International Journal of Food Properties

ISSN: (Print) (Online) Journal homepage: www.tandfonline.com/journals/ljfp20

Nutritional and phytochemical screening of

Moringa oleifera leaf powder in aqueous and
ethanol extract

Shakeela Khalid, Muhammad Arshad, Shahid Mahmood, Waleed Ahmed,

Farzana Siddique, Waseem Khalid, Mehwish Zarlasht, Turky Omar Asar &
Faten A. M. Hassan

To cite this article: Shakeela Khalid, Muhammad Arshad, Shahid Mahmood, Waleed Ahmed,
Farzana Siddique, Waseem Khalid, Mehwish Zarlasht, Turky Omar Asar & Faten A. M.
Hassan (2023) Nutritional and phytochemical screening of Moringa oleifera leaf powder in
aqueous and ethanol extract, International Journal of Food Properties, 26:1, 2338-2348, DOI:
10.1080/10942912.2023.2246685

To link to this article: https://doi.org/10.1080/10942912.2023.2246685

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Muhammad Arshad, Shahid Mahmood,
Waleed Ahmed, Farzana Siddique, Waseem
Khalid, Mehwish Zarlasht, Turky Omar Asar
and Faten A. M. Hassan

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INTERNATIONAL JOURNAL OF FOOD PROPERTIES
2023, VOL. 26, NO. 1, 2338–2348
https://doi.org/10.1080/10942912.2023.2246685

Nutritional and phytochemical screening of Moringa oleifera leaf powder in

aqueous and ethanol extract
Shakeela Khalida, Muhammad Arshada, Shahid Mahmoodb, Waleed Ahmedc,
Farzana Siddiqueb, Waseem Khalidd, Mehwish Zarlashte, Turky Omar Asarf,
and Faten A. M. Hassang
a
Department of zoology, University of Sargodha, Sargodha, Pakistan; bInstitute of Food Science and Nutrition,
University of Sargodha, Sargodha, Pakistan; cNational Institute of Food Science and Technology, University of
Agriculture Faisalabad,Faisalabad, Pakistan; dUniversity Institute of Food Science and Technology, The University of
Lahore, Lahore, Pakistan; eInstitute of Food Science and Nutrition, Gomal University Dera Ismail Khan, Pakistan;
f
Department of Biology, College of Science and Arts at Alkamil, University of Jeddah, Jeddah, Saudi Arabia; gFaculty of
Science, Department of Microbiology, Taiz University, Taiz, Yemen

ARTICLE HISTORY
Received 20 June 2023
Revised 2 August 2023
Accepted 4 August 2023
KEYWORDS
Moringa oleifera;
phytochemicals; bioactive
compounds and oxidative
stress

Introduction
Moringa oleifera due to its abundant vitamins, minerals, and protein content is considered as
a promising food plant, promoting several health benefits. M. oleifera is enriched in multiple
therapeutically active chemicals: a good applicant supplemented with effect enhancing and neutraliz-
ing side-effects combinations. It belongs to the genus Moringa, family Moringaceae, and has 14
species, also called miracle tree, nature’s gold, and sohanjna.[1] The nutrient profile of M. oleifera
mostly depends on the soil composition, climatic characteristics, cultivating conditions, processing,
and storage quality.[1] It is often recommended as a healthy plant to cure malnutrition; its leaves and
other products are promoted as nutritional supplements for its strong vitamin profile.[2] M. oleifera is
extensively consumed as food, medicines, cosmetic oil, and feed for livestock. It has been used as
a medicinal plant to provide nutrients, promote health, decrease stress, and enhance food efficiency.
Its leaves are especially beneficial in addressing chronic inflammations, including hypertension,
diabetes, insulin resistance, cancer, and hypercholesterolemia.[3]

