ASSESSMENT OF THE LIPID COMPOSITION AND ANTIOXIDANT ACTIVITIES OF TWO URHOBO SOU PS: EMUEGARI ISHAWO (EGUSI AND

OKRO), OGHWO-EVWRI(PALM OIL) (HIGH DENSITY LIPOPR OTEIN, TRIGLYCERIDES) and (NITRIC OXIDE, TOTAL PHENOL) By ICHIPI-IFUKOR, Patrick Chukwuyenum (AISLT) PATYCHYKYS RESEARCH CONSULTANCY NO. 16, COLLEGE ROAD ABRAKA. C/O P.O Box 85 Abraka, Delta State, Nigeria. E-mail: patychyky@yahoo.com Phone: +2347038268448 ABSTRACT The study investigated the assessment of the lipid composition and antioxidant a ctivities of two urhobo soups emuegari ishawo (egusi and okro), oghwo-evwri (pal m oil) (High density lipoprotein (HDL), Triglycerides) and (Nitric Oxide, Total Phenol). It employed the conventional methods of colorimetric and Singleton and Rossi, 1965. For the assay of lipid composition and antioxidant activities respe ctively. From the results, it was discovered that there was a significant differ ence on the total phenol content of both soups (P<0.05). However, there was no s ignificant difference in the nitric oxide scavenging activities, and triglycerid e levels of both soups. (P>0.05). The research concludes that the two Urhobo sou ps have a high level of antioxidant (total phenol and Nitric oxide) activities a nd lipid content with Emugari-ishawo having the highest of all parameters analyz ed. Thus it is a very rich food that may have the potency for the cure and regul ation of some lipid and antioxidant risk factor diseases. INTRODUCTION Soups are generally warm food that is made by combining ingredients such as meat and vegetables with stock, juice, water, or another liquid. Hot soups are addit ionally characterized by boiling solid ingredients in liquids in a pot until the flavors are extracted, forming a broth. Traditionally, soups are classified int o two main groups: clear soups and thick soups. Soups are similar to stews, and in some cases there may not be a clear distinction between the two; however, sou ps generally have more liquid than stews. (Goltz, 2008). The particular metabolic fate of a dietary fatty acid is determined by its struc tural characteristics which include carbon chain length (short, medium, long, or very long chain), number of double bonds or degree of saturation (saturated, mo nounsaturated, or polyunsaturated), placement of double bonds relative to the om ega carbon (omega 3, 6, or 9), and configuration of hydrogens around the double bonds (cis or trans). (Balch, 2006). Fatty acids may be oxidized for energy, inc orporated into cell membranes, utilized for synthesis of biologically active com pounds, or deposited into adipose tissue to provide an energy reserve. The essen tial fatty acids are long chain (18 carbons), polyunsaturated, with omega-6 and omega-3 double bonds, and with hydrogen around the double bonds in the cis confi guration. (Maton, et al., 1993). Polyunsaturated fatty acids are utilized as substrates for synthesis of biologic ally active compounds such as steroid hormones, prostaglandins, and leukotrienes (Ozias, et al., 2007). Saturated fat is preferentially incorporated into adipos e tissue stores because the absence of double bonds allows a higher energy yield per carbon than is obtained from oxidation of unsaturated fatty acids. Monounsa turated fatty acids are either oxidized for energy or stored as fat depending up on the demand for energy (Willett, 2007). The longer chain fatty acids are incor porated into cell membranes as phospholipids regardless of degree of saturation. However, highly excitable membranes such as in brain and nervous tissue are ric her in polyunsaturated fatty acids with 20 or more carbons and 4 or more double bonds because the bending in these fatty acids around the cis double bonds allow

