INTRODUCTION:

The COVID-19 pandemic that stemmed from the advent of novel Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), has led to consequential mortality morbidity on a global scale¹. The disease presents with various symptoms like dry cough, headache, fever, fatigue, chest pain, sore throat, shortness of breath and muscle pain among others². These symptoms can progress to severe respiratory insufficiency and adversely affect critical organs such as kidney, heart, nervous system and the liver and eventually leading to the demise death of patients, especially the ones with pre-existing health conditions³. As of July 2023, the number of deaths caused by COVID-19 stood at over 6.9 million people worldwide with Africa having over 200,000 deaths⁴. The global pandemic has had profound impacts on societies, economies, and health. Despite the current strong push for vaccination, many among the population are still hesitant to take the vaccines⁵. Additionally, despite the improvement in the dissemination of quality information from medical and scientific source, the pandemic continues to generate feelings of anxiety, fear, uncertainty, and stigmatization due to numerous narratives that have been circulated in social media as well the traditional media platforms^6,7. Furthermore, despite the availability of vaccines and the mobilization by governments to get their populations vaccinated, the continuous appearance of new strains and mutations of the SARS-CoV-2 has resulted into a continued focus on finding effective pharmacological agents that would effectively manage and control the spread and infection rates of the virus^8,9. The pandemic prompted numerous communities across the globe to search for palliative methods aimed at either preventing or easing the disease's symptoms. In Africa, for example, the fear of the disease resulted into numerous communities resorting to self-care, self-help and self- administered remedies⁸. Numerous communities utilized medicinal plants and various self-care solutions to prevent the infection or alleviate symptoms^10,11. In Africa and elsewhere across the globe, medicinal plants usage and recognition as an integrative approach in the control and management of various infections is deeply ingrained within cultural, religious and traditional customs^12,13. However, this dependence on herbal medicine rose even more during the COVID-19 pandemic due to absence of accurate and efficient treatments for SARSCoV-2 hence, the shift towards exploring plant-based methods and approaches for potential herbal medicines to combat COVID-19¹⁴. This was not different in Kenya and other African countries as steaming, herbal decoctions, herbal concoctions and infusions were predominantly used as the first line of defence in the fight against COVID-19¹⁵. Most of the plants used in these formulations were those with historical uses in management of common flu, influenza and other respiratory ailments and are known to contain bioactive ingredients that possess anti-oxidant, immunomodulatory, anti-inflammatory, and also antimicrobial activities¹⁴. In Kajiado County, in the Southern region of Kenya, there was a notable herbal formulation that was purported to have the ability to manage COVID-19. Some of the plants used to make the formulation as revealed by the source herbalist included Zingiber officinale, Curcuma longa, Croton dichogamus, Salvadora persica, Osyris compressa and Eucalyptus globulus. From previous studies, most of these plants have potentially strong anti-microbial and anti-oxidant activities and therefore their combination in a formulation could provide an avenue for new anti-microbial drugs in the wake of the rise in multi-drug resistant strains of microbes recently ignited by the unnecessary use of anti-biotics to control and manage the COVID-19 pandemic^16,17. The available evidence indicates that the COVID-19 pandemic increased the rate of anti-microbial resistance (AMR) due to the misuse and overuse of anti-biotics¹⁸. Furthermore, there has been a dearth in the development of new antibiotics which is referred to as “discovery void” which is primarily due to the scarcity of research to find new anti-microbials¹⁹. The outbreak of COVID-19 further exacerbated this situation by disrupting the production, distribution, and research related to antimicrobials. The pandemic, shifted the focus of the research community and resources toward finding new antivirals and vaccines treatments for COVID-19 instead of anti-biotics^18,20. This deficiency of specific antibiotics can lead to an increase in antimicrobial resistance, further complicating their use and the control of infectious diseases²¹. Consequently, there is an urgent demand for new lead compounds with novel modes of action against both of the Gram-positive and Gram-negative bacteria to combat the drug resistance challenges²². In this study, we aimed at assessing the chemical composition, anti-bacterial and anti-fungal properties, antioxidant potential, and cytotoxic effects of crude extracts of a herbal formulation now registered as Covidak⁴ that’s proposed for potential use in COVID-19 management.