CONTACT Faten A. M. Hassan [email protected] Faculty of Science, Department of Microbiology, Taiz University,
Taiz, Yemen; Waseem Khalid [email protected] University Institute of Food Science and Technology, The
University of Lahore, Pakistan; Muhammad Arshad [email protected] Department of Zoology, University of
Sargodha, Pakistan
© 2023 Shakeela Khalid, Muhammad Arshad, Shahid Mahmood, Waleed Ahmed, Farzana Siddique, Waseem Khalid, Mehwish Zarlasht, Turky Omar Asar
and Faten A. M. Hassan Published with license by Taylor & Francis Group, LLC.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/),
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The terms on which this
article has been published allow the posting of the Accepted Manuscript in a repository by the author(s) or with their consent.
 INTERNATIONAL JOURNAL OF FOOD PROPERTIES 2339

M. oleifera leaves are an appreciable source of alkaloids, carotenoids, saponins, isothiocya-

nates, phenolic acids, glucosinolates, and flavonoids. They present more quantity of these
compounds as compared to fruits, vegetables, and other food plants consumed as human
nutrition. This high bioactive profile is famous to boost immune systems and defeat reactive
oxygen species. They may also be used as additives on animal feed, an alternative to supple-
ments, and a remedy to many inflammations due to its relative low cost.[4] M. oleifera leaves
are enriched with tannins, the water soluble phenolic compounds that can bind to gelatin,
proteins, and alkaloids, and have antibacterial and anti-inflammatory activity.[4] Saponins are
anticancerous in nature, ranges between 64-81 g/kg of dry weight, lower the plasma choles-
terol, and decrease enterohepatic circulation of bile acids by increasing their fecal excretion.[5]
Biological activites of M. oleifera leaves and phenolic compounds depend upon the solvent
method used for the extraction of these bioactive compounds.[6] The flavonoid composition of
Moringa leaves differs significantly when collected from different varieties. The concentration of
flavonoids is dependent upon growing environment, treatment method, leaf maturity, and cultivar.
A significant positive interaction (ƿ < 0.001) is found between extraction temperature and ethanol
concentration. Total phenols decrease with the rise of temperature.[7] Polyphenols are the most
vital antioxidants present in human diet. Several researches have shown a close relationship
between many diseases such as diabetes, cardiovascular, neurodegenerative disorders, and con-
sumption of polyphenol-rich diet.[8] Moringa is a highly nutritious medicinal plant that has the
ability to cure infertility, obesity, hypertension, hyperlipidemia, bacterial infections, diabetes,
inflammation, and oxidative stress. Preventive medication has been greatly improved by the use
of natural plant. These plants contain free radical scavenging molecules such as phenols, alkaloids,
flavonoids, amines, and turpines with a great tendency of antioxidant activity.[9] Previous studies
reported that these polyphenols and antioxidants further minimizes the damages reported by
many tissues during physiological process.[10]
M. oleifera tree has an unusual variety of valuable properties saddle in nutrition, medicines, and for
many other industrial purposes.[11] M. oleifera has gained popularity in recent times due to its
phytochemical, antioxidant profile and pharmacological properties. Additionally, the enriched phy-
tochemical and antioxidant components of M. oleifera leaves also attributed to show anti-apoptotic
and anti-inflammatory effects.[12] Moringa oleifera is a chief source of minerals, proteins, folates,
phenols, and unsaturated fats, considered as a superb extract with valuable properties and hence can
be integrated into the diet as a fortifier of any kind of food.[13]
Moringa oleifera is recounted to improve a wide series of biological activities including anti-
inflammatory, antioxidant, neuroprotective, hepatoprotective, anticancer, and antidiabetic.
M. oleifera has been broadly preferred as a healthy food supplement due to its strong protection
against numerous diseases and environmental toxins.[14] The high phytochemical profile of M. oleifera
supports the usage of food plant as nutraceutical and pharmaceutical purpose to mitigate inflamma-
tory and oxidative stress in human beings at a low cost and easy access. The present research
emphasizes the health benefits and bioavailability of Moringa oleifera leaf powder and extract to
treat oxidative stress-related diseases. In the present study, we planned to study the proximate and
mineral composition of Moringa oleifera powder; the qualitative phytochemical grading in M. oleifera
leaf powder and extract; and free radical scavenging activity and total antioxidant capacity of aqueous
and 70% ethanol extract of Moringa oleifera leaf.

