INTERNATIONAL JOURNAL OF RESEARCH AND SCIENTIFIC INNOVATION (IJRSI)

ISSN No. 2321-2705 | DOI: 10.51244/IJRSI |Volume XII Issue XII December 2025

In-Vitro Antioxidant and Anti-Inflammatory Profile of the Stem

Bark Extracts of Guiera Senegalensis J. F. Gmel

Barkoma Mohammed Bashir, M.Sc. Hons¹.*, Mustapha Usman Nasir, M.Sc. Hons², Adamu

Muhammad Abdullahi, B.Sc. Hons³, Muhammed Malah Sheriff, B.Sc. Hons⁴

¹Department of Integrated Science, School of Sciences, College of Education and Legal Studies, Nguru,

Nigeria

²Department of Chemistry, Yobe State University, Damaturu, Nigeria

³Department of Dental Surgery Technician, Galtima Mai Kyari College of Health Sciences and

Technology, Nguru, Yobe State

⁴Department of Applied Chemistry, Federal Polytechnic Damaturu, Yobe State, Nigeria

^*Corresponding Author

DOI: https://doi.org/10.51244/IJRSI.2025.12120042

Received: 08 October 2025; Accepted: 14 October 2025; Published: 03 January 2026

ABSTRACT

Medicinal Plants are Plants used for traditional medicines to treat ailments. Any part of the plant could be used

as medicine. Medicinal plants have been discovered and used in traditional medicine practices since pre-

historical times. A survey of traditional medicine man in Damaturu indicated that Guiera senegalensis is a plants

used in the treatment of inflammation, wounds and cancer. The fresh samples (stem bark) of the plant was

collected and successively extracted with n-hexane, ethyl acetate, methanol and water. Phytochemical

constituents of Guiera senegalensis extracts revealed the presence of cardiac glycosides, flavonoids, saponins,

phenols, tannins, alkaloids, sterols and terpenoids. The antioxidant potential was evaluated by 2, 2-diphenyl-1-

picrylhydrazyl (DPPH) free radical assay, whereas membrane stabilization method was used to evaluate the anti-

inflammatory potential of the extracts. The IC50 values obtained for the Guiera senegalensis extracts indicated

that the methanol and water extracts exhibited higher antioxidant activities than the standard ascorbic acid. The

extracts were found effective in inhibiting the heat induced haemolysis. Diclofenac Sodium salt used as standard

drug at 100μg/ml offered 90.66% protection a significant (p<0.05) protection against damaging effect of heat

solution. The results showed that all extracts from the plant offered a significant (p<0.05) protection against the

damaging effect of hypotonic solution, Diclofenac sodium (100μg/ml), the standard drug offered a significant

(p<0.05) protection (57.52%).

Keywords: Phytochemical, Antioxidant, Anti-inflammatory

INTRODUCTION

Medicinal plants are plants used for traditional medicines or what is well known as herbal medicine. Any part

of the plant could be used as medicine, the leaves, roots, seeds, stems, etc. Medicinal plants or the medicinal

herbs have been discovered and used in traditional medicine practices since pre-historical times. Early written

reports on the use of plants as medicine appeared about 2600 BC when plants were used as medicine by

Sumerians and Akkadians (Shoeb, 2006). Since then, plants have been used to treat diseases such as headache,

toothaches, stomach aches, diarrhea, wounds, tumors and sexually transmitted diseases (van Wyk and Gericke,

2000; Khaleeliah, 2001; Von Koenem, 2001, Wuyang, 2008).

A plant is considered medicinal if it produces compounds which are therapeutically effective. Plants produce a

wide range of secondary metabolites, and the medicinal properties are attributed to the presence of these

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INTERNATIONAL JOURNAL OF RESEARCH AND SCIENTIFIC INNOVATION (IJRSI)

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metabolites such as terpenoids, steroids, saponins, tannins, flavonoids, alkaloids and phenolic compounds

(Mdlolo; 2009, Fawole 2009).