s for greater membrane fluidity. Dietary fat provides an average energy intake o f 9 kcal/gram which is twice that of carbohydrate or protein at 4 kcal/gram, A m inimum amount of dietary fat is necessary to facilitate absorption of fat-solubl e vitamins (A, D, E and K) and carotenoids. A minimal amount of body fat is also necessary to provide insulation that prevents heat loss and protects vital orga ns from shock due to ordinary activities (Ordman, 2011). Antioxidants on the other hand, are a molecule capable of inhibiting the oxidati on of other molecules. Oxidation is a chemical reaction that transfers electrons or hydrogen from a substance to an oxidizing agent. Oxidation reactions can pro duce free radicals. In turn, these radicals can start chain reactions. When the chain reaction occurs in a cell, it can cause damage or death to the cell. Antio xidants terminate these chain reactions by removing free radical intermediates, and inhibit other oxidation reactions. They do this by being oxidized themselves , so antioxidants are often reducing agents such as thiols, ascorbic acid, or po lyphones.(Sies, 1997). Although oxidation reactions are crucial for life, they c an also be damaging; plants and animals maintain complex systems of multiple typ es of antioxidants, such as glutathione, vitamin C, and vitamin E as well as enz ymes such as catalase, superoxide dismutase and various peroxidases. Low levels of antioxidants, or inhibition of the antioxidant enzymes, cause oxidative stres s and may damage or kill cells. As oxidative stress appears to be an important p art of many human diseases, the use of antioxidants in pharmacology is intensive ly studied, particularly as treatments for stroke and neurodegenerative diseases . Antioxidants however, are widely used as ingredients in dietary supplements an d have been investigated for the prevention of diseases such as cancer, coronary heart disease and even altitude sickness. Although initial studies suggested th at antioxidant supplements might promote health, later large clinical trials did not detect any benefit and suggested instead that excess supplementation is har mful (Benzie, 2003). In addition to these uses of natural antioxidants in medici ne, these compounds have many industrial uses, such as preservatives in food and cosmetics and preventing the degradation of rubber and gasoline. The present st udy was carried out to assess the lipid composition and antioxidant activities o f two urhobo soups; Emugari-Ishavwo and Oghwo-evwri. MATERIALS AND METHODS EXPERIMENTAL DESIGN The design of the experiment followed a simple process of random selection and s ampling of three pots of soups following the same procedures as described below. PREPARATION OF SOUPS PREPARATION OF OGWO-EVWRI INGREDIENTS USED The following ingredients were purchased from the local market in Abraka Ethiope East LGA of Delta-state. Red oil (Native oil) Starch Native Salt Pottash Meat/Fish/Cray-fish. Pepper. Ogiri(Native magi) PROCEDURE Water was added to a pot and allowed to boil, on boiling the native red oil was added and observed for 5 minutes. After which, the boiled meat and fish was added before adding a little quantity of starch. At the end of this, the fis h and Cray fish was added simultaneously before adding the native potash/salt an d allowed to simmer. Pepper and magi was added to taste and left for another fiv e minutes before it was ready. PREPARATION OF EMUGARI-ISHAWO INGREDIENTS USED The following ingredients were purchased from the local market in Abraka Ethiope East LGA of Delta-state. Okro

Melon (Egusi) Salt Pepper Tomatoes Onion Maggi Meat/Fish/Cray-fish. PROCEDURE The tomatoes, onion, and pepper were blended to a smooth paste using an electron ic blender. After which the melon and Cray fish was blended into powder form usi ng a local grinding stone, while the okro was grated using a hand grater. Into a medium sized pot, water was put and allowed to boil, after which the meat and f ish was added and allowed to cook. Once this was ascertained, the vegetable, sal t, and the blended mixture of pepper, tomatoes and onion paste was added and lef t for 5 minutes. To the mixture was added the melon and mixed properly for anot her two minutes before adding the Okro and allowed to simmer for 2 minutes. Magi was added to taste and the soup was ascertained ready. PREPARATION OF SOUPS FOR ANTIOXIDANT AND LIPID ASSAY The various soup samples were labeled and shared into two equal parts respective ly. The first portion was homogenized using a mortar and pestle until it was smo oth all over. The second part was labeled and stored at the refrigerator for fur ther usage. The homogenate collected was centrifuged at 3000g until a clear supe rnatant was gotten. The supernatant was however, collected and stored in the ref rigerator until it was required for the assay. Assay for the various lipid compo sitions and antioxidant activities are described below. ASSAY FOR LIPID COMPOSITION Assay for the various lipid compositions are described below. ASSAY FOR HDL The various test-tubes were washed, dried and labeled blank, standard an d test. PROCEDURE To all the test-tubes, 1ml of cholesterol was added, and to the blank, 1 0mm of water and to the tubes labeled standard and tests respectively was added 10mm of the standard reagent and sample respectively, mixed properly and allowed to stand for 15 minutes at 370c. after this the absorbance was read with a spec trophotometer against the reagent Blank. ASSAY FOR TRIGLYCERIDE PROCEDURE This was determined by colorimetric method. The triglycerides are determ ined after enzymatic hydrolysis with lipases. The indicator is a ghinonemine for med from hydrogen-peroxide, 4 aminophenazone and 4 chlorophenol under catalytic influence of peroxides. PRINCIPLE: Triglyceride + H2o Glycerol+Fatty acid Glycerol-3-phosphate + O2 Dihyroxyaetone + Pi + H2o2 2H2o2 + 4-aminophenazone + 4 Chlorophenol quinoneamine + Hcl +4H2o PROCEDURE Four test-tubes were labeled blank, 1, 2 and 3. To each of them, 1ml of trigs wa s measured and to the test-tubes labeled 1,2,3, 10ml of the soup supernatants we re added and incubated for five minutes at 37oC. After which the absorbance was read with a spectrophotometer at 500nm. ASSAY FOR ANTIOXIDANT COMPOSITION ASSAY FOR NITRIC OXIDE 10-400mg of MESB was added in the test-tubes to which 1ml of sodium nitr oprusside solution (25mM) and incubated for 370C for 2hours. An aliquot (0.5ml) of the incubated solution was removed and diluted with 0.3ml of Griess reagent ( 1% sulphanilamide in 5% H3Po4 0.1% naphthylethylenediamne dihydrochloride). The absorbance of the chromophore formed was immediately read at 570nm against reage nt blank with ascorbic acid serving as standard at 500ng. Results were expresse

