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Oseni1,2 OA, Fatokun2 DI, Amiola2 BO, Adejare3 ID



Oseni1,2 OA, Fatokun2 DI, Amiola2 BO, Adejare3 ID. Anti-microbial, In vitro Antioxidant and other Biochemical properties of Dryopteris expansa, Newbouldia laevis, and Senna hirsuta. From Anchor University, Lagos. Special Journal of Pharmacy, Pharmacology and Herbal Research. 2021, 2 (1): 1-16

High points:

  • Investigation on medicinal importance of Dryopteris expansa, Newbouldia laevis, and Senna hirsuta
  • Applications of Dryopteris expansa, Newbouldia laevis, and Senna hirsuta in drugs development and delivery
  • The results of the three plants possessed good nutritional, elemental, antioxidant, antimicrobial potentials and safety status at concentrations studied.



The leaves of Dryopteris expansa, Newbouldia laevis, and Senna hirsuta belong to different families with different genera and species. Traditionally, the leaves, stems, and roots have been used locally for treatments of various ailments.


To evaluate the anti-microbial, in vitro Antioxidant and other Biochemical properties of Dryopteris expansa, Newbouldia laevis, and Senna hirsuta. From Anchor University, Lagos

Materials and Methods:

Fresh leaves of Dryopteris expansa, Newbouldia laevis, and Senna hirsuta were collected within Anchor University, Lagos environment, air died and assessed for various parameters using standard methods.


The results obtained showed that the leaves were rich in crude fibre, protein, nutritionally valuable elements, and low anti-nutrient contents. The aqueous extracts of the leaves of the three plants contained some basic, non-toxic phytochemicals with high antioxidant potentials.  There was absence of mortality of the test animals at the oral treatment of 2000mg/kg body weight. The result of the antimicrobial activity also revealed that the aqueous extracts of the leaves showed good antibacterial and antifungal potentials when compared with some standard control antibiotics.


The study revealed good nutritional, elemental, antioxidant, antimicrobial, and safety status of the aqueous extracts of the leaves of the three plants. The results obtained therefore showed the tradomedicinal claims of the aqueous extracts of the leaves of these plants and their applicability in drugs development.


Phytochemical screening, antinutrient composition, acute toxicity, antioxidant potentials, proximate analysis.


1Department of Medical Biochemistry, College of Medicine, Ekiti State University, Ado-Ekiti, Nigeria. 2Department of Chemical Sciences, Anchor University, Lagos, Nigeria. 3Department of Biochemistry, Federal University, Dutsin-ma, Katsina State, Nigeria

Address for correspondence:

Dr Oseni O. A, mail:,, Tel: +2348035665049

Article history:

Received September 18, 2021: Accepted: November 3rd, 2021:  Published:  10th Nov 2021

Distribution and usage license:

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Plants play major roles in the treatment of many diseases as exemplified by their employment in all the major branches of medicine. Therefore, the World Health Organization supports the use of traditional medicine provided they are authenticated to be efficacious and safe for mankind (1, 2). As a result of these, plants serve presently as bases for pharmaceuticals. Antioxidants are compounds that are capable of inhibiting or stabilizing free radicals.

They could be man-made or natural which are found in foods (3). Several antioxidants are known to protect against diseases (4). There are some plants with natural antioxidant compounds that are pharmacologically potent and have little or no side effects. Newbouldia laevis also known as fertility plant is a tropical angiosperm plant belonging to the family of Bignoniaceae. It is a monotypic genus plant majorly found in tropical Africa and grows from Guinea Savannahs to dense forests (5).

Newbouldia laevis plants are used for medicinal purposes, mostly in Nigeria, it is effective in the treatment of elephantiasis, dysentery, rheumatic swelling, syphilis, constipation, pile, and as a vermifuge to round worms (6). Dryopteris expansa is commonly known as a Spreading Wood Fern. It is a species of fern, a native of cool temperate and subarctic regions of the Northern Hemisphere of Spain and Greece. Dryopteris expansa is a deciduous fern, which can be harvested from the wild for local use as food, medicine, and a source of materials. Fern is of different species but Dryopteris expansa has the same feature as Dryopteris dilatata (broad buckler fern), therefore can be easily confused.