Guiera senegalensis JF Gmel, of the family Combretaceae, commonly known as Moshi medicine (English) or

Saabara in Hausa. Guiera senegalensis is a shrubby plant and can grow to a height of 3 to 5 m depending on the

habitat (Silva et al., 2008). The leaves which are 3 to 5 cm long and 1.5 to 3.0 cm broad are arranged opposite

or sub opposite on the stem (Hutchinson and Dalziel, 1972).a survey of medicinal plants within and around

Damaturu environs indicated that the plant is used in the treatment hepatitis, inflammation, malaria, wounds and

cancer. However, there’s no scientifically backed evidence to support this claim. This poses an important

challenge to seek for more scientific studies to be carried out on each of the four plants in order to ascertain these

claims.

Herbal medicine in Nigeria is gaining more recognition and this is seen in how much inquiries people make

concerning home remedies and traditional medicine. Nigeria is richly endowed with indigenous plants which are

used in herbal medicine to cure diseases and heal other injuries. Some of these plants are used as food and/or

medicine. The extracts from this plant has shown to exhibit a wide range of biological and pharmacological

activities such as anticancer, anti-inflammatory, diuretic, laxative, antispasmodic, antihypertensive, antidiabetic,

antimicrobial activities, etc. It is generally assumed that the active medicinal constituents contributing to the

protective effects are phytochemicals, vitamins and minerals. (Okwu, Ekeke 2003 and Okwu; 2004). For this

reason, medicinal plants are considered be important to the health of the individuals and communities.

MATERIALS AND METHODS

Apparatus and Materials

Ultrasonicator (Model/AS3120) was purchased from Tianjin Automatic Science Instrument Co., Ltd. China,

analytical weighing balance (Ohaus Corp. Pine Brook, NJ USA), pestle and mortar (wooden) purchased in

Damaturu Sunday market, empty bottles purchased at Bayan Tasha market Damaturu, sieve, fume cupboard,

drying cabinet (model/FSM140)from 2 Building, Majialong Industrial Zone, Nanshen District, Shenzhen Jinly

Technology Co., Ltd. China, UV/VIS spectrophotometer (model/UV752) from Changsha, Hunan, Wincon

Company Ltd. China, Autoclave (Model/DWB-280B) and Water bath (Model/DWT-420) from Shanghai

Drawell Scientific Instrument Co., Ltd. Room 211 Building 7, sheng Yu Industrial Park No. 365 ChunHong,

Shanghai, China and other laboratory materials.

Chemicals

n-hexane, ethyl acetate and methanol were purchased from BDH Chemicals Ltd., Poole, United Kingdom. 2,2-

diphenyl-1-picryl hydrazyl (DPPH) was purchased from SIGMA-ALDRICH Company Ltd., 3050 Spruce street

St. Louis, MO63103 USA. Muller Hinton Agar was purchased from TITAN BIOTECH Ltd., A-904A, RIICO

Industrial Area, Phase-III, Rajasthan, India. Dimethyl Sulphur Oxide (DMSO) was purchased from Guangdong

Guanghua Sci-Tech Company Ltd., Add 6, Jiangyan South Road, Guangzhou, Guangdong, China. ethanol,

ascorbic acid, Hydrochloric acid (HCl), Sulphuric acid (H₂SO₄), Magnesium metal, Ferric chloride,

Dragendroffs reagent, choloroform and all other Chemicals used are of highest analytical grade and purchased

from BDH Chemicals, Poole, England.

Collection and Preparation of Plant Sample

The fresh sample of Gueira senegalensis (Voucher number 1973) (stem bark) was collected at Damaturu Local

Government Area, Yobe State, Nigeria. The herbarium specimen was identified by Mallam Salihu Abdullahi a

Taxonomist at the Department of Biological Sciences, Yobe State University, Damaturu. The stem bark of the

plant was collected two (2) meters above the ground. The sample was sorted to ensure no foreign bodies were

present. This sample was then dried under shade in the laboratory at ambient temperature. The dried sample was

then crushed into coarse particles using local pestle and mortar. It was further crushed into fine powder and

sieved with a sieve and weighed. The fine powdered sample was then weighed and stored in sealed containers

until required for further analysis.