MATERIALS AND METHODS:

Sample collection:

The study sample was a raw plant formulation consisting of various plants collected in February 2021 from a herbalist in Loodoariak, Kajiado County, Southern Kenya. This location lies at a latitude of 1°05'8.57" South, a longitude of 36°46'36.59" East, and an altitude of 1,189 meters above sea level.

Solvent extraction:

The plant powdered formulation (500g) was extracted using cold maceration with n-hexane, ethyl acetate, and methanol in increasing order of polarity at a 1:10 ratio. The extracts were concentrated using a rotary evaporator, dried at room temperature, and stored in a refrigerator for later use^23,24.

Qualitative Phytochemical Screening:

Screening for the presence of different phytochemicals was done using standard phytochemical procedures described by Faraq et al²⁵.

Quantitative Determination of Phytochemicals:

a) Estimation of Alkaloids:

2.5g of each extract was mixed with 200mL acetic acid in ethanol, left for 4hours, concentrated, filtered, and treated with ammonium hydroxide to complete precipitation. The solutions were washed, filtered, and weighed. This process was repeated three times²⁶.

b) Estimation of Total Saponins:

Experiment was carried out by the method described by Harrizul Rivai et al²⁷ was applied with modifications from Pai VR, Hegde²⁸. 2.5g of sample was heated with 100mL of 20% ethanol for 4hours at 55°C. The residue was filtered, re-extracted, and evaporated to 40mL. Diethyl ether was added, the aqueous layer was separated, and n-butanol was extracted twice with 5% sodium chloride. The solution was heated, dried, and recorded. This was done in triplicate.

c) Estimation of Terpenoids

Terpenoids were determined gravimetrically using a modified method from Oncho D.A et al²⁹. 1.5g of sample was soaked in 100mL ethanol for 24hours, filtered, and the filtrate was extracted with 100mL petroleum ether. The extract was dried, weighed, and the process repeated three times.

d) Estimation of Total Phenolic Contents:

Total phenolic content was measured using the Folin-Ciocalteu method. 50µL of extracts were mixed with 0.5mL Folin-Ciocalteu reagent and 2.5mL sodium carbonate, vortexed, and incubated for 40minutes. Absorbance was measured at 725nm using gallic acid as the standard. Results were calculated in mg of gallic acid equivalents³⁰.

e) Estimation of Flavonoids

Flavonoid content was analysed using rutin as the standard substance³¹. 500µL of extract was mixed with sodium nitrite, aluminium chloride, and sodium hydroxide, with water added to make 5µL. The mixture was vortexed, incubated for 15 minutes, and absorbance measured at 510nm. A standard curve with rutin was used to calculate flavonoid content as mg rutin equivalents per gram of sample.

Brine Shrimp Lethality Bioassay:

The brine shrimp lethality bioassay was employed to evaluate the cytotoxicity of the herbal formulation's n-hexane, ethyl acetate, and methanolic extracts³². Each extract and the positive control (vincristine sulfate) were dissolved in DMSO to prepare stock solutions (1000 mg/mL)³³. 10-fold serial dilutions were made to obtain varying concentrations (1000µg/mL to 0.01µg/mL) using distilled water. These solutions were added to vials containing 10 live brine shrimp nauplii each, and after 24 hours, the number of surviving nauplii was counted. Mortality was assessed based on the absence of controlled movement for 30 seconds. Lethality concentrations (LC50) were determined using probit analysis by plotting percentage mortality against concentration³⁴.

Anti-Oxidant Assay:

The free radical scavenging activity of the herbal formulation's extracts (methanol, ethyl acetate, and n-hexane) was assessed using the DPPH assay, with ascorbic acid as the positive control and methanol as the negative control³⁵. Stock solutions (1mg/mL) were prepared for each extract and standard, followed by 2-fold serial dilutions. A 1 mM DPPH solution was prepared and mixed with the extracts, then incubated in the dark for 15minutes. Absorbance was measured at 517nm using a UV spectrophotometer, and radical scavenging activity was calculated using the formula 1. The lower absorbance of the reaction mixture the higher free radical scavenging activity³⁶.

(Abs control- Abs sample)

% of radical = ---------------------------------------- x 100

scavenging activity (Abs control)

Equation 1:

IC50 values, representing the concentration needed to inhibit 50% of DPPH radicals, were obtained from the plotted graph of % radical scavenging activity versus extract concentration.