Though many studies have not been done on Dryopteris expansa, research has shown that the ethanolic extract of Dryopteris filix-mas (Dryopteridaceae family) has its use in the treatment of worm infections and diarrhoea (7). Senna hirsuta could be a suitable antibacterial agent against some pathogenic bacteria. It also has significant potency on bacterial-induced hepatoxicity; therefore, the antibacterial activity supports the traditional use against infectious diseases8. According to world-agroforestry website information, the leaves of Senna hirsuta are used medicinally for treating kidney disorders and herpes (9, 10)

Hence this study aimed at determining the proximate composition, elemental and anti-nutrient contents of the leaves; phytochemical constituents, acute toxicity, antioxidant and antimicrobial potentials of the aqueous extracts of the leaves of the Newbouldia laevis, Dryopteris expansa, and Senna hirsute to establish their further medicinal usefulness in the treatments of various diseases.

Materials and methods

Collection and authentication of the plant’s samples:

Fresh samples of the Dryopteris expansa (Alabama streak sorus Fern); Newbouldia laevis (Boundary tree, Ewe Akoko) and Senna hirsuta (Hairy senna, Ewuro Ijebu/Asunwon) plants were obtained from Anchor University, Ayobo, Lagos State, Nigeria. The leaves were sourced from various sites within the location between July and September 2020 by detaching large quantities of leaves from the plants.

The plants were authenticated at the Plant Science Department of Ekiti State University, Ado-Ekiti, Nigeria by the Chief Technologist with voucher numbers UHAE 2020088 Newbouldia laevis [P. Beauv] Seem. ex Bureau. [Bignoniaceae]; UHAE 2020065 Dryopteris expansa (C. Presl) Fraser-Jenkins & Jermy and UHAE 2020089 Senna hirsuta [L] H. S. Irwin & Barneby [Fabaceae]. The leaves of the plants were cleansed and air-dried in the open laboratory, crushed, and subsequently ground to powder separately with a Marlex Excella laboratory blender.

 Proximate, elemental, and antinutrient analyses of the samples:

Proximate analysis was carried out on the dried powdered plant leaves according to the procedure of the Association of Official Analytical Chemists11. The percentages of moisture content, ash content, crude fibre, crude protein, crude fat, and carbohydrate were determined and calculated accordingly. The total carbohydrate in the sample was determined by difference. The sum of the percentage moisture, ash, crude fat, crude protein, and crude fiber was subtracted from 10012.

% Total carbohydrate = 100 – (% Moisture + % Ash + % Fat + % Protein + % Fibre).

Kjeldhal apparatus was used for the estimation of nitrogen content in the leaves and protein content was calculated as N × 6.25.

The mineral element composition was determined from the obtained ash during proximate analysis which was digested and dissolved with dilute hydrochloric acid solution using Atomic Absorption Spectrophotometer (Buck Scientific, East Norwalk, CT, USA) and Flame Photometer (FP 202 PG).

The antinutrient analyses of phytate were done according to the Wheeler and Ferrel method13, oxalate by the procedure of Day and Underwood14 while the method of AOAC11 was used to determine the total cyanide content.

Phytochemical and in vitro antioxidant analyses of Dryopteris expansa, Newbouldia laevis, and Senna hirsuta samples

Preparation of plant extract:

 20 g of each plant leaf was weighed and blended in 100 mL of distilled water and filtered to obtain the aqueous solution which was used for the determination of the various parameters.

Phytochemical screening:

The qualitative phytochemical screening of [flavonoids, saponins, alkaloids, phlobatannins, and reducing sugar] was conducted using the methods described by Sofowora15 and Trease & Evans16 to identify the active constituents.

 Acute toxicity investigation:

Various concentrations of the extracts were prepared at 200, 400, 1000, and 2000 mg/kg body weight for each group of five mice weighing an average of 19.30 g respectively. A 0.2 mL each of extract of the plants was administered orally while the control group received 0.2 mL distilled water using an oral gavage feeding needle. The general, physical observations started after the mice were given the dosage, records were taken at intervals for 24 hours, physical observations like shivering, appetite, mortality were made. The mortality or otherwise of mice showed how toxic or nontoxic the leaves extract of the plants could be.

Determination of DPPH free radical scavenging ability:

The in vitro antioxidant analysis of 1,1- diphenyl-2-picryhydrazyl (DPPH) free radical scavenging ability; nitric oxide (NO) radical scavenging ability; ferric reducing antioxidant power; iron chelation, total phenol, flavonoids contents, and the antimicrobial abilities in the three leaves aqueous extracts were evaluated as described below.

