VIETNAM NATIONAL UNIVERSITY INTERNATIONAL UNIVERSITY School of Biotechnology ------ œ ¯ œ

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BSc THESIS PROJECT PROPOSAL
Student’s name: Huynh Hoai Thuong ID: BT060056 Department: Biotechnology PROJECT GENERAL INFORMATION 1. NAME OF STUDY: STUDY ON EXTRACTION OF ESSSENTIAL OIL AND PECTIN FROM GRAPEFRUIT PEELS. 2. FIELD OF STUDY: Food technology 3. SUPPERVISOR: Ms.c Le Hong Phu 4. HOST INSTITUTE IU laboratory 5. TIME From 01/09/2010 to 30/11/2011 6. BUDGET ESTIMATION 3,000,000 VND I. • PROJECT CONTENTS To make full use of cheap by-products ( grapefruit peel) to extract some valuable natural products ( pectin and essential oil). _To optimize the parameters to extract the highest obtainable essential oil yield from grapfruit peel. _To optimize the parameters to extract the highest pectin yield from treated grapefruit peel. 1. AIM

long (up to 150 mm) and thin. culture of grapefruit is similar to that of the orange.(1) These evergreen trees are around 5–6 metres tall. The condition called exanthema is caused by copper deficiency. BACKGROUND 2.[30] Grapefruit forms a core part of the "grapefruit diet". varying in color depending on the cultivars.[16][26] contains the fiber pectin. The fruit is yellow-orange skinned and largely an oblate spheroid. mottle leaf results from zinc deficiency.1.org/wiki/Grapefruit Grapefruit is an excellent source of many nutrients and phytochemicals that contribute to a healthy diet.[16][29] and there is evidence that the seeds have antioxidant properties.1 Grapefruit background 2.1.Thesis proposal • To reduce the problems of polluted environment caused by waste products.1. higher fruits are usually harvested by workers on ladders who snap the stems or clip the fruits as required. an 18th-century hybrid first bred in Barbados.1 Introduction The grapefruit (Citrus paradisi). although they can reach 13–15 metres.1. which include white.1.[16][28] Studies have shown grapefruit helps lower cholesterol.2 Vietnam Grapefruit Nutritional properties http://en. Grapefruits were formerly harvested by climbing the trees or using picking hooks which frequently damaged the fruit.1. 2.wikipedia. Nutritional experiments with grapefruit have shown that excessive nitrogen results in malformed fruit. except that wider spacing is necessary. coarser texture and less juice. It produces 5 cm white four-petaled flowers. the fruits on low branches are picked by hand from the ground. In general. Grapefruit is a good source of vitamin C.[31] 2 . the theory being that the fruit's low glycemic index is able to help the body's metabolism burn fat.[27] and the pink and red hues contain the beneficial antioxidant lycopene. The flesh is segmented and acidic. LITERATURE REVIEW 2. it ranges in diameter from 10–15 cm. 2. Today. is a subtropical citrus tree known for its bitter fruit. pink and red pulps of varying sweetness. Lack of certain minor elements is evident in symptoms often mistaken for disease. The leaves are dark green.

flu. as it shares the same properties as other citrus oils. C. cellulite. jet-lag. Phototoxicity of Grapefruit peel oil It is thought that Grapefruit peel essential oil is not phototoxic. grapefruit peel oil is dominated by (+)-limonene. muscle stiffness. As with most citrus fruits. The distinctive topnote of grapefruit juice is owing to trace amounts of 1-p-menthene-8-thiol. aurantium ssp. although Grapefruit peel oil does contain furocoumarins. The Properties of Grapefruit peel oil Grapefruit peel essential oil is anti-infectious. which are responsible for phototoxic effects. detoxifying. 2. stress and nervous exhaustion. pink or red varieties of the grapefruit have become very popular. The special grapefruit aroma is characterized by its 'bittergreen-woody' nuance. immune boosting. It is good for use in a diffuser to disinfect a room.0001 ppb).Thesis proposal 2.1. and bitter orange. but the particular grapefruit character is due to the sesquiterpene ketone nootkatone. oily skin. skin-toning and mentally rejuvenating. amara. anti-inflammatory. obesity. it also contains other chemical components which appear to neutralize the potential phototoxicity. Therefore. it is a good alternative oil to use in place of other similar citrus oils. C.1. anti-depressant. unlike most of the other citrus oils.3 Essential oil extraction methods 3 . maxima. milder. headaches. colds. depression. It is used to treat acne. water retention. New.2 Grapefruit peels essential oil background Grapefruit is believed to be a crossing of pomelo. one of the most powerful odorants known (odour detection threshold in water ~ 0. anti-septic.

thimble. there are five main components. The success of this method depends upon the difference in solubility of a compound in various solvents. Solid. but can be used whenever it is difficult to extract any compound from a solid. Soxhlet Extraction A Soxhlet extractor is a type of laboratory glassware invented in 1879 by Franz von Soxhlet.A condenser is the apparatus in which some type of condensation occurs. Chemists may conduct either a condensation reaction or a vapor-to-liquid condensation in their work. 4 . Extraction always involves two steps. Condenser. the separation or washing of the solution from the residual solid. This increases the efficiency of the extraction when compared with simply heating up the solid in a flask with the solvent. solvent extraction) is a useful method to separate components (compounds) of a mixture. The example of extraction processes are liquid-liquid extraction. Then.Thesis proposal There are many types of extraction process. In the soxhlet extractor. Liquid-liquid extraction Liquid-liquid extraction (or more briefly. solid-liquid and soxhlet extraction. The key advantage of this type of extraction. siphon arm and round boiling flask.liquid extraction Solid-liquid extraction or leaching is one process that separate the desired solute constituent or to remove an unwanted from the solid phase in the process. only clean warm solvent is used to extract the solid in the thimble. The first step is contact of the solvent with the solid to be treated so as to transfer the soluble constituent or solute to the solvent. The components are condenser. There are many substances used in modern processing industries occur in a mixture of components dispersed through a solid material. It was originally designed for the extraction of lipid from a solid test material. extraction chamber. solid-liquid extraction.

Condenser.cellulose and glass microfiber extraction thimbles are known for their purity and consistent high quality. The solvent which condense at the condenser drip down through the extraction chamber. The pectins of a plant can be water-soluble. arabinose. 5 . chelator soluble. Cellulose extraction thimbles are produced from high-quality alpha cellulose cotton linter and have excellent mechanical strength and retention.it is placed at the top of the soxhlet extractor body. 1991). Round Bottom flask. As a part of the plant structure. and glucose called ‘hairy regions’ (IPPA. 2001). convenient and efficient method of solvent extraction of solids and semi-solids. xylose.it containing a solvent that used in this extraction. The function of pectin in plants is to contribute structural integrity to the 11 cell wall and adhesion between cells. The thimbles are widely used in Soxhlet extraction units providing a safe. 2001). The general makeup of the pectin content varies with ripening of the plant and it is fairly easily brought into solution depending on the plant type (Van Buren. 1991). Pectin is primarily made up of D-galacturonic acid joined by K(1-4) glycosidic linkages (Van Buren. 2.Thesis proposal Extraction chamber. 1991).It allows the sample of solvent that used during the extraction process. Commercial pectin extraction is mainly from citrus peel and apple pomace. pectin is a complex mixture of blocks of homogalacturonic acid called ‘smooth regions’ mixed with blocks of homogalaturonic acid containing many neutral sugars including rhamnose.1. 1991). After extraction pectin consists of smooth galacturonic acid regions with a few neutral sugars still attached (IPPA. but several other sources exist such as sugar beets and sunflower heads. or protopectins (Van Buren. galactose. The capacity is 1000 ml. The methods of extraction will vary based on the actual makeup for each particular plant type. It is converted a vapor into a liquid that trickles into the extraction chamber containing the sample. A percentage of the galacturonic acid residues are generally esterified with methanol.4 Pectin background Pectin is defined as complex mixtures of polysaccharides that make up approximately one third of the cell-wall dry substance of most types of plants (Van Buren. Extraction thimble.

sulfuric.. The effects of temperature. usually hydrochloric.26% of the dried peel weight.5 Pectin extraction methods Pectin is produced commercially from citrus peel and apple pomace. (1991). The extraction conditions vary from facility to facility and are dependent on the pectin source. Optimal extraction conditions found through varying extraction time. Pectin has also been investigated for its usefulness in the pharmaceutical industry. 1999). 1984). 2. 2.53% dry basis at these extraction conditions. WHY THIS PROJECT NEED TO BE CARRIED OUT The Vietnam production of grapefruits is very high. time. or nitric acids. 84CC. Optimal extraction conditions of pH 1. Laboratory scale extractions have been conducted to determine optimal pectin extraction conditions and the feasibility of pectin extraction from a number of different plant materials.Thesis proposal Pectin is the most important gelling agent used for jam jelly and marmalade. Extraction most commonly occurs using a dilute mineral acid. and pH on pectin yield for orange pectin using nitric acid extraction was investigated by Aravantinos-Zafiris et al. however. 1988).6.1.2. and temperature for pectin extraction from sugar beet pulp were reported as the use of hydrochloric acid to adjust pH to 1. and liquid to solid ratio were optimized in a study on pectin extraction from tropical fruits (Simpson et al. pH. The initial goal is to explore the extraction of essential oil from the grapefruit 6 . Among other uses it has been considered in the class of dietary fibers known to have a positive affect on digestive processes and to help lower cholesterol (Braddock. The resulting pectin yield was 19. and 64 min resulted in yields up to nearly . It also is utilized to stabilize liquid pharmaceutical emulsions and suspensions. time. represents a significant potential for the development of valuable products. Today.5 extracted for 4 hours at 80CC (Phatak et al.. Extraction pH. and the remainder including peel is considered as wastes. This wasted peel peels. the largest use of pectin has changed to be a stabilizer for acidified milk drinks. Almost haft of these grapefruits is used to make juice.