Anti-microbial Assay:

Bacteria Test organisms (gram-negative bacteria: Pseudomonas aeruginosa, Escherichia coli, Salmonella typhi; gram-positive bacteria: Staphylococcus aureus, Bacillus subtilis, Streptococcus mutans; and fungus: Candida albicans) were cultured in appropriate media (Muller Hinton Agar for bacteria, Potato Dextrose Agar for fungi), and standardized suspensions were prepared. Extracts were dissolved in 99.9% pure dimethyl Sulfoxide DMSO to a concentration of 1mg/mL. Using the disc diffusion method, extracts and standards (Ceftriaxone, Gentamicin, Fluconazole) were applied to inoculated agar plates. Plates were incubated at 37°C for bacteria and 27°C for fungi, and the inhibition zones were measured and recorded^37–39. All tests were performed in triplicate, and results were expressed as mean±SD.

Reported Method of Administration of Covidak⁴:

To use Covidak⁴, add half a teaspoon to a half cup of hot water, mix in a pinch of eucalyptus crystals, and stir well. Inhale the resulting fumes through the nose and mouth, then gargle and swallow the warm concoction. This is administered twice a day for three or four days.

Statistical analysis:

Each experiment was conducted in triplicate, with results reported as mean values ±standard error of the mean (SEM)⁴⁰. Data were analyzed using ANOVA and Microsoft Excel 2016.

RESULTS AND DISCUSSION:

Phytochemical composition:

Phytochemical screening:

Medicinal plants contain biologically active compounds with diverse chemical structures that contribute to disease protection⁴¹, making preliminary phytochemical screening essential for identifying potential bioactive components⁴². This study revealed that flavonoids, terpenoids, phenols, alkaloids, steroids, saponins, and quinones were present in all three crude extracts, while glycosides were absent in the n-hexane extract and tannins were only detected in the methanol extract, which likely explains the methanol extract's greater potency due to its ability to extract bioactive compounds more effectively.

Table 1: Qualitative analysis of the phytochemicals in the herbal formulation extracts

Phytochemicals	Hexane extract	Ethyl acetate extract	Methanol extract
Flavonoids	+	+	+
Alkaloids	+	₊	+
Terpenoids	+	+	+
Phenols	+	+	+
Steroids	+	+	+
Saponins	+	+	+
Tannis	_	_	+
Glycosides	_	+	+
Quinones	+	+	+

(+) = Presence, (-) = Absence

When used alone or in combination, phytochemicals are essential in determining the medicinal value of a plant. Saponins, flavonoids, alkaloids, glycosides, phenolics, terpenoids, steroids, and many more represent important botanical compounds, each offering a diverse range of biological roles. The pharmacological activity of a plant can be predicted by the identification of the phytochemicals⁴³. The method of extraction, including the choice of solvent, is a key factor in determining the efficacy of extracting these bioactive compounds from plant materials. Different solvents target specific compounds, influencing the overall yield and effectiveness of the extracts.

Quantitative Analysis:

Based on the qualitative tests, quantitative analysis was conducted using standard methods to measure the concentration of key phytochemicals. The analysis was done in triplicate, and the results were processed using Microsoft Excel.

a) Total Phenols:

The total phenol content was measured using the Folin-Ciocalteu reagent and expressed as milligrams of gallic acid equivalent (GAE) per gram of the sample.

· Ethyl Acetate Extract: 238.51±2.83mg GAE/g dry weight

· Methanol Extract: 167.20±5.32mg GAE/g dry weight

· Hexane Extract: 125.30±8.63mg GAE/g dry weight

Standard Curve Equation:

y = 0.1004x-0.0056,

R² = 0.9316; Figure 1.

Figure 1: Standard Calibration Curve of Gallic acid for Total Phenolic content

Phenols, characterized by a hydroxyl group bonded to an aromatic ring, are known for their diverse biological activities, including anti-microbial, anti-viral, anti-inflammatory, and antioxidant properties⁴⁴.

b) Flavonoids

The flavonoid content was determined in terms of rutin equivalent (RuE) and the result are as shown below:

· n-Hexane Extract: 31.87±2.41mg RuE/g dry weight

· Ethyl Acetate Extract: 197.08±4.17mg RuE/g dry weight

· Methanol Extract: 84.58±2.41mg RuE/g dry weight

Standard Curve Equation: y = 0.0012x+0.0187, R²= 0.9054; (Figure 2).

