“EFFECTS OF PROCESSING ON THE AMINO ACID PROFILE, CHEMICAL AND FUNCTIONAL PROPERTIES OF JACKFRUIT (ARTOCARPUS HETEROPHYLLUS LAM)”

APPROVAL PAGE We the undersigned hereby approve that this research titled “Effects of Processing on The Amino Acid Profile, Chemical and Functional Properties of Jackfruit (Artocarpus Heterophyllus Lam)” meets the requirement for the award of Bachelor of Science (B.Sc) in Food Science and Technology, Abia State University, Uturu. Abia State, Nigeria. DR. OBIOHA, M.R. Supervisor: Sign Date EXTERNAL SUPERVISOR Sign Date DR. N. C. OGANEZI (Head of Department) Sign Date CERTIFICATION We the undersigned hereby declare that this research titled “Effects of Processing on The Amino Acid Profile, Chemical and Functional Properties of Jackfruit (Artocarpus Heterophyllus Lam)” “was conducted by, Onyeji Patra Chisom with Matriculation Number; 2017/113480/Regular, under the supervision of Dr. Obioha Okorie . To the best of our knowledge, this research has not been carried out in any other tertiary Institute in Nigeria and beyond. Onyeji Patra Chisom (Research Student) Sign. Date Dr. Obioha Okorie (Project Supervisor) Sign Date CHAPTER ONE INTRODUCTION 1.1 Background of Study Jack fruit is a fruit species found in tropical, high rainfall, coastal and humid areas of the world. People consumed it mostly as a fruit when ripe but also as vegetable in the unripe stage. The jackfruit significantly contributes to the nutrition of people as a source of vitamins, minerals and calories. Both tender and ripe fruits as well as the seeds are rich in minerals and vitamins. The fruit pulp is sweet and tasty and used as dessert or preserved in syrup (Madruga et al., 2014). Jackfruit have more protein, calcium, thiamine, riboflavin and carotene than banana but less nutritious than mango. Therefore, vitamin and mineral production per unit area are higher in jackfruit than Mango. Jackfruit is rich in dietary fiber, which makes it a good bulk laxative (Madruga et al., 2014). The seeds however, are far less utilized with occasional usage as dessert, minor ingredients in culinary recipes or eaten as a snack after boiling, steaming or roasting. Most of the time, the seeds are left untreated or discarded as wastes due to the bland taste and texture of the seeds (Kooh et al., 2016). Jackfruit seeds are potential local food source which can be processed into flour. Anneahira (2010) stated that the jackfruit seeds are known to contain a lot of carbohydrates, protein, and minerals (calcium and phosphorus). Jackfruit seeds are mostly not used as vegetable. It is eaten whole by boiling or roasting. They are easily digestible and also contain same composition as grains (Anneahira, 2010). 1.1 Statement of Problem A bulb of jackfruit contains 100 - 500 seeds (10-15% weight of the fruit), and these seeds are usually discarded during the processing of jackfruit. Jackfruit seeds are rich in protein (13.50%), carbohydrate (79.34%), dietary fiber (3.19%), fat (1.27%), calcium (3087mg/kg), sodium (60.66mg/kg), copper (10.45mg/kg), manganese (1.12mg/kg). Consequently, there is a need to harness these nutritional components by studying the effect of processing on its amino acid profile chemical, functional properties of the seed in other to ascertain the best processing method with higher nutrient retention. 1.2 Objectives of the Study The objective of this; i.. To produce flour from raw roasted, fermented and boiled jackfruit seed ii. To determine the effect of processing on its chemical properties of the jackfruit seeds flour iii. To determine the effect of processing on its functional properties of the jackfruit seed flour iv. To determine the effect of processing on its amino acid profile of the jackfruit seed flour 1.3. Justification of the Study The result of this study will enhance the knowledge of Food processors on the best processing method suitable for the processing of flour from jackfruit seeds, that will not have less or no nutrient loss. Thus, it will boost the nutrients of the consumers since it contains protein and other minerals needed for growth and development of the body. CHAPTER TWO 2.0 LITERATURE REVIEW 2.1. Origin and Distribution Artocarpus heterophyllus Lam. which is commonly known as jackfruit is a tropical climacteric fruit, belonging to Moraceae family, is native to Western Ghats of India and common in Asia, Africa, and some regions in South America (Tejpal and Amrit, 2016). It is known to be the largest edible fruit in the world. It is one of the commonly consumed foods. It is a non-seasonal fruit and had a major contribution to the food supply of the people and their livestock when there were short supplies of staple food grains. It is a monoecious tree and both male and female inflorescences are found on the same tree. The fertilization is by cross-pollination and the propagation is mostly through seeds. The complete fruit development process takes about three to seven months from the pollination, varying in different countries. Jack is a tropical fruit species found in tropical, high rainfall, coastal and humid areas of the world. It is the favorite fruit of many, owing to its sweetness (Tejpal and Amrit, 2016). The jackfruit tree is widely cultivated in tropical regions of Nigeria, India, Bangladesh, Nepal, Sri Lanka, Vietnam, Thailand, Malaysia, Indonesia and the Philippines. Jackfruit is also found across Africa, e.g., in Nigeria, Cameroon, Uganda, Tanzania, and Mauritius, as well as throughout Brazil and Caribbean nations such as Jamaica. However, India is considered to be the native of jack fruit. The name originated from its Malayalam name Chakka. It is also called kathhal (hindi and urdu), pala (tamil), halasina hannu (kannada) panasa pandu (telugu) and phanos (marathi and Konkani). The fleshy carpel which is botanically the perianth is the edible portion. Apart from its use as a table fruit, jack is a popular fruit for preparation of pickles, chips, jack leather and papad. The fruit has got good potential for value addition into several products like squash, jam, candy, halwa etc. The ripe bulbs can be preserved for one year in sugar syrup or in the form of sweetened pulp. The unripe mature bulbs can be blanched and dehydrated for further use throughout the year. Seed is a rich source of starch and a delicacy during season. The timber is highly valued for its strength and sought for construction and furniture making. The dried leaves are stitched to make disposable plates. Climate and soil requirement for jackfruit is quite different from other. It cannot tolerate cold and higher altitude also frost and drought. Jack tree mostly grown in forest and on hill slope. The jackfruit can be grown on different varieties of soil as like well drained or deep alluvial soils open texture. For Jack tree pH of soil is 5.0 to 7.5. It requires free drainage. Jackfruit also tolerates shallow, slightly saline, and infertile soils (Tejpal and Amrit, 2016). 