INTRODUCTION:

There is presence of Tannins, Phlobatannins, Saponins, Flavonoids, Steroids, Terpenoids, Cardiac glycosides, Anthraquinones and Alkaloids¹. It is also known as horse radish tree or drumstick tree or miracle tree. Moringa oleifera is the most widely cultivated species native to tropical and sub-tropical regions of the world. Many pharmacological studies have shown the ability of this plant to exhibit analgesic, anti-inflammatory, antipyretic, anticancer, antioxidant, nootropic, hepatoprotective, gastroprotective, anti-ulcer, cardiovascular, anti-obesity, antiepileptic, antiasthmatic, antidiabetic, anti-urolithiatic, diuretic, local anesthetic, anti-allergic, anthelmintic, wound healing, antimicrobial, immunomodulatory, and antidiarrheal properties^2. In fact, moringa is said to provide 7 times more vitamin C than oranges, 10 times more vitamin A than carrots, 17 times more calcium than milk, 9 times more protein than yogurt, 15 times more potassium than bananas and 25 times more iron than spinach³.

Moringa seeds can be used in cosmetics and are sources of biodiesel while the seed cakes can be used as a green manure or a fertilizer. The gum of Moringa oleifera has been used for dental cries and is astringent and rubefacient; the gum mixed with sesame oil is used to relieve headaches, fevers, intestinal complaints, dysentery, asthma, and some time used as an abortifacient, and to treat syphilis and rheumatism⁴.

MEDICINAL PROPERTIES:

1) Anticancer properties: Soluble and solvent extracts of leaves have been proven effective as anticancer agents. Furthermore, research papers suggest that the anti-proliferative effect of cancer may be due to its ability to induce reactive oxygen species in the cancer cells^5.

2) Diabetes: Diabetes leads to several complications such as retinopathy, nephropathy and atherosclerosis etc. Moringa can be used to prevent such ailments. When there is hyperglycemia, the blood glucose reacts with proteins and causes advanced glycated end products (AGEs)^6.

The aqueous extract of M. oleifera leaf protects pancreas against ROS-mediated damage by enhancing cellular antioxidant defenses and minimizing hyperglycemia in STZ- and HFD-induced diabetes might be due to the enhancement of glucose uptake in skeletal muscle, stimulating insulin secretion, inhibiting of alpha-amylase and alpha-glucosidase enzymes^.7.

Waterman et al also used moringa isothiocyanates MICs are the main anti-obesity and anti-diabetic bioactives of MC, and that they exert their effects by inhibiting rate-limiting steps in liver gluconeogenesis resulting in direct or indirect increase in insulin signaling and sensitivity^.8

3) Antioxidant: The flavonoids like quercetin and phenolics have been attributed as antioxidants that bring about a scavenging effect on ROS^.9

4) Antiepileptic activity: The ethanolic extract of Moringa concanensis leaves may produce its anticonvulsant effects via multiple mechanisms since it abolished the hind limb extension induced by MES as well as abolished seizures produced by PTZ^.10

5) Cardiovascular activity: Hypotensive activity of the ethanolic and aqueous extracts of Moringa oleifera whole pods and their parts, namely, coat, pulp, and seed was investigated^11.

6) Antiurolithiatic activity: The reduction of stone forming constituents in urine and their decreased kidney retention reduces the solubility product of crystallizing salts such as calcium oxalate and calcium phosphate, which could contribute to the antiurolithiatic property of root bark of M. oleifera¹²

7) Other diseases: Moringa can be used as a potent neuroprotectant. Moringa with its antioxidants can reduce the reactive oxygen species, thereby protecting the brain¹³.Multiple mechanisms of action, such as interfering with cell cycle progression, inducing apoptosis, promoting stabilization of microtubule, as well as several signal transduction pathways, were involved in their anticancer effects¹⁴.^.In an alternative and unconventional strategy to increase dietary VA intake, leaves from the Moringa oleifera (MO) plant were used as a VA supplement.^15.

Characterization:

1) Test for Tannins: This was done by boiling 1g of each of the dried powdered samples (separately) in 40ml of water in a test tube and then filtered. A brownish green or a blue-black coloration was observed after addition of a few drops of 0.1% ferric chloride. ii. Test for Phlobatannins An aqueous extract of the dry Moringa leaves was boiled with 1% aqueous hydrochloric acid. Appearance of red precipitate indicates the presence of phlobatannins.

