Formulation And Evaluation of An Antimalarial Syrup Using Nyctanthes Arbortristis Leaf Extract

In the last decade, 40% of the malaria-endemic countries reported reduction in the incidence of malaria by half. This has raised hopes to the stage of ‘complete eradication’ of malaria in the near future. Recently, the World Health Organization (WHO) announced an ambitious new plan to “almost” eradicate malaria by 2030; 40% by 2020 and 90% by 2030. However, alarming reports of artemisinin resistance, first from Thai-Cambodian border in 2009 and it's spread in 5 countries within 5 years have contributed to the unceasing threat of resistance development. This has put forth again an urgent need for the development of new antimalarial drugs. In spite of persistent efforts and significant investment, availability of long lasting antimalarial drugs is still a quest. Medicinal plants, being a source of diverse compounds, have been a major attraction for drug discovery scientists. The two major antimalarial drugs; artemisinin and quinine, have been discovered from plant.

MALARIA

Malaria is a life-threatening disease spread to humans by some types of mosquitoes. It is mostly found in tropical countries. It is preventable and curable. The infection is caused by a parasite and does not spread from person to person.Symptoms can be mild or life-threatening. Mild symptoms are fever, chills and headache. Severe symptoms include fatigue, confusion, seizures, and difficulty breathing. Infants, children under 5 years, pregnant women, travellers and people with HIV or AIDS are at higher risk of severe infection. Malaria can be prevented by avoiding mosquito bites and with medicines. Treatments can stop mild cases from getting worse.

Drugs used in treatment of Malaria

Cinchona alkaloids: quinine, quinidine

4 aminoquinolines: chloroquine, hydroxychloroquine, amodiaquine,

8 aminoquinolines: primaquine, tafenoquine, bulaquine

Chemical Constituents.

Many distinct compounds, including flavonoids, terpenes, steroids and alkaloids which belong to various chemical classes, have been identified and described from the various portions of the N.arbor and produces a significant number of chemicals, which fall under the category of secondary metabolites as shown in Table.

Chemical constituents present in different parts of N. arbor

Inhibition of Parasite Growth

1. Interference with parasite metabolism: The alkaloids, glycosides, and terpenoids present in Nyctanthes arbor-tristis can interfere with the parasite's metabolic pathways, such as glycolysis and mitochondrial function.
2. Inhibition of protein synthesis: The plant's constituents can also inhibit protein synthesis in the parasite, which is essential for its growth and survival.

Disruption of Parasite Cell Membrane

1. Alteration of membrane fluidity: The terpenoids and glycosides in Nyctanthes arbor-tristis can alter the fluidity of the parasite's cell membrane, making it more difficult for the parasite to maintain its structural integrity.
2. Increased membrane permeability: The plant's constituents can also increase the permeability of the

parasite's cell membrane, allowing ions and small molecules to leak out, ultimately leading to the parasite's death.

Immunomodulatory Effects

1. Enhanced immune response: Nyctanthes arbor-tristis can stimulate the host's immune system to produce more antibodies and activate immune cells, such as macrophages and T-cells, to attack the parasite.
2. Increased production of cytokines: The plant's constituents can also increase the production of cytokines, which are signaling molecules that help coordinate the immune response.

Active Compounds

Some of the active compounds present in Nyctanthes arbor-tristis that have been found to exhibit antiplasmodial activity include:

1. Nyctanthin: A quinazoline alkaloid with antiplasmodial and immunomodulatory effects.
2. Arborine: A quinazoline alkaloid with antiplasmodial and antimicrobial effects.
3. Linalool: A monoterpenoid with antiplasmodial and antimicrobial effects.

MATERIAL AND METHODS

Step 1: Collection of Plant Material

Plant Part Used: Leaves (commonly used for antimalarial properties) Time of Collection: Early morning or late afternoon

Cleaning: Wash thoroughly with water to remove dirt and dust.

Drying: Shade dry for 3–5 days, then crush to coarse powder (or use fresh leaves if immediate extraction)

Grinding: Grind the dried leaves into a fine powder using a mechanical grinder. Storage: store the powder in an airtight container further use.

Botanical name: Nyctanthes arbor-tristis L.

Family: Oleaceae (the olive family)

Synonyms: Jasminum arbor-tristis

Parts which are used  : leaves

2. EXTRACTION PROCESS

Method: Hydroalcoholic or Aqueous Extraction ( both are commonly used)

Aqueous Extraction (Decoction)

(simple and suitable for Syrup formulation)

1. Weigh 50 g of leaf powder.

2. Boil in 500 ml of distilled water for 30-45 minutes.

3. Cool and filter using muslin cloth followed by Whatman No.1 filter Paper.

4. Concentrate the extract using a water bath at 60-70 C until a thick concentrate is obtained.

5. store the extract in a refrigerator

PEPARATION AND PROCEDURE

1. Prepare Sugar Syrup

Weigh 18 g of sucrose.

Dissolve it in about 25 ml of warm purified water. Stir continuously until fully dissolved.

Cool to room temperature.

2. Hydrate Tragacanth Gum

Sprinkle 0.1 g of tragacanth gum into 5 ml warm water. Stir well to prevent lumping.

Let it hydrate for 1–2 hours (or overnight) for best results.

3. Dissolve Citric Acid

Dissolve 0.05 g citric acid in 2–3 ml purified water. Set aside.

4. Prepare Oil Blend

Mix 0.025 ml tea tree oil and 0.1 ml orange oil in 1–2 ml glycerin.

Stir thoroughly to emulsify (glycerin helps disperse the oils).

5. Combine Base Ingredients

In a clean beaker:

Add hydrated tragacanth gum, Add prepared sugar syrup,

Add remaining glycerin (totaling 4 ml), Add honey (12 ml).

Stir well to create a uniform base.

6.Add Herbal Extract

Add 2.0 ml of Nyctanthes arbortristis extract to the mixture. Stir gently and mix thoroughly.

7.Add Citric Acid and Oils

Add the citric acid solution.

Then add the oil blend (tea tree + orange oil in glycerin). Mix thoroughly to ensure even distribution.

8. Add Beetroot Coloring

Add beetroot juice/extract drop by drop.

Stir until a uniform, natural reddish color is achieved.

9. Adjust Volume

Add purified water q.s. to make exactly 50 ml. Stir the final mixture well.

10. Optional Filtration

Filter through muslin cloth or filter paper if a clearer syrup is desired.

11. Bottling and Labeling

Formulated Syrup

Result for Evaluation Parameter

CONCLUSION

The present study was undertaken to explore the potential of Nyctanthes arbor-tristis leaves in the formulation of an effective, stable, and patient-compliant antimalarial syrup. Based on traditional Ayurvedic knowledge and modern pharmacognostic principles, the leaves were selected due to their documented antipyretic and antimalarial properties. The formulated syrup using Nyctanthes arbor-tristis leaves extract is stable, palatable, and shows promising antimalarial activity in both in vitro and in vivo models. Further clinical trials are recommended to validate its efficacy in human subjects.

RESULT