2.2 Jackfruit Description Jackfruit has a green to yellow brown exterior rind that is composed of hexagonal, bluntly conical carpel apices that cover a thick, rubbery, and whitish to yellowish wall. It is a multiple aggregate fruit which is formed by the fusion of multiple flowers in an inflorescence (Mushmbusi, 2015). About 30% of the fruit weight is occupied by the flesh. There are large number of bulbs inside the fruit, which have high nutritional value. The sweet yellow sheaths around the seeds are about 3-5cm thick and have a milder and lesser juice. Seeds are separated horny endocarpus enclosed by sub-gelatinous exocarpus(1mm thick) a thin whitish membrane. The fruit is made up of three main regions. They are (1) The fruit axis; (2) The persistent perianth; (3) The true fruit. Due to the presence of laticiferous cells that produce latex, which helps to hold the fruits together, the axis and the core of the fruit are inedible. The perianth is made up of three regions: (1) The bulb (the lower fleshy edible region); (2) The middle-fused region that forms the rind of the syncarp; (3) Horny non edible region commonly known as the spikes. The fruit colour changes from yellowish green to yellow due to the conversion of chlorophylls, anthocyanin’s, and carotenoids like pigments during ripening (Tiwari and Vidyarthi, 2015). Depending on the variety, the colour of the bulb can be cream, white, light yellow, yellow, deep yellow,lemon yellow light saffron, saffron, deep saffron, or orange. There is a widespread belief that excessive consumption of jackfruit flakes can lead to certain digestive disorders. The utilization of jackfruit as a commercial crop is limited due to its wide variations in fruit quality and long seed dormancy. The fruits must be utilized as early as possible when it reached the maturity as very sharp off flavours can be developed. Therefore, it is practiced to harvest the fruit when it is firm and in a semi ripen condition before ripening on the tree and then store until becoming soft and fit for processing (Tiwari and Vidyarthi, 2015). 2.2.1 Varieties of Jackfruit There are two main varieties of jackfruit, 1. Firm jackfruit 2. Soft jackfruit In the firm variety, the perianth remains firm even at full ripeness, while in the soft variety, the perianths become soft and fleshy on ripening. The soft variety has fruit with small, fibrous, soft, and spongy flakes with very sweet carpels, whereas, the firm variety is crunchy with crisp carpels and not sweet as the soft variety. The firm variety is considered to be of high quality. 2.2.2 Chemical Composition and Nutritional Value The chemical composition of jackfruit varies depending on the variety. When compared with other tropical fruits jackfruit flesh and seeds contain more protein, calcium, iron, and Thiamine. A study has reported that the ripe jackfruit is richer than apple, apricot, avocado, and banana in some minerals and vitamins (Tiwari and Vidyarthi, 2015). The caloric content of jackfruit is low, where 100 g of jackfruit only contains 94 calories. Jack fruit is rich in several nutrients (Table 2.1). It can act as source of complete nutrition to the consumers. The fruit is equivalent to Avocado and olive in terms of the healthier mix of nutrients for human dietary needs, almost having the exact nutrient equivalents of mother’s milk. It is rich in vitamin B and C, potassium, calcium, iron, proteins and high level of carbohydrates, affordable and readily available supplement to our staple food. Its seeds are rich in proteins and can be relished as a nutritious nut. The fruit is also the source of chemical “Jacalin” useful in preventing colon cancer, AIDS etc., Table 2.1. Nutritive value of jackfruit (100 g) Constituent Average value Moisture (%) 76.20 Energy (cal) 88.00 Protein (g) 1.90 Fat (g) 0.10 Fibre (g) 1.10 Carbohydrates (g). 19.80 Potassium (mg). 107.00 Calcium (mg) 20.00 Phosporous (mg) 41.00 Iron (mg) 0.56 β Carotene (mg) 175.00 Thiamine (mg). 0.03 Riboflavin (mg) 0.13 Niacin (mg) 0.40 Vitamin C (mg) 7.00 Source: Tiwari and Vidyarthi (2015) 2.2.3 Reasons for Underutilization of Jackfruit Despite its nutritional values and enormous health benefits, the jackfruit is underutilized and not classified as a commercial crop and is rarely grown on regular plantation scale due to its short shelf life and insufficient processing facilities in the regions where they are grown. About 60% of the whole jackfruit consists of inedible parts such as outer prickly rind, inner perigones, and central core (Xu et al., 2018) and only around 35% of the whole fruit consist of edible flesh. The jackfruit flesh is highly perishable and often undergoes flavour loss, tissue softening, and cut surface browning (Mondal et al., 2013). The softening of the fruit makes it more susceptible for bruising and mechanical injury. The separation of jackfruit edible bulbs from jackfruit rind is a difficult and more labour intensive task and consumes more time, which makes it unattractive to urban population where most of them have a busy life style. Also, the intense flavour of the fresh fruit makes it unacceptable to some consumers and there is a widespread belief that excessive consumption of jackfruit can lead to problems of digestion (Baliga et al., 2011). 