The 1,1- diphenyl-2-picryhydrazyl (DPPH) free radical scavenging ability of the extract was determined using the modified method of Gyamfi et al17.

Briefly, 1.0 mL of different concentrations (20, 40, and 80 mg/mL) of the extracts were placed in respective test tubes. A 1.0 mL of 0.1 mM methanolic DPPH solution was added to the samples. These samples were vortexed and incubated in dark at room temperature for 30 minutes. The respective solutions were thoroughly mixed and incubated in the dark for 30 minutes before measuring absorbance at 516 nm. Decreased absorbance of the sample indicated DPPH free radical scavenging capability.

Distilled water was replaced for the extract in the control. Percentage radical scavenging ability was calculated using the following expression:

% DPPH radical scavenging ability    = 1  –  Abs Sample /    Abs Control   X      100

Determination of Nitric Oxide (NO) radical scavenging ability:

The modified methods of Baliga et al18 were used to determine the nitric oxide radical scavenging ability. Sodium nitroprusside in aqueous solution at physiological pH 7.0 spontaneously generated NO, which interacted with oxygen to produce nitrite ions that were estimated by the use of Greiss reagent [1.0 mL sulfanilic acid reagent (0.33%) prepared in 20% glacial acetic acid at room temperature for 5 minutes with 1 mL of naphthyl ethylenediamine dichloride (0.1% w/v)].

The reaction mixture contained 2.0 mL of 5mM sodium nitroprusside in phosphate-saline solution, 0.2 ml of the extract, and 0.5 mL of the Greiss reagent. The absorbance of the blank, test, and control solutions were measured at 546 nm with a spectrophotometer and radical scavenging ability was determined.

Nitric oxide radical scavenging ability = (AbsControl – AbsSample /AbsControl) x 100.

 Determination of ferric reducing antioxidant power:

The reducing property of the three aqueous extracts of the leaves was determined by the modified method of Pulido et al19. This method is based on the reduction of (Fe3+) ferricyanide in stoichiometric excess relative to the antioxidants. Different concentrations of the aqueous extract of the sample and its various fractions (10-50 μg/mL) were added to 1.0 mL of 200mM of sodium phosphate buffer pH 6.6 and 1.0 mL of 1% potassium ferricyanide.         [K3Fe (CN)6].

The mixture was incubated at 50 oC for 20 min, thereafter 1.0 mL of freshly prepared 10% TCA was quickly added and centrifuged at 2000 rpm for 10 min, 1.0 mL of the supernatant was mixed with 1.0 mL of distilled water and 0.25 mL of 0.1% of FeCl3 solution was added. Distilled water was used for blank without the test sample while the control solution contained all other reagents except the 0.1% potassium ferricyanide. Absorbances of these mixtures were measured at 700 nm using a spectrophotometer. Decreased absorbance indicated ferric reducing power capability of the sample.

The percentage ferric reducing antioxidant power (%) was subsequently calculated =

[(Abscontrol – Abssample) /Abscontrol] x 100.

Determination of iron chelation antioxidant power:

The ability of the three aqueous extracts of the leaves to chelate Fe2+ was determined using a modified method of Minotti and Aust20 and Puntel et al21. Freshly prepared 500 ?M FeSO4 (150 ?L) was added to a reaction mixture containing 168 ?L of 0.1 M Tris-HCl (pH 7.4), 218 ?L saline and different concentrations of the extracts (0 –25 ?L). The reaction mixture was incubated for 5 min, before the addition of 13 ?L of 0.25% 1, 10-phenanthroline (w/v). The absorbance was subsequently measured at 510 nm in a spectrophotometer.

The percentage of Fe (II) chelating ability was subsequently calculated concerning the control (which contained all the reagents without the test sample).

The % Fe2+ chelation ability =  (Abscontrol – Abssample) /Abscontrol) x 100

 Estimation of Total Phenolic Content:

The extractable phenol content was determined on the three aqueous extracts using the method reported by Singleton et al22. 0.2 mL of the extract was mixed with 1.5 mL of 10% Folin ciocalteau’s reagent and 2 mL of 7.5% sodium carbonate. The reaction mixture was subsequently incubated at 45 oC for 40 mins, and the absorbance was measured at 700 nm in the spectrophotometer, garlic acid was used as standard phenol.

The concentration of total phenolic compounds in the extract was determined by using the formula:

Total phenolic contents were expressed in terms of gallic acid equivalent, GAE (standard curve equation:

Y = 0.005x + 0.464 (R2=0.961) mg of GA/mg of dry extract.