2) Test for Saponins: To 10ml of distilled water, 1g of the powdered dry Moringa leaves (separately) was added and boiled in a water bath. The mixture was then filtered and to resultant 5ml of filtrate, 2-3ml of distilled water was added and shook vigorously for attainment of a stable persistent froth. Then, followed by a mixture of frothing with 1-2 drops of olive oil and shook vigorously, then observed in the formation of emulsions.

3) Test for Flavonoids: This was determined through heating of 0.5g of the dry powdered moringa leaves extract sample (separately) with ethyl acetate (10ml) over a steam water bath for 3 min. To 1ml of dilute ammonia solution, 4ml of filtrate from the filtered mixture mixture was added and shook. Appearance of yellow coloration is an indication of presence of flavonoids

Test for Steroids: This was carried out by addition of 4ml of acetic anhydride to 1g of each of the crude extract (separately) with further addition of H2S04(2ml). The presence of steroids was indicated by change of colour from violet to blue or greenzation:

4) Test for Terpenoids: This was carried out by Salkowski‟s test described by Parekh and Chands (2008), To 4ml of chloroform, 10ml of the crude extract was added, followed by the careful further addition of 5ml concentrated (H2SO4). Formation of the reddish brown coloration at the interface is an indication of a positive result for the presence of terpenoids.

5) Test for Cardiac Glycosides: The Keller-Killani test method was used for Cardiac Glycosides determination. To 2ml of glacial acetic acid containing one drop of ferric chloride (FeCl3) solution, 5ml of the plant extract was added, this was followed by addition of 1 ml concentrated Sulfuric acid. Brown ring was formed at the interface which indicated the presence of deoxy sugar of cardenolides. A violet ring may appear below the brown ring, though in the acetic acid layer, a greenish ring may also form just progressively throughout the layer.

6) Test for Anthroquinones: 5ml of each of the plant extracts was boiled with 10ml of sulfuric acid (H2SO4) and was filtered while hot. The filtrate was shook with 5 ml of chloroform. The chloroform layer was pipette into another test tube and 1ml of dilute ammonia was added. The resulting solution was observed for color changes

7) Test for Alkaloids: 5ml of the Moringa leaves extracts were added to 8ml of 1% HCl mixed, warmed and later filtered. Maeyer‟s and Dragendorff's reagents were added to the 2ml of the filtrate, then alkaloids absence or presence were determined based on the turbidity or precipitate development

Qualitative analysis of the phytochemicals^29,30:

i. Test for Tannins: This was done by boiling 1g of each of the dried powdered samples (separately) in 40ml of water in a test tube and then filtered. A brownish green or a blue-black coloration was observed after addition of a few drops of 0.1% ferric chloride.

ii. Test for Phlobatannins: An aqueous extract of the dry Moringa leaves was boiled with 1% aqueous hydrochloric acid. Appearance of red precipitate indicates the presence of phlobatannins.

iii. Test for Saponins: To 10ml of distilled water, 1 g of the powdered dry Moringa leaves (separately) was added and boiled in a water bath. The mixture was then filtered and to resultant 5ml of filtrate, 2-3ml of distilled water was added and shook vigorously for attainment of a stable persistent froth. Then, followed by mixture of frothing with 1-2 drops of olive oil and shook vigorously, then observed in the formation of emulsions.

iv. Test for Flavonoids: This was determined through heating of 0.5g of the dry powdered of moringa leaves extract sample (separately) with ethyl acetate (10ml) over a steam water bath for 3 min. To 1ml of dilute ammonia solution, 4ml of filtrate from the filtered mixture mixture was added and shook. Appearance of yellow coloration is an indication of presence of flavonoids.

v. Test for Steroids: This was carried out by addition of 4 ml of acetic anhydride to 1g of each of the crude extract (separately) with further addition of H2S04(2ml). The presence of steroids was indicated by change of colour from violet to blue or green.

vi. Test for Terpenoids: This was carried out by Salkowski‟s test described by Parekh and Chands (2008), To 4ml of chloroform, 10ml of the crude extract was added, followed by the careful further addition of 5ml concentrated (H2SO4). Formation of the reddish brown coloration at the interface is an indication of a positive result for the presence of terpenoids.

vii. Test for Cardiac: Glycosides The Keller-Killani test method described by Parekh and Chands (2008) was used for Cardiac Glycosides determination. To 2ml of glacial acetic acid containing one drop of ferric chloride (FeCl3) solution, 5ml of the plant extract was added, this was followed by addition of 1 ml concentrated Sulfuric acid. Brown ring was formed at the interface which indicated the presence of deoxy sugar of cardenolides. A violet ring may appear below the brown ring, though in the acetic acid layer, a greenish ring may also form just progressively throughout the layer.