Materials and methods

The M. oleifera leaves were collected from the lawn of botanical garden, University of Sargodha. The
leaves were washed with fresh water, and the inedible parts were removed.[15] After that, the experi-
mental work was performed at the soil and water testing laboratory, Institute of Food Science and
Nutrition, University of Sargodha, Pakistan.
2340 S. KHALID ET AL.

Aqueous and ethanol (70%) extract formation

M. oleifera leaf extracts (MoLE) aqueous and ethanol 70% were prepared according to the process
discussed in detail in the previous manuscript.[15,16]

Determination of yield %
The Yield % of MoLE was calculated by following formula:
wt. of extract(g)
Yield (%) = × 100
wt. of simple(g)

Proximate analyses of Moringa oleifera leaf powder (MoLP)

The proximate analysis of MoLP was done to find the moisture content (%), crude proteins, crude fats,
crude fiber, ash carbohydrates, and nitrogen free extract (NFE). The proximate analysis was performed
by following the procedure of Association of Official Analytic Chemist.[17]

Estimation of Minerals of Moringa oleifera Leaf Powder

The analyses of MoLP for N, P, K, Ca, Mg, Zn, Cu, Mn, Cd, Fe, Ni, and Pb were performed by
following the methods of digestion, filtration, dilution, and reading according to the procedure of
Association of Official Analytic Chemist.[17] N by Kjeldahl’s method (Fahey, 2005), P by Colorimeter,
Phospho-Molybdate Vonadate, K by Flame Photometer, and Ca, Mg, Cd, Ni, Pb, Zn, Cu, Fe, and Mn
by Atomic Absorption Spectrophotometer.[18]

Phytochemical screening of Moringa oleifera leaf extract

Phytochemical examinations of flavonoid, phenols, alkaloid, tannins, saponins, glycosides, steroids,
triterpenoids, phytosterols, reduced sugars, carbohydrates, proteins, amino acids, fats, and oil present
in the MoLE were carried out as per standard tests.[19,20]

Presence of flavonoids. Flavonoids were detected by Ferric Chloride test. The formation of dark
brown or blackish red colored precipitates depicted flavonoids.

Presence of phenols. Phenols were detected by Folin-Ciocalteu test. The formation of gray or blackish
colored precipitates showed phenols.

Presence of alkaloids. Wagner’s test was carried out to confirm alkaloids in M. oleifera leaf extract.
Reddish brown precipitates developed when 2 ml extract was mixed thoroughly in 1% HCl solution
with 0.5 ml of Wagner’s reagent.

Presence of saponins. Extract (0.5 g) was taken in 2 ml water and continuously shaken; foaming
occurred for 10 min, which indicated the saponins’ concentration.

Presence of tannins. Add 1% gelatin reagent containing few drops of NaCl in 1 ml of extract; white
precipitates appear to confirm tannins.

Presence of glycosides. MoLE and dilute HCl were mixed and legal’s reagent was used to dissolve
sodium nitropruside in pyridine and NaOH. Glycosides were confirmed due to the appearance of
blood-red color.
 INTERNATIONAL JOURNAL OF FOOD PROPERTIES 2341

Presence of steroids. Steroids were detected by Salkowski’s test in which MoLE was dissolved in
chloroform and filtered, mixed with concentrated H2SO4. Brown color appeared to indicate the
presence of steroids.

Presence of protein and amino acids. Protein was confirmed by following Xanthoproteic test. MoLE
was treated with few drops of concentrated HNO3, which resulted in the formation of yellow colored
precipitates. Amino acids were indicated by Ninhydrin test. Extract was mixed with 0.25% Ninhydrin
reagent and boiled for few minutes; blue colored precipitates appeared.