Figure 2: Standard Calibration Curve of Rutin for Flavonoid content

Flavonoids are secondary metabolites that are predominantly found in vegetables, seeds, nuts, fruits, cereals, stems and herbs with potential health benefits, including antioxidant, anti-bacterial, anti-fungal and anti-inflammatory activities⁴⁵^,⁴⁶. Several flavonoids have been explored to determine their potential as anti-viral agents, with some demonstrating significant antiviral effects in both in-vitro and even in-vivo studies⁴⁶.

c) Saponins:

The highest total saponins content was observed in the methanol extract as shown below:

· Methanol Extract: 95.56±3.85mg DE/g dry weight

· Ethyl Acetate Extract: 57.78±3.85mg DE/g dry weight

· n-Hexane Extract: 48.89±3.85mg DE/g dry weight

Saponins represent a diverse group of chemical compounds that contain a triterpene or steroid aglycone group combined with at least one sugar chain and are abundant in dicotyledonous plants and certain marine animals⁴⁷. They are known for their antimicrobial properties and various biological functions, including hypoglycemic effects, antimicrobial properties, lowering serum cholesterol levels and exhibiting anti-viral and anti-inflammatory activities⁴⁸.

d) Alkaloids:

Quantity of alkaloid was lowest in the methanol extract and highest in the n-hexane extract as shown below, which can be attributed to its low polarity⁴⁹.

· n-Hexane Extract: 171.11±3.85 mg DE/g dry weight

· Ethyl Acetate Extract: 137.78±3.85mg DE/g dry weight

· Methanol Extract: 124.44±3.85mg DE/g dry weight

Alkaloids, characterized by their heterocyclic nitrogen-containing structures, are known for their medicinal properties, including anti-proliferative, antioxidant, anti-viral, anti-bacterial, and anti-cancer effects⁵⁰^,⁵¹.

e) Terpenoids:

Terpenoids, derived from the mevalonic acid pathway, exhibit a wide range of bioactivities, including antimicrobial, anti-cancer, anti-malarial, and anti-inflammatory effects⁵²^,⁵³. Its variation in the extracts is as show below:

· n-Hexane Extract: 77.78±3.85mg DE/g dry weight

· Ethyl Acetate Extract: 100.00±6.67mg DE/g dry weight

· Methanol Extract: 120.00±6.67mg DE/g dry weight

The quantitative analysis indicated that the ethyl acetate extract showed the highest concentration of total phenols and flavonoids, suggesting it has potent antioxidant activity. The methanol extract exhibited the highest saponin content, while the n-hexane extract had the highest alkaloid content. These findings highlight the diverse phytochemical profile of the herbal formulation and its potential for various therapeutic applications.

Brine Shrimp Lethality Bioassay:

The brine shrimp lethality assay, a cost-effective method to evaluate cytotoxicity⁵⁴, revealed that all the herbal formulation crude extracts exhibited toxicity, with LC₅₀ values ranging from 0.01μg/mL (ethyl acetate extract) to 0.545μg/mL (n-hexane extract), indicating high cytotoxicity according to Clarkson’s toxicity scale^55,56. The methanolic extract (LC₅₀ = 0.2287μg/mL) also showed significant toxicity, though all extracts were less toxic compared to the positive control, Vincristine sulphate (LC₅₀ = 0.0000772μg/mL) as shown in table 2.

The difference in toxicity observed among the three crude extracts of the herbal formulation could be attributed to the phytochemical distribution such as alkaloids, phenols, terpenoids, tannins and flavonoids within the extracts, which are influenced by their different polarities^55,57,58.

Anti-oxidant Assay:

The current study used DPPH assay as the exclusively chosen model because of its stability and reproducibility of the results. The basis of the DPPH assay is the capacity of extract or compound to donate a hydrogen atom, which then turns DPPH from a purplish color to a yellowish or colorless state⁵⁹. Table 3 displays the respective data of the DPPH free radical scavenging activity of the crude extracts. IC₅₀ is used to determine the concentration of the sample that is capable of scavenging 50% of the DPPH free radicals and the lower the IC₅₀ value the higher the antioxidant activity of the sample³⁵.