The extraction of orange juice yields 55% juice with 45% wet mass residues left over.3. resulting in a large amount of waste material for disposal (Braddock. In addition. THE CONTRIBUTION OF THIS PROJECT INTO SCIENCE This study will help to reduce a waste of grapefruit peel in order to protect the environment. It is of interest to explore pectin extraction from grapefruit peels. The basic pectin extraction procedure for orgrane peels and other procedures that have been successfully applied to pectin extraction from a number of other plant materials will be applied to grapefruit peel. 7 . this also contribute to optimize the best conditions to produce essential oil and pectin form grapfruit peel. 2. 2004).Thesis proposal Because of the success found in the organe industry for extraction of pectin from peel.

2.Thesis proposal II. CONTENTS 3.1. HYPOTHESIS What are the best conditions to get the high yield of extracting essential oil and pectin form grapefruit peel ? 3. PROCESS OF PROJECT 8 .

9 . After cutting. Method: Grapefruit peels were cut into small pieces of about 1-2 cm.2. the peels were milled to fine powder (0.1.5 mm) with a grinder. DETAIL CONTENTS: 3.acid hydrolysis Pectin recovery and analysis 3.1 Grapefruit peel preparation Material: grapefruit peels. grinder.1.Thesis proposal Grapefruit peels preparation Sohxlet extraction of essential oil Isolation of essential oil Treated peels Dilute. knife.2. Part I: Extraction of essential oil from grapefruit peels 3.2.

The extraction procedures were described above. samples were extracted with distilled water.2. • Single factor experiments were performed to determine the appropriate range of conditions for grapefruit peels essential oil extraction. solvent concentration. The best solvent concentration was selected according to the percentage of essential oil yield.1. Method: • The conventional Soxhlet extraction apparatus consisting of a condenser. 3. The best solvent type was selected according to the percentage of essential oil yield. The best extraction time was selected according to the percentage of essential oil yield.3 Isolation of essentail oil by Rotary evaporator Place flask on the rotary evaporator. The solvent was recovered on the condenser 10 . solvents. namely. 60% (v/v) ethanol. 60% (v/v) acetone.Thesis proposal 3. 60% (v/v) n-hexan and 60% (v/v) methanol respectively. The extraction procedures were repeated by varying the extraction time from 60 to 420 min. b) Selection of solvent concentration Using the best solvent type selected in (a). a) Selection of solvent type By fixing extraction time (180 min) and solvent concentration. solvent type.2. and an extraction flask were used. extraction time by varying one independent variable at a time while keeping the others constant. All experiments with Soxhlet were caried out at room temperature and were triplicate for statistical evaluation. c) Effect of extraction time Samples were extracted using the best solvent type and the best solvent concentration selected in single factor experiments sections (a) and (b). 20 grams of milled peels were placed in a Soxhlet apparatus and extracted with 100ml of an appropriate solvent.2 Soxhlet extraction Material: milled grapefruit peels. samples were extracted with solvent ranging from 20% (v/v) to 100% (v/v) by fixing the extraction time at 180 min. Putting the flask into a water bath with the appropriate boiling point of the solvent. respectively.1. a Soxhlet chamber. Soxhlet extractor.

Method: • The hydrolyzate supernatant was vacuum filtered as using filter paper and the solids were resuspended in 400 mL of 600C disstilled water for 5 min.3 Pectin recovery and analysis Material: equipments: vaccum filter. 3.2.2. pH meter. 11 . magnatic stirrer. grinder. The pH was then increased to the appropriate value.2.2. centrifuegator.the flask contained the essential oil and distilled water so anhydrous sodium slufate ( Na2SO4) was added to dry essential oil.2. The centrifugation and filtration steps were repeated.2 Dilute.Thesis proposal and drip into the receiving flask.2. 3. sulfuric acid. The mixture was agitated at a constant temperature until the desired extraction time had elapsed. The mixture was centrifuged at 11500g for 10 minutes to get hydrolyzate supernatant. 3. chemicals: 5N NaOH.4 Analysis the esseential oil yield The percent yield of the essential oil was determined as the essential oil voulme divided by the weight of the milled peel. filter paper.2.1 Preparartion of treated grapefruit peels Treated gapefruit peels were dried at 50 0 C for 48h.1. alcohol. Once all the solvent has evaporated. Part II: Extraction of pectin from treated peels 3. 3. Method: A weighed portion of pressed peel was added to the distilled water. oven.2.2. Measured amounts of acid sulfuric were added to the peel-water mixture until the desired concentration of acid (v/v) was obtained.acid hydrolysis Material: dried-treated peels.