2.2.4 Health Benefits of Jack Fruit There are many health benefits gotten from jackfruit like; Aids in Healthy Digestion Jackfruit is also known to contain anti-ulcer properties which help cure ulcers and digestive disorders. In addition, the presence of high fiber in the jackfruit prevents constipation and helps in smooth bowel movements. These fibers also offer protection to mucous membrane by driving away the carcinogenic chemicals from the large intestine (Priya et al., 2014). Protection against Cancer In addition to vitamin C, jackfruit is also rich in phyto-nutrients such as lignants, isoflavones and saponin which have anti-cancer and anti-aging properties. These phyto-nutrients may help eliminate cancer causing free radicals from the body and slow down the degeneration of cells that can lead to degenerative diseases (Priya et al., 2014). . Maintenance of Healthy Eye and Skin Jackfruit contains vitamin A, a powerful nutrient which is known to maintain a healthy eye and skin. It also helps prevent vision related problems such as macular degeneration and night blindness (Priya et al., 2014). Maintenance of Healthy Thyroid Jackfruit is loaded with copper, an important mineral, which plays a key role in the thyroid metabolism, especially in hormone production and absorption (Priya et al., 2014). Prevention of Anemia Jackfruit also contains iron which helps to prevent anaemia and also helps in proper blood circulation in our body (Priya et al., 2014). Strengthening of Bones Jackfruit is rich in magnesium, a nutrient which is important in the absorption of calcium and works with calcium to help strengthen the bone and prevent bone related disorders such as osteoporosis (Priya et al., 2014). . Controls Asthma The root of jackfruit has been found to help those who suffer from asthma. Extract of boiled jackfruit root has been found to control asthma (Priya et al., 2014). . Lowering of High Blood Pressure Potassium contained in Jackfruit has been found to be helpful in the lowering of blood pressure and thus reducing the risk of heart attack as well as strokes (Priya et al, 2014). . Strengthening of Immune System Jackfruit is an excellent source of vitamin C, which helps to protect againstviral and bacterial infections. Vitamin C helps to strengthen the immune system (Priya et al., 2014) Energy Booster Jackfruit is considered as an energy generating fruit due to the presence of simple sugars like fructose and sucrose which give you an almost immediate energy boost. Although Jackfruit is an energy rich fruit, it contains no saturated fatty oil and cholesterol making it a healthy fruit to savour (Priya et al., 2014). 2.2.5. Value Addition in Jackfruit Jack fruit has got great potential for value addition. More than 100 items can be prepared from jack fruit right from immature stage to well ripened stage. Each item has its own virtues in terms of taste, preference, keeping quality etc. Jack fruit is a potential raw material for value addition. Several delicious products can be made out of jack fruit right from immature fruit to seeds. There are various items that can be prepared out of immature jackfruit which includes jackfruit pickle, jackfruit cutlet, jackfruit papad, masala papad, jackfruit chips, jackfruit bajjas, jackfruit pakodas, jackfruit xacuti, jackfruit halwa, jackfruit pudding, jackfruit fritters, jackfruit custard, jackfruit wine, tender jack bhaji, jackfruit jam, jackfruit sweet vada, jackfruit kheer, jackfruit squash, etc. 2.3 Jackfruit Seeds Jackfruit seed is a potential local food source, which can be derived as flour. However, this seed contains several oligosaccharides such as raffinose and stachyose, which can cause flatulence for humans, and these substances will also create a darker colour during flour processing. Fully mature sweet bulb contain seeds which is 10 to 15 % of fruit weight. Colour of this is light brown, oval, oblong ellipsoid or rounded in shapes length of this is 2-3 cm(0.8-1.2inch) and diameter is 1-1.5 cm(0.4-0.6inch). Up to 500 to 550 seeds can be found in each jackfruit (Abraham and Jayamuthunagai, 2014). Jackfruit seeds have some pasting, antioxidant properties. It is eaten whole by boiling or roasting. They are easily digestible. Also it contains same composition as like grains. The two lectins found in jackfruit seeds; jaclin and artocarpin which have been proved to improve the immune status of HIV infected patients. It possesses immunological properties (Gupta et al., 2011). Jaclin has been seen to inhibit the herpes simplex virus type 2, it is also used for the isolation of human plasma glycoproteins, the investigation of lgA nephropathy, the analysis of o- linked glycoproteins and the detection of tumors. It is necessary to eliminate the oligosaccharides from the jack bean seed before using it as food. An alternative technique to reduce oligosaccharides from the bean is fermentation process (Gu et al., 2013). Several microbes that have been widely used in the fermentation of foodstuffs are lactic acid bacteria (LAB), such as L.plantarum SMN 25, L.plantarum pentosus SMN 01 and L. plantarum pentosus FNCC 235 isolated from the fermentation of traditional foods, showing the ability to produce galactosidase which is able to degrade oligosaccharides. According to Lee et al. (2011), LAB can ferment various types of carbohydrates include raffinose and stachyiose, as these oligosaccharides are found in many plants, especially ingrains. Jackfruit seeds contain starch 22% and dietary fiber 3.19%. It contains lignans, isoflavones, saponins, all phytonutrients and their health benefits are wide ranging from anticancer to antihypertensive, antiaging, antioxidant, antiulcer. Protein, and carbohydrate contents of jackfruit seeds has 13.50%, and 79.34%, respectively (Ocloo et al., 2010). Various triterpenes and flavonoids can be isolate from this fruits it also contain alkaloids, phenols, tannins, saponins, steroids (Hari et al., 2014). The antinutritional factors present in jackfruit such as tannin and trypsin inhibitors, result in digestive ailment when eaten raw (Swami et al., 2012). The Jackfruit seed contains very high amount of calcium that is 3087 mg/kg, iron 130.74 mg/kg, potassium content 14781 mg/ kg, sodium 60.66 mg/kg, copper 10.45 mg/kg and manganese 1.12 mg/kg. The pH and titratable acidity values is 5.78 and lactic acid is 