Detection of fats and oils. Extracts were pressed under the folds of filter paper and results were
observed.

Detection of reducing sugars. Brick red colored precipitates appeared to show the availability of
reducing sugars in MoLE when 1 ml Fehling’s reagent was added in 2 ml of each extract.

Detection of carbohydrates. The aqueous solution and 70% ethanol MoLE was treated with
Molicsh’s reagent. After dissolving in 5 ml distilled water, a purple ring appeared. Now mix
few drops of alcoholic α naphthol solution in a test tube to show the presence of
carbohydrates.

Presence of phytosterols. Liebermann-Burchard’s test was effective to indicate phytosterols in MoLE.

Add Chloroform in MoLE and filter. A few drops of acetic anhydride was mixed, boiled, and cooled
rapidly. Finally, H2SO4 was mixedtill brownish ring appeared.

Detection of triterpenoids. Triterpenoids were detected by Salkowski’s test in which MoLE was
dissolved in chloroform. H2SO4 (few drops) was added after filteration and mixed thoroughly.
Golden color appeared to indicate the presence of triterpenoids.

Bioactive compounds
The bioactive compounds were determined from MoLE by using the different methods. For measur-
ing the bioactive compounds in MoL extract, different tests were performed including total poly-
phenols, total flavonoids, total saponins, total tannins, DPPH, FRAP, and ABTS.

Total polyphenols. TPC (Total polyphenols content) of MoLE was estimated by Folin-Ciocalteu
methodology as described by.[11,21] About 3 ml distilled water and 50 µl MoLE were mixed in test
tube and put on vortex for 15 min. Then, 750 µl of Na2CO3 and 950 µl of distilled water was added,
followed by incubation for 30 min at 25°C. UV-visible spectrophotometer was used to measure
absorbance at 765 nm. Total polyphenols content was represented as mg Gallic acid equivalent per
100 ml.[11]

Total flavonoids. TFC (Total flavonoid content) was calculated by following AlCl3 method as
described by.[11,21] About 1 ml MoLE and 4 ml distilled water were filled in test tube and added to 5%
NaNO2 (300 µl), put aside for 15 min, then a methanol solution of 10% AlCl3 (300 µl) by vortex was
added. 1 M NaOH (2 ml) was mixed with distilled water to make 10 ml of total volume. UV- visible
spectrophotometer was used to measure absorbance at 415 nm. TFC was represented as mg quercetin
equivalent per 100 ml.[11]

Total saponins. TSC (Total saponins content) was found as described by.[11,22] A sample of MoLE
(250 µl) and 8% vanillin reagent (250 µl) were mixed with 72% H2SO4 (2.5 ml) for 10 min, and
absorbance at 544 nm was measured by UV-visible spectrophotometer. Total saponin content was
represented as mg diosgenin equivalent per 100 ml.[11]
2342 S. KHALID ET AL.

Total tannins. TSC (Total tannins content) was calculated by following the method as described
by.[11,23] A sample of MoLE (0.5 ml), 4% vanillin reagent (3 ml) and 36% HCl (1.5 ml) were mixed
thoroughly and kept for 15 min in darkness at room temperature. Absorbance at 500 nm was
calculated by UV-visible spectrophotometer. Total tannins content was represented as mg catechin
equivalent per 100 ml.[11]

Radical scavenging assay

DPPH (1,1-diphenyl-2-picryl-hydrazyl) assay and ABTS (2,2-azinobis-3-ethylbennzothiazoline-6-sul-
phonnnic acid) assays were conducted to assess the antioxidant activity of MoLE by the method
of.[11,23] The absorbance was noted at 515 nm for decline in DPPH and 734 nm for ABTS. The
graduation curve for DPPH and ABTS was set by following Trolox (standard); data were represented
as mM Trolox equivalents/100 ml.[11]

Ferric reducing antioxidant power assay

Ferric Reducing Antioxidant Power (FRAP) was assayed to calculate the antioxidant activity of MoLE
by the method of.[24] .About 1 ml extract and varying amounts of FeSO4 were taken, 6 ml of FRAP
reagent was added, and kept for 30 min in a water bath at 37°C. The absorbance was determined at 593
nm, and a decline in values was found.[25]

Data analysis and statistical application

All results were statistically analyzed by applying One-way analysis of variance (ANOVA),and data
were organized as mean ± SEM. Statistical analysis was performed by using MiniTab software 18.1.