 Determination of total flavonoid:

 The total flavonoid content of the three aqueous extracts was determined using a colorimeter assay method developed by Bao23. A 0.2 mL of the extract was added to 0.3 mL of 5% NaNO3 at zero time. After 5 min, 0.6mL of 10% AlCl3 was added and after 6 mins, 2 mL of 1M NaOH solution was added to the mixture followed by the addition of 2.1 mL of distilled water. Absorbance was read at 510 nm against the reagent blank and flavonoid content was expressed as mg gallic acid equivalent:

Y = 0.005x + 0.464 (R2 = 0.961) mg of GAE/mg of dry extract

 Antimicrobial analysis of the samples:

 The antimicrobial ability of the three aqueous crude extracts was determined using selected bacteria and fungi were grown in nutrient agar (28 g of nutrient agar in 1000 mL of distilled water and autoclave at temperature 121 ºC for 15 minutes) and potato dextrose agar (39 g of potato dextrose agar in 1000 mL of distilled water and autoclave at temperature 121º C for 15 minutes) respectively. The respective agar was brought out of the autoclave to cool a bit before pouring about 15 mL of the agar into the petri dish that was allowed to gel.

Absorbing the extract into the disc absorbent:

2 mL of each extract was placed into respective sterile evaporating dishes and a few adsorbents discs were poured into the evaporating dishes for the extract to be adsorbed.

Introducing the organism into the media in the petri dish:

About 0.1 mL from the slant of the pure organism was introduced with an inoculating loop by streaking into the petri dish. Afterward, the extract adsorbed discs were introduced into the respective agar plates and incubated at 37 ºC for 18 to 24 hours for bacteria in nutrient agar and fungi in potato dextrose agar at 26 ºC to 28 ºC for 18 to 24 hours.

Standard antibiotics for both gram-positive and gram-negative bacteria (PEF = Pefloxacin, GN = Gentamycin, APX = Ampliclox, Z = Zinnacef, AM = Amoxicillin, R = Rocephin, CP/CPX = Ciprofloxacin, STN = Streptomycin, SXT = Septrin, E = Erythromycin and PEF = Pefloxian, CN = Gentamicin, CH = Chloramphenicol, AU = Augmentin were also used. The plates were observed for antimicrobial activity as presented in Tables 11.0a-c. The results obtained from the various analyses were expressed as Mean ± standard deviation of three determinations.

 Results and Discussion

The proximate composition of the leaves of the plants was investigated to estimate the crude fiber, crude protein, ash contents, crude fat, and carbohydrate contents as observed in Table 1.0.

Table 1 Proximate composition of the three leaves
Sample Moisture

Content %


Fibre %


Protein %


Content %


Lipid %





Dryopteris expansa  














Newbouldia laevis 29.92±0.20 26.30±0.15 6.52±0.07 2.12±0.05 16.78±0.03 18.35±0.08 99.99±0.10
Senna hirsuta 24.38±0.16 7.04±0.08 13.86±0.57 2.56±0.12 12.32±0.21 39.82±0.10 99.98±0.21

CHO = Carbohydrate

From the results obtained, the aqueous extracts of the leaves of the three plants showed a relatively high percentage of crude fibers, proteins, fats, and ash in different proportions. The importance of these parameters in plants cannot be overemphasized for use by humans and animals alike. Many plants of nutritional and medicinal importance have been reported previously by many researchers (24)

The Newbouldia laevis leaves contained the highest percentage of crude fiber (26.30%) among the three plants studied which is also far higher than that reported by Iqbal et al (24) for leaves of three varieties of mulberry (8.17 to 12.32). The three studied plants contained a higher percentage of crude lipid (12.32 to 19.32) which were far higher but of lower protein (6.52 to 13.86) than what was reported for the three varieties of mulberry. Other plants of nutritional importance have also been studied by other researchers to contain comparable proximate constituents as observed in the studied plants(25-27)

The composition of the elemental minerals in the studied plants is reported in Table 2.0. The plants contained essential minerals like iron, copper, zinc, and manganese which play important roles in biological systems.