1.12 % (Ocloo et al., 2010). Diarrhea and dysentery are treated by using jackfruit seeds. Seeds are helpful in digestion. The excess amount of free radicals disrupted the biomolecules and their prevention is vital for cytoprotective effects. All past studies proved that polyphenols, carotenoids, anthocyanins and flavonoids present in this fruits and this works as excellent scavengers of the free radicals and the human body protects from cellular damage (Baliga et al., 2011). Jackfruit seeds have all phytonutrients and their health benefits are wide-ranging from anticancer to antihypertensive, antiaging, antioxidant, antiulcer. The jackfruit seeds worked as a cooling as well as a nutritious tonic (Swami et al., 2012). For good hair growth jackfruit seeds are very beneficial besides healthy blood circulations and also healthy digestion. A class of glycoprotein that is lectin present in jackfruit seeds which shows some antibacterial, antifungal and anticarcinogenic properties (Chowdhury et al., 2012). Dietary fibre present in jackfruit is high in amount, which makes it an excellent bulk laxative. Fibre has ability to protect the colon mucous membrane by reducing exposure time and also binding to cancer causing chemicals in the colon (Mondal et al., 2013). The jackfruit seeds contain around 55% moisture content, thereby lessening the possibility of being kept for a long time (Mahanta and Kalita, 2015). 2.3.1 Chemical composition of jackfruit seeds Jackfruit seeds contain starch is 22% and dietary fiber 3.19%. It contains lignans, isoflavones, saponins, all phytonutrients and their health benefits are wide ranging from anticancer to antihypertensive, antiaging, antioxidant, antiulcer. Protein, and carbohydrate contents of jackfruit seeds as 13.50%, and 79.34%, respectively (Ocloo et al., 2010). The protein content is very different, because the seeds were collected from different region because different region sample have slightly different nutrition content (Swami et al., 2012). Various triterpenes and flavonoids can be isolate from this fruits it also contain alkaloids, phenols, tannins, saponins, steroids (Hari et al., 2014). The antinutritional factors present in jackfruit such as tannin and trypsin inhibitors, result is digestive ailment when eaten raw (Swami et al., 2012). When seed converted into seed flour then the ash and fat contents were 2.70 % and 1.27 %, respectively. The Jackfruit seed contains very high amount of calcium that is 3087 mg/kg, iron 130.74 mg/kg, potassium content 14781 mg/ kg, sodium contain 60.66 mg/kg, copper 10.45 mg/kg and manganese 1.12 mg/kg. The pH and titratable acidity values are 5.78 and lactic acid is 1.12 % (Ocloo et al., 2010). Compared with other tropical fruit seeds, jackfruit seeds packed many important nutrients. They contain high levels of starch, protein, vitamin, minerals and antioxidants. A 1ounce (28grams) serving of jackfruit seeds contain Calories : 53grams Protein : 2grams Carbs : 11grams Fats : 0gram Fiber : 0.5grams Riboflavin : 8 % of the reference daily intake( RDI) Thiamine : 7% of RDI Magnesium : 5% of RDI Phosphorus : 4% of RDI Furthermore, jackfruit seeds offer fiber and resistant starch, both of which pass through the body undigested and act as food for the beneficial gut bacteria. Fiber and resistant starch have been linked to many powerful health benefits, including hunger control, reduced blood sugar levels and improved digestion and insulin sensitivity. 2.3.2. Nutritional and Health Benefits of Jackfruit Seeds Jackfruit seeds have been used in traditional Chinese medicine as an aphrodisaiac and treatment for digestive issue among other applications. Anti-microbial effects Diarrhea and dysentery are treated by using jackfruit seeds. Seeds are helpful in digestion. The excess amount of free radicals disrupted the biomolecules and their prevention is vital for cytoprotective effects. All past studies proved that polyphenols, carotenoids, anthocyanin’s and flavonoids present in this fruits and this works as excellent scavengers of the free radicals and the human body protects from cellular damage (Baliga et al., 2011). Anti-cancer properties Jackfruit seeds have lignans, isoflavones, saponins, and all phytonutrients and their health benefits are wide-ranging from anticancer to antihypertensive, antiaging, antioxidant, antiulcer. The seeds content starch which is considered as useful in relieving biliousness, besides the roasted seeds are considered as an aphrodisiac. The jackfruit seeds worked as a cooling as well as a nutritious tonic (Swami et al., 2012). For good hair growth jackfruit seeds are very beneficial besides healthy blood circulations and also healthy digestion. High amount of starch, low in calcium and iron and very good source of vitamin B2 (Arpit and John, 2015). A class of glycoprotein that is lectin present in jackfruit seeds which shows some antibacterial, antifungal and anticarcinogenic properties (Chowdhury et al., 2012). Aids in digestion Dietary fibre present in jackfruit in high amount, which makes it an excellent bulk laxative. Fibre have ability to protect the colon mucous membrane by reducing exposure time and also binding to cancer causing chemicals in the colon (Mondal et al., 2013). Strengthening of bone This seeds are also helpful in bone health because rich in magnesium which is an essential mineral necessary in absorption of calcium and works along with calcium for strengthening the bones and prevents disease like Osteoporosis (Mourya, 2016). Cholesterol reduction Elevated levels of LDL (bad cholesterol) are associated with high blood pressure, diabetes, and an increased risk of heart disease. Contrarily, higher levels of HDL (good cholesterol have been found to have heart protective effects. Eating more jackfruit seeds have reduced levels of LDL and increased HDL. 2.3.3. Processing Technology of Jackfruit Seeds After ripening of the fruit, processing is done on seeds in jackfruit growing area. The fruit is opened with knife, seeds are separated from the bulb that embedded in fruit. Every fruit contains about 100-500 seed, there is no relation between fruit size and the quantity of seeds that fruit contains. There