الفهرس 
ACKNOWLEDGEMENT
NutritionOnly 14 pages are availabe for public view
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Abstract
6- Summary and Conclusion:
6.1- Summary:
The present study was conducted to study the physical and chemical properties of Moringa seed oil and its leaves extracts. The study also aimed to verify the effect of oil and Moringa oleifera leaves extract on chemical and biological evaluation of the experimental rats. It was created two main groups. The first group (n = 9 rats) used as a comparison group (negative control-ve) fed on basal diet, the other group (n = 51 rats) fed a basal diet containing (1% cholesterol+ 0.5% cholic acid) for ten days to raise the level of cholesterol in the blood. Used one group of them (n = 9 rats) as a positive control (+ve), while were fed three groups of (5%, 10% and 15%) of Moringa oleifera seed oil and injected through the mouth and the other three groups with 200, 400 600 mg/g of b. wt. of Moringa oleifera leave extract.
6.1.1-Chemical composition of Moringa oleifera seeds:
The protein, ash and lipids contents of Moringa oleifera seed were recorded 32.8%, 3.81% and 35.75% respectively. Concerning the high percentage of oil represent the major constituent of Moringa oleifera seed makes this seed a distinct potential for the oil industry.
6.1.2- Physical analysis of Moringa oleifera seeds oil:
The viscosity, refractive index at 25 ̊ C and color of oil extracted from Moringa oleifera seeds were 70, 1.462 and red (1) / yellow (35) respectively. Viscosity is generally considered as an adequate indicator of resistance to the oil flow affecting by the reactions possible occur in it and by its composition. Refractive index is one of the most important physical parameters which used in the identification of oils as it is useful for estimation the degree of saturation of oils. Results could be concluded that refractive index was increased by increasing the degree of unsaturation. The color of the edible oils is considered one of the most considerable commercial importance physical characteristics. The obtained results showed that, red unit, yellow unit were 1 and 35 respectively. There upon, Moringa oleifera color index are more suitable for edible and many purposes.
6.1.3- Chemical composition of Moringa oleifera seeds oil:
Acidity: The acidity of Moringa oleifera oil was (0.56 %) as oleic acid. Free fatty acids content is a measure of the acidic components in the oil. So, acidity used as a good indicator for the hydrolysis extent before and during extraction procedures.
Iodine value: The iodine value of Moringa oleifera oil was (66.04 g of I2 / 100 g of oil). Iodine value is a measure of overall unsaturation and is widely to characterize oils and fats. The higher the iodine value the increased the unsaturation of the oil. There for, Moringa oleifera seeds oil could be classified as non-drying oil. Lowering iodine value of the oil reflect its characteristics as higher resistance to oxidation.
Peroxide value: The peroxide value of Moringa oleifera oil was (0.68 meq / Kg). Peroxide value is a measure of the amount of peroxides formed in fats and oils through autoxidation processes. Indirectly, it is a measure of the degree of initial oxidation of fats and oils. Obtained results could concluded that peroxide value was (0.68 meq I2 / kg of oil).
Thiobarbituric acid (TBA): The TBA value of Moringa oleifera oil was (0.156 mg malonaldehyde/Kg of oil). TBA is considered a good chemical quality measure to identify the oxidation state of fresh edible oils and fats and to measure secondary oxidation extent has been done in these lipids. Moringa oleifera seeds oil was characterized with a good oxidative state and healthy safe quality. Saponification number: The saponification number of Moringa oleifera oil is (168.16 mg/g). The values of saponification are indicative of the average molecular weight of fatty acid content as a glyceride in the oil.
Conjugated dienes and trienes:
Dienes: The absorbance value at 232 nm of Moringa oleifera oil was (1.785). The (Σ1% 1cm) at 232 nm which is an indicator of the formation of conjugated diene. The specific extinction coefficient at 232 nm (K232) is related to the degree of primary oxidation of the oil and thus directly correlated to the amount of hydroperoxides and it is also an indicator of polyunsaturated FA conjugation (Carla et al., 2016).