Table 2.0: Mineral elements composition of the leaves of the plants in parts per million (ppm)
Samples Na K Ca Cu Cd Co Cr Fe Mn Pb Zn Se
Dryopteris expansa 19.30 41.40 18.33 0.25 0.015 ND 0.015 0.23 0.016 ND 0.38 ND
Newbouldia laevis 16.40 34.70 12.94 0.12 0.016 0.004 ND 0.38 ND ND 0.20 ND
Senna hirsuta 12.70 30.60 10.48 0.04 ND ND ND 0.29 ND ND 0.16 ND

ND = Not detected

The results in the table showed the presence of Na, K, and Ca as macro-elements. Zn, Cu, and Fe were also obtained as micro-elements while Se and Pb were not detectable in the three studied plants. Co, Cr, and Mn were scantly detected in the three plants as reported in Table 2.0. These minerals are inorganic chemical elements supplied to the body in varying amounts and required in either macro or micro concentrations from external sources as they cannot be synthesized within the body.

The three studied leaves contained these elements in varying amounts with Dryopteris expansa containing the highest concentration of Na, K, Ca, Cu, and Zn with Cr and Mn not detected in the other two leaves. Co was detected in Newbouldia laevis which was not present in the other two studied plants. This plant also contained the highest amount of Fe when compared with the other two plants.

Mineral elements have been reported previously by various researchers from the studied plants and animal sources products28. Arika et al.29 also reported the presence of K, Ca, Mn, Fe, Zn, Cr, Cu, Pb, and other elements in five plants traditionally used in the management of diabetes mellitus in Kenya.

Antinutrients in plant foods are responsible for deleterious effects related to the absorption of nutrients and micronutrients. For example, phytic acid, oxalate, cyanide, lectins, tannins, saponins, amylase inhibitors, and protease inhibitors have been shown to reduce the bioavailability of nutrients and cause growth inhibition. Some of the antinutrients studied in this work are shown in Table 3.0

 Table 3.0: Antinutrient composition of the leaves
Sample Cyanide content


Oxalate content


Phytate content (mg/100g)
Dryopteris expansa    5.74±0.01 0.28±0.01 0.20±0.01
Newbouldia laevis    8.15±0.01 0.43±0.01 0.19±0.01
Senna hirsuta    8.11±0.01 3.07±0.01 0.21±0.02

The three plants studied contained cyanide, oxalate, and phytic acid in varying amounts, though Newbouldia laevis contained the highest amount of cyanide (8.15 mg/100 g), Senna hirsuta (8.11 mg/100 g), and Dryopteris expansa (5.74 mg/100 g). Table 3.0 also revealed oxalate and phytate contents of Senna hirsuta to be higher than in others.

The results obtained for the antinutrient in this study were comparable to what was reported by Ogunka-Nnoka and Mepba27 on antinutrients content of some raw common spices in Nigeria. It has also been reported that plant processing like boiling, abrasion, and dehulling reduced antinutrients content of plants27,30,31.

Some basic phytochemicals of importance like flavonoids, phlobatannins, alkaloids, saponins, and reducing sugars were screened in the three plants as represented in Table 4.0. The presence of these and other phytochemicals in plants play prominent roles in their medicinal usefulness.

Table 4.0: Some phytochemical composition of aqueous extract of the leaves
Plant species Flavonoids Phlobatannins Alkaloids Saponins Reducing Sugars
Dryopteris expansa ++ ++ ++ +
Newbouldia laevis ++ ++ +
Senna hirsuta ++ + + +

 Present in the aqueous extracts of the three plants were flavonoids, saponins, and reducing sugars. Whereas only Newbouldia laevis aqueous extract did not contain alkaloids, phlobatannins were not present in any of the three plants’ leaves aqueous extract. The results obtained were similar to what was reported by Ekpa and Sani32 in their study on the phytochemical contents of some commonly consumed fruits in Nigeria.

Oral administration of each of the extracts did not produce any clinical signs of toxicity or deaths of the mice as seen in Table 5.0 except in Senna hirsuta at 2000 mg/kg body weight where one of the mice in a group of five died as there were no mortality and clinical signs of toxicity in each of the other tested doses.

Table 5.0: Acute toxicity study of the aqueous extract of the three leaves 
mg/kg body weight dosage of each plant extract after 48 body weight dosage of each plant extract after 48 body weight dosage of each plant extract after 48 hrs.2,000 mg/kg body weight dosage after 48 hours2,000 mg/kg body weight dosage after 48 hours2,000 mg/kg body weight dosage after 48 hours
Shivering ++++---
Cuddling together+++++--
Water consumption+++++++
Food intake+++++++
Sound response+++++++
Skin color+++++++
Gripping strength+++++++
Mortality++++++ - 1/5

+ = Normal;  − = Absent.

