Isolation and Characterization of Ficus benghalensis Linn Milk Latex by using UV Spectrophotometry

Biswanath Prusty*, Santanu Kumar Hotta, Rabindra Padhy, Ganesh Kumar Sahoo,

Samir Kumar Pradhan

Department of Pharmaceutical Analysis, College of Pharmaceutical Sciences, Mohuda, Berhampur, Ganjam, Odisha, Pin - 760002, India.

*Corresponding Author E-mail: biswanath065@gmail.com

ABSTRACT:

Herbal medicines play a significant part in human healthcare. Traditional medical systems including Ayurveda, Unani, Siddha, and homoeopathy have all used Ficus benghalensis. The fruits, aerial roots, bark, and leaves of the tree are all known to have historical associations with menorrhagia, dysentery, diarrhoea, anti-hemorrhoidal, neurological diseases, and tonic qualities. On the other hand, Ayurveda is mostly recognized for its therapies for diabetes. There are no publications that provide a thorough analysis of the phytochemicals found in F. benghalensis milk latex. The goal of the study is to assess its bioactive components and apply conventional knowledge to understand its biological actions. Extracts of hexane and chloroform from the bark milk latex of F. benghalensis were assessed using a range of physiological and phytochemical tests. Additionally, the spectrophotometric UV-2400PC Series instrument with measurement properties was used to characterize the extract. Wavelength Range (200.00 to 800.00nm) with Medium Scan Speed. And the reaction was enthusiastic and extremely fulfilling.

Fig- Graphical Abstract.

KEYWORDS: Ficus benghalensis linn milk latex, UV spectrophotometry, Phytochemical study, Physiological study, medicinal uses.

INTRODUCTION: 

Ficus benghalensis is a member of the Moraceae family. This species is native to several Asian countries, such as Pakistan, Malaysia, China, India, and Nepal. Various parts of F. benghalensis, including its bark, roots, stems, buds, leaves, fruits, and latex, have been utilized in traditional medicine to address neurological issues like anxiety, insomnia, and seizures. Numerous neuropharmacological potentials of F. benghalensis have been reported by various studies^1–3. There have been reports of great cognitive boosting potential for the bark aqueous extract.

Different parts of the banyan tree are used in traditional medicine to treat a range of illness. The bark can be used to treat burns, bleeding, diarrhea, dysentery, ulcers, diabetes, and skin disorders. The leaves help with leprosy, ulcers, skin allergies, etc.^4,5. Diarrhea and dysentery are treated with the buds. Latex can help with skin diseases, inflammation, rheumatism, and hemorrhoids. The wood consumption and termite infestation were considerably decreased by F. benghalensis plant latex and its combinatorial mixes^6–8.

Figure 1. Biological profile of Ficus benghalensis.

Plant description:

Taxonomic Rank	Classification
Kingdom	Plate
Subkingdom	Tracheobinote
Super division	Spermatophyta
Division	Magnoliophyta
Class	Magnoliopsida
Subclass	Hamamelidae
Order	Urticales
Family	Moraceae
Genus	Moraceae
Species	Benghalenses, indica
Synonyms	Ficus banyana Oken, Ficus chaveiri G.Nicholson, Ficus cotoneifolia Vahl
Common name	Vata, Bara, Raktaphala, Skandaja
Habitat	Tropical, lowland area

Morphological Description:

A towering evergreen tree, characterized by its expansive branches and multiple pillar-like aerial roots for support, can grow up to 20 meters tall. It features a prominent, smooth, oily gland at the top and leaves that are ventrally compressed, hairy, ovate-cordate in shape, with entire margins, rounded tips, measuring 8 – 20 × 6 – 15cm, leathery, striking on the upper side, finely pubescent beneath, and having 3 – 5 basal veins with 4–6 pairs of lateral veins. The inflorescence is a hypanthodium, appearing in axillary pairs, sessile, spherical, 1.5–2cm in size, hairy, and supported by three tiny hairy bracts. It matures from green to red. The tree bears small flowers of three types: male, female, and gall. Male flowers are numerous, pedicellate, located near the fig's ostiole, with three tepals and a single stamen; female flowers are sessile; gall flowers are pedicellate and host an emerging insect. The fruit is an achene, globose-ellipsoid, and creamish-brown^9–12.

Figure 2. F. benghalensis milk latex.

Figure 3. F. benghalensis adventitious root.

MATERIALS AND METHODS:

Collection and authentication of plant material:

 From our renowned Herbal Garden at the College of Pharmaceutical Sciences, Mohuda, Berhampur, 100ml of fresh Ficus benghalensis Linn milky latex from aerial buds were harvested on March 23, 2024, and verified by a botanist (department of botany, Khalikote college, Berhampur). The plant was categorized using a number of sources, including pertinent taxonomic literature and the certification of our pharmacognosy department.