are about 50–90 seeds/kg of fruit. The thin, slimy coating of bulb around the seed which should be removed and thoroughly rinsed in water to remove any remaining pulp juice or sugary residue. Seeds when shade dried for an hour for handling its slimy layer but should not get dried more time in open place because there may be loss of nutrients. The jackfruit seed has been utilized for processing more than the peel. Nowadays the extensive processing of jackfruit seeds is done and it is becoming new trend to utilize the jackfruit seeds for various purposes. About 75% of the Jackfruit seeds are converted into flour (Cheok et al., 2016). For processing of seeds into flour, the seeds are first sorted and then cleaned. Cleaning is followed by roasting; roasting is done at appropriate temperature. When roasting it is done at 1600C, there is good retention of nutrients like protein, minerals due to reduction of moisture content. The roasted seeds are then dried in tray drier or in cabinet drier and milled in flour mill. Flour is also prepared as it is after drying (Eke- Ejiofor et al., 2014). 2.3.4. Applications of Jackfruit Seed Flour The jackfruit seed have higher nutrition content so nowadays it is used in bakery products like bread by using 10 and 20% jackfruit seed flour with wheat flour for reducing gluten content and for digestion aid (Butool and Butool, 2013). Cake making is also done by incorporation of jackfruit seed flour 5 to 15 % and result is increased protein and reduction in fat content (Arpit and John, 2015). Extruded product also prepared by using jackfruit seed flour like noodles, upto 20% seed flour was incorporated in it and its protein and dietary fibres increased (Nandkule et al., 2015). Jackfruit seed flour up to 25 per cent level, can also be utilized in chapatti preparation. Addition of preservatives in such chapatti extends its shelf life up to 3 to 4 days for ambient and up to 30 days at refrigeration temperature (Sultana et al., 2014). Also seeds are used in starch extraction, starch was isolated from the flour because its amylose content is high, and also it shows good paste stability during heating. CHAPTER THREE MATERIALS AND METHODS 3.1 Source of Raw Materials The Jackfruit seed (Artocarpus heterophyllus Lam) seeds were obtained from Ubani main market, Umuahia North Local Government Area in Abia State. The laboratory analysis was carried out at the Department of Food Science and Technology, Abia State University, Uturu, Abia State, Nigeria. 3.2 Preparation of Jackfruit Seed Flours 3.2.1 Seed Pretreatment The method described by Akaerue and Onwuka (2010) was used. The jackfruitseeds were sorted, washed and drain off. The jackfruit seeds were divided into four portions weighing 200g each; one waskept raw (untreated), while the second set was roasted, third portion was fermented. The fourth portion was boiled. 3.2.2 Preparation of Raw Jackfruit Seed Flour The method described by Akaerue and Onwuka (2010) was used for preparation of raw jackfruit seed flour. About 200g of the soaked jackfruit seeds was drained and rinsed three times with 600ml distilled water before drying in the oven (DHG Oceanmed, England) at 65oC for 9 hours and then cooled in a desiccator. The dried jackfruit seeds were ground using an attrition mill and sieved with 80μm mesh sieve and packaged in air-tight plastic container for analysis thereafter. The processing scheme for the production of raw jackfruit seed flour is presented in figure 1. Soaking (in distilled water for 12 hours) Drying (in hot-air oven at 65oC for 9 hours) Milling (in attrition mill) Sieving (using a 80 μm mesh sieve) Packaging (in air-tight plastic container) Figure 1: Processing Scheme for the production of raw jackfruit seed flour 3.2.3 Preparation of Roasted Jackfruit Seed Flour The method described by Miftakhussolikhah et al. (2015) was used for preparation of roasted jackfruit seed flour with slight modifications. About 200g of the soaked jackfruit seeds was drained and rinsed three times with 600ml distilled water followed by oven drying (DHG Oceanmed, England) at 65°C for 9 hours. The jackfruit seeds were roasted using a roaster at 160oC for 15 minutes with the help of an electric induction heater. The beans were stirred continuously for avoiding any phenomenon or uneven roasting. The roasted seeds were ground using a attrition mill and then sieved with 80μm mesh sieve and packaged in air-tight plastic container for analysis thereafter. The processing Scheme for production of roasted jackfruit seed flour is presented in figure 2. Soaking (in distilled water for 12 hours) Drying (in hot-air oven at 65oC for 9 hours) Roasting (at 160oC for 15 minutes) Milling (in attrition mill) Sieving (using a 80 μm mesh sieve) Packaging (in air-tight plastic container) Figure 2: Processing Scheme for production of roasted jackfruit seed flour 3.2.4 Preparation of Fermented Jackfruit Seed Flour The method described by Abasiekong (2008) and Ariahu et al. (1999) was used for preparation of fermented jackfruit seed flour with slight modifications. About 800 g of the soaked jackfruit seeds was drained and rinsed three times with 600 ml distilled water followed by soaking in 1500 ml of distilled water for 4 hours, then decanting a portion of the water and allowing the seeds to ferment at room temperature (25 - 30°C)for 48 hours. The fermented jackfruit seeds were dried in an oven (DHG Oceanmed, England) at 65°C for 9 hours. The dried seeds were ground using an attrition mill and then sieved with 80μm mesh sieve and packaged in air-tight plastic container for analysis thereafter. The Processing Scheme for production of fermented jackfruit seed flour is presented in figure 3. Clean jackfruit seeds Raw jackfruit seed flour Soaking (in distilled water for 4 hours) Fermentation (at 25 - 30oC for 48 hours) Drying (in hot-air oven at 65oC for 9 hours) Milling (in attrition mill) Sieving (using a 80 μm mesh sieve) Packaging (in air-tight plastic container) Figure 3: Processing Scheme for production of fermented jackfruit seed flour 3.2.5 Preparation of Boiled Jackfruit Seed Flour The method described by Akaerue and Onwuka (2010) was used for preparation of boiled jackfruit seed flour. About 200g of the jackfruit seeds was drained and rinsed three times with 600 ml distilled water. The jackfruit seeds was boiled at 1000C for 1hrs, followed by oven drying (DHG Oceanmed, England) at 65oC for 9 hours and cooling in a dessicator. The dried seeds was subjected to dry milling and passed through 80μm mesh sieve. The flour was cooled and packaged for analysis thereafter in air-tight plastic container. 