Table 2: Effect of n-hexane, ethyl acetate and methanol extracts of herbal formulation and positive control vincristine sulphate on brine shrimp

Conc. (µg/mL)	% Mortality				LC₅₀(µg/mL)
Conc. (µg/mL)	Hexane	Ethyl acetate	Methanol	Vincristine Sulphate	Hexane	Ethyl acetate	Methanol	Vincristine Sulphate
0.01	20	40	23.33	60	0.545	0.01	0.2287	0.0000772
0.1	30	63.33	33.33	73.33
1	53.33	80	70	93.33
10	80	100	93.33	100
100	100	100	96.67	100
1000	100	100	100	100

Table 3: Anti-oxidant activity [DPPH free radical scavenging] of Ascorbic acid and the three extracts of the herbal formulation

Extract	IC₅₀μg/mL
n-Hexane	48.43±0.39
Ethyl acetate	19.87± 0.34
Methanol	28.23±0.38
Ascorbic acid	11.52±0.08

Table 4: Anti-microbial activity (zone of inhibition in mm) of extracts and controls

Conc. (µg/mL)	*E. coli*	*S aureus*	*P. aeruginosa*	*C. albicans*	*S. typhi*	*B. subtilis*	*S. mutans*
Hexane extract
1000	9.33±0.58	13.00±1.00	8.33±0.58	9.33±0.58	8.33±1.15	9.33±0.58	11.67±0.58
100	8.67±0.58	8.67±0.58	8.00±1.0	9.33±0.58	7.00±0.00	7.67±0.58	7.67±0.58
10	8.33±0.58	7.67±0.58	7.67±0.58	8.33±0.58	7.00±0.00	7.33±0.58	-
1	7.67±0.58	7.33±0.58	7.00±0.00	7.67±1.15	-	-	-
0.1	7.00±0.58	7.00±0.00	-	7.33±0.58	-	-	-
Ethyl acetate extract
1000	8.67±0.58	13.00±1.00	11.00±1.00	12.00±1.00	15.00±1.00	10.67±1.52	12.00±1.00
100	7.33±0.58	11.00±1.73	11.00±1.00	11.00±1.00	10.33±1.52	9.33±0.58	11.67±0.58
10	7.33±0.58	10.00±1.00	9.67±0.58	10.00±1.00	8.67±0.58	8.67±1.15	11.00±1.73
1	-	9.00±1.00	9.33±1.52	10.00±1.00	7.33±0.58	7.67±0.58	10.33±0.58
0.1	-	8.67±0.58	8.67±0.58	8.33±0.58	-	7.33±0.58	8.33± 0.58
Methanol extract
1000	10.33±0.58	14.00±1.00	11.67±1.00	10.33±1.53	12.33±0.58	12.33±1.53	10.00±1.00
100	10.00±1.00	12.67±0.58	11.00±0.58	10.00±1.00	10.67±0.58	10.00±1.00	8.00±1.00
10	8.33±0.58	10.00±1.00	10.00±1.00	9.33±0.58	10.33±1.53	10.00±1.00	7.33 ±0.58
1	8.00±1.00	10.00±1.00	10.00±1.00	7.33±0.58	9.33±0.58	9.67±2.08	7.00±0.58
0.1	7.33±0.58	9.67±1.15	8.67±0.58	7.33±0.58	7.33±0.58	8.33±0.58	-
Positive Controls (Anti-biotics)
CTR	32.67±1.53	12. 33±1.53	23.67±0.58	28.67±0.58	26.67±0.58	16.00±1.00	16.67±0.58
GEN	-	-	19.00±1.00	-	17.00±1.00	-	19.33±0.58
FCZ	-	-	-	27.00±0.00	-	-	-
Negative Control
DMSO	7.00±0.00	7.00±0.00	7.00±0.00	7.00±0.00	7.00±0.00	7.00±0.00	7.00±0.00

KEY: GEN – Gentamicin, CTR – Ceftriaxone, FCZ—Fluconazole, DMSO – Dimethyl Sulfoxide

Among the results, the ethyl acetate extract showed the highest antioxidant activity with an IC₅₀of 19.87±0.34 μg/mL, followed by methanol (28.23±0.38μg/mL) and n-hexane (48.43±0.39μg/mL), while ascorbic acid had an IC₅₀ of 11.52±0.08μg/mL. The strong antioxidant activity of the ethyl acetate extract, potentially due to its high content of alkaloids, glycosides, tannins, and flavonoids, is supported by its higher phenolic and flavonoid content in phytochemical analysis. Flavonoids and phenolics possess considerable anti-oxidant activity ^60,61. These compounds, known for scavenging free radicals, offer protection against oxidative stress and related diseases such as cancer, diabetes, and cardiovascular conditions, and may also help strengthen the immune system, especially relevant in managing COVID-19^62,63.

