Evaluation of Phytochemical Compounds Present in Horned Melon Seed Extract Using Different Solvents (Research Proposal Sample)

EVALUATION OF PHYTOCHEMICAL COMPOUNDS PRESENT IN HORNED MELON SEED EXTRACT USING DIFFERENT SOLVENTS
MICHELLE WANJIRU
2014
DECLARATION
This research proposal is my original work and has not been presented for a degree in any other university.
Kamau Michelle Wanjiru (S20-A07-0004/2007)
Signature………………………………… Date………………………
This research proposal has been submitted for examination with our approval as university supervisors.
Dr. Leonard Gitu
Chemistry Department, JKUAT, Kenya
Signature………………………………… Date………………………
Table of Contents
TOC \o "1-3" \h \z \u Title page PAGEREF _Toc396229847 \h i
Declaration PAGEREF _Toc396229848 \h i
Table of Contents PAGEREF _Toc396229849 \h ii
List of Figures PAGEREF _Toc396229850 \h iv
1.0 INTRODUCTION AND LITERATURE REVIEW PAGEREF _Toc396229851 \h 1
1.1 Historical perspective PAGEREF _Toc396229852 \h 1
1.2 Characteristics of melon seed oil PAGEREF _Toc396229853 \h 3
1.3 Phytochemical Analysis PAGEREF _Toc396229854 \h 4
1.4 Constituents of melon seed PAGEREF _Toc396229855 \h 4
1.5 Analytical Techniques PAGEREF _Toc396229856 \h 5
1.5.1 Soxhlet Extractor PAGEREF _Toc396229857 \h 5
1.5.2 Rotor Evaporator PAGEREF _Toc396229858 \h 5
3.0 Statement of the problem PAGEREF _Toc396229859 \h 5
4.0 Justification PAGEREF _Toc396229860 \h 6
5.0 Objectives: PAGEREF _Toc396229861 \h 6
5.1 Overall objective: PAGEREF _Toc396229862 \h 6
5.2 Specific objectives: PAGEREF _Toc396229863 \h 6
6.0 MATERIALS AND METHODS PAGEREF _Toc396229864 \h 7
6.1 Materials and Reagents PAGEREF _Toc396229865 \h 7
6.2 Sample Collection and Sampling PAGEREF _Toc396229866 \h 7
6.3 Extraction using Soxhlet Extractor PAGEREF _Toc396229867 \h 7
6.4 Evaporation of solvents using rotor evaporator PAGEREF _Toc396229868 \h 8
6.5 Analysis of horned melon seed for phyto-chemicals PAGEREF _Toc396229869 \h 8
6.5.1 Detection for Alkaloids PAGEREF _Toc396229870 \h 9
6.5.2 Detection for Flavanoids PAGEREF _Toc396229871 \h 9
6.5.3 Detection for Terpenoids PAGEREF _Toc396229872 \h 10
6.5.4 Detection for Saponins PAGEREF _Toc396229873 \h 10
WORK PLAN PAGEREF _Toc396229874 \h 11
BUDGET PAGEREF _Toc396229875 \h 12
REFERENCES PAGEREF _Toc396229876 \h 13
List of Figures
TOC \h \z \c "Figure" Figure 1: Water melon PAGEREF _Toc396229895 \h 2
Figure 2: Water melon seeds PAGEREF _Toc396229896 \h 2
Figure 3: Linoleic Acid PAGEREF _Toc396229897 \h 3
1.0 INTRODUCTION AND LITERATURE REVIEW
1.1 Historical perspective
Melons (Figure 1) or culinary vegetables are fleshy fruit which originated from America and Central America. Three major melon species produced in the world are water melon (Citrullus anatus), honey dew (Cucumis melo indorus) and rockmelon (Cucumis melo reticulates). When ripe the sweety juicy pulp is eaten and seed can be roasted as a snack or ground into an ingredient used in oil and sauces. Melon plants are grown as a ground vine and depending on the cultivar, the shape of the fruit may change from round to ablong, light green to dark green and the flesh from red to yellow and the seeds from white to yellow, brown to black (McGregor, 1976). Boriss et al., 2006 stated that, in the past melon have been considered to be seasonal delights because of their limited availability throughout the year but due to the production flexibility and increased import availability, melons have become available year around and gained popularity.