viii. Test for Anthroquinones: 5ml of each of the plant extracts was boiled with 10ml of sulfuric acid (H2SO4) and was filtered while hot. The filtrate was shook with 5 ml of chloroform. The chloroform layer was pipette into another test tube and 1ml of dilute ammonia was added. The resulting solution was observed for color changes

ix. Test for Alkaloids: 5ml of the Moringa leaves extracts were added to 8ml of 1% HCl mixed, warmed and later filtered. Maeyer‟s and Dragendorff^„s reagents were added to the 2ml of the filtrate, then alkaloids absence or presence were determined based on the turbidity or precipitate development.

Quantitative Analysis of the Phytochemicals:

i. Estimation of Alkaloids: To a 500ml of beaker, 10g of the dry Moringa leaves and 400ml of 10% acetic acid in ethanol were added; the beaker was then covered and allowed to stand for 4 hours. This was then filtered, extracted and concentrated on a water bath to one-quarter of the original volume. This was preceded by dropwise addition of concentrated ammonium hydroxide to the extract until the precipitation is completed. The whole solution was allowed to settle and the precipitate was collected, washed with dilute ammonium hydroxide and then filtered; the residue is the alkaloid, which was dried and weighed to a constant mass. Formula = B - A × 100/S where, B = Weight of Whatman filter paper. A = Weight of Whatman filter paper, after drying. S = Sample weight^31.

ii. Estimation of Saponins: 50ml of 20% aqueous ethanol was added to 10g of the dry Moringa leaves in a conical flask. At about 55°C, for 4 hours with continuous stirring, the mixture was heated using a hot water bath after which the mixture was filtered and the residue re-extracted with a further 100ml of 20% ethanol. The combined extracts were reduced to 20ml over a water bath at about 90°C. The concentrate was transferred into a 100ml separatory funnel and 10ml of diethyl ether was also added and then shaken vigorously. The aqueous layer was recovered while the ether layer was then discarded. The purification process was repeated three times. 30ml of n-butanol was added. The combined n-butanol extracts were washed twice with 5m1 of 5% aqueous sodium chloride. The remaining solution was heated in a water bath. After evaporation, the samples were dried in the oven to a constant weight; the Saponin content was calculated as the percentage of the starting material. Formula = B – A × 100/S Where, B = Weight of Whatmann filter paper. A = Weight of Whatmann filter paper with sample. S = Sample weight^32.

iii. Estimation of Phenols: To 0.5ml of freshly prepared plant extracts in test tubes, 8ml of distilled water and 0.5 ml of Folin‟s Ciocalteau reagent were added to all the tubes. All the tubes were kept in the Biological Oxygen Demand chamber for 10 minutes at 40°C for incubation. This was followed by addition of 1ml of sodium carbonate solution to all the test tubes, subsequently; the tubes were incubated in the dark for an hour. The colour so developed was read spectophotometrically at 660nm. Standard curve was drawn using tannic acid as standard. Different concentrations of tannic acid were prepared and O.D was read at 660nm in a shimadzu UV-1650 spectrophotometer. The concentrations of sample were calculated based on the standard curve.³³

iv. Estimation of Total Flavonoids: 200ml of 80% aqueous methanol was used for recurrent extraction of 20g of the dry moringa leaves at room temperature. Whatman filter paper No 42 was then used for filtration of the whole solution. The filtrate was then transferred into a crucible and evaporated to dryness over a water bath; the dry content was weighed to a constant weight³⁴.

v. Estimation of Tannins: 100mg of tannic acid was dissolved in 100ml of distilled water. 5ml of stock solution was diluted to 100ml with distilled water. 1ml containing 50μg tannic acid. Extraction of Tannin: 0.5 gm of the powdered material was weighed and transferred to a 250ml conical flask and 75ml water was added. The flask was heat gently and boiled for 30 min centrifuge at 2,000rpm for 20 min and the supernatant was collected in 100ml volumetric flask and make up the volume. 1ml of the sample extract was transferred to 100 ml volumetric flask containing 75ml water. 5ml of folin denis reagent, 10ml of sodium carbonate solution were added and diluted to 100ml with water. Shake well. The absorbance was read at 700nm after 30 min. If absorbance is greater than 0.7 make a 1 + 4 dilution of the sample. A blank was prepared with water instead of the sample. A standard graph was prepared by using 100 mg tannic acid. The tannins content of the sample was calculated as tannic acid equivalents from the standard graph^.32,33,34.

