INTRODUCTION:

Throughout history and in all cultures, herbal medicines have been well-recognized for their illnesses. Herbal medicine has its origins dating back to the dawn of human existence. The term "herbal medicine" refers to a broad range of therapeutic modalities used in conjunction with conventional and non-traditional therapies. The methods used in herbal treatment differ greatly from those in contemporary orthodox medicine¹.

Plant compounds with anti-disease or protective qualities are called phytochemicals. The body does not require these nutrients to thrive; so they are non-essential. Although these chemicals were protected them. New research indicates that they can also serve as disease-fighting agents². The process of identifying phytoconstituents found in various plant components that are useful for finding new drugs through scientific analysis, examination, extraction, experimentation, and subsequent research on the active components is known as phytochemical screening. The method used, known as phytochemical evaluation, was preliminary qualitative. Such research findings may be useful in the development of many plant-derived drugs to treat a range of illnesses. Flavonoids, quinine, terpenoids, and other plant-derived substances perform specific biological roles that augment medicinal actions including antioxidant, anti-inflammatory, anti-carcinogenic, and anti-mutagenic qualities³.

Baccaurea courtallensis (Wight) Muell. Arg., the medium-sized unexplored fruit tree, known locally as "mooty pazham," belongs to the Phyllanthaceae family. Tribal people in Kerala have traditionally utilized the leaves to reduce inflammation by drinking water that has been boiled with fruit, bark, and leaf fragments. In addition to being used for swellings, the leaf paste has other anti-inflammatory uses. Many tribal peoples of Kerala use the fruits of B. courtallensis and fry its leaves to make side dishes to take with rice soup⁴.

The aim of the present study was to compare the extraction nature of dried leaves of Baccaurea courtallensis (Wight) Muell. Arg using maceration and soxhlet extraction method, by the use of several solvents with different polarity such as petroleum ether, chloroform, methanol, ethyl acetate and water, as well as qualitatively evaluating extracts and estimating the total flavonoid content present in the extract. Other extraction procedures, such as maceration and soxhlation, are included in the argument.

MATERIALS AND METHODS:

Materials:

The fresh leaves of Baccaurea courtallensis were collected from Nanniyode, Thiruvananthapuram district, Kerala, India. The plant specimen (leaf and flower) identified by botanists and authenticated by Dr. EA Siril, Professor and Head of the department of Botany, Kariavattom, Thiruvananthapuram. A voucher specimen (Voucher Number- KUBH 11372) has been deposited.

Sample preparation:

The fresh leaves were washed thoroughly in distilled water to clean and remove adhering mud and other solid particles. Dried at room temperature under shade about three weeks and then mechanically pulverised, and sieved.

Preparation of the extract:^5-10

Maceration:

The powdered leaves of Baccaurea courtallensis were subjected to successive maceration using several solvents named pet ether (60–80^oC), chloroform, ethyl acetate, methanol, and water. The prepared coarse powder of leaves of Baccaurea courtallensis (50g) was taken into a maceration jar, and a sufficient quantity of various solvents was added. Then it was kept around 7 days. Shake occasionally for better extraction and filtered the resultant solvents. The extract was evaporated to dryness and noted for its nature, colour, and percentage yield.

Soxhlation:

The weighed quantity (50g) of powdered leaves undergo successive soxhlet extraction using various solvents of distinct polarity namely, petroleum ether (60-80⁰C), chloroform, ethyl acetate, and methanol. The filtrate obtained was evaporated, and the colour, nature, and percentage yield were noted.

Preliminary phytochemical analysis:^11-18

A preliminary phytochemical analysis of all extracts was conducted for the identification of various phytoconstituents.

Quantitative estimation of total flavonoid content:^19-24

Total flavonoid content in the extract was estimated spectroscopically by the aluminium chloride colorimetric method. 0.2 millilitre of 10% solution of aluminium chloride, 0.1 millilitre of 1M solution of potassium acetate, and double distilled water (8.6 ml) were added to 1 millilitre of plant extract, thoroughly mixed, and allowed to stand at room temperature for 30 minutes. Different concentrations of quercetin (standard) were prepared per the above-prescribed method, and UV absorbance was taken at 415 nm. All the measurements were taken three times, and the total flavonoid content was calculated and reported as quercetin equivalent per mg of dried extract.

Statistical analysis:

All the results in this study were expressed as Mean ± SEM (standard error mean). Linear correlation analysis was done using MS office Excel 2010. Statistical analysis was carried out with SPSS Program, version 26.