In this study, no morbidity or mortality was observed in many mice in the majority of the groups, which showed survival throughout the 48 hours of observation. This finding, therefore, suggests that the extract at the limit dose tested is essentially non-toxic and safe after oral administration. Similarly, no major differences occured in food and water intakes as observed between control and treated groups during this period.

Although acute toxicity helps to determine the toxicity nature of substances, it is important to determine the food intake and water consumption during the investigation of the safety of a product or extract with medicinal purpose, as proper intake of supplements is important to the physiological status of the animal and the achievement of best possible response to the compounds under investigation.

In this study, the food intake and water consumption were not affected by the administration of the extracts because they did not promote any appetite suppression and harmful effects. There were no serious differences in body weight gain between control and treated groups, hence it could be concluded that the extracts of plants under investigation at each dose were almost non-toxic.

The results observed however showed some resemblance in observations to those reported in earlier studies by Ali et al44 in the acute toxicity, brine shrimp cytotoxicity, and relaxant activity of fruits of Callistemon citrinus Curtis; Ndarubu-Tsado et al45 in acute toxicity studies and anti-plasmodial potentials of Newbouldia laevis and Crateva adansonii in Plasmodium berghei-infected mice as well as Roy et al46 in acute and sub-acute toxicity studies on the effect of Senna alata in Swiss Albino mice.

Flavonoids are a group of naturally occurring polyphenolic compounds ubiquitously

found in plants and are increasingly thought to be responsible for the longer life expectancy of populations with well-balanced and healthy diets. The flavonoids contents of the three studied plants showed increased amounts in mg quercetin equivalents/g as the concentration of the extract increased as shown in Table 6.0.

Table 6.0: Total flavonoids contents of the leaves (mg quercetin equivalents/g)
SAMPLE 20 mg (QE/g)   40 mg (QE/g)    80 mg (QE/g)
Dryopteris expansa 36.30±0.30 74.40±0.40    87.20±0.60
Newbouldia laevis 48.50±0.50 54.50±0.30    57.20±1.60
Senna hirsute 35.10±0.30 40.60±0.60    49.30±0.10

 Newbouldia laevis (48.50 mg/g) showed highest content of flavonoids than the other two studied plants, closely followed by Dryopteris expansa (36.30 mg/g) and Senna hirsuta (35.10 mg/g) quercetin equivalents in that order. The flavonoids contents in these plants were observed higher than what Di Carlo et al33 reported in antioxidant activity of some selected traditional Indian medicinal plants for Cyperus rotundus and Vitex negundo.

In humans, phenolic compounds have been reported to exhibit a wide range of biological effects including anti-bacterial, anti-inflammatory, anti-tumour, anti-mutagenic and antioxidant properties. The higher the phenolic content of any plant, the more powerful antioxidant property it contained. The total phenolic contents of the studied plants are expressed in Table 7.0, with Dryopteris expansa having highest phenolic content among the three plants.

 Table 7.0: Total phenol contents of the leaves (mg garlic acid equivalents/g)
SAMPLE 20 mg (GAE/g) 40 mg (GAE/g) 80 mg (GAE/g)
Dryopteris expansa 52.80±0.10 73.10±0.10 80.10±0.30
Newbouldia laevis 36.60±0.06 36.60±0.10 41.50±0.10
Senna hirsute 37.60±0.30 38.00±0.20 55.60±0.10

Table 7.0 also showed an increase in total phenol amount as the concentration of the aqueous extract increased ranging from the highest in Dryopteris expansa to Senna hirsuta and Newbouldia laevis in total phenol contents. The results obtained for the flavonoids in this study for the three plants were higher than those reported by Song et al35 for antioxidant capacities of selected Chinese medicinal plants as also corroborated by Umaru et al34.

Scavenging ability of 1,1- diphenyl-2-picryhydrazyl (DPPH) free radical assay is one of the most commonly employed methods for determining antioxidant capacity. The DPPH assay measures the ability of a compound to scavenge DPPH radicals and is widely used to screen antioxidant activity of different plants, fruits, and vegetables. Table 8.0 reveals the DPPH radical scavenging ability of the studied plants.

