A Comprehensive Review Of Water Quality Analysis: Parameters, Analytical Methods, And Emerging Perspectives

Fig.1: Mangifera indica L.

2. Common Names:

Aam, Manga, Ma, Mango, Maavu, Mangoo

3. Taxonomical classification:

• Kingdom            :  Plantae
• Subkingdom      :  Tracheobionta
• Superdivision    : Spermatophyta
• Division              :  Magnoliophyta

• Class                   :  Magnoliopsida
• Subclass             :  Rosidae
• Family                : Anacardiaceae
• Order                 :  Sapindales
• Genus                 : Mangifera
• Species               : indica

4. Morphological characters:

• Root        : Deep tap root with well-developed lateral roots.
• Stem        : Woody, erect trunk with rough, dark grey-brown bark.
• Leaves     : Simple, alternate, lanceolate, glossy dark green; young leaves are reddish.
• Flowers   : Small, fragrant, yellowish-white flowers arranged in terminal panicles.
• Fruits      : Fleshy drupe with sweet, juicy pulp and a hard stone.
• Seeds       : Single large, flattened seed enclosed in a hard fibrous endocarp.
• Crown     : Broad, dense, rounded crown with spreading branches.

     5. Chemical Constituents:

• Polyphenols    
• Flavonoids      
• Phenolic acids   
• Carotenoids     
• Vitamins           
• Terpenoids and triterpene
• Essential fatty acids
• Sugars, proteins, carbohydrates, dietary fibre, and minerals

6. Pharmacological activities:

• Anticancer activity
• Antidiabetic activity
• Anti-inflammatory activity
• Hepatoprotective activity
• Anti-haemorrhagic activity
• Anti-tetanus activity
• Analgesic and antipyretic activity
• Anti-ulcer activity
• Lipid-lowering (Hypolipidemic) activity
• Antioxidant activity
• Antimicrobial activity

7. Part used:

Flowers

MATERIALS AND METHOD:

Plant material:

The flowers of Mangifera indica L. were collected from vandavasi, Tiruvannamalai district in Tamil Nadu. which was authenticated by Dr. J. Sureshkumar, M.Sc, M.Phil, Ph.D, PGDCA.,

Chemicals:

Ethanol, n-Hexane, Tween 80 were obtained from GREAT SCIENTIFIC INDUSTRIES Seriyenthal, Tiruvannamalai, Tamil Nadu.

Methods of Extraction:

Mangifera indica L. (Flower) were collected and shade dried at room temperature for 3 weeks. The dried flowers were powdered and passed through sieve no: 22# and 42#.

Requirements:

Plant: Dried flower powder (Mangifera indica L.)

Solvent: Ethanol, n-Hexane

Fig.2: Mangifera indica L. Flower

Extraction process of Mangifera indica L.:

Soxhlet Process:

35 g of dried coarse powder was packed into a cellulose thimble, which was placed in the chamber of a Soxhlet apparatus. The extracting solvent in the flask was heated, and its vapours were condensed in the condenser. The condensed solvent dripped into the thimble containing the crude drug, allowing it to come into contact with the sample. When the liquid level in the chamber rose to the tip of the siphon tube, the liquid contents of the chamber were siphoned back into the flask. This process was continuously carried out until a drop of the solvent from the siphon tube left no residue.

Fig.3: Soxhlet process.

PHYTOCHEMICAL SCREENING:

All the Mangifera indica L. (Ethanol and n-Hexane) extracts were analysed for preliminary phytochemical screening for identification of various phytoconstituents.

TEST FOR ALKALOIDS

HAGER’S TEST

To 1mL of the extract, add 3mL of Hager’s reagent (saturated aqueous solution of picric acid), yellow coloured precipitate indicates the presence of alkaloids.

TEST FOR CARBOHYDRATES

FEHLING’S TEST

1mL of the extract, add equal quantities of Fehling’s solution A and B. Upon heating formation of the brick red precipitate indicates the presence of sugar.

TEST FOR PROTEIN AND AMINO ACIDS

MILLION’S TEST

1mL of the test solution is made acidic with sulphuric acid and add million’s reagent (Mercuric nitrate in nitric acid) and boil this solution. A yellow precipitate is formed indicates the presence of protein.

TEST FOR FLAVONOIDS

LEAD ACETATE

1mL of plant extract add few drops of 10% lead acetate solution. Formation of yellow precipitate indicates the presence of flavonoids.

TEST FOR PHENOLIC COMPOUNDS

FERRIC CHLORIDE TEST

1mL of plant extract add few drops of 5% ferric chloride solution. Formation of dark green/bluish black indicate presence of phenolic compound.

TEST FOR TANNINS

GELATIN TEST

To a few mL of extract, add 1% gelatin solution containing 10% sodium chloride. Formation of white precipitate indicates the presence of tannins.

TEST FOR SAPONINS

FOAM TEST

Take a small quantity of  extract and add 20mL of distilled water and shake in a graduated cylinder for 15 mins lengthwise. A 1cm layer of foam indicates the presence of saponins.

TEST FOR STEROIDS

SALKOWSKI TEST

Dissolve the extract in chloroform and add equal volume of concentrated sulphuric acid Formation of bluish red to cherry red colour in chloroform layer and green fluorescence in the acid layer represents the steroidal components in the tested extract.

TEST FOR TERPINOIDS

SALKOWSKI TEST

Dissolve the extract in chloroform and add equal  volume of concentrated sulphuric acid Formation of bluish red to cherry red colour in chloroform layer indicates presence of terpenoids.

TEST FOR GLYCOSIDES

BORNTRAGER’S TEST

1g of plant extract and add 5 to 10mL of dilute HCL boil on water for 10 mins and filter  Filtrate was extracted with CCL4/benzene. Then add equal amount of ammonia solution to filtrate and shake. Formation of pink or red colour in ammonical layer is due to presence of glycosides.

IN VITRO ASSAY TO EVALUATE THE ANTI-INFLAMMATORY ACTIVITY:

PROTEIN DENATURATION INHIBITION ASSAY:

Egg albumin  is used as the protein in this assay.

PRINCIPLE OF THE ASSAY

The process by which proteins lose their secondary and tertiary structures as a result of external pressures or chemicals like heat, an organic solvent, a concentrated inorganic salt, or strong acid or base is known as protein denaturation.

Proteins lose their biological functions when they become denatured.  Protein denaturation is a key feature of inflammation. Therefore, the ability of a drug or a plant product to stop or reduce protein denaturation may have the ability to stop or control inflammation. Heat induced egg albumin/bovine serum albumin denaturation inhibition assays are frequently used as protein denaturation inhibition assays.

 When the reaction mixture is exposed to heat, protein denatures and increases the turbidity of the mixture. This leads to an increase in the absorbance of the mixture. If a potential drug/plant product has anti-inflammatory properties, it will inhibit protein denaturation, lowering the progressive increase in absorbance due to protein denaturation.

Method Of Heat-Induced Egg Albumin Denaturation Inhibition Assay

Making a 1% egg albumin solution: 1% egg albumin solution can be prepared with fresh hen eggs or easily found egg albumin powder from stores.

To  make an egg-albumin solution with a fresh hen's egg, crack the egg carefully. Then add 1mL of the translucent part to 100mL of w/v distilled water and stir until well mixed. Egg albumin is the transparent part of the egg. While making the solution, the water should be cold. When water is brought to boil, it will coagulate.

Test Mixture: For each test mixture, 2.8 mL of Phosphate-Buffered Saline (PBS) adjusted to pH 6.4, 0.2 mL of egg albumin solution, and 2 mL of the respective plant extract at concentrations of 100, 200, 300, 400, and 500 µg/mL were mixed to obtain a final volume of 5 mL.

Standard Mixture: For the standard mixture, 2.8 mL of PBS (pH 6.4), 0.2 mL of egg albumin solution, and 2 mL of the reference drug, aspirin, at concentrations of 100, 200, 300, 400, and 500 µg/mL were mixed to obtain a final volume of 5 mL.

For positive control, mix 2.8mL of PBS (pH6.4), 0.2mL of egg albumin solution, and 2mL of distilled water to obtain a volume of 5mL.

As the negative control, 5mL of distilled water can be used.

Incubate all the mixtures (tests, standards, controls) at 37 ± 2 °C for 15 minutes. Then, increase the temperature gradually to 70°C. Once it reaches 70 °C, keep the mixtures at 70°C in water bath for another 5 minutes. Take out the mixtures from the water bath and allow them to cool down to room temperature. After cooling, measure the absorbance of each mixture at a wavelength of 660nm using a spectrophotometer. Carry out the test in triplicates.

Finally, calculate the percentage of protein denaturation inhibition using the formula below,

(Vc- absorbance of the positive control, Vt -absorbance of the test)

Fig.4: Reaction mixture of  Standard     

Fig.5: Reaction mixture of Ethanol

Fig.6: Reaction mixture of n-Hexane

Statistical Analysis

Results are expressed as Mean ± SEM (n = 3). Statistical analysis was performed using one-way analysis of variance (ANOVA) followed by Dunnett's multiple comparison test (Control vs test groups) using GraphPad Prism. Differences were considered statistically significant at P < 0.001           

 RESULTS AND DISCUSSION:

Table 1: Extractive Values Of Various Extracts.

Fig.7: n-Hexane Residue       

Fig.8: Ethanol Residue

Table 2: Phytochemical Screening Of Various Extracts

Fig.9:Test for Ethanolic extract   

Fig.10: Test for n-Hexane extract

Table 3: In Vitro Anti-Inflammatory Activity Of Mangifera Indica L. Extracts By Protein Denaturation Assay.

Each value represents Mean ± SEM (n = 3). Experimental groups were compared with the control by one-way Variance ( ANOVA ) followed by Dunnett's multiple comparison test.            P < 0.001 was considered statistically significant.

Fig.11: Percentage inhibition of protein denaturation by ethanol extract, n-hexane extract, and aspirin.