Triene: The absorbance value at 232 nm of Moringa oleifera oil was (0.146). The (Σ1% 1cm) at 270 nm which is an indicator of the formation of conjugated triene. The specific extinction at 270 nm (K270) related to the secondary oxidation products α-unsaturated ketone and α-diketone (Carla et al., 2016).
Total tocopherols and phenolic content: The Moringa oil was found to be rich in total phenolic content and tocopherols contents, with values of (38.75 mg/100g) and (14.45 µg/ 100 g of oil), respectively. Phenolic and tocopherols compounds have been proved to be responsible for antioxidant activity on many vegetable seeds oils.
Antioxidant activity: Moringa oleifera seeds oil exhibited a good scavenging abilities against DPPH radicals (68.32%). The Moringa oleifera oil exhibited a high radical scavenging activity than the DPPH radicals at the same concentration.
Induction period of Moringa seeds oil: The induction period of Moringa oleifera, oil (24.67 h at 100ºC). High induction period values of Moringa oleifera oil as exhibited in the present analysis indicate the presence of a high level of monounsaturated fatty acids, particularly, C18:1, which was less prone to oxidation than polyunsaturated. Consequently, Moringa oleifera seeds oil was characterized with a good oxidative state and stability.
Phenolic compounds identified and fractionation: The total phenolic compound of Moringa oleifera seed oil was fractionated and identified by HPLC and it is apparent that, there were 22 identified compounds in Moringa oleifera seeds oil. The identified compounds percentage ranged from (4.33 µg/100g) for reversetrol to (3188.66 µg/100g) for e-vanillic. Therefore, this analysis revealed that, the major constituents of the Phenolic compounds from Moringa oleifera seeds oil were e-vanillic 3188.66 µg/100g (79.38%), followed by Salycilic 172.65 µg/100g and 3-OH Tyrosol 114.98 µg/100g.
Fatty acid compositions of Moringa oleifera oil: The main fatty acids were oleic (C18:1 n-9), linoleic (C18:2 n-9.12), palmitic (C16:0), arachidic (C20:0), stearic (C18:0), behenic acid (C22:0), palmitoleic (C16:1 n-9) and linolenic acid (C18:3 n-9, 12, 15). Moringa oleifera oil was found to contain a high level of oleic acid (C18:1 n-9), which for 73.87% of the total fatty acid. Thus, Moringa oleifera oil is belongs to the oleic acid oil category (Sonntag, 1982). In addition, great amounts of palmitic (C16:0, 5.47%) and linoleic (C18:2, 0.82%) acids were detected. Minor amounts of palmitoleic (C16:1, 1.35%), stearic (C18:0, 4.61%), and behenic (C22:0, 7.31%) was also detected.
6.1.4- Moringa oleifera leaves:
6.1.4.1- Chemical composition of Moringa oleifera leaves:
Moisture, Ash, total fats and Protein of Moringa oleifera leaves were 6.18%, 11.83%, 14.87% and 29.5% respectively that means Moringa oleifera leaves are a good source of minerals elements and protein.
Total phenolic content: Moringa oleifera leaves contained of (99.04 mg GAE/g) total phenolic content. Phenols are antioxidants that can scavenge biological free radicals, and chemo prevents diseases with biological oxidation as their main etiological factor.
Antioxidant activity of Moringa oleifera leaves:
Spectrophotometric electron transfer based assays which were employed to measure the capacity of an antioxidant in the reduction of an oxidant, which changes color when reduced. The degree of color change (either an increase or decrease of absorbance of the probe at a given wavelength) is correlated to the concentration of antioxidants in the sample. Data showed the antioxidant activity of Moringa oleifera leaves were 85%.
Amino acid composition of Moringa oleifera leaves: The dried Moringa oleifera leaves contained 18 amino acids. Moringa oleifera contains essential amino acids essential amino acids, 49.32 % of total amino acids. The higher percent was Lucien (9.28%) followed by phenylalanine (6.79%), lysine (6.27%), arginine (6.19%), Valine (6.10), isoleucine (4.94%), therionine (4.73%), histidine (3.26%) and Methionine (1.76), was the lower percent. The nonessential amino acids percentage comprise 50.51% of total amino acids, the higher was glutamic (12.46%) followed by aspartic (10.14%), alanine (6.57%), glycine (5.58%), proline (4.90%), serine (4.64%), Tyrosine (4.42) and the lowest is methionine (1.76%). Moringa is reported to have high quality protein which is easily digested and that is influenced by the quality of its amino acids.