According to AMA (2006), China and Australia are the leading producer of water melon, rock melon and honey melon. In 2004, China accounted for 50% of the world’s production by weight, Turkey 6.1% and Iran 4.4%. In Africa, melon is amongst the most popular African indigenous vegetable crops produced on large scale in Nigeria. The crop occupies a vital role in income generation ability. According to Ojo et al., (2002), the seed yield in Nigeria anges between 131 to 1005kg/ha. Bankole and Adebanjo (2004), acknowledged that melon seeds (Colocynthis citullus L.) (Figure 2) are important indigenous seed oil consumed by many rural communities in West Africa. Dehulled seeds contain oil, protein, amino acids, and some amounts of vitamins. Because of the nutritional and economic importance of melon, it is imparative that adequate studies should have been made to improve on its seed production practices.
Figure SEQ Figure \* ARABIC \s 1 1: Horned water melon
Figure SEQ Figure \* ARABIC \s 1 2: Water melon seeds
1.2 Characteristics of melon seed oil
Horned melon seed oil, rich in linoleic acid (Figure 3) (~64.5%), is used for frying and cooking in some African and Middle Eastern American countries owning to its unique flavor (Akoh, C.C, and C.V. Nwosu, 1992). Much research has been published on the oxidative stability of vegetable or fruit oils, but a little has been reported on the stability of melon seed oil. The modification of melon seed oil fatty acid composition by incorporation of oleic acid (18:1) has been explored (Charment et al., 1997). The modified melon seed oil was produced with the better balance of monounsaturate (18:1) and essential fatty acids (18:2), and also improved the seed oil oxidative stability and nutritional value (C h a r m e n t et al., 1998).
Figure SEQ Figure \* ARABIC \s 1 3: Linoleic Acid
1.3 Phyto-chemical Analysis
History of herbal remedies is very old as there are many herbs and spices which have their uses. The plants extracts have authentic effect on particular human disorder since they have the potential for antioxidant and antimicrobial activity. Plants and herbal extracts have formed important position in modern medicine due to their chemical and medicinal content found in natural form. Their secondary metabolites represent a large reservoir of structural moieties which wok together together exhibiting a wide range of biological activities. Microorganisms have the genetic ability to transmit and acquire resistance to antibiotics and have become a major global health problem.
Medicinal plants contain some organic compounds which provide definite physiological action on the human body and these bioactive substances including tannins, alkaloids, carbohydrates, terpenoids, steroids and flavanoids and these can be derived from melon seed (Edogo et al., 2005; Kumar et al., 2010).
1.4 Constituents of melon seed
Analysis made on melon seeds by Ajilola et al., 1990 indicated that melon seeds consist of 50% oil by weight, 37.4% protein, 2.6% fibre, 3.6% ash and 6.4% moisture. Immature fruits of some types of melon are often cooked as vegetables. Matured seeds are sometimes are often roasted and eaten. Processing of melon includes fermentation, coring, washing, shelling and oil extraction.
1.5 Analytical Techniques
1.5.1 Soxhlet Extractor
Soxhlet method will provide a fundamental method of extraction for gravimetric quantification of fats and oil in the horned seed melon. These fats and oils are groups of substances that will dissolve in organic solvents like ethanol and acetone but will relatively be soluble in water. Therefore, an accurate and precise quantitative analysis of these lipids in the horned melon seed soxhlet extractor is best qualified. The soxhlet procedure will allow for the calculation of total lipid (fat) content in the horned melon seed.
1.5.2 Rotor Evaporator
This technique will be applied for efficient and gentle removal of remaining ethanol and acetone solvents from the horned melon seed extract by evaporation. This machine will utilize a lower pressure than atmospheric pressure which will allow these solvents to boil at lower temperatures. Moreover, the rotations will increase the surface area and thus evaporation process will proceed more rapidly.
3.0 Statement of the problem
Drugs used in Kenya today are chemically synthesized during their production. These chemicals are highly harmful to the human health and also they are not environmental friendly. Natural drugs for instance extracts from water melon seeds are environmental friendly and low toxicity to humans.
4.0 Justification
Water melons are locally grown crops by Kenyan farmers. Water melon fruits have a worldwide market. The water melons seeds are locally available and cost efficient. Naturally extracted flavanoids, terpenes, and sappins have medicinal values which are very efficient in human health treatment.
5.0 Objectives:
5.1 Overall objective:
The overall objective will be evaluation of phyto-chemical presence in horned melon seed extract using different solvents
5.2 Specific objectives:
The specific objectives will be;
1 Extraction of the different compounds present in the melon seed using water, acetone and ethanol
2 Phyto-chemical examination for flavanoids, tannins, terpenes for all the extracts
6.0 MATERIALS AND METHODS
6.1 Materials and Reagents
All organic solvents will be purchased from Sigma-Aldrich and all the reagents to be used in this project will be of analar qual...