Figure 5. F. Benghalensis collected milk latex.

Figure 6. Collection of milk latex.

Figure 7. F. benghalensis collected milk latex after storing in Refrigerators.

Extraction and Isolation of Plant Material:

The collected around 100ml plant latex was stored in an airtight glass container in the refrigerator at a temp. 2-8° C the next day it was extracted by using Continuous extraction using chloroform and n-hexane solvents. After 2-3hrs of extraction from chloroform and n-hexane extracted milk latex in the form of solution ^4,13,14.

Figure 8. F. benghalensis Chloroform extraction.

Figure 9. F. benghalensis Hexane extraction.

SOLUBILITY STUDY:

The collected plant materials were proceeding to solubility study by using different solvents (DMSO, Benzene, Chloroform, Toluene, Methanol, Ethanol, Water, and n-hexane) at different temperatures. The current study was observed that Chloroform and n-hexane gave excellent solvent for the extraction and isolation of natural crude medicinal product from the milk latex ^15–17.

Figure 10. F. benghalensis milk latex solubility studies.

Phytochemical Screening:

Qualitative Analysis of Alkaloids

Test Name	Reagent Used	Procedure	Inference	Result
Dragendorff’s Test	Dragendorff’s reagent (Potassium bismuth iodide)	1ml of extract + 1ml of reagent. Observe for precipitate formation.	Formation of orange-red precipitate indicates presence of alkaloids.	Chloroform extract and hexane fraction showed positive result.
Mayer’s Test	Mayer’s reagent (Potassium mercuric iodide)	1ml of extract + 1ml of reagent. Check for precipitate.	Cream or whitish-yellow precipitate confirms alkaloid presence.	Hexane and chloroform fractions produced cream-colored precipitate.
Wagner’s Test	Wagner’s reagent (Iodine in potassium iodide)	1ml of extract + 2ml of reagent. Look for precipitate formation.	Reddish-brown precipitate signifies the presence of alkaloids.	Chloroform fraction gave a reddish-brown precipitate.

Qualitative Phytochemical Screening of Various Extracts:

Phytochemical	Test Name	Reagent/Procedure	Observation/Inference	Result
Carbohydrates	Molisch’s Test	2ml extract + 1 ml α-naphthol → add conc. H₂SO₄ along test tube wall. Observe color at junction.	Purple or reddish-violet ring at the interface indicates carbohydrates.	Chloroform fraction showed deep violet coloration.
	Fehling’s Test	1ml extract + equal volumes of Fehling’s A and B → heat.	Brick-red precipitate denotes reducing sugars.	No color change observed in chloroform and hexane fractions.
Proteins and Amino Acids	Biuret Test	1ml extract + 2 ml Biuret reagent (alkaline copper sulfate).	Development of deep purple color suggests proteins.	Chloroform fraction gave deep violet coloration.
	Ninhydrin Test	1ml extract + 2 ml 5% Ninhydrin → heat in water bath.	Purple coloration after heating indicates amino acids.	Chloroform fraction turned deep purple; two layers observed.
Glycosides	Legal’s Test	Dissolve extract in pyridine → add sodium nitroprusside → make alkaline.	Pink to red coloration shows presence of glycosides.	No pink/red coloration observed in any fraction.
Tannins and Phenolics	Lead Acetate Test	Mix extract with basic lead acetate solution.	White precipitate confirms presence of tannins.	Chloroform and hexane fractions gave white precipitates.
Steroids	Libermann–Burchard Test	1 gm sample + few drops chloroform → add 3 ml each of acetic anhydride and glacial acetic acid → heat → cool → add conc. H₂SO₄ along wall of test tube.	Bluish-green color indicates presence of sterols/steroids.	Hexane and chloroform fractions showed bluish-green color.
Saponins	Foam Test	Mix small amount of extract with 20 ml distilled water in a cylinder → shake for 15 mins.	Persistent foam (≥1 cm) layer confirms saponins.	Foam layer observed in both chloroform and hexane extracts.

Table 1. Phytochemical screening of Chloroform and n-hexane extract of F. benghalensis milk latex.

Name of phytoconstituents	Chloroform extract	n-Hexane extract
Alkaloid	+++	+++
Carbohydrate	++	+
Reducing sugar	--	--
Protein and Amino acid	++	+
Glycoside	--	--
Tannins and Phenolic compound	+++	+++
Steroids	++	++
Saponin	+++	+++

SPECTRAL ANALYSIS AND STRUCTURAL ELUCIDATION:

The Chloroform extract was characterized by using spectrophotometric UV-2400PC Series instrument with Measurement Properties Wavelength Range (nm.): 200.00 to 800.00 Scan Speed: Medium. The absorbance of phytochemical constituents was anticipated from resources of different journals. ^1,17,19–22

Figure 11. F. benghalensis Chloroform extract Phytochemical screening.