Results and discussion

Weight of MoLP. Weight of Moringa oleifera Leaves (MoL) fresh, dried, and powder form were
measured and a considerable decrease in the weight was found.

Extraction rates of MoLP. MoL were dried and powdered @ 11.50%. The extraction rate of MoLP in
distilled water and 70% ethanol was 13.23, and 12.65%, respectively.

Proximate analysis of Moringa oleifera leaf powder. Proximate composition of MoLP is showed in
Table 1. The moisture, crude protein, fat, fiber, ash, and NFE of M. oleifera leaves powder were 6.30,
26.23, 2.82, 10.25, 8.24, and 46.14, respectively. The moisture content of current study was similar to
the previous study.[26] Another study showed the similar results of moisture to our outcomes.[27]
Research data suggested that the low content of moisture in MoLP that can reduce the microbial
activity and enhance storage value. However, the discrepancies are augmented by processing and
different storage methods.[28]
The protein contents of MoLP as fall in the same range as of the previous study (22.99–29.36.[27]
Fats content of MoLP is less than form from different plant leaves of M. oleifera.[26,27] However, the fat
value of the present research is also parallel to the Kenaf leaves.[29] High proteins and fats highlighted
the energy value of MoLP and its’ efficacy as a main source of energy for human body.[28]

Table 1. Physicochemical composition (%) of

MoLP.
Parameter Results
Moisture 6.30 ± 0.15
Crude Protein 26.23 ± 0.41
Fat 2.82 ± 0.02
Fiber 10.25 ± 0.02
Ash 8.24 ± 0.01
NFE (Carbohydrates) 46.14 ± 0.26
Data is presented in means ± SEM
 INTERNATIONAL JOURNAL OF FOOD PROPERTIES 2343

Table 2. Mineral composition of MoLP.

Minerals Ppm Minerals ppm
Zn 25 ± 2.0 P .2 ± .1
Cu 8.0 ± 1.0 K 2.2± .2
Fe 410 ± 5.0 Ca 2.5 ± .2
Mn 80 ± 3.0 Mg .6 ± .1
Ni 4.0 ± 1.0
Cd .1 ± .2
Pb 10 ± 1.0
Data is presented in means ± SEM

Fiber contents of MoLP is more than the previous study.[26,27] The ash content in MoLP is similar
to the soybean and corn meal.[30] The high value of ash indicated that MoLP is a good source of
mineral elements. NFE is similar to M. oleifera leaves of different plants.[26] .[31] The little bit variation
in carbohydrates values noted in this work to previous study due to variation in soil composition and
climatic conditions.[28]

Mineral composition of Moringa oleifera leaf powder. The mineral profile of MoLP are showed in
Table 2. The contents of Cd, Ni,, Pb, Mg, Zn, and Cu (±1.0) are 0.1, 0.5, 10, 0.6, 25 and 8.0, respectively.
The current outcomes are similar to the range of the previous study.[32] However, the slight variations
in the mineral content may be due to soil nature, weather conditions, and growth pattern of leaves.
The outcome of K is similar to previous result (1.3–2.1).[25] The Fe value of MoLP is more as
compared to Acer pictum thumb.[33] The current research agreed with the findings of Kumar et al.[34]
that the leaves of the particular plants are a rich source of minerals. The study suggested that plants
contain freely accessible phytonutrients and restocked nutrients. Though, the nutrients of the plants
may be influenced owing to the growth level of leaves and by the abiotic and biotic settings of the
environment.[28]