6.1.5- Biological evaluation and blood serum analysis:
6.1.5.1- Biological evaluation:
6.1.5.1.1- Body weight gain and organs weight:
Rats fed on the diets containing Moringa oleifera leaves extract and oil for fifty days compared with rats fed on the diets containing cholesterol and cholic acid for ten days:
Body weight gain: The decreases are in all treatments either the (leaves extract or oil) comparable with hypercholesterolemic group (positive control +ve).
Liver weight: The decreases are in all treatments either the (leaves extract or oil) comparable with hypercholesterolemic group (positive control +ve).
Kidney weight: The increases are in all treatments either the (leaves extract or oil) comparable with hypercholesterolemic group (positive control +ve).
6.1.5.1.2- Blood serum analysis:
Rats were fed on basal diets by orally Moringa oleifera leaves extract (200, 400 and 600 mg/g of b.wt) and Moringa oleifera seeds oil (5%, 10% and 15%) a markedly decreased in both of total cholesterol, triglycerides, low density lipoprotein (LDL) and atherogenic index observed, while high density lipoprotein (HDL) increased as compared to hypercholesterolemic rats that fed on diets content of 1% cholesterol and 0.5% colic acid.
6.1.5.1.3- Liver and Kidney functions:
Glutamate oxaloacetate transferase (GOT) and glutamate pyruvate transferase (GPT) activities are most frequently measured for diagnosis of liver diseases particularly infective hepatitis, alcoholic cirrhosis, biliary obstruction, toxic hepatitis and liver cancer.
The study recorded that GOT, GPT, creatinine and uric acid were decreased in all treatments of Moringa oleifera either the leaves extract or oil as compared to hypercholesterolemic group.
6.1.5.2- Histopathological observation:
The Histopathological changes in comparison with the control group in liver and kidney of the experimental animals were in form of fat degeneration, hydropic degeneration and congestion. Due to the important role, the liver plays in biotransformation of drugs and toxins, drug-induced Hepatotoxicity should be a major concern in drug development and chronic drug therapy.
6.1.5.2.1-Pathological changes in Liver:
Treatment 2, 3, 5 and 6 recorded mild changes of Hydropic, fat degeneration and Congestion as compared with hypercholesterolemic group.
Treatment 1 and 4 recorded mild changes of Hydropic and congestion while recorded moderate changes of fat degeneration as comparable to hypercholesterolemic group.
6.1.5.2.2- Pathological changes in kidney:
Treatment 1, 3 and 4 recorded modenit of Hydropic, fat degeneration while recorded mild in Congestion.
Treatment 2, 5 and 6 recorded mild changes of Hydropic, fat degeneration and Congestion as compared with hypercholesterolemic group.
6.2- Conclusion:
from the former results and discussion, it could be concluded that:
• Oils and leaves of Moringa oleifera have definite hypocholesterolemic activity and hepatoprotective effect of liver and kidney that, there is valid pharmacological basis for employing them for this purpose.
• The Moringa oil made it recommendable for commercial use with the help of the physical and chemical characterizations of Moringa oleifera seeds oil which justify as a nondrying oil, in other words it is good for human.
• The data derived from nutrient characterization of Moringa oleifera are clear indications that, the plant leaves are rich nutrients and has potential to use as a feed additive with multiple purposes. These include serving as a protein, fatty acid, phenols, antioxidant and vitamin resources for human feed formulation.
• Histopathological examination was improved with increasing Moringa oleifera seeds oil or its leaves extract levels as compared with hypercholesterolemic group (positive control +ve).
• Results obtained suggest that Moringa oleifera seeds oil and/or its leaves extract had adequate lowering action of hypercholesterolemic.