Plants have been used to produce medicines for millennia [1]. The hyacinth plant was a hydrophilic plant that floated freely and was commonly found in local water bodies in tropical and subtropical region [2].Water hyacinth (Eichhornia crassipes), is a perennial aquatic plant that floats freely and is indigenous to tropical and subtropical regions of South America [3].This perennial plant, which grows quickly and floats freely, is native to Ecuador and the Amazon basin in Brazil [4]. Hence, the hyacinth plant was the hydrophilic plant that floated freely and was primarily found in local water bodies in tropical and subtropical areas [5]. It is a vascular free-floating plant that has been shown to have a significant impact on the environment and society [6]. Plants naturally contain phytochemicals, which offer good health benefits [6]. It has been demonstrated that higher plants may include sources of novel antimicrobial compounds [7]. In settings without floating plants, the plant gives the upper part of the water column a complex structure that may be unique [8].

Fig 1: Water hyacinth

A great range of chemical compounds are synthesized by plants, and these can be categorized into primary and secondary metabolites based on their chemical class, origin of biosynthesis, and functional groups [9]. Alkaloid has antibacterial activities by disrupting virulence genes, bacterial membranes, inhibiting respiration and enzymes, and suppressing bacterial cell division [10].By preventing light from entering the river, water hyacinth reduced the productivity of the waterway [11]. The only effect of the weed's control efforts has been the temporary removal of the water hyacinths due to the high labor and cost requirement [12]. The gallic acid equivalents (mg gallic acid/g extract) that quantify the total phenolic content of five extracts were computed using gallic acid (0.2-1.0 mg/mL in methanol) as the standard [13].

It is critical to investigate alternative antioxidants produced from nature rather than synthetic antioxidants that cause cancer. Water hyacinth was tested for its antioxidant and other biological characteristics [14].

Antioxidants can scavenge free radicals and active oxygen species, suppress lipid peroxidation, stop hydrogen peroxides from breaking down into free radicals, or chelate heavy metal ions to demonstrate their antioxidant activity [15]. Environmental and genetic factors may influence the antioxidant capacity of fruits and vegetables [16]. These molecules help the body's antioxidant and detoxification functions break down and eliminate cancer-causing substances [17]. With a few modest modifications, a Winston et al. technique was employed to calculate the antioxidant value of each chemical studied. Integration was used to calculate the area under the kinetic curve. The total antioxidant capacity (TAC) of each molecule under investigation was then determined using equation 1. Vitamin C and phenolic were compared using the % increase in integrated area [18].On the other hand, it has been shown that peel has a high antioxidant content [19]. Gram-positive [Bacillus cereus (MTCC-1369)] and gram-negative [Escherichia coli (MTCC-739)] pathogens were utilized to examine the antibacterial impact, with little changes to previously defined parameters [20].

Flavonoids are hypothesized to work by rupturing the cell wall of bacteria bacteria, since alkaloids can combine with soluble and extracellular proteins to generate complex molecules. These secondary metabolites are classified into multiple groups by phytochemical screening, including terpenoids, compounds, essential oils, alkaloids, lectinic and polypeptidic, phenolic and polyphenolic, quinonic, flavonic, flavonolic, tannic, and coumarinic [21].

The purpose of this study was to determine the phytochemical elements of certain water hyacinth extracts (ethanol and distilled water), as well as to investigate their antibacterial and antioxidant capabilities.

2. Materials and Methods

2.1 Sample preparation

In the present study, Sample preparation Water hyacinth (Eichhornia crassipes) plants were collected from a pond in Birgaon. After giving the plants a good rinse with deionized water, they were allowed to air dry. After that, the dried leaves were ground into a fine powder in a mortar pestle.

2.2    Preparation of water extract

Distilled water the extraction of E. crassipes leaves was prepared by dissolving 1000 mg of leaves in 1000 ml of distilled water. Allowed to macerate for 60 min. in an ultrasonic bath. The suspension was filtered and stored in airtight containers.