Phytochemical profiling of MoLE. In the present study, the phytochemical analysis of MoLP was
conducted by using an aqueous and organic solvent (70% ethanol). Results depicted that phenolic
acids are frequently miscible in nonpolar solvents.[34] Table 3 depicted the results of the basic
phytochemicals of the aqueous and 70% ethanol of MoLE.
The aqueous extract showed the moderate abundance of flavonoids, steroids, carbohydrates,
and amino acids, while least availability of phenols, alkaloid, saponins, glycosides, proteins sugars,
fats, and oils was detected. Triterpenoids, tannins and phytosterol were not detected in aqueous
MoLE. The data are in accordance with results mentioned by Sudha et al.[19] More abundance of
flavonoids, phenols, alkaloids, and carbohydrates were reported in 70% ethanol extract. Moderate
abundance of proteins, glycosides, steroids, amino acids, triterpenoids, and tannins and least
availability of phytosterols, reducing sugars, and fats were found in 70% ethanol extract. The
phenols and flavonoids found in MoLE might be active as usual antioxidant supplements.[34]
Leone et al.[4] attributed the medicinal worth of M. oleifera to the higher yield of phenolic acids,
saponins, alkaloids, flavonoids, polyphenols, and tannins.
The ferric chloride test for flavonoids indicated its moderate availability in aqueous while more
abundant availability in ethanol extract. The Folin-Ciocalteu test showed the presence of grayish black
precipitates to indicate the more abundance of phenols in alcoholic solvent than water. The less
concentration of alkaloid in aqueous MoLE and higher in 70% ethanol MoLE was proved by reddish
brown precipitates appeared by using Wagner’s reagent. Less foaming appears in aqueous whereas rich
foaming was observed in organic solvent that revealed the presence of saponins in MoLE. All
phytochemicals are supported by the previous study conducted by Sudha et al..[19]
Gelatin test conducted for tannins was negative in aqueous and positive in 70% ethanol
extract. Legal’s test showed pink to red colored precipitates for glycosides existence. Moderate
abundance of steroids in aqueous and more in ethanol extract was confirmed by the appearance
2344 S. KHALID ET AL.

Table 3. Qualitative phytochemical grading in Moringa oleifera leaf aqueous and 70% ethanol extract.
Presence of Presence of
phytochemical phytochemical
Sr. in Aqueous in 70% ethanol
no Phytochemical Reagent used Method Observation extract extract
1 Flavonoids Ferric Chloride 2 ml MoLE + few drops of Blackish red or ++ +++
reagent FeCl3 (10%) brown color
2 Phenols Folin-Ciocalteu 0.5 ml Folin-ciocalteu Grey or black color + +++
reagent solution+ 1 ml MoLE +
few drops of Na (aq)
3 Alkaloid Wagner’s 0.5 ml wagner’s solution + 2 Reddish brown + +++
reagent ml MoLE + 1 ml HCl (1%) precipitation
4 Saponins Froth reagent 5 ml water + 1 ml MoLE Foaming + +++
5 Tannin Gelatin 1% gelatin solution+ 1 ml No precipitation in - ++
reagent MoLE + few drops of NaCl aqueous
(aq) White in ethanol
6 Glycosides Legal’s test 2 ml pyridine and sodium Pink to red color + ++
nitropruside + 1 ml MoLE
+ 0.5 ml lead acetate +
NaOH 5 ml water
7 Steroids Salkowski’s 1ml chloroform + 2 ml MoLE Brownish ring ++ ++
reagent +.H2SO4(few drops)
8 Proteins Xanthoproteic 1 ml conc. HNO3 +few drops Yellow color + ++
reagent of 40% NaOH + 2 ml MoLE
9 amino acids Ninhydrin 2% Ninhydrin solution (few Blue color ++ ++
reagent drops) + 2 ml MoLE
10 Reducing sugars Fehling’s 1 ml Fehling’s reagent +2 ml Brick red + +
reagent MoLE precipitation
11 Carbohydrates Molicsh’s 2 ml MoLE + 2 ml conc. H2 purple ring ++ +++
reagent SO4+2 drops of molicsh’s
reagent
12 Triterpenoids Salkowski’s 2 ml MoLE +2 ml H2SO4 No precipitation in - ++
reagent +1ml chloroform aqueous
golden in
ethanol
13 Oils & fats Filter paper MoLE (few drops) + press in Oil mark found + +
folds of filter paper
14 Phytosterols Liebermann- 2 ml MoLE + 0.5 ml. H2SO4 Brown circle - +
Burchard’s +acetic anhydride (few appear
reagent drops
MoLE = Moringa oleifera leaf extract
+++ More abundant
++ Moderate abundant
+ Least abundance
- Absent