130. respectively. 100. 0.0 (v/v) with fixing extraction conditions of 3000 C for 9 min at pH 1.Thesis proposal • The filtered solutions were combined and the approximate volume of alcohol was added for overnight precipitation.154. Sulfuric acid concentration was tested at 0. extraction time. acid concentration. 0.3 Methods for optimization of extraction parameters Single factor experiments were performed to determine the appropriate range of conditions for pectin extraction. 0. temperature and pH by varying one independent variable at a time while keeping the others constant.6. All experiments were triplicate for statistical evaluation.108.The specific extraction conditions were 3000 C for 9 min at pH 1. namely. .(0.200.3. The precipitate was separated by centrifugation at 180g for 60 min and washed five times with ethanol (45%) according to a previous procedure (Faravash and Ashtiani.8 and sulfuric acid concentration 0. b) Sulfuric acid concentration Using the best ratio of solid to liquid in (a). The procedure outlined above was followed. solid to liquid ratio. 0. c) Extraction time 12 .292g/mL.2.5 (v/v). 0.5.246 and 0. which were 70.1. Ground samples were stored in small plastic sample bags. following the equation: P=W. a) Solid to liquid ratio Solid to liquid ratio was tested using measured amounts of treated peel in 650 mL of distilled water. 160 and 190g of peel with corresponding solid to liquid ratios of 0.92). • The quantity of pectin was measured by precipitating with calcium chloride. 2007).2. weighed and ground using a mortar and pestle. 0.7 and 1. 100 B While w: the quantity of precipitated calcium pectinate (g) B: the quantity of pectin was used to precipitate with calcium chloride (g) 3. The resulting pectin was dried under vacuum at 500C.

1999.J. Rouse. 60: 127-9. 14(5): 310-13. 12. 3. PROGRESS AND EXPECTED RESULTS 13 . b.info/commercial_production_of_pectin. 2. R. 6.4.G. 3. Crandall.J. Journal of Food Engineering. IV. Journal of Food Technology.2 and 2.b. Braddock. 2001. sulfuric acid and extraction time in (a.c and d).K. S. Extraction and characterization of sunflower pectin. Importance of byproducts to citrus juice processing.G.8. IFT Committee.htm. Available online at URL http://www. Handbook of citrus by-products and processing technology. e) pH extraction Using the best solid to liquid ratio. Braddock. J. pH was tested at 1. 4. G.3.Effect of drying on pectin made from lime and lemon pomace.8. extraction temperature was tested at 150. Journal of Science in Food and Agriculture. 81: 983-7.0. 43:1680-2.E. 62: 215-23. sulfuric acid. Procedures of the Florida State Horticulture Society. Pair. A. IPPA: International Pectin Producers Association. 13:496-500. c). Journal of Food Science.D. Oreopoulou. R..6. P. Collins. P.J. A. What is pectin?. 1978a. (Accessed September 2004) 9. R.Thesis proposal Using the best solid to liquid ratio and the best sulfuric acid concentration in (a and b). 5. Aravantinos-Zafiris. 1978b. Braddock. Braddock. REFERENCES 1. 6. Inc. New York. Crandall. R. 250.H. 2001. 8. 1991. Pectin standardization: final report of the IFT committee. 2004. 200. 7. W Roberts. 2. 191-7. Journal Science Food Agriculture. Perkins-Veazie. d) Extraction temperature Using the best solid to liquid ratio. and 15 minutes with fixing extraction conditions of 3000 C for 9 min at pH 1. P. The effect of nitric acid extraction variables on orange pectin. 2. J. 2004. extraction time was tested at 3. 300 and 3500 C at pH 1.H. Lozano. 1959. M. John Wiley & Sons. extraction time and extraction temperature in (a. Iglesias. V. Rouse.ippa.. 9. Determining the yield and quality of pectin from fresh peel and pectin pomace.J. 1. 91: 109-11.8.T. Fruit Processing..

........................................ 14 .. SUPERVISOR’S APPROVAL . .......30/11 1/10 – 7/10 7/10 – 30/10 V...................................................................................................................................................................... ............................................... ................Thesis proposal Contents Part I Extraction of essential oil from grapefruit peels Grapefruit peels preparation Soxhlet extraction Isolation evaporator Analysis of essential oil yield of essentail oil by Rotary Expected results Time 01/09 – 7/09 7/09 – 30/09 Part II Extraction of pectin from treated peels Preparation of treated peels Dilute......................................................................................................................................................................................................................acid hydrolysis Pectin recovery and analysis Complete the thesis 1/11...............

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