Phytochemical Investigation and Herbal Cream Development from Acalypha Fruticosa Forssk Ethanolic Extract

Medicinal plants have served as the foundation of traditional healthcare systems since antiquity and continue to play a vital role in modern therapeutics. The use of plant-derived materials, crude extracts, or isolated phytoconstituents for the treatment and prevention of diseases represents a time-tested therapeutic modality. Even today, a significant proportion of modern drugs are derived directly or indirectly from natural products, underscoring their pharmacological relevance. Botanicals such as Ispaghula, Garlic, Ginkgo biloba, St. John’s Wort, and Saw Palmetto have gained widespread acceptance among contemporary physicians due to their proven efficacy and safety profiles. Despite this, only a small fraction of the world’s terrestrial plants—approximately 35%—have been scientifically investigated for their bioactive constituents, leaving vast potential for research and drug development from medicinal plants.

Ethnopharmacology, which involves the scientific study of traditional medicinal practices, provides valuable leads for the discovery of novel therapeutic agents. Validation of traditional knowledge through systematic scientific evaluation not only supports the development of new herbal formulations but also promotes sustainable utilization of indigenous medicinal resources. Furthermore, integrating quality control, standardization, and conservation strategies with herbal research can yield substantial socioeconomic benefits, particularly for developing countries where medicinal plants form an integral part of primary healthcare.

Acalypha fruticosa Forssk, belonging to the family Euphorbiaceae, is an aromatic shrub widely distributed in tropical regions of Africa, Southern India, Sri Lanka, and Myanmar. Traditionally, the plant has been used for the treatment of various ailments such as stomach disorders, dyspepsia, rheumatism, dermatitis, wounds, scabies, and inflammatory conditions. Leaves of A. fruticosa are commonly applied externally for skin infections and sores, indicating its dermatological significance. Previous phytochemical investigations have reported the presence of biologically active constituents including alkaloids, flavonoids, tannins, saponins, steroids, triterpenoids, anthraquinones, and sugars. Advanced analytical studies such as GC–MS analysis have identified compounds like 1,2-benzenedicarboxylic acid dioctyl ester and n-hexadecanoic acid, which contribute to the plant’s pharmacological properties.

Among the various phytoconstituents, alkaloids represent a particularly important class of secondary metabolites due to their diverse biological activities. Alkaloids are nitrogen-containing compounds that exhibit antimicrobial, anti-inflammatory, analgesic, and antioxidant properties. These activities support the traditional use of A. fruticosa in the management of skin disorders, wounds, and inflammatory conditions. In topical formulations, alkaloid-rich extracts can help reduce microbial load, alleviate inflammation, neutralize free radicals, and promote skin healing, making them suitable candidates for herbal creams.

Extraction plays a crucial role in isolating these bioactive compounds from plant materials. Various extraction techniques such as maceration, Soxhlet extraction, supercritical fluid extraction, microwave-assisted extraction, and ultrasound-assisted extraction are employed depending on the nature of the phytoconstituents and the desired yield. Among these, Soxhlet extraction remains one of the most widely used and reliable methods for exhaustive extraction of phytochemicals, particularly when the compounds of interest have limited solubility and require controlled heating.

Characterization of plant extracts is essential to ensure their quality, safety, and efficacy. Preliminary phytochemical screening helps identify the major classes of phytoconstituents present in the extract, while chromatographic techniques such as Thin Layer Chromatography (TLC) and High-Performance Thin Layer Chromatography (HPTLC) are widely used for qualitative and quantitative analysis of natural products. These techniques aid in confirming the presence of specific bioactive compounds and provide a basis for standardization of herbal formulations.

Herbal creams are topical semisolid emulsions containing both aqueous and oily phases, formulated to deliver active ingredients directly to the skin. Depending on the nature of the emulsion, herbal creams may be classified as oil-in-water (O/W) or water-in-oil (W/O) systems. The selection of suitable excipients such as emulsifiers, preservatives, emollients, and fragrances plays a critical role in determining the stability, spreadability, penetration, and patient acceptability of the formulation. An ideal herbal cream should possess good skin penetration, non-irritating properties, ease of application, acceptable pH, and stability during storage.

In view of the traditional medicinal importance of Acalypha fruticosa, its rich phytochemical composition, and the growing demand for safe and effective herbal dermatological products, the present study was undertaken to extract, characterize, and formulate a herbal cream incorporating the ethanolic extract of Acalypha fruticosa. The formulated cream was evaluated for its physicochemical properties to assess its suitability for topical application, thereby providing scientific validation for its traditional use in skin-related conditions.

METHODLOGY

The collected aerial parts were washed with running water and subjected to shade-dried at room temperature for 7–10 days. The dried material was homogenised using a grinder to obtain moderately coarse (22/60) powder. Moderately coarse powder of size (22/60) is that powder which all the particles pass through sieve NO: 22 and not more than 40% pass through a sieve NO: 60) The powder is stored in an airtight container until extraction.

Preparation of ethanolic extract : The powder weighing 30g of dried aerial parts was subjected to Soxhlet extraction using 200ml of 70% ethanol. The extraction was carried out until the extractive solution became Colourless. The solvent was then removed by distillation to separate it from the extract. The remaining extract was concentrated using an electrical water bath. The obtained semi-solid was weighed and percentage yield was Calculated. The concentrate was stored in a desicator for further analysis.