Anti-microbial Assay:

The results of the antimicrobial assay (Table 4) showed different levels of antimicrobial activity for each extract against various test organisms. The antimicrobial activities of the crude extracts were measured in terms of the diameter of zones of inhibition⁶⁴.

Candida albicans exhibited the highest susceptibility to the ethyl acetate extract, showing a zone of inhibition of 12.00±1.00mm at 1000µg/mL, indicating the presence of bioactive compounds that inhibit fungal growth. These bioactive compounds are likely phenols, flavonoids, and terpenoids, which contribute to the extract's antifungal properties⁶⁵.

The methanolic extract showed the strongest antibacterial activity against E. coli, S. aureus, P. aeruginosa, and B. subtilis, with zones of inhibition of 10.33±0.58mm, 14.00±1.00mm, 11.67±1.00mm, and 12.33±1.53mm, respectively, at 1000µg/mL. These bacteria are generally known to give rise to a wide range of infections and these results suggest the presence of potent antibacterial compounds, likely secondary metabolites such as saponins, alkaloids, and tannins^66,67. For Salmonella typhi and Streptococcus mutans, the ethyl acetate extract demonstrated the highest antibacterial activity, with inhibition zones of 15.00±1.00mm and 12.00±1.00mm, respectively, indicating its effectiveness against pathogens causing typhoid fever and dental caries⁶⁸. The study found that the antimicrobial activity was concentration-dependent, suggesting potential optimization through dosage control, although further research is required to establish the minimum inhibitory concentration (MIC) to prevent toxic effects^69,70.

The hexane extract exhibited the lowest antimicrobial activity compared to the methanol and ethyl acetate extracts, likely due to hexane being a non-polar solvent, which may not efficiently extract polar antimicrobial compounds. This difference in activity suggests that the formulation’s key bioactive components are polar, such as flavonoids, phenols, anthraquinones, and glycosides, known for their antimicrobial properties⁴⁹. The study indicates that methanol and ethyl acetate extracts are more effective, highlighting the herbal formulation’s potential as a source of novel antibiotics, particularly against multi-drug-resistant organisms that increased during the COVID-19 pandemic^71,72.

CONCLUSION:

In conclusion, the herbal formulation demonstrated significant anti-microbial activity, strong antioxidant potential, and an acceptable cytotoxicity profile compared to the standards used. These findings suggest its potential as a source of novel antibiotics, especially in light of increasing multi-drug-resistant organisms during the COVID-19 pandemic. However, further research is needed to explore its mechanisms of action, optimize dosage, assess safety and efficacy in vivo and clinical trials, and identify the active compounds responsible for its effects.

CONFLICT OF INTEREST:

The authors declare no conflict of interest.

ACKNOWLEDGEMENTS:

We are grateful to Jomo Kenyatta University of Agriculture and Technology for providing facilities and the support accorded to K. F. Tumelei.

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Received on 18.04.2024 Revised on 13.09.2024

Accepted on 16.01.2025 Published on 10.05.2025

Available online from May 14, 2025

Res. J. Pharmacognosy and Phytochem. 2025; 17(2):83-90.

DOI: 10.52711/0975-4385.2025.00014

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

MATERIALS AND METHODS:

Sample collection:

Solvent extraction:

Qualitative Phytochemical Screening:

Quantitative Determination of Phytochemicals:

a) Estimation of Alkaloids:

b) Estimation of Total Saponins:

c) Estimation of Terpenoids

d) Estimation of Total Phenolic Contents:

e) Estimation of Flavonoids

Brine Shrimp Lethality Bioassay:

Anti-Oxidant Assay:

Anti-microbial Assay:

Reported Method of Administration of Covidak4:

Statistical analysis:

RESULTS AND DISCUSSION:

Phytochemical composition:

Phytochemical screening:

Quantitative Analysis:

Brine Shrimp Lethality Bioassay:

Anti-oxidant Assay:

Anti-microbial Assay:

ACKNOWLEDGEMENTS:

Reported Method of Administration of Covidak⁴: