In Vitro Anthalmentic Activity and Phytochemical Screening of Methanolic Extract of Duranta Erecta Leaves

Anthelmintics are a class of medications used to treat infections in humans and animals caused by parasitic worms. These worms, which include nematodes (roundworms), cestodes (tapeworms), and trematodes (flukes), are responsible for a wide range of diseases globally. Anthelmintics work by Targeting various physiological processes of the worms, such as their nervous systems, metabolism, or structural integrity, leading to their immobilization or death [9]. In emerging nations, people and communities are increasingly using therapeutic herbs. Due to its effectiveness, affordability, accessibility, cultural acceptance, and alleged lack of adverse effects when compared to synthetic pharmaceuticals, the use of herbal medicines has increased [1,3]. Plants are a great source of medications because they produce a variety of bioactive molecules with therapeutically valuable components. In USA, about 25 percent medications are made from herbal resources [2, 3]. Various elements impart distinct characteristics and qualities to plants. The Verbenaceae family, to which Duranta belongs, comprises 35 species [3] This plant is native to regions of South and Central America, Africa, and Asia. Commonly known as "golden dewdrop," Duranta erecta (synonymous with D. repens) is an upright, sprawling shrub that typically grows to a height of one to three meters[4] .In many Ghanaian houses, it is cultivated as an ornamental plant or hedge [5,6].Angels Whisper, Brazilian Sky Flower, Duranta, Golden Dew Drop, Golden Tears, and Pigeon Berry are some of its other names. Chemical compounds found in its leaves, bark, fruit, and flowers exhibit antioxidant, antibacterial, antifungal, anthelmintic , and anticancer properties ,This properties are due to phytochemicals such as alkaloids ,flavonoids ,terpenoids and saponins etc.[7].            

       
           
       
Fig- Leaves of Duranta erecta.

1.1] Scientific identification of plant:

Scientific name: Duranta erecta

Family:   verbenaceae

Kingdom:   Plantae

Genus:   Duranta                        

Species:   Duranta erecta

        
           
       
1.2] Properties of Duranta erecta:

Anticancer, Antioxident, Antihelmentic, Antifungal, Larvicidal and Antibacterial.

1.3] Botanical identification:

The botanical description is as follows:

Overall Qualities Growth Habit:

A small tree or sprawling shrub with a maximum height of 3–6 meters (10–20 ft). Its branches arch and its growth habit is dense and spreading.

Habitat: Usually found in hedges, roadsides, and gardens in tropical and subtropical areas.

Type of Leaves: Oval to elliptic, simple. Arrangement: Whorled or opposite. Dimensions: 2–8 cm in length and 1-4 cm in width.

Texture: a little shiny and smooth. Entire to slightly serrated margin.

Blooms Color: Light blue with a white throat, lavender, or purple. Shape: Tubular, with thin racemes of five petal lobes.

Dimensions: Compact, with a diameter of 1-2 cm. Arrangement: Inflorescence clusters that hang from the branches.

Fragrance: Not very strong. In warm climates, the blooming season is nearly year-round.

Fruit Type: Drupe, which resembles berries. Color: When fully grown, bright orange or yellow. Dimensions: tiny, round, with a diameter of roughly 1 cm. Toxicity: If consumed, poisonous to humans and certain animals.

Bark and Stem Stem: Soft and green at first, it becomes woody as it ages. Bark: rough and light brown. The root system is expanding and fibrous. Function: Prefers well-drained soils but is tolerant to a variety of soil types. The significance of ecology and ornamentation Pollinators: Draws hummingbirds, butterflies, and bees. Uses: Well-liked as a container plant, hedge, or decorative shrub. Although Duranta erecta is prized for its colorful blooms, ornamental foliage, and fruit, caution should be exercised because of its toxicity.

1.4] Chemical constituents:

Alkaloids ,Flavonoids such as luteolin and derivatives, Saponins, Tannins ,Terpenoids, Essential oils, toxic compounds and other compounds such as Carbohydrates ,Fats, Proteins

2] MATERIALS AND METHODOLOGY

2.1] Solvents: methanol, chloroform, conc. sulphuric acid,0.1?rric chloride, Ethylacetate,  Glacial acetic acid, Hexane,

2.2] Collection of plant leaves : The leaves of Duranta erecta are collected from Bramhanal , Sangli 416416,Maharashtra,India ON 29/10/2024.Authenticated by dr. Patangrao Kadam college, Sangli.

2.3] Preparation of extract:

1. The leaves are separated from stems. And dried in shade for 8 to 10 days.
2. The dried leaves are powdered.
3. The extraction was prepared by cold maceration process .
4. 70  gm of leaves powder kept into  200   ml methanol for 24 hrs.
5. The extract was evaporated on hot plate to concentrate the extract .

3] Thin layer chromatography:

3.1] TLC plate preparation: slurry of  silica gel G poured on to a glass slide to make a thin layer .and then dried into the hot air oven for 10 min. Preparation of developing chamber: The TLC developing chamber may consist of a glass jar with a lid or a beaker topped with a watch glass . The solvent is poured into the chamber and then solvent and chamber saturation is carried out with a filter paper by putting it inside the beaker with the covered lid Sample application in TLC plate: Samples are diluted and then applied on the plate using a micro syringe or by a micro capillary tube for qualitative work.  Detection of the spots: The compounds are detected by the UV light or by treating the plate with visualizing agents such as iodine chamber. Evaluation of chromatogram: The evaluation of the detected spots on the plate is done by qualitative and quantitative methods. Qualitative analysis such as; Rf, Hrf and values are detected.

3.2] TLC for Flavonoids:    

Mobile phase: Ethyl acetate: formic acid: glacial acetic acid: water

Stationary phase: silica gel G.

Visualizing agent: Iodine chember

3.3] TLC for Alkaloid:

Mobile phase: Methanol: chloroform (3:7)

Stationary phase: silica gel G

Visualizing agent: Iodine chember

3.4] TLC for Terpenoid:

Modible phase: chloroform: hexane (9:1)

Stationary phase: silica gel G

Visualizing agent: Iodine chamber.

4]Anthelmintic activity of Duranta Erecta

Helminth infections are among the most common diseases in developing countries, primarily caused by intestinal nematodes. These infections often lead to gastroenteritis, which results from mixed infections involving various species of stomach and intestinal worms. Such infections can cause symptoms like weakness, loss of appetite, reduced feed efficiency, and weight loss. Anthelmintic drugs are used to eliminate or expel parasitic helminths from the body.

4.1] Collection of worms: for the anthalmentic activity the worms of Eisenia Fetida (redworms) are collected from Earth green organics from Khatav, Sangli 416416. Maharashtra ,India. They are carried to the laboratory in a properly ventilated bag with ample nourishment.

4.2] Biological description: Biological name of warm is Eisenia Fetida , known under another  common names Redworm, brandling worm, panfishworm, trout worm. Length of worm increases upto 1-1.5 foot.  1500-2000 species of Eisenia Fetida are available .Life span is about 20 years. For their survival conditions like moisture maintaining is important.

4.3] In Vitro Anthelmentic activity :

 Procedure :

1. The worms and clean with distilled water
2.  Three clean petridish  is taken as control, test ans standard and put the worms individually into  each petridish.
3. In control , 10ml normal saline solution is added.
4. In test ,10mg/ml Duranta erecta extract is added.
5. In standard, 10mg/ml Albendazole stock solution is added.
6. Additionally, each worm plate's paralysis and death time are recorded.

RESULT AND DISCUSSION

The average yield of dried powder obtained from duranta ertecta leaves  was 7.69% w\w. the powder was  dark greenish colour .

5.1] Phytochemical Tests:

5.2] Thin Layer Chromatography:

Flavonoid:  for detection of flavonoid By using mobile phase ethyl acetate ,formic acid ,glacial acetic acid and water in 10:1.1:1.1:2.7 ratio respectively and the  RF value was found 0.32 and HRF value 32%.

       
           
       
Fig: TLC of flavonoid

Alkaloid: For detection of alkaloid by using mobile phase chloroform and methanol 7:3 respectively and the RF value was found 0.432 and HRF valur 43%.

       
           
       
Fig: TLC of alkaloid

Terpenoid: For detection of terpenoids by using mobile phase chloroform and hexane 9:1 respectively and the RF value was found 0.40 and HRF value 40%

       
           
       
Fig : TLC of terpenoid

5.3] Anthalmentic Activit of Duranta Erecta

In the present experiment comparison between methanolic extract of duranta erecta leaves and Albendazole is shown below:

       
           
       
Fig: Control Group