of brown ring with Salkowski’s reagent. Proteins and amino acids gave yellow and blue
precipitates by using reagents Xanthoproteic and ninhydrin, respectively, in both extracts of
M. oleifera. Fehling’s reagent, Molicsh’s reagent, and filter paper pressing was used to perform
decisive tests for the indication of reducing sugars, carbohydrates, and fats and oils, respectively,
discussed in Table 3.
The Salkowski’s test was only positive in 70% MoLE but negative in aqueous MoLE.
Phytosterols were detected in ethanol extract by performing Liebermann-Burchard’s test,
while they were absent in aqueous extract. The observations noticed in the current research
were similar to the previous research Sudha et al.,[19] and emphasized the nutritional worth of
M. oleifera being enriched with carbohydrates, proteins, and amino acids. Data in our study
revealed the high yield of some phytochemicals including amino acids, proteins, alkaloids,
saponins, steroids, and flavonoids, which is quite related to the study of Okiki et al.,[35]
phytochemical screening of MoL from Nigeria showed more amount of flavonoids, alkaloids
and saponins.
 INTERNATIONAL JOURNAL OF FOOD PROPERTIES 2345

Table 4. Free radical scavenging activity and total antioxidant capacity of aqueous and 70% ethanol mole.
Ttotal
Total tannin saponin Ferric reducing Radical Radical
Total phenol Total flavonoid content content antioxidant scavenging scavenging
content (tpc) content (TFC) (TTC) (TSC) power assay (DPPH) assay (ABTS)
(mgGAE/100 (mg QE/100 mg CE/100 mg DE/100 (FRAP) mmol (mM trolox/ (mM trolox/
MoLE ml) ml) ml Ml FeSO4 100 mL) 100 mL)
AQ 128.60 ± 1.67 3.91± 0.24 10.09 ± 0.25 36.26 ± 3.1 .87 ± .03 33.42±1.92 265.76 ± 22.86
70% 136.57 ± 1.78 4.02 ± 0.35 12.58 ± 0.64 25.14 ± 1.02 1.24±.05 28.48±1.02 328.57 ± 19.68
Ethanol
Data is presented in means ± SEM

Bioactive compounds in MoLE. MoLE are enriched with vital phenolic compounds, flavonoid
compounds, saponins, and tannins. Table 4 represents total flavonoid contents, total phenolic con-
tents, total saponin content,s and total tannin contents in aqueous and 70% ethanol MoLE. TPC was
measured in aqueous and 70% ethanol extract are similar to the aqueous and 50% ethanol extract of
Moringa oleifera.[11,36]
Further, MoLE outcomes showed TPC concentration similar to the leaf extract [37, whereas higher
concentrations in moringa infusion and methanolic extract.[38] Du Toit et al.[36] reported that
moderately reaped moringa leaves contain high amount of TPC from spring to summer climates.
Food plant contains physiologically and therapeutically active flavonoid, tannins, and saponins along
with phenolic acids.[14] In this research, TFC was found in aqueous and 70% ethanol MoLE as 3.91 and
4.02 mg QE/100 ml, respectively; the values are in accordance with aqueous and 50% ethanol extract of
Moringa oleifera.[11]
Total tannins content of aqueous and 70% ethanol was 10.09 and 12.58 mg CE/100 ml, respectively.
The outcome of current study is similar to the aqueous and 50% MoLE.[11] The TSC of liquid samples
showed highest extraction in aqueous and alcoholic extract.[39,40] The current results of tanning are
similar to previous study.[41] Total Saponin content was calculated in the range from 36.26 to 25.14 mg
DE/100 ml.

