Moringa Oleifera belongs to the family of Moringaceae included 14 species which growing fast in the tropics and subtropics areas is native to India, Africa, Asia Minor and sub-Himalayan tracts, drought tolerant, the height of the tree ranged from 5 to 10 m in three years. It is spread in North and South America, Cambodia, The pacific and Caribbean islands (Morton, 1991; Anwar and Bhanger, 2003; Anwar et al., 2005; Crosby, 2007; Oluduro, 2012 and Rockwood et al., 2013).
Moringa called as the miracle tree which contain 36 anti-inflammatories, more than 539 bio-chemical activities and 46 antioxidants natural components. Its leaves a rich source of high nutrition value such as various phenolics, essential minerals like calcium and potassium, vitamins like B,C and A amino acids and β-carotene, so it can used in modern science to prevent many diseases and cultivated in Malnutrition areas and remote countries for medical and nutrition values benefits (Debnath and Guha, 2007; Toba et al., 2010; Mishra et al., 2011; Oluduro, 2012 and Misra and Misra, 2014). Moringa leaves act as antimicrobial, antidiabetic, antioxidant, anti-atherosclerotic and hypertensive agents and anticancer, which a cure liver diseases, cardiovascular and hypoglycemic actions, asthma, malaria, eye and ear infections, headaches, skin diseases, hyperglycemia, diarrhea, scurvy, heart burn, bronchitis, syphilis, Strengthens the immune and digestion systems, hypocholesterolemic and regulate thyroid hormone, also enhance the shelf life of fat containing food to contain antioxidant compounds like phenolics, ascorbic acid and carotenoids (Dillard and German, 2000; Sreelatha and Padma, 2010; Mbikay, 2012 and Rockwood et al., 2013).
Moringa leaves is eaten raw as a salad green or combined with other vegetables and grains or cooked as spinach and other greens, introduced as a side dish with other food or as a nutrition main course, the dried powder from leaves added to soups, sauces or sprinkled on other food to raise the nutritional value, and is used the dried powder in supplement form or as a healthful drink like tea or mixed with cold or hot drinks ( Fahey, 2005 ). So, the objective of this work was to study the possibility of preparing a new beverages formula from Moringa Oleifera leaves rich in natural phenolic compounds and antioxidants in compare with other common beverages.
MATERIALS AND METHODS
Raw materials: Moringa Oleifera (M. Oleifera) trees have been cultivated to ensure the plants were healthy and uninfected, leaves was collected from Faculty of Agriculture farm, Mansoura University, El-Mansoura City, El-Dakahleia Governorate, Egypt.
Commercial beverages: Commercial Green tea, Peppermint, Anise and Cinnamon were purchased from local Market (Sun Mall), El-Mansoura City, El-Dakahleia Governorate, Egypt.
Chemicals: All chemicals were purchased from El-Gomhouria pharmaceutical company, El-Mansoura City, El-Dakahleia Governorate, Egypt.
Preparation of Moringa Oleifera leaves: Leaves of M.Oleifera were collected and washed under running tap water to eliminate dust and other foreign particles, then dried in the greenhouse (30- 45ºC) for one week then crushed to fine powder using domestic blender (BRAUN). Powdered was stored in polyethylene plastic bags at 5±1ºC until analysis.
Preparation of Moringa Oleifera beverages: Powder of M.Oleifera leaves was added to green tea, peppermint, anise and cinnamon as shown in Table (1), each packet contain 2 gm, then 200 ml hot water was added, finally prepared beverage were panel tested at Food Industries Dept., Fac. of Agric., Mans.University.
Proximate chemical analysis:
Moisture content, crude oil, crude protein, crude fiber and ash of M.Oleifera leaves was estimated according to (A.O.A.C. 2000) .
Identification and fractionation of phenolic compounds:
Phenolic compounds of dried M.Oleifera leaves was determined using HPLC at Food Tech. Res. Institute, Agric. Res. Center, El-Giza, Egypt, according to (Goupy et al., 1999).
Determination of total phenolic compounds (TPC):
The Folin-Ciocalteu method was used for determining of total phenolic compounds of dried M.Oleifera leaves and other combined beverages by using standardized spectrophotometric method at Food Tech. Res. Institute, Agric. Res. Center, El-Giza, Egypt according to (Ivanova et al., 2010 ).