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Extraction of Phytochemicals

About 550g of the powdered plant material of Guiera senegalensis was separately extracted successively with

2.5 L portions of n-hexane, ethyl acetate, methanol and water in that order using ultrasonicator for two hours at

room temperature. The solvent containing the extracts was allowed to settle after the extraction, then the mixture

was separated from the residue by filtering with Whatmann No. 1 filter paper and then kept in a clearly labelled

container ready for solvent recovery. The residue of the sample was then mixed with the next solvent for the

further extraction. The procedure was repeated for the remaining solvents namely; ethyl acetate, methanol and

water in that order.

Phytochemical Screening

Phytochemical screening to detect the presence of phytochemicals from the samples was carried out using the

procedures outlined by Tiwari et al. (2011); Sabri et al. (2012) and Solomons et al. (2013).

Test for alkaloids

The extract (0.5g) was dissolved in 5ml of 2N HCl and filtered. The filtrate was treated with Dragendroff”s

reagent (Solution of potassium Iodide and bismuth Iodide). Formation of red precipitate indicates the presence

of alkaloids.

Test for flavonoids

The extract (0.5g dissolved in 2ml of methanol) was treated with few drops of sodium hydroxide solution.

Formation of intense yellow colour, which becomes colourless on addition of dilute acid, indicates the presence

of flavonoids.

Test for saponins

Frothing test: The extracts (0.5g) were diluted with distilled water to 20ml and this was shaken in a graduated

cylinder for 15minutes over a vortex mixer. Formation of 1cm layer of foam indicates the presence of saponins.

Test for cardiac glycosides (Keller Kelliani’s test)

To 5ml of each extract (0.5g dissolved in 5ml methanol) was treated with 2ml of glacial acetic acid in a test tube

and a drop of 2%ferric chloride solution was added to it. This was carefully underlayed with 1ml of concentrated

sulphuric acid. A brown ring at the interface indicates the presence of deoxysugar characteristic of cardenolides.

A violet ring may appear below the ring while in the acetic acid layer, a greenish ring may form.

Test for oxalate

To 3ml portion of the extract (0.2g in 3ml of methanol) were added a few drops of glacial acetic acid. A greenish

black colouration indicates the presence of oxalates.

Test for quinones

A small portion of the extract was treated with concentrated hydrochloric acid. The formation of yellow

precipitate/colouration indicates the presence of quinones.

Test for terpenoids (Salkowski’s test)

To 1ml of chloroform was added to 2ml of the extract followed by a few drops of concentrated sulphuric acid.

A reddish brown precipitate produced immediately indicates the presence of terpenoids.

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Test for tannins (Braymer’s test)

To 2ml of the extract was treated with 10% alcoholic ferric chloride solution. Formation of blue/greenish

colouration indicates the presence of tannins.

Test for sterols (Libermann-Burchard test)

To 1ml of the extracts was treated with few drops of chloroform, acetic anhydride and concentrated sulphuric

acid. The formation dark pink or red colour indicates the presence of sterols.

Test for phenols

A fraction of the extract was treated with aqueous 5% ferric chloride solution. The formation of deep blue or

black colour indicates the presence of sterols.

Measurement of Antioxidant Activities

The antioxidant activities of Guiera senegalensis extracts were determined on the basis of their scavenging

activity of stable 2, 2-diphenyl-1-picrylhydrazyl (DPPH) free radical as follows; to 1 ml of each solution of

different concentrations (10, 25, 50. 100, 125, 250, 300, 500µg/ml) of the extracts, 3 ml of 0.004% ethanolic

DPPH free radical solution was added. After 30 minutes, the absorbance of the preparations were taken at 517nm

by UV spectrophotometer. This was then compared with the corresponding absorbance of standard ascorbic acid

concentrations (10, 25, 50. 100, 125, 250, 300, 500 µg/ml) as described by Hatano et al. (1988), with some

modifications.

Then the % inhibition was calculated by the following equation;

% Radical scavenging = (absorbance of blank – absorbance of sample) x 100%

Activity (absorbance of blank)

The blank was prepared by adding 3 ml of 0.004% ethanolic DPPH to 1 ml of the ethanol.