COMMERCIAL APPLICATIONS:

1. Moringa seeds are used to extract oil called the Ben oil. This oil is rich in oleic acid, tocopherols and sterols. It can also withstand oxidative rancidity. The oil can be used in cooking as a substitute for olive oil, as perfumes and also for lubrication^35.

2. M. oleifera seed is a natural coagulant, containing a cationic protein that can clarify turbid water. This property of M. oleifera seeds is attracting much research as other coagulants such as alum, activated carbon and ferric chloride are expensive and rare.³⁶

3. M. oleifera functionalized with magnetic nanoparticles such as iron oxide were found beneficial in surface water treatment by lowering settling time.³⁷.

4. Moringa flowers are used to manufacture a tea that has hypocholesterolemic qualities. When fried, moringa blooms are said to taste like mushrooms^.38.

5. The growth hormone from the leaves, called Zeatin, is an excellent foliar and can increase the crop yield by 25%–30%^39.

6. It also used moringa as a fortificant and produced cream and butter crackers with moringa and Ipo-moea batatas as fortificants, with the hope of adding additional nutrients to snacks. The sensory evaluation proved the cream crackers to be widely accepted^40.

7. M. oleifera leaves can be incorporated in the diet of hens and layers thereby providing an excellent protein source, substituting other expensive ingredients such as soybean meal and ground nut cake.⁴¹⁴²

8. The moringa flowers are great sources of nectar and are used by beekeepers. The root bark has medicinal values and is used for dyspepsia, eye diseases and heart complaints.^43.

9. Moringa's tap root is used as a spice. The tree's gum can be used to create calico prints. Additionally, the gum and roots have anti-inflammatory, anti-bacterial, and antifungal effects^.44.

It is suggested that this addition can be made to other snacks as well after taking into account the opinions of various such fortifications. The addition of moringa to the snacks can increase their nutritional content. The majority of snacks are composed of cornmeal, and multiple studies have shown that adding a small amount of moringa to maize flour can increase the snack's nutritional value by adding protein, energy, and minerals.

CONCLUSION AND FUTURE PROSPECTS:

The key objective of this review was to unfold and explore the pharmacological and medicinal values of Moringa oleifera. The proper use of Moringa Tree solve a lot of problems arising from nutrition, health and general wellbeing of the masses Moringa oleifera is a multi-purpose herbal plant used as human food and an alternative for medicinal purposes worldwide. For instance, in this paper, it views the general nutrition constituents and its medicinal used of the specific part of the Moringa oleifera and also its medicinal properties and commercial applications furthermore it also aims to review the qualitative and quantitative analysis of the phytoconstituents.

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Received on 05.12.2022 Modified on 09.02.2023

Res. J. Pharmacognosy and Phytochem. 2023; 15(2):155-160.

DOI: 10.52711/0975-4385.2023.00024

Part of plant	Constituents	Use
Root	Tannins alkaloids sterols saponins minerals like magnesium and sodium	Antidiabetic ani-atherosclerotic antioxidant neuroprotective ¹⁶
Bark	Saponins flavonoids Alkaloids like morphine, moriginine, minerals like calcium,	The alkaloid helps the bark to be antiulcer, a cardiac stimulant and helps to relax the muscles The alkaloid helps the bark to be antiulcer, a cardiac stimulant and helps to relax the muscles ¹⁷
Flower	It contains calcium, potassium and amino acids.	The alkaloid helps the bark to be antiulcer, a cardiac stimulant and helps to relax the muscle The presence of nectar makes them viable for use by beekeeper^{. 18, 19}
Pods	Lipids, non-structural carbohydrates,	The presence of PUFA in the pods can be used in the diet of obese^{. 20}
Leaves	Phytochemicals like tannins, sterols, saponins, terpenoids, phenolics, alkaloids and flavonoids	Anticancer agents.The presence of flavanoids gives leaves the antidiabetic and antioxidant properties. The isothiocyanates are anticancer agents. Flavanoids like quercetin and others are known for anti-proliferative, anticancer agent. The presence of minerals and vitamins help in boosting the immune system and cure a myriad of diseases ^{21, 22, 23, 24}
Seeds	Phytochemicals like tannins, saponin, phenolics, phytate, flavonoids, terpenoids and lectins	Flavanoids are what give it its anti-inflammatory properties. Antimicrobial activities are brought on by the antibiotic pterygospermin. The additional phytochemicals aid in the treatment of many diseases^{. 25, 26, 27, 28}