABLE OF CONTENTS Declaration.
ii Table of contents. iii List of abbreviations. v List of tables. vi List of figures.
vii List of figures. vii Thesis abstract. Origin and distribution of tomato. Tomato processing waste.
Source of lycopene. Role of lycopene in the human health. Physical and chemical properties of lycopene. Solvent extraction method.
Other methods of lycopene extraction. Materials and methods. Sample collection and preparation. Results and discussion.
Selection of the suitable organic solvent for lycopene extraction. Selection of treatment regime of tomato waste for lycopene extraction. Response surface methodology for optimization of lycopene extraction. Conclusions and recommendations.
48 iv c LIST OF ABBREVIATIONS Abbreviation Description DPPH 1,1-diphenyl-2-picrylhydrazyl_C18H12N5O6 DW Dry weight HPLC High performance liquid chromatography w/v Weight/ volume v c LIST OF TABLES Table 2. World tomato area, production and productivity, 2013. World leading tomato producing countries in the world. Tomato area, production and productivity of some region in Viet Nam, 2009.
Typical composition in 100 gram of a ripe tomato fruit. Carotenoid composition of tomato fruit, tomato processing wastes and tomato paste (mg/100g wet sample). Lycopene content of common fruit and vegetables. Lycopene content in common tomato –based food.
Physical properties of lycopene. Total lycopene and Cis-isomer content in the dehydrated tomato. Effect of solvent system on lycopene extraction from tomato waste. Experimental design for drying of tomato waste.
Box- Behnken experimental design for lycopene extraction. Results of optimization treatment regimens for tomato waste. Summary of effect of independent factors to the output variables. Results of the analysis of variance on lycopene content.
Result of the analysis of variance antioxidant capacity of lycopene extract. Results of optimization condition for lycopene extraction. Summary of effect of independent factors to the output variables. Results of the analysis of variance of lycopene content.
Result of the analysis of variance of antioxidant capacity of lycopene extract. 39 vi c LIST OF FIGURES Figure 2. Structure of trans and cis isomeric forms of lycopene. Tomato ‘Chanoka F1’ fruit, B.
Fresh tomato waste, C. Dried tomato waste. HPLC chromatogram of (A) lycopene analytical standard at 0.25mg/ml and (B) CT5 sample in section 3. HPLC calibration curve for lycopene standards dissolved in n-hexan and dichloromethane (1:1).
Trolox calibration curve. Effect of solvents systems on lycopene concentration. Effect of solvents systems on antioxidant capacity of lycopene extract. Profiler showing the optimal drying conditions of tomato waste.
Profiler showing the optimal extracting conditions of lycopene extraction. 40 vii c THESIS ABSTRACT Master candidate: Nguyen Thi Kim Thanh Thesis title: Optimization of some factors influencing lycopene extraction from tomato processing waste using response surface methodology Major: Food technology Code: 24180557 Educational organization: Vietnam National University of Agriculture (VNUA) Research Objectives: The aim of this research is to optimize some factor (solvent/material ratio, temperature and time) influencing lycopene extraction process from tomato waste which could be used to produce functional foods. Materials and Methods: - Materials: The red ripe tomato cv. Chanoka F1 was harvested in Bac Ninh province.
Tomato waste was obtained by removing the juice. Tomato paste was passed through a fruit pulper to obtain waste. Tomato waste was dried by a convective oven after that they were ground to use as material for lycopene extraction - Methods: Suitable organic solvent for lycopene extraction from tomato waste was studied ranging from single solvent (acetone, ethanol, ethyl acetate), double solvent (acetone: ethanol) and triple solvent system (acetone: ethanol: ethyl acetate). The treatment regime (moisture content and drying temperature) of tomato waste for lycopene extract also investigated.
Tomato waste, which was dried in the oven at the optimal temperature and moisture content, was used to optimize of several factors (ratio of solvent/dried tomato waste, temperature, time) influencing extraction of lycopene with the most suitable solvent by response surface methodology. - Analytical methods: Moisture content of tomato waste (%) was measured using fast moisture detector (MA37, Germany). Lycopene content was quantified by HPLC. Antioxidant capacity of lycopene content was quantified by DPPH radical scavenging test.
Main findings and conclusions The results of the present study indicated that ethyl acetate solvent proved to be the most efficient compared to other solvents for lycopene extraction. The optimal conditions for drying of tomato waste is temperature of 65oC until the moisture content of the material reached 23%. The optimal extraction conditions for lycopene were: + Ratio of solvent/waste 40/1 (v/w), + Temperature 55oC and viii c + Extraction time 120 min. Under this optimization condition lycopene content in the extract was 7.391 mg/g DW and antioxidant capacity of extract was 10.384 µmol TE/g DW.