TLC analysis

TLC was used to detect alkaloid constituents from the ethanolic extract of Acalypha fruticosa. Separation was performed on silica gel using n butanol: acetic acid: water (4: 1: 5) as mobile phase and alkaloids were visualized with Hager’s reagent.

Procedure for TLC :

1. Preparation and Activation of Plates: Glass plates were thoroughly cleaned, dried, and coated uniformly with a slurry of silica gel 60 prepared in distilled water. The coated plates were air-dried and activated in a hot-air oven at 100 °C for 1 hour. After activation, the plates were cooled in a desiccator until use.

2. Preparation of Mobile Phase: The mobile phase consisting of n-butanol : acetic acid : water (4 : 1 : 5) was freshly prepared and transferred to the development chamber, which was closed and allowed to saturate for 10–15 minutes.

3. Sample Application: A baseline was marked 1.5 cm from the lower edge of the plate. Small, concentrated spots of the ethanolic extract were applied using a capillary tube and allowed to dry.

4. Development of Plate: The spotted plate was placed in the chamber ensuring the sample line remained above the solvent level. Ascending development was carried out until the solvent front reached approximately three-fourths of the plate length. The plate was removed, solvent front marked, and air-dried.

5. Visualization: The dried plate was sprayed with Hager’s reagent, and yellow-colored spots appeared within 1–3 minutes. Rf values were calculated using the formula: 

Rf=Distance travelled by the spotDistance travelled by the solvent front

Method of preparation of acalypha fruticosa herbal cream

The herbal cream was prepared by a conventional emulsification method. Beeswax, mineral oil, soft paraffin, and cetyl alcohol were accurately weighed and melted together at approximately 70 °C in a water bath to form the oil phase. Separately, borax was dissolved in distilled water and heated to the same temperature (70–75 °C) to obtain the aqueous phase. The hot aqueous phase was gradually added to the oil phase with continuous stirring until a uniform emulsion was formed. The ethanolic extract of Acalypha fruticosa, previously dissolved in a small quantity of ethanol or warm water, was incorporated into the emulsion after cooling to about 40 °C, followed by continuous stirring to ensure uniform distribution. After cooling to room temperature, rose oil was added for fragrance. The final formulation was transferred into sterile, airtight containers and stored at room temperature for further evaluation.

Evaluation of Herbal Cream

The prepared herbal cream was evaluated for various physicochemical parameters to assess its quality, stability, and suitability for topical application, following standard procedures.

1. Physical appearance: The formulation was visually examined for colour, odour, physical state, and consistency. A small quantity of cream was spread on a glass slide to detect the presence of coarse particles or lumps.
2. Type of cream (Dye test): The emulsion type was determined using the dye test. A small quantity of cream was mixed with methylene blue and observed under a microscope. Uniform coloration of the continuous phase indicated an oil-in-water (O/W) emulsion, whereas coloration of the dispersed phase indicated a water-in-oil (W/O) emulsion.
3. pH determination: Approximately 1 g of cream was dispersed in 100 mL of distilled water, and the pH was measured using a digital pH meter at room temperature. The acceptable pH range for topical formulations is 4.5–7.0 to ensure skin compatibility.
4. Viscosity: Viscosity was measured using a Brookfield viscometer at 25 °C with spindle no. 64 at 12 rpm. This parameter reflects the consistency and ease of application of the formulation.
5. Spreadability: Spreadability was evaluated using the parallel plate method. About 0.5 g of cream was placed between two glass plates, and a 50 g weight was applied for 1 minute at room temperature (25 ± 2 °C). The diameter of the spread cream was measured, and the average value was taken as the spreadability.
6. Phase separation: The cream was stored in a closed container at temperatures ranging from 30–80 °C for 24–48 hours and examined for any signs of phase separation or oil separation. Absence of separation indicated good physical stability.
7. Stability test: Mechanical stability was assessed by centrifugation. The cream sample was centrifuged at 3000 rpm for 15 minutes and observed for phase separation. The absence of separation indicated formulation stability.

RESULT AND DISCUSSION

1. Authentication

The aerial parts of Acalypha fruticosa Forssk were authenticated by Dr. V. S. Hareesh, confirming its identity in the Euphorbiaceae family. A voucher specimen (No. 7795) was deposited at MBGIPS, ensuring accuracy and reproducibility of the study.

2. Morphological characterization

The plant is an erect, much-branched shrub with a cylindrical, woody, greenish-brown stem. Leaf characteristics are summarized below:

Table 1 : Morphological characters of Acalypha fruticosa leaf

3. Preparation of extract

The dried aerial parts of Acalypha fruticosa were coarsely powdered and extracted using Soxhlet apparatus with 200 mL of 70% ethanol. The extraction was maintained at approximately 60?°C for 2 days to ensure complete recovery of ethanol-soluble constituents.

Table 2 : Summarizes the colour, consistency, and yield of ethanolic extract

4. Phytochemical screening 

Table 3 : Results of Phytochemical Analysis of Ethanolic Extract

5. TLC analysis

Table 4 : Result of TLC analysis

6. Formulation of herbal cream

Table 5 : Observations of Developed Herbal Cream

7. Evaluation

1. Physical Appearance / Visual Inspection

Table 6 : Evaluation of physical characteristics