Procedure

1. Eight 8 test tubes were taken to prepare the different concentration (10, 25, 50. 100, 125, 250, 300, 500µg/ml)

plant extracts and ascorbic acid standard.

2. Extracts and ascorbic acid were accurately weighed and dissolved in ethanol to make the required

concentrations by dilution technique.

3. First 0.004g of DPPH was weighed and dissolved in 100ml of ethanol to make 0.004% (w/v) homogenous

solution using a vortex mixer.

4. 3ml of 0.004% DPPH solution was added to each of the 8 test tubes by means of a auto pipette, after preparing

the desired concentrations

5. The room temperature was recorded and the test tubes were kept for 30 minutes to complete the reactions.

6. DPPH was also added to the blank test tube at the same time where only ethanol was taken as blank.

7. The absorbance of each test tube was measured using a UV spectrophotometer.

8. IC_50’swere measured from % Inhibition vs. Concentration graph using Microsoft excel

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INTERNATIONAL JOURNAL OF RESEARCH AND SCIENTIFIC INNOVATION (IJRSI)

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Anti-Inflammatory Activity

Membrane stabilization method

The Human Red Blood Cell (HRBC) membrane stabilization has been adopted as a method to study in vitro

anti-inflammatory activity since the erythrocyte membrane is comparable to the lysosomal membrane (Gandasan

et al., 1991, Shenoy et al., 2010) and it is believed that its stabilization indicates that the extract may capably

stabilize lysosomal membranes. The stabilization of lysosomal membrane is important as it helps in limiting the

inflammatory response by stopping the release of lysosomal constituents of activated neutrophils, such as

enzymes like proteases and bacterial products which can be the cause of further tissue inflammation and harm

upon extra cellular release. The lysosomal enzymes released during inflammation generate various disorders.

The extra cellular activity of these enzymes are believed to be connected to acute or chronic inflammation. The

Non-steroidal Anti-inflammatory Drugs (NSAIDs) produce their effects either by inhibiting the lysosomal

enzymes or by stabilizing the lysosomal membranes (Rajendran et al., 2008).

Preparation of red blood cells (RBC’s) suspension

The blood sample was collected from healthy human volunteers of postgraduate students of the Department of

Biochemistry of Bayero University Kano, Kano State who have not taken Non-steroidal Anti-inflammatory

Drugs (NSAIDs) for the past two weeks preceding the experiment and the samples were transferred to centrifuge

tubes. The tubes containing the blood were centrifuged at 3,000 rpm for 10 min and were (blood) washed three

(3) times with an equal volume of the normal saline. The volume of blood was weighed and re-constituted as

10% v/v suspension with normal saline.

Heat induced haemolysis

The reaction mixture (2 ml) consisting of 1ml of the test extracts of different concentrations (100, 300 and 500

µg/ml) and 1ml of 10% suspension, instead of test sample only the saline was added to the control test tube.

Dichlofenac sodium salt was used as a standard drug and was weighed (0.0025g in 5ml of water). All the

centrifuge tubes containing reaction mixture were incubated in a water bath for 30 min at 56ºC. At the end of

the incubation the tubes were cooled under a running tap water. The reaction mixtures were further centrifuged

at 2500 rpm for 5 min and the absorbance of each supernatant was taken at 560nm using UV/VIS

spectrophotometer. The experiment was performed in triplicates for all the test samples. The percentage

inhibition of haemolysis was calculated as follows:

Percentage inhibition = Absorbance (control) – Absorbance (sample) ×100

Absorbance (control)

Hypotonic solution induced haemolysis

Different concentrations of the plant extracts were prepared (i.e. 100, 300 and 500 µg/ml), reference sample and

control were both separately mixed with 1 ml of phosphate buffer pH7.4, 2 ml of hyposaline and 0.5 ml of HRBC

suspension. Dichlofenac sodium salt was used as a standard drug. All the reaction mixtures for the assay were

incubated at 37ºC for 30min and thereafter centrifuged at 3000rpm for 10/min. The supernatant liquid was

decanted and haemoglobin content was determined by spectrophotometer at 560nm. The percentage of Red

Blood Cell membrane stabilization of defensive was calculated by the following equations:

% protection = 100 - Optical density of drug treated sample

X 100

Optical density of control

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RESULTS AND DISCUSSION

Extraction Yield

The powdered stem bark of Guiera senegalensis was extracted successively with n-hexane, ethyl acetate,

methanol and water using ultrasonicator. The results of the extraction yield are shown in the table below:-

Table 1. Percentage yield of crude stem bark extracts of Guiera senegalensis

S/No.

Solvents used

n-Hexane

Ethyl Acetate

Methanol

Water

Weight of plant part used (g)

550 (Guiera senegalensis)

Weight of extract(g)

Percentage yield (%)

1

2

3

4

8.65

10.73

8.19

8.30

1.57

1.95

1.49

1.51

The results above revealed that in the 550g of Guiera senegalesis extracted, all the four extracts of Guiera

senegalesis exhibited similar pattern of phytochemical yield with ethyl acetate found to be the highest 10.73g

(1.95%), followed by n-hexane 8.65g (1.57%), water 8.30g (1.51%) and methanol 8.19g (1.49%). These results

suggested that crude phytochemicals for each accounted for less than 10% of the plant sample and that ethyl

acetate extracted more than the other solvents.

Phytochemical Screening

Guiera senegalensis

Phytochemical constituents of the Gueira senegalensis n-Hexane, Ethyl acetate, methanol and Water extracts

were drtermined. The results (Table 2) revealed the presence of alkaloids, flavonoids, saponins, tannins and

cardiac glycosides in all the extracts. However, terpenoids were detected only in the n-Hexane and water extracts

while phenols were present in n-hexane, ethyl acetate and methanol extracts. Quinones, Sterols and Oxalate were

not detected in all the extracts.

Table 2. The Results of the Phytochemical Screening of Gueira senegalensis n-Hexane, Ethyl acetate, Methanol

and Water Extracts

Phytochemicals

Alkaloid

n-Hexane extract Ethyl acetate Extract

Methanol

Extract

Water Extract

+

++

+

-

+

++

+

-

+

-

Flavonoid

Saponins

++

+

-

Cardiac glycoside

Oxalate

Quinones

-

Terpenoids

Tannins

+

-

+

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Sterols

-

Phenols

+

Key: ++ = Highly present, + = Present, - = Absent

The results also suggested the presence of more flavonoids in ethyl acetate and methanol extracts and saponins

in the n-hexane and ethyl acetate extracts. It also showed that only non-polar and extremely polar terpenoids

were present in Gueira senegalensis. The presence of these secondary metabolites may be very essential for the

biological and pharmacological activities of this plant.

Antioxidant Studies

The antioxidants are mainly derived from food and medicinal plants such as fruits, vegetables, cereals,

mushrooms, beverages, flowers, spices and traditional medicinal herbs (Cai et al., 2004). Natural antioxidants

from plant materials are mainly Polyphenols (comprising mainly of Phenolic acids, Flavonoids, Tannins,

Anthocyanins, Lignans and Stilbenes), Carotenoids (Xanthophylls and Carotenes) and Vitamins (Vitamin C and

E), Baiano et al., (2015). These natural antioxidants, especially the Polyphenols and Carotenoids are reported to

exhibit a wide range of biological effects, such as anticancer, antibacterial, anti-inflammatory, antiviral and anti-

aging Fang, et al., (2014).

The antioxidant studies in the present work were carried out on the n-hexane, ethyl acetate, methanol and water

extracts of Guiera senegalensis using the free radical scavenging activities of the samples on 2,2-diphenyl-1-

picryldihydrazyl (DPPH) radical and ascorbic acid as standard. From the caliberation curves obtained, the 50%

inhibitory concentration (IC₅₀) values were determined. IC₅₀value denotes the concentration of the sample

required to scavenge 50% of the DPPH free radicals measured at 517nm as reported by Gupta et al., (2003). The

absorbance and percentage inhibition of the standard and the extracts were shown in Tables 3, respectively.

Table 3. UV Absorbance measured at 517nm of antioxidant activity of Standard Ascorbic Acid, n-hexane extract,

ethyl acetate extract, methanol extract and water extract of Guiera senegalensis.

