To Evaluate the Antioxidant, Anti-Inflammatory and Antimicrobial Activities of Tridax Procumbens

Tridax Procumbens is medicinal plant commonly known as tridax daisyor kansari (Hindi) or Ghamara (in local language) or Jakhamjudi and Tuntuni (Marathi) belongs to family Asteraceae. It is Found mainly in Maharashtra and Madhya Pradesh as a weed. Plant widely used in Indian traditional medicine system. It has long stalked, yellow comparative flower. In old time Tridax Procumbens has been used in Ayurvedic system in India. By using these plant various creams, oils and skin product like skin poultices are manufactured. Tridax Procumbens also called as Coat Button which is a small, creeping, and fast-growing herb. It is widely found in tropical and subtropical regions, especially in India, Africa, and the Americas. The plant grows easily on roadsides, fields, and wastelands, making it a common and economically important weed with medicinal value. Originally native to Central and South America, it has been recognized and utilized since ancient times. Local communities have traditionally crushed its leaves to apply on fresh wounds and cuts to promote rapid healing. The plant is also used in the treatment of fever, diarrhea, stomach disorders, cough, cold, and hair fall, demonstrating its importance in indigenous healthcare systems. Tridax procumbens is a semi-prostrate or procumbent herb that typically grows 30 to 60 cm tall, with a firm taproot. Its branches are ascending, brittle, green or purplish, and covered with long white hairs. The leaves are opposite, simple, ovate to ovate-lanceolate in shape, typically 2 to 6 cm long and 1.5 to 4.5 cm wide, with coarsely toothed margins and a strongly prominent midrib on the underside. The leaves are often hairy on both surfaces. The plant produces daisy-like flower heads that are terminal and about 1.5 to 2 cm in diameter, borne on long peduncles ranging from 11 to 40 cm. The outer involucral bracts are foliaceous and green, while the inner bracts are membranous and purplish. The flower heads consist of a few female ray florets with pale yellow or white corollas, surrounding numerous bisexual yellow disc florets. The fruit is a dark brown to black achene, 1.6 to 3 mm long, densely covered with white long hairs and topped with a tuft of 15-20 feathered bristles. The plant’s stem is cylindrical, solid, and very hairy with multicellular hairs about 1 mm long. The leaves have a pinnate venation pattern with 2-3 lateral veins on each side of the midrib and a cuneate base. Seedlings have glandular-haired cotyledons and first true leaves that are ovate to lanceolate.

Fig.2 Tridax Proccumbens with flower

2. Plant Profile

Present days, the therapeutic value of medicinal plant is increasing at significant rate for developing new drugs and combatting emerging diseases. Drug developers are targeting new sources of active materials to cope up with multi drug resistance (MDR) of different microorganisms. Tridax Procumbens was targeted in this study as it is widely spreaded as a common weed in Indian subcontinent as well as all over the world Tridax Procumbens is native to many parts of Africa, Asia, America, Australia and some part of Europe. They are given many names depending on the regions. Botanically, Tridax Procumbens is a creeping perennial herb characterized by hairy stems, opposite leaves with serrated margins, and small daisy-like flowers. The plant’s simple morphology often leads to its underestimation; however, its pharmacological significance has been increasingly validated by modern scientific studies. Numerous preclinical studies have reported properties including anti-inflammatory, antimicrobial, antioxidant, hepatoprotective, immunomodulatory, and wound-healing activities, supporting many of its traditional uses. Moreover, the plant contains a rich spectrum of phytocompounds such as flavonoids, alkaloids, tannins, carotenoids, saponins, and terpenoids, which contribute to its biological activities. Notable bioactive compounds like quercetin, luteolin, and β-sitosterol have been identified and are believed to play crucial roles in tissue repair, immune modulation, and oxidative stress reduction.

• Common names

Geographical Source

• Found throughout India, Tropical Africa, Australia, and America.
• Commonly grows as a weed along roadsides, wastelands, and fields.