3.3 Determination of Proximate Composition 3.3.1 Determination of Moisture Content Moisture content was determined by the gravimetric method described by AOAC (2006). Five (5) g of the samples was placed in previously weighed moisture cans and the weight was taken. The can and its content was dried in the oven at 105˚C for 3 hours in the first Clean jackfruit seeds Roasted jackfruit seed flour instance and then cooled in desiccators and re-weighed. The weight was recorded and the dish was placed back into the oven to be further dried. The drying, cooling and weighing at intervals of 1 hour will continue until a constant weight is obtained. The weight of moisture loss was determined by difference and expressed as a percentage of the sample analyzed. It was calculated using the formula: % Moisture Content= Where: W1= Weight of empty dish, W2= Weight of empty dish + sample before drying, W3= Weight of empty dish + sample after drying 3.3.2 Determination of Ash Content Ash content was determined by furnace incineration gravimetric method described by AOAC (1990). Two (2) g of the samples was placed into previously weighed porcelain crucible. It was allowed to ash in a digital muffle furnace (model no: SX2-2.5-12; made in China) at 550˚C for 2 to 3 hours. After cooling in desiccators, it was weighed and the weight of the ash was obtained by difference and expressed as a percentage of the weight of the sample analyzed. It was calculated using the formula: % Ash Content= Where: W1= Weight of empty crucible, W2= Weight of crucible + ashes, W3= Weight of sample 3.3.3 Determination of Crude Protein Content Protein content of the samples was determined by Kjeldahl method described by James (1995). Total nitrogen was determined and multiplied with a factor (6.25) to obtain the protein content. Two (2) g of sample was mixed with 10 ml of concentrated sulphuric acid (H2SO4) in a kjeldahl apparatus and heated under a fume cupboard (digestion) until a clear solution is obtained. The digest was diluted to 100 ml and used for analysis. Ten (10) ml of the digest was mixed with equal volume of 45 % sodium hydroxide (NaOH) solution in a kjeldahl distillation apparatus. The mixture was distilled and the distillate was collected in 10 ml of 4 % boric acid solution containing 3 drops of mixed indicator (methyl red and bromocressol green) was added. A total of 50 ml of distillate was collected and titrated against 0.02N H2SO4 solution. Not more than 40 ml of the distillate was used. A reagent blank will also be digested and titrated. The nitrogen (N2) content was calculated using the formula: %N2= The percentage protein content was calculated using the formula: % Protein Content= % N2 x 6.25 3.3.4 Determination of Fat Content Fat content of the samples was determined by the continuous solvent extraction method using a soxhlet apparatus. The method described by James (1995) was used. Two (2) g of each sample was wrapped in thick paper (Whatman 42 filter paper) and put in a thimble. It will then be placed in a soxhlet reflux flask and mounted into a weighed extraction flask containing 200 ml of petroleum ether. The upper end of the reflux flask was connected to a condenser. When the heated solvent condense into the reflux flask, it will cover the sample until the flask is filled up and siphoned over carrying oil (fat) extract down to the boiling flask. The process was allowed to go on repeatedly for about 4 hours before the defatted sample was removed and kept for crude fiber determination. The solvent was removed and the flask with its oil extract was dried in the oven at 60˚C for 30 minutes, cooled in a desiccator and reweighed to obtain the weight of the oil extract (fat) which will then be expressed as a percentage of the sample analyzed. The percentage fat content was calculated using the formula: % Fat Content= Where: W= Weight of sample, W1= Weight of empty extraction flask, W2= Weight of extraction flask + oil (fat) extracted 3.3.5 Determination of Crude Fiber Content Crude fiber content was determined by the gravimetric method described by AOAC (2006). Two (2) g of each sample (defatted) was boiled in 20 ml of 1.25 % H2SO4 for 30 minutes under reflux. The mixture was washed in hot water using a two-fold muslin cloth to retain the particles. The retained particles was boiled again in 1.25 % NaOH solution for another 30 minutes, washed as before and allowed to dry in the oven at 105˚C for 3 hours. The samples was cooled in Fermented jackfruit seed flour Clean jackfruit seeds desiccators and weighed. Lastly, the samples was burnt to ashes in a digital muffle furnace (model no: SX2-2.5-12; made in China), cooled and weighed. The crude fiber content was calculated using the formula: % Crude Fiber Content= Where: W1= Weight of sample, W2= Weight of crucible + sample after boiling, washing and drying, W3= Weight of crucible + sample after ashing 3.3.6 Determination of Carbohydrate Content The carbohydrate content of the samples was determined using the method described by Pearson (1976) and Udoh and Ogunwale (1986). The carbohydrate content was calculated by difference. % Carbohydrate Content= 100 – (% Moisture + % Ash + % Protein + % Fat + % Crude Fiber) 3.4 Determination of Functional Properties 3.4.1 Determination of Water Absorption Capacity The method as described by Abbey and Ibeh (1988) was adopted. One gram of each sample was weighed separately and placed in a clean dry centrifuge tube. Distilled water was mixed with the samples to make up to 10ml dispersion. The sample and 10ml distilled water was mixed thoroughly for 30 seconds, then the sample was allowed to stand for 5minutes at room temperature and then centrifuged at 1000 rpm for 5minutes, then the volume of free water (supernatant) was