2.3    Soxhlet Extraction

Load the 10 g sample material containing the desired compound into the thimble. Place the thimble into the main chamber of the soxhlet extractor. Add the 110 ml of distilled water to a round bottom flask and place onto a heating mantle. Attach the Soxhlet extractor above the round bottom flask. Attach a reflux condenser above the extractor with cold water entering at the bottom and exiting above. No, the apparatus is set up to heat the solvent to reflux and leave to extract for the required amount of time.

2.4    Phytochemicals Analysis

Plant samples were analyzed following standard methods to demonstrate the presence of the different pharmacologically active compounds.

1.       Test of Alkaloids

Mayer’s test: Take a few ml of filtrate and add 1-2 drops of Mayer’s reagents (along the sides of the test tube). A white, creamy, or yellowish precipitate forms; the test is positive otherwise absence of alkaloids.

Iodine test: Take 3 ml of extract solution and add a few drops of iodine solution. Occurrence of a reddish-brown precipitate shows presence otherwise absence of alkaloids.

2.       Test of Carbohydrates

Molish's test: take 2 ml of filtrate, add 2 drops of alcoholic naphthol, and add 1 ml of conc. H₂SO₄ (along the sides of the test tube). Development of a purple ring at the layer formed by the concentrated acid is a positive indicator otherwise absence of carbohydrates.

3.       Test of reducing Sugars:

Bendict's test: Take 0.5 ml of filtrate; add 0.5 ml of Bendict’s reagent, and boil for 2 min. Brick-red precipitate indicates the presence of reducing sugars otherwise absence of reducing sugar.

Fehling’s test: Take 1 ml of each of Fehling’s solutions A and B, add 1 ml of filtrate, and boil in a water bath. Reddish-brown precipitate indicates the presence of reducing sugars otherwise absence of reducing sugar.

4.       Test of Phytosterols

1 ml of sample extract, 3 ml of chloroform, and 1 ml of sulfuric acid were added. The presence of sterol is confirmed if the lower layer appears red in color.

5.       Test of Glycosides

Take 0.5 ml of extract, 2 ml of glacial acetic acid, 1-2 drops of ferric chloride, and add 1 ml of H₂SO₄. The presence of glycosides is confirmed by the interface brown ring color.

6.       Test of Quinones

Take a little ml of plant extract and add Conc. HCl. The presence of quinones is confirmed by the green color.

7.       Test of Phenolic Compound

Take plant extract dissolved in 5 ml of distilled water and add 3 ml of lead acetate solution. The presence of a phenolic compound is confirmed by a white precipitate.

8.       Test of Protein and Amino Acid

Take 1 ml of filtrate and 10% NaOH, 1 drop of 1% (0.5 ml) copper sulfate solution, and 1 ml of ethanol (95%). If the solution turns purple or violet, the sample contains protein otherwise absence of protein and amino acid.

9.       Test of Tannins

Take 0.4 ml plant extract, 4 ml 10% NaOH, and shake well. The presence of tannins is confirmed by the formation of an emulsion (hydrolysable tannins).

10.    Test of Terpenoids

Take 2.5 ml extract, 1 ml chloroform (CHCl₃), and 1.5 ml conc. H₂SO₄ (boiled in a water bath). The presence of terpenoids is confirmed by a reddish brown-colored solution.

2.5   Total antioxidant:

To create a DPPH solution, measure 0.025 mg of the drug, dissolve it in methanol, and dilute it at a 1:9 ratio. Create a blank solution, prepare a test sample, and add 0.1 ml of the desired extract. Place the sample in a dark setting for an hour to stabilize and determine the antioxidant capabilities of the extract.

   ………(i)                                            

Where,

A₀ = absorbance of dilute DPPH solution

A = absorbance of plant extract 

Table 1: Absorbance of samples

2.6 Total Phenolic Content (TPC)

The DPPH solution method is a widely used method for assessing the total phenolic content and antioxidant activity of various samples. It involves combining a sample extract with a DPPH solution and measuring absorbance changes at 517 nm after a 30-minute incubation time. This indicates the sample's antioxidants' ability to effectively scavenge free radicals. Researchers can compare this percentage suppression to a standard curve of known antioxidants like ascorbic acid or gallic acid, indicating the sample's phenolic content and antioxidant properties. The DPPH test is a reliable tool for researchers, allowing them to make informed conclusions about the antioxidant content and activity of their experimental materials.

2.7 Antibacterial analysis

To obtain pure bacterial cultures like E. coli, inoculate them into nutrient broth and incubate overnight at 37°C. Prepare PDA agar plates by placing them on sterile Petri plates and spreading them uniformly. Each stage ensures the purity and isolation of cultures, which are essential for future studies. Researchers can achieve high-quality pure cultures by following precise protocols and adherence to sterile techniques.

The antimicrobial testing process involves creating wells in agar with sterile tools and adding extracts like water hyacinth, ethanol, E.coli, or antibiotic tablets. Alternatively, disk diffusion involves soaking paper disks with the extract. After incubating at 37°C for 24 hours, assess zones of inhibition for antibacterial properties. This method collects reliable data for analyzing the efficacy of tested extracts or compounds.

3. Result and discussion

1.       Phytochemicals screening:

The study examined the nutritional and phytochemical components of water hyacinth, focusing on its ethanol and distilled water extracts. Results showed glycosides, flavonoids, tannins, phytosterol, and quinone in the ethanol extract, while alkaloids, proteins, amino acids, and carbohydrates were absent in the water extract.

Table 2: Phytochemicals analysis

2.       Total Phenolic contents:

Phenolic chemicals, found in fruits, vegetables, and herbs, are known for their strong antioxidant capabilities. This study examines the hydroethanolic extract of E.crasspies, which has high polyphenol and flavonoid content. Gallic acid and quercetin are two potent natural antioxidants found in the plant extracts. The phenolic content of the ethanolic leaves and water extract was determined, and their hydroxyl groups aid in their ability to scavenge free radicals. Flavonoids, plant secondary metabolites, depend on free OH groups, particularly 3-OH, for their antioxidant activity. The ethanolic extract of E.crasspies was found to be higher than the water extracts, which had lower concentrations of various phenolic acids. The leaves of water hyacinth showed strong ability to chelate iron and prevent lipid peroxidation in fish oil and liposomes.

Table 3: Absorbance of total phenolic content sample

3.       Total Antioxidant activity:

The study reveals that Eichhornia crassipes has antioxidant activity that can be beneficial for both the food and pharmaceutical industries. Its ant oxidative capacity is demonstrated through its ability to reduce DPPH radicals, a stable free radical. The DPPH assay reveals a strong association between phenolic content and antioxidant activity. Higher concentrations of polyphenols and flavonoids are linked to each extract's IC50 value. Water hyacinth leaves contain antioxidant glutathione, which can prevent or manage illnesses linked to oxidative stress. The study suggests that E. coli can be used as an antioxidant and preservative in both food and non-food systems.

Table 4: Absorbance of total antioxidant sample

…..(ii)                                                           

                                 =0.941-0.785÷0.941×100

                                 =0.156÷0.941×100 = 16.57%

4.       Antibacterial Test

The water hyacinth extract showed potent antibacterial activity against various pathogenic and fecal indicator bacteria, with a significant difference in performance compared to E. coli. The extract had an inhibition zone of 2.2 mm, while E. coli had a zone of 2.0 mm. This highlights the extract's potential as a natural, effective antibacterial agent.

Conclusion

E.crasspies demonstrated the existence of primary and secondary metabolites in the current investigation. The presence of phenolics appears to be responsible for the antioxidant properties of plant extracts. This work encourages further research into spectroscopic approaches for isolating and identifying active compounds. This study found that the damaging aquatic plants could be used to generate alternative substances to treat infectious disorders caused by bacterial infections. Water hyacinth (Eichhornia crassipes) ethanol extracts showed significant antibacterial and antioxidant activity. This could be due to a variety of biologically active chemicals, such as glycosides, flavonoids, and phenols.