Naturalized Distribution

It has become widely naturalized in tropical and subtropical regions around the world, including: -

• Asia: India, Sri Lanka, Bangladesh, Nepal, Thailand, Malaysia, Indonesia, the Philippines
• Africa: Widely spread across tropical and subtropical Africa
• Australia and Pacific Islands
• Southern United States: Especially in Florida, Texas, and Hawaii

Botanical Classification

OBJECTIVES: -

1. Antioxidant Activity - To evaluate the antioxidant potential of the selected sample by employing standard in vitro assays (such as DPPH,) and to quantify its free radical scavenging capacity
2. Antimicrobial Activity - To assess the antimicrobial efficacy of the sample against selected microbial strains (bacterial and/or fungal) using standard methods like agar well diffusion or broth dilution, and to determine its inhibitory effect

Fig 3. Tridax Procumbens with flowers and daisy-like flowers and sprawling

3. Anti-inflammatory Activity - To evaluate the anti-inflammatory activity of the selected sample using the Human Red Blood Cell (HRBC) membrane stabilization method, by assessing its ability to inhibit hemolysis and stabilize erythrocyte membranes under stress conditions.

MATERIALS AND METHOD

MATERIALS:

Chemicals:

• Methanol (70% or 90%)     

METHODS:

Collection of Materials:

Fresh leaves of Tridax Procumbens was collected from local area of loha and aside of harbal near sonkhed Nanded. It kept under dried shaded area in mid dark room. That were dried for 07-10 days.

• Authentication of plant material:

Sample of fresh leaves of Tridax Procumbens taken for the authentication and were authenticated by Dr. Vishal R Marathe (Associate Professor and head of botany department) on 20/02/2026 in N.E.S.

Image no.1 Authentication Letter

Science College Nanded. Authentication of plant was done. Collection, Authentication, Identification, Processing and Storage had been done according at standard procedure for the plant.

Procedure for Extraction of Tridax Procumbens by maceration method:

Principle: -

Maceration is a simple extraction technique in which the coarsely powdered plant material is soaked in a suitable solvent for a specific period to dissolve the active constituents.

Materials Required

Dried leaves of Tridax Procumbens

• Methanol (70% or 95%)
• Conical flask or glass container
• Whatman filter paper
• Funnel
• Rotary evaporator / water bath
• Grinder
• Analytical balance

Image no. 02 Filtering the Extract

Procedure: -

Collection and Preparation of Plant Material

1. Fresh leaves of Tridax Procumbens were collected from aside of road.
2. Then leaves were washed with distilled water to remove dust particles and impurities also somewhere insecticides are also sprayed.
3. The leaves were kept for shade dried for 7–10 days at normal room temperature, Room with no high sunlight and not too dark.
4. The dried leaves were taken and convert into coarse powder using a grinder or morter and pestle till fine particles (Powder form) are not found.

2. Maceration Process

1. Take powdered plant material was accurately weighed to 50gm in weighing balance.
2. Take the powder and was transferred into a clean conical flask.
3. Then 500 ml of Methanol was taken and added to the conical flask with powder as extraction a solvent.
4. The mixture was shaken for a particular period kept it for maceration for 72 hours at room normal temperature. The conical flask was shaken occasionally after hourly to improve extraction.

3. Filtration

1. After 72 hours completion, the mixture was ready to filter using Whatman filter paper.
2. The filtrate containing the plant extract was collected in a clean container (Beaker).

4. Concentration of Extract

1. Then filtrate was evaporated using a water bath to remove the solvent.
2. After filtration semi-solid crude extract was obtained.

5. Storage

1. The extract was transferred to a sterile airtight amber color glass container and keep away from sunlight.
2. It was stored at 4°C in a refrigerator until further use for antimicrobial activity testing.

• Antioxidant Activity of Tridax procumbens Extract

Method: DPPH Radical Scavenging Assay

Principle: -

The DPPH (2, 2-Diphenyl-1-picrylhydrazyl) radical is a stable free radical having a deep purple color. Antioxidants present in the plant extract donate hydrogen atoms to DPPH, converting it into a yellow colored compound (DPPH-H). The decrease in absorbance is measured using a UV-Visible spectrophotometer at 517 nm, which indicates antioxidant activity.

Materials Required

· DPPH reagent

· Methanol

· Plant extract (Tridax procumbens)

· Ascorbic acid (standard antioxidant)

· Test tubes

· Micropipette

· UV-Visible spectrophotometer

· Volumetric flasks

Procedure: -

1. Preparation of DPPH Solution

Weigh 3.94 mg of DPPH. Dissolve it in 100 mL methanol to prepare 0.1 mm DPPH solution. Keep the solution in a dark bottle to prevent light degradation.

2. Preparation of Extract Solution     

Dissolve the plant extract in methanol. Prepare different concentrations such as 20, 40, 60, 80 and 100 µg/ml

Antioxidant Activity of Tridax procumbens Extract

Method: DPPH Radical Scavenging Assay

Principle: -

1. The DPPH (2, 2-Diphenyl-1-picrylhydrazyl) radical is a stable free radical having a deep purple color.
2. Antioxidants present in the plant extract donate hydrogen atoms to DPPH, converting it into a yellow colored compound (DPPH-H).
3. The decrease in absorbance is measured using a UV-Visible spectrophotometer at 517 nm, which indicates antioxidant activity.