RESULTS AND DISCUSSIONS:

Successive solvent extraction:

The successive solvent extraction of powdered leaves of Baccaurea courtallensis by maceration and soxhlation was carried out and colour, nature, and percentage yield were noted in Table 1. The extraction by soxhlation gives more yield than maceration. The comparison of the percentage yield of maceration and soxhlation shown in the Figure 1.

Preliminary phytochemical evaluation:

The preliminary phytochemical screening helps to identifying the different classes of numerous primary and secondary metabolites that the plant extract contains. Many authors have reported phytochemical screening of leaves of Baccaurea courtallensis in various solvents.^{4, 25-27} The present study proved that the plant contains flavonoids, phenolics, tannins, steroids, proteins, carbohydrates, saponins, mucilages, glycosides, coumarins, and anthraquinones in various extracts (Table 2).

Table 2: The preliminary phytochemical screening of extracts of maceration and soxhlation.

Extract	Maceration	Soxhlation
Petroleum ether	Steroids	Steroids
Chloroform	Steroids	Steroids
Ethyl acetate	Steroids, proteins	Steroids, proteins
Methanol	Steroids, proteins, tannins, phenols, carbohydrates, flavonoids, saponins, mucilage, glycosides, coumarins, anthraquinones	Steroids, proteins, tannins, phenols, carbohydrates, flavonoids, saponins, mucilage, glycosides, coumarins, anthraquinones
Chloroform water	proteins, tannins, phenols, carbohydrates, flavonoids, saponins, mucilage, glycosides	proteins, tannins, phenols, carbohydrates, flavonoids, saponins, mucilage, glycosides

Quantitative estimation of total flavonoid content:

Total flavonoid content was determined spectroscopic technique by the Aluminium chloride colorimetric method using quercetin as standard reference. The absorbance of the standard and sample is shown in Table 3 and the calibration graph were plotted in Figure 2. Methanolic leaf extract has the highest flavonoid content, 64.8483μg/ml compared to aqueous extracts (Table 4).

Table 3: Absorbance of standard and extracts

Sr. No.	Concentration (μg/ml)	Absorbance at 415 nm
1	20	0.072 ± 0.0003
2	40	0.162 ± 0.0004
3	60	0.248 ± 0.0007
4	80	0.344 ± 0.0002
5	100	0.458 ± 0.0009
6	Baccaurea courtallensis Methanolic Extract	0.28 ± 0.0003
7	Baccaurea courtallensis Aqueous Extract	0.186 ± 0.0002

Values were expressed in Mean ± Standard mean, n=3

Figure 2: Calibration curve of standard quercetin at different concentrations

Table 4: Total flavonoid content of various leaf extracts of Baccaurea courtallensis

Sr. No.	Extract	Total flavonoid content (quercetin equivalent/ mg of extract)
1	Baccaurea courtallensis Methanolic Extract	64.84833 ± 0.0091
2	Baccaurea courtallensis Aqueous Extract	45.2042 ± 0.064

Values were expressed in Mean ± Standard mean, n=3

CONCLUSION:

Comparative study of maceration and soxhlet extraction were performed successfully along with qualitative and quantitative phytochemical screening. The comparative study indicates that soxhlet extraction is preferable than maceration method. Phytochemical studies are helpful for understanding the valuable phytochemicals with disease-modifying ability.

ACKNOWLEDGMENTS:

The authors are thankful to Dr. (Prof) E. A. Siril, Professor of the Botany Department, Kerala University campus, Karyavattom for identifying and authenticating the herbarium.

CONFLICT OF INTEREST:

In relation to this inquiry, the writers have no conflicts of interest.

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Received on 07.08.2024 Revised on 06.09.2024

Accepted on 27.09.2024 Published on 20.12.2024

Available online from November 25, 2024

Res. J. Pharmacognosy and Phytochem. 2024; 16(4):203-206.

DOI: 10.52711/0975-4385.2024.00038

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.

Sr. No.	Extract	Colour	Nature	Percentage yield (%w/w)
Sr. No.	Extract	Colour	Nature	Maceration	Soxhlation
1.	Petroleum ether	Dark green	Sticky	1.81	2.50
2.	Chloroform	Dark green	Sticky	2.65	4.89
3.	Ethyl acetate	Dark green	Sticky	1.24	2.8
4.	Methanol	Reddish brown	Gummy	6.02	17
5.	Chloroform Water	Reddish brown	Gummy	8.18	10.16