Extract

Absorbance

S/No.

Concentration

(µg/ml)

Ascorbic

Acid

n-Hexane

Ethyl Acetate

Methanol

Water

1

2

3

4

5

6

7

8

9

0

0.675

0.433

0.354

0.249

0.122

0.093

0.084

0.075

0.073

0.675

0.526

0.365

0.253

0.174

0.124

0.096

0.088

0.086

0.675

0.513

0.322

0.206

0.128

0.097

0.088

0.073

0.675

0.494

0.296

0.189

0.095

0.086

0.073

0.062

0.059

0.675

0.542

0.388

0.281

0.193

0.138

0.097

0.094

0.091

10

25

50

100

125

250

300

500

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The Table 3 above was used to plot a graph of Absorbance of standard ascorbic acid and n-Hexane, Ethyl

Acetate, Methanol and Water extracts of Guiera senegalensis against concentration using MS. Excel.

0.8

0.7

0.6

0.5

ascorbic

0.4

0.3

0.2

0.1

0

n-hexane

ethyl acetate

methanol

water

0

100

200

300

400

500

600

Concentration

Figure 1. Graph of Absorbance at 517nm Against Concentration of Standard Ascorbic Acid, n-Hexane extract,

Ethyl Acetate extract, Methanol extract and Water extract of Guiera senegalensis.

The above graph in Fig. 1 is showing the DPPH scavenging activity of n-Hexane, Ethyl Acetate, Methanol and

Water extracts of Guiera senegalensis compared to that of the standard ascorbic acid. Because of difficulties of

interpretation Table 3 was converted to % inhibition as shown in Table 4.

Table 4. Percentage Inhibitions of Ascorbic acid, n-Hexane, Ethyl Acetate, Methanol and Water extracts of

Guiera senegalensis.

S/No.

Concentration

(µg/ml)

Ascorbic acid n-Hexane

Ethyl Acetate

Methanol

Water

1

2

3

4

5

6

7

8

9

0

10

35.9

47.5

63.1

81.9

86.2

87.5

88.9

89.2

22.1

45.9

62.5

74.2

81.6

85.8

86.9

87.3

24.0

52.3

69.5

81.0

85.6

86.9

89.2

26.8

56.1

72.0

85.9

87.3

89.2

90.8

91.3

19.7

42.5

58.3

71.4

79.6

85.6

86.1

86.5

25

50

100

125

250

300

500

From this table, a graph of % inhibition of n-hexane, ethyl acetate, methanol and water extracts of Guiera

senegalensis against concentration were plotted separately.

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120

100

80

60

40

20

0

y = 0.1361x + 40.137

R² = 0.5201

%inhibition

Linear (%inhibition)

0

100

200

300

400

500

600

Concentration (µg/ml)

Figure 2. %inhibition of n-hexane extract of Guiera senegalensis

The IC₅₀value of n-hexane extract of Guiera senegalensis was calculated from the Fig. 2. The Inhibitor

Concentration against the percent activity is plotted using the linear equation provided in the graph (y = 0.1361x

+ 40.137), for y = 50 value becomes IC₅₀value of n-hexane extract of Guiera senegalensis which determined to

be 72.5 µg/ml, a value much lower than that shown by ascorbic acid.

120

y = 0.1311x + 44.376

100

R² = 0.4615

80

60

%inhibition

Linear (%inhibition)

40

20

0

100

200

300

400

500

600

Concentration (µg/ml)

Figure 3. %inhibition of ethyl acetate extract of Guiera senegalensis

The IC₅₀value of ethyl acetate extract of Guiera senegalensis was calculated from Fig. 3. The IC₅₀value of ethyl

acetate extract of Guiera senegalensis was calculated to be 42.9µg/ml using the straight line equation (y =

0.1311x + 44.376) obtained from the graph. The value was slightly in activity than ascorbic acid.