Determination of radical scavenging activity (DPPH%):
2,2 diphenyll-picrylhydrazyl (DPPH%) assay of dried M.Oleifera leaves and beverages were carried out according to the method of ( Brand-Williams et al., 1995 ) at Food Tech. Res. Institute, Agric. Res. Center, El-Giza, Egypt.
Calcium, potassium and iron were determined using Sens AA ″GBC scientific equipment″ model ″Sens AA Dual″ made in Dandenong, Victoria, Australia at Atomic absorption, Micro-Analysis unit, Faculty of Science, Mansoura University, Egypt.
Data were statistically analyzed according to the technique of analysis variance (ANOVA), the least significant difference (L.S.D) and Duncan’s methods was used to compare the differences between the means of treatment values to the methods described by (Gomez and Gomez, 1984). All statistical analyses were performed using analysis of variance technique by means of Co STATE computer software.
RESULTS AND DISCUSSION
Proximate chemical composition of Moringa Oleifera leaves powder:
The proximate chemical composition of M.Oleifera leaves powder was determined, results in Table (2), revealed that M.Oleifera leave powder could be considered as a good source of crude protein, crude ash and fibers . From data presented in Table (2), it could be noticed that the moisture content of M.Oleifera leaves powder (MOLP) was 8.16%, the protein content reached to 21.40 %, which consider M.Oleifera leaves as a good and cheap source of protein supplement in countries suffering from malnutrition. These obtained results were lower than those obtained by (Ilyas et al., 2015 and Ismael et al., 2016) who mentioned that the crude protein of (MOLP) were 28.11 and 38.1 % respectively .
Also results presented in Table (2), showed that fiber content of (MOLP) was 7.46 %, this higher fibers content aids indigestion and prevention of many diseases ( Saldanha, 1995 ). These results was similar to those reported by ( Ismael et al., 2016 ) who found that the crude fibers of (MOLP) was 7.40% but was lower than those mentioned by ( Ilyas et al., 2015 ) who reported that the crude fibers of (MOLP) was 19.61 ± 0.38 % .
As shown in the same Table (2), it could be observed that the oil content of (MOLP) was 10.02 %. The ash content of (MOLP) was 5.33% which indicated the presence of suitable quantity of minerals in the leaves, these results was lower than those mentioned by ( Ilyas et al., 2015 and Ismael et al., 2016) who found that ash content of (MOLP) was 10.50 and 6.80 % respectively. So the differences in chemical composition would be attributed to the differences in the stage of maturity of the plants as well as the soil fortified with different chemical fertilizers and geographical location of the plants as reported by ( Ilyas et al., 2015 ) .
Identification and fractionation of phenolic compounds content (ppm) in Moringa Oleifera leaves powder:
Phenolic compounds are known as antioxidants which have long been recognized to have protective function oxidative damage in diet and may provide health benefits with reduced risk of chronic diseases (Karppinen et al., 2004). Therefore phenolic compounds were determined and identified in M.Oleifera leaves powder and results presented in Table (3), it can be observed that M.Oleifera leave powder contained 24 fractionated and identified phenolic compounds. The predominant phenolic compound being e-vanillic was (2022.38 ppm) followed by syringic acid (1011.28 ppm) and benzoic (939.99 ppm), while moderate amounts of pyrogallol, catechol, protocatchuic and caffeine recorded (515.68, 468.86, 289.70 and 111.11 ppm) respectively.
Also, an adequate amounts of cinnamic, 3,4,5- methoxy-cinnamic, 4-amino-benzoic, gallic, p-coumaric, alpha-coumaric, ferulic, Coumarin, iso-ferulic, chlorogenic, salycilic, caffeic, catechein, vanillic, p-oH-benzoic, epicatechein and ellagic were registered.
The presence of these compounds in M.Oleifera leaves powder can also modulate the lipid peroxidation involved in atherogenesis, coagulation and carcinogenesis in humans ( Siddhuraju and Becker, 2003 ).
Sensory evaluation of Moringa beverages:
Sensory evaluation considered as an important indicator of potential consumer preferences. In spite of its short comings, it will remain the most serious quality indicator. Results of sensory evaluation include appearance, taste, aroma, colour and overall-acceptability are presented in Table (4).