INTRODUCTION Lycopene is one of 750 carotenoids found in nature and is responsible for the red color of fruits. It is present in high concentration in red fruit and vegetable, such as tomato, gac, carrot, watermelon… (Britton, 2004). In the food industry, lycopene is used as a natural pigment in the dyeing of food product. Besides, lycopene is also known as a potential antioxidant which is believed to be responsible for protecting cell against oxidative damage and thereby decreasing the risk of chronic diseases (Rao et al.
Thus, lycopene demands on using in pharmaceutical, food, feed and cosmetic industries calls more attention nowadays. Tomato, Lycoperisicon esculentum, is one of the most widely cultivated vegetable in worldwide and known as one of fruits which are rich in lycopene. World tomato production in 2013 was about 163 million tons of fresh fruit from an estimated 4. Tomato contains a wide variety of antioxidants including vitamin E, ascorbic acid, carotenoids, flavonoids, phenolic compounds (Sathish et al.
Lycopene represents about 80-90% of total carotenoids in tomato. Lycopene is located in different fractions of tomato such as tomato skin, water insoluble fraction, and fibrous fraction including fiber and soluble solids. Tomato processing industry produces large amounts of solid waste. It is about 10–40% of the total tomato processed in the facility and includes 33% seeds, 27% skin and 40% pulp (Topal et al.
Toor and Savage (2005) indicated that 70–90% of the lycopene was associated with the water insoluble fraction and the skin. In Vietnam and other countries, the waste is usually used for animal feed or for organic fertilizer but it is not used for human consumption (Knoblich et al. Therefore, large quantity of carotenoids is lost as waste. In addition, this waste has a high moisture content that makes it susceptible to microbial proliferation and spoilage.
Therefore, it can be preserved by drying or other methods and then for lycopene extraction. Tomato carotenoids are liposoluble. Recently, there are several methods used for lycopene extraction. Sabio et al.
(2003) studied a lycopene extraction process based on supercritical CO2, which allows the extraction of over 60% of 1 c the lycopene from tomato waste. Xi (2006) reported that the lycopene yield from high pressure processing treatment of tomato paste waste for 1 min was much higher than from solvent extraction for 30 min. However, lycopene is commonly extracted with organic solvents due to the cheap cost of technology and better recovery as compared to other methods. There are a lot of organic solvents, which are usually used in several studies to non-polar carotenoid extraction such as ethyl acetate, ethanol, acetone, etc.
However, their parameters (solvent/material ratio, temperature and time) are largely influence on lycopene extraction. Therefore in the current study, we conduct research entitled “Optimization of some factors influencing lycopene extraction from tomato processing waste using response surface methodology”. General objective The aim of this research is to optimize some factors (solvent/material ratio, temperature and time) influencing lycopene extraction process from tomato waste which could be used to produce functional foods. Specific objectives - To select suitable organic solvent for lycopene extraction process from dried tomato waste; - To optimize the moisture content and drying temperature by convective drying of tomato waste for lycopene extraction; - To optimize several factors (ration of solvent/material, temperature, time) influencing on extraction of lycopene from dried tomato waste; - To characterize the extracted lycopene in term of lycopene content and anti-oxidant activity.
Origin and distribution of tomato Tomato (Lycopersicon esculentum Mill.) is one of the most widely cultivated vegetable worldwide. Tomatoes are members of the family Solanaceae (also known as the nightshade family), genus Lycopersicon, subfamily Solanoideae and tribe Solanceae (Taylor, 1986). It was originated in the coastal highlands of Andean region that includes parts of Chile, Colombia, Ecuador, Bolivia and Peru (Sims, 1979). The Spanish introduced tomato into Europe in the early 16th century (Harvey et al.
European acceptance of tomato as a cultivated crop and its inclusion in the cuisine were relatively slow. Tomatoes were initially grown only as ornamental plants: the fruits were considered to be poisonous, because of the closely related deadly nightshade (Solanum dulcamara). Since the mid-16th century tomatoes have been cultivated and consumed in southern Europe, though they only became widespread in north- western Europe by the end of the 18th century (Harvey et al. In 17th century, European took the tomato to South, Southeast Asia and China.
In the 18th century, tomato came to Japan and the USA. The production and consumption of tomato expanded rapidly in the USA in the 19th century, and by the end of that century, processed products such as soups, sauces and ketchup were regularly consumed (Harvey et al. World tomato area, production and productivity, 2013 Area Production Productivity Location (1000 ha) (1000 tons) (tons/ha) Africa 902.79 Source: FAOSTAT (2014) 3 c Nowadays, tomatoes become the most important vegetable in the world.