INTRODUCTION:

In fact, all plant components are screened for phytoconstituents^1-3 that may be evident; further separation, purification and identification of the plant may be attributable to the presence of a phytoconstituents of interest.^4,5. Then it can be used for a new pharmaceutical⁶ product as the basis. Caesalpinia bonducella (CB) is a large, thorny, straggling shrub that belongs to the Fabaceae or Leguminosae plant family (Pea family) and is also known as Bonduc nut, Fever nut, or Gray nicker.

The major uses and benefits of fever nut are to suppress fever^7,8 and malaria. Leaf and twig paste were used to relieve toothaches, as well as boiled leaves are used for gargling to alleviate sore throat and also used to treat abdominal pain⁹, ruminant liver fluke. Fresh juice of tender leaves is administered to avoid mucous secretions along with the honey. Leaves and twigs are historically used in tumour therapy¹⁰, inflammation¹¹and liver disorders^12,13. Elephantiasis and smallpox have traditionally been treated with leaf juice.

MATERIALS AND METHODS:

Plant collection and authentication:

CB leaf was collected from puthupalayam village, Villupuram district, Tamilnadu, India. It was authentified by Dr. N. Srinivasan, Assistant Professor, Department of Pharmacy, Annamalai University.

Preparation of ethanolic extract of Caesalpinia bonducella:

By the hot percolation method using Soxhlet¹⁴ apparatus, coarse powder of CB leaves was extracted with ethanol. For 24 hours, the extraction was carried out. The solvent was extracted out to obtain a concentrated extract following extraction¹⁵. Then the concentrated extract was vacuum dried and the dry extract was kept in air tight jar for further phytochemical studies.

Phytochemical screening of ethanolic extract of Caesalpinia bonducella:

The EECB was subjected to preliminary phytochemical screening¹⁶ for the identification of various active constituents.

Carbohydrates¹⁷

Separately, the extract was dissolved in 5 mL of distilled H₂O and filtered. The filtrate was used in the following test:

i. Molisch test: Filtrate was treated in the test tube with 2 drops of alcoholic alpha-naphthol solution; the occurrence of carbohydrate is indicated by the formation of a violet ring at the junction.

ii. Benedict's test: The filtrate was treated and gently heated with Benedict's reagent, and the presence of orange red precipitate implying the existence of sugar reduction.

iii. Fehling test: 2 ml of extract was hydrolysed with dilute HCl, neutralised with alkali, and heated with Fehling's A and B solutions; red precipitate formation confirms the presence of reducing sugar reduction.

iv. Iodine test: 2 mL of extract treated with 5 drops of I₂ solution turns blue, indicating the presence of carbohydrates.

Alkaloids¹⁸

A quantity (3 ml) of concentrated extract was taken into the test tube and 1 ml HCl was added to the test tube, the mixture was gently heated and cooled and filtered for 20 minutes, and the filtrate was used for the subsequent test.

i. Mayer's Test: Mayer's reagent was applied to a fraction of the extract to determine the occurrence of alkaloids was observed for white precipitate or creamy-coloured precipitate formation.

ii. Wagner test: Filtrate has been treated with Wagner reagent; the presence of alkaloids is suggested by the formation of brown reddish precipitate.

iii. Hager's test: A yellow precipitate was formed when the filtrate was treated with Hager's reagent, indicating the existence of alkaloids.

iv. Dragendroff Test: The extract fraction was treated with the Dragendroff reagent, and the presence of alkaloids was determined by the formation of yellow precipitate.

Flavonoids¹⁹

i. Alkaline reagent test: The extract was treated with 10% NaOH solution and the presence of flavonoids is indicated by the creation of an intense yellow colour.

ii. Mg turning test: After treating the extract with Mg turning and adding conc. HCL to this solution, 5ml of 95 percent ethanol was added, and the formation of crimson red colour shows the existence of Flavonoids²⁰.

iii. Zn test: 2 ml extract was treated with Zn dust and concentrated HCl; the development of red colour proves the existence of Flavonoids.

Saponin:

5 ml extract was mixed with 20 ml of distilled H₂O and stirred in a graduated cylinder for 15 min of foam formation, suggesting saponin²¹.

Tannins

i. 2ml extract has been added to 1 percent lead acetate. A yellowish precipitate suggests the presence of tannins.

ii. With about 10 ml of distilled water, approximately 0.5 g of each component was stirred and then filtered. The presence of tannins suggests a few drops of 1% ferric chloride solution is applied to 2 ml of the filtrate occurrence of a blue-black, green or blue-green precipitate²².

iii. Bromine water test: The extract was dissolved and diluted with 50 percent alcohol. 3-4 drops of Br₂were used to react with the filtrate. The presence of buff colour indicates the presence of tannins in the sample of the plant test, while hydrolysable tannins did not give such an indication²³.

Proteins and Amino acids²⁴

i. Xanthoproteic test: Extract treated with a few drops of Conc.HNO3. Yellow formation shows protein presence²⁵.

ii. Biuret Test: Sodium hydroxide solution (40 percent) was applied to 0.5 mg plant extract solution in equal quantity, followed by a few drops of CUSO₄ solution (1 percent). In the research samples, the presence of violet colours marked the existence of proteins.

iii. Millon's test: A few drops of a million reagent were applied and heated to the extract. The red color means that proteins and free amino acids are present.

iv. Ninhydrin test: When the test solution was boiled with a 0.2 percent solution of Ninhydrin, it turned purple, indicating the presence of amino acids.

Steroids

i. Libermann-Buchard’s test: After chloroform treatment, the plant extract was filtered. The filtrate was treated with a few drops of acetic anhydride at room temperature before being boiled and cooled. The presence of steroids in the test samples was demarcated by the creation of brown-colour rings at the junction using concentrated H₂SO₄.

ii. Salkowski's test: The extract was chloroform-treated and purified. The filtrate was treated with a few drops of Conc. H₂SO₄ and shaken, resulting in a favourable outcome with a golden red colour.

Phenol:

Ferric Chloride Test: 4 drops of alcoholic FeCl₃ solution are treated with a test extract. The presence of Phenol suggests the formation of a bluish black colour²⁶.

Glycosides

i. Legal Test: The extract was dissolved in pyridine and added to make it alkaline with a sodium nitroprusside solution. The formation of pink-red to red coloration indicates the existence of glycosides.

ii. Baljet Test: 1 ml of ethanol extract was added with 1 ml of sodium picrate solution. Glycosides were noticed in the yellow to orange hue.

iii. Bontrager's test: A few ml of dilute HCl was added to 1 ml of extract solution. The chloroform filtrate was then removed after it was boiled and filtered. The chloroform layer was then treated with 1 ml of ammonia. The presence of anthraquinone glycosides was shown by the formation of red colour²⁷.

iv. Keller-Killani Test: Plant extract treated with a drop of FeCl₃ containing 2 ml of CH3COOH. The existence of a positive test is indicated by a brown ring.

Catechin:

2 ml of test solution in alcohol was mixed with Ehrlich reagent and a drops of Conc. HCl. The pink colour formation suggests the presence of catechins.

Coumarin:

3 ml of 10% NaOH was mixed with 2 ml of aqueous extract. The presence of coumarins causes the formation of a yellow colour.

Terpenoid:

In ethanol, a little of each portion has been dissolved. Adding to it was 1 ml of acetic anhydride followed by the addition of Conc. H₂SO₄. The presence of terpenoids was shown by a change in colour from pink to violet²⁸.

Oils and resins detection

i. Filter Paper Test: An extract sample was pressed between two filter papers. The oil-stained filter paper was evidence of fixed oils.

ii. Saponification test: With a few drops of 0.5 N alcoholic KOH and a drop of phenolphthalein were added to 1 ml of the extract. In a water bath, the mixture was heated for 1-2 hours. Soap formation or partial neutralization have both been used to demonstrate the presence of fixed oils.

RESULTS:

Preliminary phytochemical screening of Caesalpinia bonducella

Preliminary Phytochemical screening of Caesalpinia bonducella revealed the presence of following phytoconstituents showed in Table 2.

Table 2: Phytochemical screening of Caesalpinia bonducella

S. No	Test	EECB
1	Carbohydrates	+
2	Alkaloids	+
3	Flavonoids	+
4	Saponin	+
5	Tannins	+
6	Proteins & Amino Acids	+
7	Steroids	+
8	Phenolic Compound	+
9	Glycosides	+
10	Catechin	_
11	Coumarin	_
12	Terpenoid	_
13	Oils and resins	_

(-) indicates the absence of compound; (+) indicates the presence of compound

CONCLUSION:

Most of the remedies were taken from plants and proved to be beneficial, but, except for a few plants and some patented composite herbal drugs, the reason behind their use is not well known and they are stated to be successful without side effects. Phytochemicals play vital roles in protecting various organs against various toxic compounds. The presence of alkaloids, carbohydrates, proteins & amino acids, hormones, phenols, tannins, flavonoids, glycosides, and saponins was reported by preliminary phytochemical screening of Caesalpinia bonducella ethanolic extract. The needs of the human body can be supplemented by plants containing beneficial phytochemicals. This preliminary analysis of phytochemicals provides an idea of isolating the plant's most active constituents and also helps to establish new pharmacologically active compounds.

CONFLICT OF INTEREST:

The authors declare no conflict of interest.

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Received on 28.12.2020 Modified on 18.03.2021

Research J. Pharm. and Tech 2021; 14(11):5891-5894.

DOI: 10.52711/0974-360X.2021.01024