read directly from the graduated centrifuge tube and calculated as follows: WAC (ml/g) = V1-V2/W, Where W=weight of flour, V1=initial volume before water absorption, V2=final volume after water absorption 3.4.2 Determination of Oil Absorption Capacity The method as described by Abbey and Ibeh (1988) was adopted. One gram of each sample was weighed separately and placed in a clean dry centrifuge tube. Oil was mixed with the samples to make up to 10ml dispersion. The sample and 10ml oil was mixed thoroughly for 30 seconds, then the sample was allowed to stand for 5minutes at room temperature and then centrifuged at 1000 rpm for 5minutes, then the volume of free oil (supernatant) was read directly from the graduated centrifuge tube and calculated as follows: OAC (ml/g) = V1-V2/W, Where W=weight of flour, V1=initial volume before oil absorption, V2=final volume after oil absorption 3.4.3 Determination of Bulk Density This was done according to the method described by Onimawo and Akubor (2005). Ten milliliter (10ml) capacity graduated measuring cylinder was filled with the sample and the initial volume was recorded. The cylinder was tapped continuously to displace air and vacuum until the volume becomes constant. The final volume was noted after it is leveled, and bulk density of the samples was calculated as follows: The bulk density (g/ml) = weight of samples / volume of sample 3.4.4 Determination of Emulsion Capacity The Method of Palmashted (1987) was adopted. Emulsion capacity was estimated by blending 2g of flour sample with 25ml distilled water at room temperature for 30seconds in a warring blender. After complete dispersion, 25ml of vegetable oil (groundnut oil) was added and the blending will continue for another 30seconds, then it was placed into a centrifuge tube and centrifuged at 1000rpm for 5minutes. The volume of oil separated from the sample after centrifuging was read directly from the tube. Emulsion capacity was expressed as the amount of oil emulsified and held per gram of sample. Emulsion capacity (%) = (H1 / H2) x 100; where H1 = height of emulsified layer and H2 = height of whole solution in the centrifuge tube 3.4.5 Determination of Swelling Index This was determined as described by Appiah et al. (2011). One gram of the flour was mixed with 10 ml distilled water in a centrifuge tube and heated in a hot water bath at 80°C for 30 minutes while continuously shaking the tube. After heating, the suspension was centrifuged at 1000 rpm for 15 minutes. The supernatant was decanted and the weight of the paste taken. The swelling index was calculated as: Swelling index = weight of the paste / weight of dry flour 3.5 Determination of Mineral Content The mineral content of the samples was determined by the dry ash extraction method followed by specific mineral content. Two gram (2 g) of the samplewas dried and burnt to ash in a muffle furnace. The resulting ash was dissolved in 100 ml of dilute hydrochloric acid (2M HCl), heated for 30 minutes and made up to 100 ml with distilled water. The digest obtained was used for various analyses. 3.5.1 Determination of Calcium and Magnesium Calcium and magnesium content of the samples was determined by the Versenate EDTA complexiometric titration method described by James (1995). Twenty milliliter (20) ml of each of the extract was dispersed into a conical flask; then pinches of the masking agent hydroxyamine hydrochloride, potassium cyanide was added followed by 20 ml of ammonia indicator solution pH 10.0. A pinch of the indicator, Erichrome black was added and the mixture was shaken very well and titrated against 0.02N EDTA solution. Titration was from mauve colour to permanent blue colouration. A reagent blank consisting of 20 ml distilled water will also be treated as described above. The titration gave a reading for combined calcium and magnesium complexes in the samples. A separate test was conducted for calcium. Titration for calcium alone was a repeat of the previous one with slight change. Ten percent (10 %) NaOH solution at pH 12.0 was used in place of the ammonia buffer while solochrome dark-blue (calcon) was used as indicator in place of Eriochrome black T. Calcium and magnesium contents was calculated separately using the formula below: Calcium or magnesium (mg/100g) = Where: W= Weight of the sample analyzed, EW= Equivalent Weight, VF= Total volume of extract, VA= Volume of extract titrated, N= Normality of EDTA (0.02N) and T= Titre value less blank 3.5.2 Determination of Phosphorous Content The phosphorous content of each sample was determined by Molybdovanadate (yellow) spectrophotometric method described by James (1995). One milliliter (1 ml) extract of each sample was dispensed into a test tube. Similarly, the same volume of standard phosphorous solution as well as water was placed into other test tubes to serve as standard and blank, respectively. The content of each test tube was mixed with equal volume of molybdovanadate colour reagent. They was allowed to stand at room temperature before their absorbance was read using Jenway electronic spectrophotometer at a wavelength of 420 nm. The reading was taken with the blank at zero. The phosphorous content of the sample was calculated using the formula: P (mg/100g) = X X C X Where: W= Weight of sample, Au= Absorbance of the standard solution, C= Concentration of the standard, Vf= Total volume of the filtrate and Va= Volume of filtrate analyzed 3.5.2 Determination of Sodium and Potassium Potassium (K) and Sodium (Na) in the sample was determined by flame photometry method as described AOAC (2006). The instrument was set up according to the manufacturer’s instruction. The equipment was switched on and allowed to stand for about 10 minutes. The gas and air lets of the knob was turned on. The equipment was adjusted to a non-luminous level (i.e. blue flame). Meanwhile, standard potassium (k) and sodium (Na) solutions wasprepared separately and each was diluted to concentration of 2, 4, 6, 8 and 10 ppm. Each mineral was analyzed using the appropriate filter and the instrument was flushed with distilled water. The highest concentrated standard solution was in place and the reading adjusted to 100 