Free radical scavenging and total antioxidant capacity in MoLE. Reactive oxygen species accumu-
lated in biological systems and cause various degenerative disorders due to oxidative damage. There is
an equilibrium required for the scavenging activity of antioxidants and production of free radicals and
oxidative stress due to minor changes in this equilibrium.[42] Ahmed et al.[34] justified that antioxidant
constituents present in dietary sources are comparatively safe. They suggested that M. oleifera leaf
extracts contribute antioxidant property against free radicals and provide defense against oxidative
mutilation.
Table 4 revealed that MoLE (70% ethanol) proved to be more functioning than aqueous MoLE.
This is perhaps due to high content of phenols, flavonoids, tannins, and saponins in MoLE. MoLE is
blessed with more free radical scavenging activity, reducing power, and anti-oxidant capacity.[43,44] It
was mentioned that polyphenols contain phenol units considered to be a class of biologically active
compounds in MoLE, further represent a group of secondary metabolites efficacious in curing chronic
diseases owing to their strong anti-oxidant potential.[34]
The anti-oxidative activity of MoLE varied with different solvents used for extraction. The MoLE
aqueous has more anti-oxidative potential with the ABTS assay than the MoLE 70% ethanol. The
results are in agreement with the values in water and ethanol extract respectively.[11]
The anti-oxidant activity estimated with the DPPH assay was higher in 70% ethanol MoLE (28.48
mM trolox/100 mL) and aqueous (33.42 mM trolox/100 mL). Moreover, DPPH inhibition in methyl
and aqueous extracts of M. oleifera are similar to the current study. Ahmed et al.[34] investigated that
M. oleifera leaf extract significantly decrease DPPH radicals. They narrated that anti-oxidants donate
an electron or hydrogen to DPPH, hence balances its free radical property. Data provided in this
research suggested the relationship between phenolic acids and anti-oxidative response. 70% ethanol
2346 S. KHALID ET AL.

MoLE contain more phenolic acids due to organic in nature justified the high anti-oxidative action as
compare to aqueous MoLE. This particular behavior was also supported by the study in the leaves of
Morus nigra and Morus alba L..[45]
Ferric Reducing Antioxidant Power (FRAP) assay was performed to evaluating the total anti-
oxidative activity of MoLE in aqueous and 70% ethanol extract. The solution of FeSO4 was used and
concentration of extract was noted as 0.87 and 1.24 mmol FeSO4 in aqueous and 70% ethanol MoLE
respectively. These results clearly indicate that M. oleifera leaves are potentially more beneficial than
seeds.[43]

Conclusion
It is concluded that 70% ethanol extract of Moringa oleifera leaves are contained appreciable amount of
flavonoids, alkaloids, glycosides, steroids, phenols, and proteins. Based upon radical scavenging
activities, total reducing power and total antioxidant capacity measured by ABTS, DPPH, and FRAP
assayed in Moringa oleifera leaves depicts its worth as a natural effective. However further research is
required to isolate individual components to develop antioxidants for pharmaceutical and food
industries.

Author contribution
Shakeela Khalid: Writing original draft-Equal, Muhammad Arshad: supervision and Validation-Equal, Shahid
Mahmood: Validation-Equal, Farzana Siddique: review and editing-Equal, Waseem Khalid: Supervision-Equal, Turky
Omar Asar: Formal Analysis-Equal and Faten A. M. Hassan: Writing, review and editing-Equal.

Disclosure statement
No potential conflict of interest was reported by the author(s).

Ethical approval
This article does not involve any study with animal and human subjects.

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