120

y = 0.1301x + 46.937

R² = 0.4412

100

80

60

40

20

0

%inhibition

Linear (%inhibition)

0

100

200

300

400

500

600

Concentration (µg/ml)

Figure 4. %inhibition of methanol extract of Guiera senegalensis

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The IC₅₀value of methanol extract of Guiera senegalensis was calculated from the Fig. 4. The IC₅₀value of

methanol extract of Guiera senegalensis was calculated to be 23.5µg/ml using the straight line equation (y =

0.1301x + 46.937) obtained from the graph. This value was much higher in activity than ascorbic acid. The water

extract of Guiera senegalensis was subjected to similar analysis as shown in Fig. 5.

120

y = 0.1404x + 37.644

100

R² = 0.5511

80

60

%inhibition

Linear (%inhibition)

40

20

0

100

200

300

400

500

600

Concentration (µg/ml)

Figure 5. %inhibition of water extract of Guiera senegalensis for its IC₅₀Value.

The IC₅₀value of the aqueous extract of Guiera senegalensis was then calculated to be 88.0µg/ml using the

straight line equation (y = 0.1404x + 37.644) obtained from the graph. This value was more than twice lower

activity than ascorbic acid.

Table 5. The summary of the IC₅₀values of Guiera senegalensis extracts as compared to standard ascorbic acid.

IC₅₀(µg/ml)

S/No. Extract/*Standard

G. S

38.9

87.3

42.9

23.5

88.0

1

*Ascorbic acid

n-hexane

2

3

4

5

Ethyl acetate

Methanol

Water

Key: G. S - Guiera senegalensis

The IC₅₀results of Guiera senegalensis samples showed that only methanol sample exhibited higher antioxidant

activity (23.5 µg/ml) than the standard ascorbic acid. These results were taken to indicate that Guiera

senegalensis stem bark can be used as the reliable antioxidant which is very important component of wound

healing.

RBC Membrane Stabilization

Membrane stabilization is a process of maintaining the integrity of biological membranes such as the erythrocyte

and lysosomal membranes against osmotic and heat-induced lysis as reported by Sadique et al., (1989). In this

study the effects of the various extracts on RBC membrane stabilization against haemolysis induced by heat and

hypotonicity were determined. The results of each test were expressed as mean ± SD using Graph Pad prism

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(version 4), using a one-way analysis of variance (ANOVA). The statistical method applied in each analysis was

described in each Table. Results were considered to be significant when p-values were less 0.05 (p<0.05)

Heat induced haemolysis.

The results of the effect of extracts on heat induced haemolysis of RBC Are represented in Table 6. The extracts

were found to be effective in inhibiting the heat induced haemolysis at different concentrations. The results

showed that n-Hexane extract of Guiera senegalensis at concentration 500µg/ml protect significantly (p<0.05)

the erythrocyte membrane against lysis induced by heat Table (4.31). Diclofenac Sodium salt used as standard

drug at 100µg/ml offered 90.66% protection a significant (p<0.05) protection against damaging effect of heat

solution.

Table 6. Effect of n-Hexane, Ethyl acetate, Methanol and water Extracts of Gueira senegalensis on Heat Induced

Haemolysis

Extract

Treatment(S)

Absorbance at 560nm

1.264±0.0012^a

0.1180±0.0000^b

0.2780±0.0020^c

0.2530±0.0015^c

0.2440±0.0006^c

1.264±0.0012^a

0.1180±0.0000^b

0.5397±0.0097^c

0.3500±0.0023^d

0.3407±0.0012^d

1.264±0.0012^a

0.1180±0.0000^b

0.7333±0.0037^c

0.4617±0.0009^d

0.3430±0.0051^e

1.264±0.0012^a

0.1180±0.0000^b

0.4017±0.0487^c

0.3990±0.0020^c

0.3923±0.0003^c

% Inhibition

0

Negative Control (Normal Saline)

n-hexane

Positive Control (Diclofenac Sodium)

90.66

78.01

79.98

80.69

0

100

300

500

Negative Control (Normal Saline)

Ethyl acetate

Methanol

Water

Positive Control (Diclofenac Sodium)

90.66

57.30

72.31

73.04

0

100

300

500

Negative Control (Normal Saline)

Positive Control (Diclofenac Sodium)

90.66

41.98

63.47

72.86

0

100

300

500

Negative Control (Normal Saline)

Positive Control (Diclofenac Sodium)