Table 2. Chloroform extraction (Aqueous phase) of F. benghalensis milk latex.

Sl. No	Wavelength (nm)	Absorbance	Description	References
1	775	0.054	Friedelin	Gopukumar and Praseetha, 2015; Rao et al., 2014
2	704	0.049
3	641	0.050
4	626.5	0.052	Theaflavin-3,30-digallate
5	532	0.070	Pelargonidin (leucopelargonidin –3–O–β–D-glucopyranoside)
6	526	0.070
7	495	0.079	Beta glucoside, alpha-D-glucose	Naquvi et al., 2015; Murti et al, 2011; Joseph and Raj, 2010; Babu et al., 2010
8	472	0.079
9	455	0.079
10	439	0.082	Rhamnoside (leucopelargonidin– 3–O–α–L-rhamnopyranoside)	Gopukumar and Praseetha, 2015; Rao et al., 2014
11	399	0.079
12	380	0.079
13	366.5	0.083	quercetin-3-galactoside
14	350	0.081
15	295	0.175	Leucocyanidin
16	280	0.100	Leucodelphinidin
17	242	0.291	Bengalenoside

Table 3. Chloroform extraction (oil phase) of F. benghalensis milk latex.

Sl. No	Wavelength (nm)	Absorbance	Description	References
1	725	0.049	Friedelin	Gopukumar and Praseetha, 2015; Rao et al., 2014
2	681	0.037
3	650	0.035
4	612	0.036	Theaflavin-3,30-digallate
5	589	0.038	lupeol	Naquvi et al., 2015; Murti et al, 2011; Joseph and Raj, 2010; Babu et al., 2010
6	579	0.039	beta-progesterone	Verma et al., 2015
7	554	0.040
8	481	0.043	20-traxasten-3-ol, Teraxosterol	Gopukumar and Praseetha, 2015; Rao et al., 2014
9	463	0.045
10	427	0.050
11	404	0.049	Psoralen
12	381	0.051
13	345	0.048
14	293	0.083	leucocyanidin- 3 – O –β–D-glucopyrancoside
15	257	0.116	B- amyrin	Naquvi et al., 2015; Murti et al, 2011; Joseph and Raj, 2010; Babu et al., 2010
16	257	0.116
17	232	0.091	B- sitosterol
18	212	0.089

Figure 12. UV Spectroscopy of F. Benghalensis, Chloroform extraction (Aqueous phase)

Figure 13.  UV Spectroscopy of F. Benghalensis, Chloroform extraction (Oil phase).

Figure 14. UV Spectroscopy of F. Benghalensis, n-hexane extraction

The n-hexane extraction has contained minute quantity of volatile Flavonoids which was showed in the above graph.

RESULT AND DISCUSSION:

In this study the Ficus benghalensis linn milk latex was evaluated or screened by various phytochemical analysis method by using ultraviolet spectrophotometry, taking their absorbance and wavelength from different literature it was confirmed that the milk latex contains volatile Flavonoid and terpenoid which exhibit various pharmacological activity.

The chloroform extract shows more activity due to presence of maximum number of phytoconstituent whereas n-hexane extract has least number of constituents.

CONCLUSION:

The phytochemical investigation of the latex extracted from Ficus benghalensis confirmed the presence of diverse bioactive constituents, including tannins, saponins, waxes, lupeol derivatives, β-sitosterol, and flavonoid glycosides such as leucocyanidin and leucopelargonidin derivatives. The identification and characterization of these compounds were successfully validated using UV spectroscopic analysis. These findings suggest that the latex of Ficus benghalensis may serve as a potential source of pharmacologically active compounds, warranting further studies for its therapeutic applications and mechanism of action.

FUTURE STUDY OR SCOPE OF THIS RESEARCH:

As the milk latex contains maximum number of phytochemicals and exhibits a potent Anti-hemorrhoidal and a potent hepatoprotective properties was the future study of this research.

ACKNOWLEDGEMENTS:

The authors sincerely thank the Principal and the Head of the Department of Pharmaceutical Chemistry, College of Pharmaceutical Sciences, Berhampur, for providing the necessary facilities. The support and cooperation of the research team are also gratefully acknowledged.

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Received on 15.05.2024      Revised on 09.11.2024

Accepted on 25.04.2025 Published on 08.11.2025

Available online from November 13, 2025

Research J. Pharmacy and Technology. 2025;18(11):5502-5508.

DOI: 10.52711/0974-360X.2025.00793

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