View of PHYTOCHEMICAL SCREENING OF GUAVA (PSIDIUM GUAVAJAVA L.) LEAVES EXTRACT AND ITS MEDICINAL IMPORTANCE

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VAOLUME: 09, Special Issue 06, (IMC-RMSPE-2022) Paper id-IJIERM-IX-VI, August 2022 7

PHYTOCHEMICAL SCREENING OF GUAVA (PSIDIUM GUAVAJAVA L.) LEAVES EXTRACT AND ITS MEDICINAL IMPORTANCE

Zahoor Ahmad Lone and N.K. Jain

Department of Botany, Govt. Holkar Science College, Indore (MP) India

Abstract - The purpose of the current study was to look into the therapeutic benefits and phytochemical content of Psidium guajava L. leaves. Plants chosen for the study are Psidium guajava L..The leaves powder was successively extracted with water, methanol, ethyl acetate and petroleum ether. Aklaloid, tannins and sterols are all present, according to phytochemical study. In tropical and subtropical nations, Psidium guajava is a significant food crop and medicinal plant.Guava (Psidium guajava L.) preparations have historically been used to treat a variety of illnesses. Most indigenous peoples around the world include traditional medicine into their culture and view of health. Guava (Psidium guajava L.) leaf tea has been used for centuries to treat a variety of ailments, including cough, rheumatism, diarrhea, and diabetes mellitus. The findings of this study proved that the guava plant's leaves contain certain significant bioactive chemicals that can impede the growth of microorganisms, making them a potent source of antibiotics.

Keyword: Guava, Phytochemical,guava leaves extract and Soxhlet extraction.

1 INTRODUCTION

Plants have been a great source of medical agents for many years.Natural medicines have gained popularity in the treatment of numerous ailments as a result of public perceptions that these phyto-medicines are safe, easily available, inexpensive, and have fewer side effects. Compared to traditional pharmaceuticals, many plant-based remedies are more affordable and accessible to the majority of people. After use, there are less negative effects.

These factors may be responsible for their widespread use and attention. The primary source of novel pharmaceutical and healthcare products is medicinal plants. These "green factories" have produced some high-profile medications through the extraction and characterisation of some potent phytocompounds[1]. Psidium guajava is a plant that thrives in tropical and subtropical climates. Other documented uses include dysentery, gastroenteritis, stomach, and antibacterial colic pathogenic germs of the intestines, with the primary traditional usage being as an anti-diarrheal[2]. A small tree that can grow to a height of 20 feet, Psidium guajava L. has spreading branches. The guava tree is known for its smooth, thin, copper-colored bark that peels off to show the layer of greenish tissue underneath it and for the attractive, "bony" appearance of its trunk, which over time may expand to a diameter of 10 inches[3]. The plant Psidium guajava, which is thought to be indigenous to Mexico, is also found in South America, Africa, Europe and Asia. It can be found primarily in tropical and subtropical areas. Bihar, Maharashtra, Uttar Pradesh, Assam, West Bengal, Andhra Pradesh and Haryana are among the Indian states where it is distributed[4]. The leaves are opposite, shortpetioled, opposite, oval or oblong-elliptic, somewhat uneven in shape, leathery, with noticeable parallel veins, and more or less downy on the underside, they are 7-15 cm long to 3-5 cm wide[5]. White flowers are produced in the leaf axils either singly or in tiny clusters. Flowers are 2-3 cm broad, have 4 or 5 white petals that fall off fast, and a noticeable tuft of 250 or so white stamens with pale yellow anthers at the tips[6].When ripe, the fruit has a strong, sweet, musky smell and is round, ovoid, or pear-shaped. Its thin, light-yellow skin is frequently tinged pink, and it has 4 or 5 protruding floral remnants (sepals) at the fruit's apex. A layer of somewhat granular flesh that is 1/8 to 1/2 inch thick, white, yellowish, light or dark pink, or nearly red, juicy, acidic, subacidic, sweet, or delicious is found next to the skin. Although some uncommon species contain soft, chewable seeds, the core pulp, which is slightly deeper in tone, is juicy and typically filled with very hard, yellowish seeds that are 1/8 inch long. However, some guavas are either seedless or almost seedless. Actual seed counts have ranged from 112 to 535. The fruit is green, hard, sticky within, and quite astringent while young and up until just before maturity. Smooth, light pinkish brown bark that is frequently tinted with chlorophyll[7]. Psidium guajavaL. is utilized in many parts of the world as an anti- inflammatory, for diabetes, hypertension, caries, wounds, pain relief, and lowering fever, according to more recent ethno pharmacological investigations. Guava has a long history of

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use as a traditional medicine, particularly in Mexico and other Central American nations like those in the Caribbean, Africa, and Asia[8].

Taxonomic Classification Kingdom - Plantae

Sub-division - Spermatophyte Division - Magnoliophyta Class - Magnoliopsida Sub-class - Rosidae Order - Myrtales Family - Myrtaceae Genus - Psidium Species - Guajava

The different names of Guava (Psidium guajava) are as given below:

Common Name : Guava

Botanical Name : Psidium guajava L.

English Name : Guava, Abas, Govavier, Kautonga, Kuahpa

Hindi Name : Amrud

Sanskrit Name : Amratafalam, Perala Punjabi name : Amrut

Tamil name : Segapu, Koyyo, Koyapalam Telugu name : Goya-pandu, Jam-pandu, Jama 2 MATERIALS AND METHODS

2.1 Collection of plants:

Psidium guajava L. belongs to the family Myrtaceae. Guava (Psidium guajava L.)leaves are gathered from a kalibit forest in the khandwa district (madhya pradesh). The plant leaves was identified in Govt. Model autonomous Holkar science college indore (M.P) india by Dr.

Navin Kumar Jain (Professor, Department of Botany). Psidium guajava L. dry leaf powder was utilized in the current investigation. A study was conducted utilizing several solvent extracts to examine various phytochemical characteristics. The shade dried Psidium guajava leaves were powdered mechanically and stored in an airtight container.

2.2 Prepration of Plant Extract:

Psidium guajava leaves from the kalibit forest in the khandwa district (madhya pradesh) were collected, shade dried, powdered, and then extracted in a soxhlet apparatus with water, methanol, ethyl acetate and petroleum ether respectively.

2.3 Prepration of Extract by Soxhlet Extractor

To prepare the extracts, soxhlet extraction was performed on dried powder. Methanol, ethyl acetate and petroleum ether were used as solvents. Each sample's dry powder (35 gm) was placed in the thimble of the Soxhlet apparatus. Each sample was given 400 ml of a different solvent, one at a time. The extraction was carried out until the thimble contained clear solvent. It requires roughly 25 hours. The concentrated extract was then heated to room temperature and evaporated until a black residue was produced. For a preliminary phytochemical study, weighed extracts were stored in airtight containers.

2.4 Preliminary Phytochemical Analysis

These extracts were tested in order to find out the presence of active compounds by use of following standard methods.

2.5 Test for Carbohydrates:

Extracts were dissolved individually in 5ml of distilled water and filtered. The filtrate was used for the following test.

(a) Molisch’s test: filtrate were treated with 2 drops of alcoholic ∝ −naphthol solution, formation of violet ring at the junction indicates the presence of carbohydrate.

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(b) Benedict’s test: 1 millilitre of plant extract and 1 millilitre of Benedict's reagent were heated for 5 minutes. The presence of carbohydrates (disaccharides) was shown by the formation of an orange precipitate.

(c) Fehling’s Test: 2 ml of extract was hydrolyzed with dilute HCl and neutralized with alkali and heated with fehling’s solution A and B, formation of red ppt indicates the presence of reducing sugar.

2.6 Test for Proteins:

(a) Xanthoprotic test: Extract was treated with few drops of conc. HNO3. Formation of yellow indicates the presence of proteins.

(b) Biuret test: 1ml of plant extract was taken in a test tube followed by 4% NaOH and 1% CuSO4. Violet pink colour development indicated the presence of proteins

2.7 Test for Lipids:

(a) Solibility test: 1 ml of plant extract was evaporated to dried powder. Few drops of petroleum ether were poured into the test tube and shaken well. Complete dissolution of extract identified the presence of lipids.

(b) Glycerol test: To 1 ml of 1% CuSO4.5H2O solution, 5 drops of the plant extract were added and mixed thoroughly. Then it received 5 drops of a 10% sodium hydroxide solution. A clear blue solution was obtained which indicates the presence of glycerol.

(c) Sudan III test: To 1ml of plant extract, few drops of Sudan III solution were added.

Appearance of red color indicated the presence of lipids.

2.8 Test for Alkaloids

(a) Mayer Test: 1 ml of guava leaf extract, 2 drops of chloroform, and 2 drops of meyer reagent were added. A positive alkaloid reaction resulted in the production of white deposits.

(b) Wagners test: 1 ml extract was treated with Wagner’s reagent, formation of brown reddish precipitate indicates presence of alkaloids.

2.9 Test for Tannins:

(a) Lead acetate test: To the filtrate, 5ml of 10% lead acetate solution was added.

Formation of white precipitation indicates the presence of tannin.

(b) Ferric chloride test: In this test, a Ferric chloride solution plus 5% ferric chloride solution will be added drop by drop, 2-3ml in the solution of leave of guava extract in order to observed the appearance of Tannins.

2.10 Test for Saponin:

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Foam test: 1 ml of plant extract was taken in a test tube with small amount of water. Sodium bicarbonate was added to it and shaken vigorously for 5 min.

Formation of foam indicated the presence of saponins.

2.11 Test for Resins:

In a dry test tube, to 0.5 ml of acetic acid and 2 drops of conc. Sulphuric acid. A yellow colouration that disappeared on standing indicated the presence of flavonoids.

2.12 Test for Sterols (Salkowski test)

2 millilitre chloroform and 2 millilitre concentrated sulphuric acid were applied successively from the side of the test tube to 0.5 ml of the plant extract. A few minutes were spent gently shaking the test tube. Sterols were present because the chloroform layer turned red as they developed.

2.13 Test for Cardiac Glycosides (Keller Killiani test)

Approximately 5 ml of each plant extract was evaporated at 40°C and the residue was collected. A few mg of residue was suspended in 5 ml water. 2 ml of glacial acetic acid containing one drop of ferric chloride solution was added to it. This solution was underplayed with 1 ml of conc. sulphuric acid. A brown ring at the interface indicated the presence of deoxy sugar, a characteristic of cardiac glycosides

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VAOLUME: 09, Special Issue 06, (IMC-RMSPE-2022) Paper id-IJIERM-IX-VI, August 2022 10 2.14 Test for Triterpenes:

Few milligrams of plant extract residue was mixed with 5 ml of chloroform and warmed for 30 min at 40°C. Few drops of conc. sulphuric acid were added and mixed well. The appearance of red colour indicated the presence of triterpenes.

2.15 Test for Anthraquinones:

2ml of each plant extract was shaken with 10ml benzene, and 5ml of 10% ammonia solution was added. The mixture was shaken in order to obatained the coulur of antraquinones.

3 RESULTS AND DISCUSSIONS

The phytochemical screening of Guava extractrevealed the presence of lipids, alkaloids, sterols and tannins. While carbohydrates, proteins, saponins, resins, cardiac glycosides, Triterpenes and anthraquinones absents stated in (Table 1).

Table 1 Result of the Phytochemicals analysis of Guava leaves extract

S. No. Phytochemicals Extracts

Aqueous Methanol Ethyl acetate Petroleum ether 1 Carbohydrates

a Molisch’s test - - - -

b Benedict’s test - - - -

c Fehling’s test - - - -

2 Protein

a Xanthoprotic test - - - -

b Biuret test - - - -

Lipid

a Solubility test - - + +

b Glycerol test - - - -

c Sudan III test - - - -

3 Alkaloids

a Mayer’s test - - - -

b Wagner’s test + + - -

4 Saponins

a Foam test - - - -

5 Resins test - - - -

6 Tannins test

a Lead Acetate test - - + +

b Ferric chloride test - - - -

7 Sterols

a Salkowski test + - - -

8 Cardiac

Glucosides a Keller – Killiani

test - - - -

9 Triterpenes test - - - -

10 Anthraquinone

test - - - -

Note + represents the presence of the constituents - represents the absenence of the constituents

Carbohydrate test used three methods namely Molisch’s test, Benedict’s test and Fehling;s test, all methods show negative results of guava leaf extract. In protein test also two methods are used namely Xanthoprotic test and Biuret test, both methods show negative result. In Lipid test three methods are used Solubility test, Glycerol test and Sudan III test, all methods show negative results. In alkaloids test used two methods namely Mayer’s test and Wagner’s test, In Mayer’s test it shows negative result and Wagner’s test it shows positive results in aqueous extract and methanol extract. In saponins test only one method is used namely foam test and this test show negative result. Resin test also shows negative result. In Tannin’s test two methods are used namely Lead acetate test and Ferric chloride test. Lead acetate test shows positive result in (Ethyl acetate and petroleum ether) and Ferric chloride test shows negative result. In sterols test only one method used namely Salkowski method, it shows positive result in aqueous extract. Cardiac glycosides,

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Triterpenes test and Anthraquinone test all three test show negative result. According to the findings of this study's investigation, guava leaf extract included phytochemicals with established pharmacological action.This investigation indicated the existence of numerous medically significant phytochemicals in guava leaf extract.

This study revealed the presence of various phytochemicals in guava leaves extract.

According the result obtained in the determination of lipids in a guava leaf extract complete dissolution of extract indicates the presence of alkaloids. Alkaloids are the most effective and important plant compound for healing[9].

However in the determination of Tannin in guava leaf extract, a curdy white precipitation indicates the presence of tannin. The presence of tannin in guava leaves suggests that guava leaf can be used as antidiabetic,antioxidant, antimicrobial, antifungal, antidiarrheal, antioxidant and anti-inflammatory [10]

Similarly in the determination of sterols in guava leaf extract, a red color in the chloroform layer indicated the presence of sterols. Steroids interact with sex hormones like oxytocin, which controls the start of labor and subsequent milk production in pregnant women. Due to their interactions with other substances, such as sex hormones, steroidal chemicals are particularly important and fascinating to the field of pharmacy [11].

In many countries, preparations of the leaves have been used traditionally in folk medicine, primarily as an anti-diarrhea treatment[12]. Psidium guajava leaf extracts show strong antimicrobial effects against Propionibacterium skin disease and should be effective in treating acne, especially as they become more well known to have medicinal properties [13]. In many parts of the world, the fast rising incidence of diabetes mellitus is starting to seriously threaten human health. Traditional medicine offers good clinical potential and predicts a bright future in the treatment of diabetes mellitus and its consequences thanks to its unique traditional medical viewpoints and natural medicines that are primarily derived from herbs. Several authors have investigated how guava bark, leaves, and fruit can act as antidiabetic drugs [14].In many parts of the world, the fast rising incidence of diabetes mellitus is starting to seriously threaten human health. Traditional medicine offers good clinical potential and predicts a bright future in the treatment of diabetes mellitus and its consequences thanks to its unique traditional medical viewpoints and natural medicines that are primarily derived from herbs. Several authors have investigated how guava bark, leaves, and fruit can act as antidiabetic drugs [15]. The application of conventional medication to treat childhood diarrhea. Guava leaves were one of the typical traditional remedies recommended by respondents as a first line treatment for diarrhea [16].

CONCLUSION

Phytochemical screening of aqueous, methanol, ethyl acetate and petroleum ether extracts revealed the presence of alkaloids, tannins and sterols by positive reaction with the respective test reagent. The aqueous and methanol extracts contained the most phytoconstituents, according to phytochemical screening. The effectiveness of Psidium guajava L. It has been proven to be successful in treating conditions including diarrhoea, dysentery, gastroenteritis, hypertension, diabetes, caries, pain alleviation, cough, mouth ulcers, and liver damage inflammation and other widespread illness. Guava possesses antibacterial, antidiabetic, antiviral, antioxidant and anti-inflammatory activities. Due to these biological activities it can be quite helpful for the preventions and treatments of diseases.

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