90.66

68.21

68.43

68.96

100

300

500

Values are expressed in mean ± SD (n = 3). Data analysed using one way Anova;

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Values along same column differently superscripted differ significantly (P<0.05)

The results showed that all extracts of Guiera senegalensis have membrane stabilizing effects. The n-hexane

extract showed the highest values of % inhibition (78.01 – 80.69%, (P<0.05) and no much differences were

observed with increase in concentration. This followed by ethyl acetate extract with the values of (57.30 –

73.04%, (P<0.05). The water extract which also showed very little difference as concentration increases (68.21

– 68.96%, (P<0.05). Methanol extract showed the lowest stabilizing effect (41.98 – 72.86%, (P<0.05).

Hypotonicity induced haemolysis of RBC.

Table 6. Effect of n-Hexane Extract of Gueira senegalensis on Hypotonicity Induced Haemolysis

Extract

Treatment(S)

Absorbance at 560nm

0.5690±0.0000^a

0.2417±0.0003^b

0.3843±0.0007^c

0.2238±0.0007^b

0.1897±0.0003^d

0.5690±0.0000^a

0.2417±0.0003^b

0.5583±0.0007^a

0.4973±0.0009^c

0.2487±0.0007^b

0.5690±0.0000^a

0.2417±0.0003^b

0.3653±0.0322^c

0.3350±0.0150^c

0.2877±0.0027^d

0.5690±0.0000^a

0.2417±0.0003^b

0.2053±0.0003^c

0.2047±0.0003^c

0.1453±0.0003^d

% Inhibition

0

Negative Control (Hyposaline)

n-hexane

Positive Control (Diclofenac Sodium)

57.52

32.46

60.66

66.66

0

100

300

500

Negative Control (Hyposaline)

Ethyl acetate

Methanol

Water

Positive Control (Diclofenac Sodium)

57.52

1.88

100

300

12.60

56.29

0

500

Negative Control (Hyposaline)

Positive Control (Diclofenac Sodium)

57.52

35.79

41.12

49.43

0

100

300

500

Negative Control (Hyposaline)

Positive Control (Diclofenac Sodium)

57.52

63.91

64.02

74.46

100

300

500

Values are expressed in mean ± SD (n = 3). Data analysed using one way Anova;

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Values along same column differently superscripted differ significantly (P<0.05)

The results of the study on the extracts of Guiera senegalensis (Table 6) have revealed variable levels of

membrane stabilizing effects against haemolysis induced by hypotonic solution. The water extract showed the

highest significant values of 63.91 – 74.46%, (P<0.05), even at lower concentration it showed a significant value

higher than the standard drug. The next extract with the high protective effect was n-hexane which showed a

significant values of 32.46 – 66.66%, (P<0.05). The methanol extract follows the n-hexane extract but it showed

lower inhibitions (35.79 – 49.43%, (P<0.05)) than the standard drug it was also observed that as the concentration

increases the inhition of haemolysis also increases. The lowest anti haemolytic effect of Guiera senegalensis

came from the ethyl acetate extract with a significant values of 1.88 – 56.29%, (P<0.05), but showed a very little

effects at lower concentrations. These results suggest that Guiera senegalensis extract, especially the aqueous

and hydrophobic, can protect RBC from hypotonic induced haemolysis.

CONCLUSION

These plants extracts showed good antioxidant activity and anti-inflammatory activity. The therapeutic potential

of the stem barks of Guiera senegalensis could be attributed to classes of active components present in the stem

barks such as alkaloid, flavonoids, tannins and phenolic etc. which may be acting in synergy or individually. The

antioxidant and anti-inflammatory activities carried out in this study lend credence to the traditional claim about

the wound healing property of the stem barks of G. senegalensis.

Secondary metabolites such as alkaloids, flavonoids, phenols, tannins, terpenoids, etc. may be responsible for

the positive antioxidant, anti-inflammatory and antibacterial activities reported in this study thus comparing with

report of Nandagoapalan et al., (2016). Such activities as antimicrobial, anti-inflammatory, antioxidant,

anticancer, among others, may be principal indicators of a plant's value in medicine.

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