ml. Thereafter, starting with the least concentration i.e. 2 ppm, all the standard solutions was sucked into the instrument and caused to spray over the non-luminous flame. The readings was recorded and plotted into a standard curve and was used to extrapolate the potassium level in the samples. After the standard, the sample digest was siphoned into tarns into the instrument and their readings was recorded. The samples was repeated with sodium standard and sodium filter. The concentration of the test mineral in the sample was calculated with reference to the graphs and obtained as follows: Na or K (mg/100g) = Where: W= Weight of sample used, X= Concentration of the test element from the curve, Vf= Total extract volume, Va=Volume of extract analyzed and D= Dilution factor where applicable 3.6 Determination of Vitamin Content 3.6.1 Determination of Thiamine (Vitamin B1) Thiamine content was determined using the method described by AOAC (2006). Two (2) g of the samples was homogenized with ethanolic sodium hydroxide solution in a 50 ml conical flask and filtered. Ten (10) ml of the filtrate was pipetted and the colour was developed by addition of 10 ml of potassium dichromate and read at 360 nm in a Jenway electronic spectrophotometer (model no: 7305; made in England). A blank sample was prepared and the colour was also developed and read at the same wavelength. Standard solution will also be prepared using thiamine acid to get 100 ppm and serial dilutions of 0.0, 0.2, 0.4 and 0.6 ppm was made and used to plot the calibration curve. The thiamine content of the samples was calculated using the formula: Thiamine (mg/100g) = X X C X Where: W= Weight of sample, Au= Absorbance of sample, As= Absorbance of standard thiamine solution, C= Concentration of standard thiamine solution, Vf= Total volume of filtrate (extract), Va= Volume of extract analyzed 3.6.2 Determination of Riboflavin (Vitamin B2) The riboflavin content of each sample was determined using the method described by AOAC (2002). Two (2) g of each sample was extracted with 100 ml of 50 % ethanol solution and shaken for one hour. This was filtered into a 100 ml flask. Ten (10) ml of the extract was pipetted into 50 ml volumetric flask. Ten (10) ml of 5 % potassium permanganate (KMNO4) solution and 10 ml of 3 % hydrogen peroxide (H2O2) was added and the solution was allowed to stand over hot water bath for about 30 minutes. Two (2) ml of 4 % sodium sulphate (Na2SO4) solution was added and a yellowish-pale colour is formed. This was made up to 50 ml mark and the absorbance was measured at 510 nm in a Jenway electronic spectrophotometer (model no: 7305; made in England). The riboflavin content of the samples was calculated using the formula: Riboflavin (mg/100g) = X X C X 3.6.3 Determination of Niacin (Vitamin B3) Niacin content of the samples was determined using the method described by AOAC (2002). Two (2) g of each sample was treated with 50 ml 1N sulphuric acid and shaken for 30 minutes. Three (3) drops of ammonia solution was added to the sample and filtered. Ten (10) ml of the filtrate was pipetted into 50 ml volumetric flask and 5 ml of potassium cyanide was added. The mixture was acidified with 5 ml of 0.02N sulphuric acid and absorbance was measured in Jenway electronic spectrophotometer (model no: 7305; made in England) at 470 nm. A standard (100 ppm) was prepared using niacin acid and a serial dilution of 0.0, 0.2, 0.4, 0.8 ppm was made from it. This was used to calibrate the curve. The niacin content of the samples was calculated using the formula: Niacin (mg/100g) = X X C X Where: W= Weight of sample, Au= Absorbance of sample, As= Absorbance of standard niacin solution, C= Concentration of standard niacin solution, Vf= Total volume of filtrate (extract), Va= Volume of extract analyzed 3.6.4 Determination of Vitamin A The method described by AOAC (2006) was used. Two (2) g of each sample was dispersed in 30ml absolute alcohol; Three (3) ml of 50 % potassium hydroxide was added to it and boiled under reflux for 30 minutes. After cooling rapidly in running water, 30ml of distilled water was added and the mixture was transferred to a separation funnel. Fifty (50) ml of petroleum ether was used to wash the mixture thus extracting the vitamin A. The lower layer (aqueous) was discarded while the vitamin A extract was washed four times with 50 ml distilled water. The extract was then evaporated to dryness and re-dissolved in 10 ml of isopropyl alcohol and its absorbance was measured at 325 nm using Jenway electronic spectrophotometer (model no: 7305; made in England) at a wavelength of 325 nm. The vitamin A content was calculated using the formula: Vitamin A (mg/100g) = X X C Where: W= weight of sample, As= Absorbance of sample, Au= Absorbance of standard solution, C= concentration of standard solution. 3.6.5 Determination of Vitamin C Vitamin C content of the samples was determined using the Brakat titrimetric method described by AOAC (2002). Two (2) g of each sample was homogenized in 100 ml EDTA extraction solution for 5 minutes. The homogenate was filtered and the filtrate was used for analysis. The filtrate of each sample was passed through a packed cotton wool containing activated charcoal to remove the colour. The volume of the filtrate was adjusted to 100ml by washing with more of the extraction solution. Twenty (20) ml of each filtrate was measured into a conical flask. Ten (10) ml of 20 % potassium iodide solution was added to each flask followed by 5 ml of starch solution (indicator). The mixture was titrated against 0.01M CuS04 solution. Titration was done until the endpoint marked by black specks at the brink of the mixture. The vitamin C content was given by the relationship that 1ml of 0.01M CuSO4 equals 0.88mg vitamin C. Therefore, the vitamin C content of the sample was calculated using the formula: Vitamin C (mg/100g) = X 3.8 Statistical Analysis The mean score of the laboratory analysis results was subjected to one way Analysis of Variance (ANOVA), using the Statistical Package for the Social Sciences (SPSS) version 21. The mean scores were separated using Duncan’s Multiple Range Test (DMRT). Significant difference was determined at P