 Table 8.0: DPPH radical scavenging ability of aqueous extract of the leaves (%)
SAMPLE 20 mg 40 mg 80 mg
Dryopteris expansa 38.70±0.00 85.2± 0.00 97.10±0.00
Newbouldia laevis 13.00±0.24 58.60±0.00 69.60±0.00
Senna hirsuta 8.70±0.00 34.10±0.00 48.40±0.00

Among the three plants, Dryopteris expansa possessed the highest radical scavenging ability followed by Newbouldia laevis and Senna hirsuta in that order; though, all the plants showed increased scavenging ability as the concentration of the extract increased. Miliauskas et al36 also reported similar observations in their study on the radical scavenging activity of some medicinal plants extracts.

Another measure of antioxidant capacity of any plant is the ability of any of its extract solutions to reduce ferric (III) ions to ferrous (II) ions. The percentage ferric reducing antioxidant power (FRAP) of the three plants studied is seen in Table 9.0

Table 9.0: Ferric reducing antioxidant power of aqueous extract of the leaves (%)
SAMPLE 20 mg 40 mg 80 mg
Dryopteris expansa 33.71±0.00 34.42±0.00  39.28±0.00
Newbouldia laevis   76.71±0.00 93.07±0.07   98.42±0.42
Senna hirsuta  61.00±0.00 75.78±0.21 85.35±0.35

The results obtained in this study showed Newbouldia laevis with the highest ferric reducing antioxidant power followed by Senna hirsuta and Dryopteris expansa in that order. A concentration-dependent increase in ferric reducing antioxidant power was also observed. The results obtained in this study for the three plants showed resemblance to what was obtained in earlier studies reported by Koksal et al37 in antioxidant activity of Melissa officinalis leaves and Song et al35 in antioxidant capacities of selected Chinese medicinal plants.

The primary goal of iron chelation therapy is to prevent the accumulation of iron from reaching harmful levels in the body as the transition metal ion Fe2+ possesses the ability to perpetuate the formation of free radicals by the gain or loss of electrons. Table 10.0 shows the percentage iron II chelation ability of the three studied plants.

Table 10. % Fe2+ Chelation antioxidant potential of aqueous extract of the leaves
SAMPLE 20mg 40mg 80mg
Dryopteris expansa 50.00±7.00 79.00±1.00 88.00±5.00
Newbouldia laevis 71.00±1.00 84.00±2.00 95.00±0.00
Senna hirsute 76.00±0.00  89.00±3.00 98.00±1.00

The results of the study showed that the three plant aqueous extracts are good chelators for the Fe2+. Senna hirsuta, Newbouldia laevis, and Dryopteris expansa showed chelation with the highest activity to the lowest ranging from 76.00 to 50.00% in that order. The iron chelation power also showed dependence on extract concentration. The results of the study corroborated earlier studies reported by Fai-Chin et al38 in their work on antioxidant, metal chelating, anti-glucosidase activities, and phytochemical analysis of selected tropical medicinal plants.    

The anti-bacterial and anti-fungi potentials of the three aqueous extracts of the plants were carried out using Gram-positive (Staphylococcus aureus and Streptococcus pyrogenes) and Gram-negative (Escherichia coli) bacteria and two species of fungi to assess the possibilities of using the aqueous extracts as antimicrobial agents to suppress bacterial and fungi diseases as seen in Tables 11.0a-c.

The results for the extracts were compared with the standard gram positive and gram-negative antibiotic discs on Streptococcus pyrogenes, Staphylococcus aureus, and Escherichia coli growths. The results showed that, while Streptococcus pyrogenes was susceptible to Dryopteris expansa aqueous extract but resistant to Newbouldia laevis and Senna hirsuta extracts Staphylococcus aureus and Escherichia coli showed resistance to the three extracts studied. The three bacteria also showed susceptibility and resistance to the various standard antibiotics used as obtained in Tables 10.0a-b.

Sample Organisms Plant


Dryopteris expansa Staphyloccus aureus R S S S R R S S S S S
Streptococcus pyrogenes S R S R R R R R R R R
Newbouldia laevis Staphyloccus aureus R S S S R R S S S S S
Streptococcus pyrogenes R R S R R R R R R R R
Senna hirsute Staphyloccus aureus R S S S R R S S S S S
Streptococcus pyrogenes R R S R R R R R R R R
Table 11.0a: Antibacterial activity of the aqueous extract of the leaves on Gram-positive bacteria

S = Susceptible/Sensitive, R = Resistant, PEF = Pefloxacin, GN = Gentamycin, APX = Ampliclox, Z = Zinnacef, AM = Amoxicillin, R = Rocephin, CP = Cirpofloxacin, STN = Streptomycin, SXT = Septrin, E = Erythromycin

Though the antibacterial activities of many Dryopteris species have been reported, the literature was scanty on the antimicrobial activity of Dryopteris expansa which was observed in this study to be active against Streptococcus pyrogenes. Other Scientists (Femi-Adepoju et al39 and Mandal and Mondal40 have also reported the antimicrobial activity of ethanolic, methanolic, and acetone extracts of Dryopteris filix-mas.