d as percentage radical scavenging activity (RSA). ASSAY FOR TOTAL PHENOL This was carried out as described by Singleton and Rossi, 1965. Na2CO3 s alt was added to 0.5ml of the soup sample, after 3minutes 0.5ml of saturated NaC O3 solution was added and solution was made up to 5ml with distilled water. The reaction mixture was kept in the dark for 90mins and the absorbance was read at 725nm against the blank. STATISTICS AND DATA ANALYSIS All values obtained were analyzed using descriptive statistics and the p aired t-test in the data tool pack of Microsoft excel 2007. Level of significanc e was determined at P<0.05 RESULTS From the result, there was a significant difference on the total phenol content of both soups (P<0.05). However, there was no significant difference in the nitr ic oxide scavenging activities, and triglyceride levels of both soups. (P>0.05) the results of the lipid and anti-oxidant contents of both soups are presented i n tables 1 and 2 below. Table 1 lipid content and antioxidant content of two urhobo soups. Parameter Emuegari-ishawo Oghwo-evwri Total Phenol 14.87 0.410** 11.86 0.304** Triglyceride 6.67 0.29 6.25 0.35 High density Lipoprotein 8.1 0.042 6.94 0.297 All values are expressed as Mean SD in mg/ml, all values followed by ** differ s ignificantly. That is P > 0.05 Table 2 Showing the Nitric Oxide radical scavenging activities(RSA) of the soups . Concentration 50mg/ml 100mg/ml 150mg/ml Soups Emuegari-ishavwo 95.777 2.828 95.4421.417 17.6664.242 Ogwho-evwri 67.6660.157 40.4444.399 35.2771.828 All values are expressed in %age RSA as Mean SD.

DISCUSSION From the result presented above, it was discovered that there was a sign ificant difference in the level of total phenol in both soups (P<0.05) with Emug ari-ishawo having the highest mean value of 14.87mg/ml. The triglyceride and HDL level however had no significant difference (P>0.05) but Emugari-ishawwo had th e highest mean values of 6.67mg/ml and 8.81mg/ml respectively for triglyceride a nd HDL. In another development, the radical scavenging activity of nitric oxide in the soups also showed a level of significant difference (P<0.05) with 95.77mg /ml of Nitric oxide in the 50mg/ml category. It is worthy of note that this diff erence is concentration dependent. Previous studies on the nutritional evaluation of foods such as the work of Craig, et al., 2009 that analyzed the nutrient content and health implicatio ns of okinawan diets showed that food rich in antioxidants have great potentials for the reduction and control of various diseases such as diabetes, ulcer and n eurodegenerative diseases. This study thus gave credence to the work of Janeway, 2005 who submitted that the consumption of antioxidant rich diets is a major wa y of managing neurodegenerative and reactive oxygen implicated diseases. Foods high in lipid sources have also been noted for their high level contribution to energy while a high level intake could also raise some risk factors such atheros clerosis (Kwiterovich, 2000). Particularly for triglycerides, while those of HDL are more of a protective function. Atherosclerosis is a chronic disease that re mains asymptomatic for decades leading to the depositions of fatty substances on and fibrosis of the inner lining of the arteries. Although the role is not ful

ly understood, high triglycerides play a part in the development of this disease that involves the hardening of the arterial walls and the buildup of plaque. At herosclerosis increases the risk of high blood pressure, heart disease and strok e. CONCLUSION In conclusion, the two Urhobo soups have a high level of antioxidant (to tal phenol and Nitric oxide) activities and lipid content with Emugari-ishawo ha ving the highest of all parameters analyzed. Thus it is a very rich food that ma y have the potency for the cure and regulation of some lipid and antioxidant ris k factor diseases.

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