Data in Table (4), indicate that control beverage sample contained only M.Oleifera have an acceptable level up to 40.400 while the other beverage formula which containing (M.Oleifera : green tea) with the ratio of 1.25 : o.75 and the formula containing (M.Oleifera: anise) with the ratio of 1.25 : 0.75 nearly showed the same overallacceptability which recorded 40.025 and 39.700 respectively.
Also, from the same Table it can be observed that addition of (M.Oleifera : green tea) at the ratio of 1.75:0.25 enhanced the taste to be more accepted in compared with control one. Also results showed that non significant differences observed in aroma was found in the beverage formula M and G2. Results in the same Table showed that addition of (M.Oleifera : pepperiment) at the ratio of 1:1 and 1.75 : 0.25 could improve the taste in compared to control formula with score 7.550.
Also in the same Table (4), it could be noticed that formula contained (M.Oleifera : anise) at ratio of 1.25 : 0.75 enhanced taste to be more accepted in compared with control with score 7.550, and addition of the (M.Oleifera : anise) at ratio of 1.25 : 0.75 and 1.50 : 0.50 also improved colour in compare with control one .
Total phenolic compounds, antioxidant activity and some minerals content of prepared beverages with or without Moringa Oleifera:
Amount of phenolic compounds could be considered as a good preventative tool against many diseases. From data presented in Table (5), it can be noticed that The highest content of total phenolic compounds (TPC) was observed in G2 beverage formula which contained 1.25 Moringa : 0.75 green tea which recorded (31.5 mg/g) followed by Moringa beverage formula (17.17 mg/g). Results also in the same Table (5), indicated that prepared extract from Moringa beverage had the highest level of antioxidant activity (92.58%), these results were higher than those obtained by ( Ilyas et al., 2015 ) who reported that the antioxidant activity of Moringa oleifera leaf powder was up to (87.02%), while the G2 beverage which contained 1.25 Moringa : 0.75 green tea recorded (89.13%) followed by A2 beverage which contained 1.25 Moringa : 0.75 anise (87.72%) .
Minerals content of Moringa beverages, namely (Ca, K and Fe) is also presented in Table (5). Calcium (Ca) considered as essential element for transport of oxygen and cellular activity, for blood clotting, stabilizes blood pressure, contributes to normal brain function and bone health as reported by ( Antia et al., 2006 ). Our obtained results showed that calcium exhibited the highest amount of minerals content in Moringa beverage being (61.10 ppm) in compared with formula G2 beverage (32.26 ppm) and formula A2 beverage (45.61 ppm) . Potassium (K) is an essential nutritional element for transmission of nerve impulses and electrolyte balance. From data presented in Table (5), it can be noticed that the potassium (k) of Moringa beverage reached to (13.33 ppm), while in the formula A2 beverage 1.25 Moringa: 0.75 anise reached to (12.89 ppm) followed by G2 beverage 1.25 Moringa: 0.75 green tea (8.80 ppm). Finally iron (fe) play an important role in energy metabolism, gene regulation, cell growth, enzyme reaction and treatment and prevention of anemia. Deficiency of iron could resulted in decreased work capacity, depressed immune, increased rates of infection, increased lead and cadmium absorption and fetal growth retardation ( Antia et al., 2006 ). Obtained results in Table (5), indicate that the iron was higher in Moringa beverage (2.455 ppm) than G2 beverage (1.512 ppm) and A2 beverage (1.356 ppm). So, These obtained results indicated that Moringa Oleifera is a good source of some essential minerals for human health and could be recommended as daily used beverage .
So, from above mentioned data it could be observed that addition of Moringa Oleifera could particularly enhanced some sensorial properties of prepared beverages and could be accepted alone or combined with other commercial beverages .
Results of present study indicated that Moringa Oleifera leaves is a rich source of nutritional value, phenolic compounds and antioxidant activity. Beverages prepared from M.Oleifera leaves also showed agood minerals content, strong antioxidant properties and a rich source of phenolics. So, the study recommended M.Oleifera leaves as a daily beverage alone or combined with some other commercial beverages.
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