Usman and Osuji41 also reported the antimicrobial activity of methanolic extract of Newbouldia laevis through the results obtained in this work showed that all the microorganisms studied were resistant to the aqueous extract of Newbouldia laevis. Adedayo et al42 and Idu et al43 also reported the antimicrobial activity of Senna alata in various solvents while Escherichia coli, Staphylococcus aureus, and Streptococcus pyrogenes were resistant to the aqueous extract of Senna alata as observed in this work.

Table 11.0b: Antibacterial ability of the aqueous extract of the leaves on Gram-negative bacteria
Samples Organism Antibiotics
Dryopteris expansa Escherichia coli R R R R R R R S R R
Newbouldia laevis Escherichia coli R R R R R R R S R R
Senna hirsute Escherichia coli R R R R R R R S R R

Antimicrobial activity of the extracts on the organism:  S = Susceptible; R= Resistant, PEF = Pefloxacin, GN = Gentamycin, CH = Chloramphenicol, AU = Augmentin, AM = Amoxicillin, OFX = Tarivid, CPX = Ciprofloxacin, SP = Sparfloxacin, SXT = Septrin

In this study, apart from Streptococcus pyrogenes that were susceptible to Dryopteris expansa aqueous extracts, all the other studied organisms were resistant to the three plants’ extracts. This observation was similar to what was observed for the various standard antibiotics except for gentamycin. Staphylococcus aureus was susceptible to pefloxacin, gentamycin, ampiclox, Rocephin, ciprofloxacin, streptomycin, Septrin, and erythromycin except for zinnacef and amoxicillin.

The results of anti-fungal ability of the three aqueous extracts on Aspergillus candidius and Penicillium chrysogenum in Table11.0c showed that the organisms were resistant to the three extracts of the plants.

Table 11.0c: Antimicrobial ability of the aqueous extract of the leaves on selected fungi
Samples Organism Plant Extract
Dryopteris expansa Aspergillus candidius Resistant
Penicillium chrysogenum Resistant
Newbouldia laevis Aspergillus candidius Resistant
Penicillium chrysogenum Resistant
Senna hirsuta Aspergillus candidius Resistant
Penicillium chrysogenum Resistant

However, the resistance of these organisms to the three extracts might not nullify the susceptibility of other organisms to the extracts.  Besides, other solvents’ extracts of the same plants might cause susceptibility of these same organisms to the other solvents’ extracts.

Conclusion and Recommendations

The study of the three plants’ aqueous extracts; Dryopteris expansa, Newbouldia laevis, and Senna hirsuta revealed the good status of the nutritional, elemental, phytochemical compositions, safe dosage, antioxidant potentials, and antimicrobial abilities of the plants leaves.

The results obtained therefore showed the traditional medicinal claims of these plants’ leaves and their possible applications in drugs development. It is however recommended that these plants species be domesticated and preserved for the use and further investigations embarked for structural elucidation of the bioactive compounds.

Ethics approval and consent to participate

The study was carried out under strict laboratory precautions and standard processes to arrive at the results obtained. There was no experimental animal used in the course of this study.

 Consent for publication

The authors hereby declare that the work presented in this article is original and that any liability for claims relating to the content of this article will be borne by them. We also give our consents for the manuscript publication.

Availability of supporting data

All the data and materials for the study are available in the authors’ possession.

 Competing interests

The authors declare no conflict or competing interests exist.


This research project was not sponsored by any grant or organisation. It was self-funded work.

 Author contributions:

All the authors read and approved the manuscript for submission.  Author OAO designed, executed, and wrote the draft manuscript. Authors FDI, ABO, and AID were involved in the laboratory analyses.


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Anti-microbial, In vitro Antioxidant, and other Biochemical properties of Dryopteris expansa, Newbouldia laevis, and Senna hirsuta. from Anchor University, Lagos

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