Materials Required

· DPPH reagent

· Methanol

· Plant extract (Tridax procumbens)

· Ascorbic acid (standard antioxidant)

· Test tubes

· Micropipette

· UV-Visible spectrophotometer

· Volumetric flasks

Procedure: -

1. Preparation of DPPH Solution

1. Weigh 3.94 mg of DPPH.
2. Dissolve it in 100 mL methanol to prepare 0.1 mm DPPH solution.
3. Keep the solution in a dark bottle to prevent light degradation.

2. Preparation of Extract Solution   

1. Dissolve the plant extract in methanol.
2. Prepare different concentrations such as 20, 40, 60, 80 and 100 µg/ml

3. Preparation of Standard Solution

1. Prepare ascorbic acid solution in methanol.
2. Make different concentrations similar to the extract.

4. Reaction Mixture

1. Take 1 ml of plant extract solution in a test tube.
2. Add 1 ml of DPPH solution.
3. Mix the solution properly.

5. Incubation

1. Incubate the reaction mixture in the dark at room temperature for 30 minutes.

6. Measurement of Absorbance

1. Measure the absorbance at 517 nm using a UV-Visible spectrophotometer.
2. Use methanol as blank.

Result: -

The extract showed concentration dependent antioxidant activity.

Antimicrobial Activity of Tridax procumbens Extract

Method: Agar Well Diffusion Method

Materials Required

• Plant extract (Tridax Procumbens)
• Test microorganisms (e.g., Staphylococcus aureus, Escherichia coli)
• Nutrient agar medium
• Sterile Petri plates
• Sterile cotton swabs
• Cork borer (6 mm diameter)
• Micropipette
• Incubator
• Sterile saline solution
• Standard antibiotic (positive control)
• Distilled water/solvent (negative control)

Procedure

1. Preparation of Nutrient Agar

1. Weigh 28 gm of nutrient agar powder.
2. Dissolve it in 1000 ml distilled water.
3. Heat the mixture until the agar completely dissolves.
4. Sterilize the medium by autoclaving at 121°C for 15 minutes.
5. Allow the medium to cool to about 45–50°C.
6. Pour 20–25 ml of molten agar into sterile Petri plates.
7. Allow the agar to solidify at room temperature.

2. Preparation of Microbial Culture

1. Obtain pure cultures of test microorganisms.
2. Inoculate the bacteria into nutrient broth.
3. Incubate at 37°C for 18–24 hours.
4. Adjust the turbidity using sterile normal saline to match approximately 0.5 mc and standard.

2. Inoculation of Agar Plates

1. Dip a sterile cotton swab into the bacterial suspension.
2. Spread the culture evenly over the entire surface of the nutrient agar plate.
3. Rotate the plate while swabbing to ensure uniform bacterial growth.

3. Formation of Wells

1. Use a sterile cork borer (6 mm) to punch wells in the agar.
2. Remove the agar plugs carefully using sterile forceps.
3. Maintain sufficient distance between wells to avoid overlapping zones.

5. Addition of Plant Extract

1. Fill each well with 50–100 μl of plant extract using a micropipette.
2. Add standard antibiotic in one well as positive control.
3. Add solvent (distilled water or ethanol) in another well as negative control.

6. Incubation

1. Keep the plates at room temperature for 30 minutes to allow diffusion of extract into agar.
2. Incubate the plates inverted at 37°C for 24 hours.

7. Measurement of Zone of Inhibition

1. After incubation, observe the plates for clear zones around the wells.
2. Measure the diameter of the zone of inhibition (mm) using a ruler or caliper.
3. Record the results for each microorganism.

 8. Interpretation of Results

1. Larger zone of inhibition indicates stronger antimicrobial activity.
2. Compare results with standard antibiotic.

Result: -

1. Preparation of Test Solution

Prepare the reaction mixture in test tubes as follows:

1. Incubation

1. Incubate all test tubes at 37°C for 30 minutes.
2. Temperature should 37°

2. Centrifugation

1. After incubation, centrifuge the mixture
2. This centrifuge at 3000 rpm for 10 minutes.

3. Measurement of Absorbance

1. Measure the absorbance of the supernatant using a UV-Visible spectrophotometer at 560 nm.

Result: