Preliminary Phytochemical Characterization and Micromeritic Properties evaluation of Azadirachta indica and Syzygium aromaticum Powder

 

Kriti Sethi*, Nayyar Parvez

Department of Pharmacy, School of Medical and Allied Sciences, Galgotias University, Greater Noida, India.

*Corresponding Author E-mail: ngraghu@rediffmail.com, drngraghu@gmail.com

 

ABSTRACT:

From history, human societies have kinetically got round plants which have affected a great amount to the all persons as a group. Plants have the building to put in danger different variety of phytochemical and biochemical compounds which can be acclimated to act different biological functions. Many of these phytochemicals have well, with good effect effects on in the long run being healthy when destructed by the human and can be effectively used to treat human diseases. The current research paper deals with the various Phytochemical Characterization and Micromeritic Properties evaluation of Azadirachta indica and Syzygium aromaticum Powder. The analysis was carried out utilizing standard methods and protocols. The result suggests that the Azadirachta indica extracts contain plenty of phytochemicals with antimicrobial, anti-inflammatory and antioxidant properties and Syzgium aromaticum showed antibacterial properties and used in disparate dental creams, tooth pastes, mouth washes, and esophagus sprays to remove bacteria. However, from the aggregation of all our outcomes, we arrived at a reasoned opinion that the aforementioned herbs definitely have as owner possible unused quality therapeutic properties of changing degrees of existence of several phyto-active compounds in them.

 

KEYWORDS: Medicinal Plant, Syzygium aromaticum, Azadirachta indica, FT-IR, Micromeritic Properties, Phytochemical Characterization.

 

 


INTRODUCTION:

Plants are the primary source of medicine. Medicinal plants are considered to be rich sources of secondary metabolites and oils which are beneficial. The most important advantages of medicinal plants in various treatments are: their safety besides being less expensive and easily available around the world. Herbal remedies prepared from various herbal drugs have been used against various diseases since from ancient times.

 

Over past few years compounds from natural origin have been gaining importance due to the presence of the vast phyto-constituents present in it and the vast chemical diversity it offers. This has resulted in an increased demand in herbal preparations. The increased demand is because of the relatively safe use of the herbal drugs and its ease of availability. [1]

 

Herbal and natural products of population of society use medicines form centuries in all culture far and wide in the world. “Let food be your medicine and let medicine be your food” was investigated by Hippocrates, over two million years ago. It’s likewise true today that “you are what you eat. [2] Herbal medicines have minority side-effect in analogy with timid medicines, yet side-effects do occur. The intuitive products derived from medicinal plants one as neem, tulsi, amla, dhatura, nimbu, clove, asafotida etc., have proven to be abundant source of biologically active compounds, many of which have adopt the essence for the development of new leads of chemicals for pharmaceuticals. Use of herbal and dietary supplements (HDSs) is a well-documented area among consumers of all age groups. Many complacent properties a well known as anti-inflammatory, antioxidants, antibacterial, and astringent transpire have been bottom in either plants or their parts. [3]

 

Azadirachta indica It is popularly known as the miracle tree. It is known as Nimba in India. The Sanskrit name of Neem is Arishtha meaning the reliever of the sickness. Margosa tree Neem tree characterizes the family Meliaceae which is found in abundance in scorching and semitropical regions like India, Bangladesh, Pakistan, and Nepal.

 

Neem equivocates shows the no end in sight number of medicinal uses. The non-wood products of neem appreciate flowers, leaves, whoop, fruits, seeds (oil, cake), and gum furthermore find contrasting uses. The Neem shows many biological activities which are antibacterial, antifungal, insecticidal and disparate activities. [4]

 

Syzygium aromaticum Cloves are the aromatic dried buds of a tree (Eugenia caryophyllata also mostly Syzgium aromaticum belong to family Myrtaceae used as spices in virtually the entire world’s cuisine. The order ‘Clove’ is derived from the French style ‘Clou’ and the English style ‘Clout’, both meaning ‘nail’ from the likeliness of the establishment bud of the Clove tree to a broad- headed nail. Clove is supported to engross antibacterial properties and used in disparate dental creams, tooth pastes, mouth washes, and esophagus sprays to remove bacteria.

 

Taking into consideration all the above mentioned qualities of the two herbal ingredients namely, Syzygium aromaticum, and Azadirachta indica, the present study was carried out to assess their Phytochemical Characterization along with the Micromeritic Properties and FTIR study. [5]

 

MATERIAL AND METHODS:

Plant Material (Neem Azadirachta indica):

Neem leaves were collected from city park, Greater Noida, Uttar Pradesh. It was ensured that the plant was healthy and uninfected. Leaves were washed under running tap water to remove any traces of soil particles and other dirt. Then washed with distilled water, air dried and cut into small pieces and dried for 25 days in shade. Then the leaves were grind and sieved to get fine powder.

 

Plant Material (Clove Syzygium aromaticum):

Clove was collected from the local market as “laung” and dried in an oven for 24hrs. It was ensured that the spices were original and fresh, and then clove spices was crushed in pastel and mortar and sieved with sieve no 100 to get fine powder.

 

PHYSICOCHEMICAL CHARACTERIZATION:

Organoleptic properties of extracts:

It was characterized for various parameters, like color, taste, odor, and smell. These properties are shown in table no 2.

 

Preparation of extract of Neem:

Five gram of grinded powder was percolated with200 ml of solvent (Methanol and aqueous) for extraction and was kept at soxhlet apparatus at 150°C temperature for 34 hours. After extraction, the extracts obtained were filtered and concentrated. Then the extract was used to be taken for phyto-chemical evaluation.

 

Phyto-chemical screening:

The extract was analyzed through following procedures to test foe the presence of the alkaloids, glycosides, flavonoids, reducing sugars, Terpenoids, saponins, tannins. The powdered leaf was extracted with the required solvent and the obligatory reagent was integrated to the right quantity of the extract. All observations were taken. [6]. The tests performed are shown in table no 3.

 

Test for alkaloids:

The extract of Azadirachta indica was evaporated to dryness and the residue was heated on a boiling water-bath with 2%Hydrochloric acid. After cooling, the mixture was filtered and treated with a few drops of Meyer’s reagent. The samples were then observed for the presence of turbidity or yellow precipitation.

 

Test for glycoside:

2 ml of test solution, 3 ml of glacial acetic acid and 1 drop of 5% ferric chloride were integrated in a test tube. 0.5 ml of concentrated sulphur acid was integrated by the sides of the test tube. Formation of blue color in the acetic acid layer indicates the presence of cardiac glycosides.

 

Test for flavonoid:

5 mg of extract solution was treated with 1.5ml of 50% methanol solution. The solution was warmed and metal magnesium was integrated. To this solution, 5 6 drops of concentrate dhydrochloric acid was integrated and orange or red color was observed for presence of flavonoids.

 

 

 

Test for reducing sugars:

To 0.5 ml of extract solution, 1 ml of dihydrogen monoxide and 5-8 drops of Fehling’s solution was integrated at boiling and observed for brick red precipitate.

 

Test for saponins:

5 ml of extract was shaken vigorously with 5 ml of distilled water in a test tube and heated. The formation of stable foam was accepted as an indication of the presence of saponins.

 

Preparation of extract of Clove:

For the initial extraction, five gm of each of the powdered spice were added to petroleum ether in separate conical flasks in the ratio of 1:4 and placed in a mechanical shaker for 24hrs. The solution obtained was filtered and the filtrate was stored for the upcoming experimentations. The next solvent was added to the residue for further extraction. The order of addition of solvents was as follows: petroleum ether (pet ether), benzene, chloroform, acetone, alcohol and finally water. The filtrates obtained from the above extractions were further subjected to phytochemical, biochemical and anti-microbial analyses.

 

Phytochemical analyses:

Various extracts of the three spice samples were tested for Tannins, Saponins, Cardiac glycosides, Flavonoids and Alkaloids. These tests are shown in table no 6.

 

Test for Tannins: [7]

2ml of each extract was added separately to 4ml of water and a few drops of 0.1% FeCl3 were added to the extracts to form a blue coloured solution.

 

Test for Terpenoids: [8]

Salkowaski test was used. 5ml of various extracts was taken in different test tubes. To each of them 2ml of chloroform was added, along with it 3ml of concentrated sulphuric acid was added slowly to form a layer.

 

Test for Saponins: [9]

1ml of the extract was added to 20ml of distilled water in a test tube and was shaken vigorously for 15 minutes. Formation of the foamy layer indicated the presence of saponins.

 

Test for Cardiac Glycosides: [8]

Keller-Killani test was used. 2ml of glacial acetic acid was added to 5ml of the extracts containing a drop of FeCl3 solution followed by the addition of 1ml of concentrated sulphuric acid.

 

 

 

Test for Flavonoids: [8]

a. Aqueous filtrate along with concentrated sulphuric acid was taken in a test tube; 5ml of dilute ammonia solution was added.

 

b. To all the other filtrates few drops of 1% aluminium solution was added. The presence of flavonoids was indicated by the development of yellow colour.

 

MICROMERITIC PROPERTIES:

Bulk density and bulkiness:

As described by the author, bulkiness is the inverse of bulk density. For determination of bulk density accurately weighed quantity of 5g was introduced into a graduated measuring cylinder and the cylinder was fixed on the bulk density apparatus. The volume occupied by the powder was noted down. Bulk density is calculated by the formula given in equation 1 respectively.[10]

 

Mass

Bulk density = --------------------------------------------------------------------------- Equation 1

Bulk volume

 

Tapped density:

As described by the author elsewhere, tapped density was calculated by tapping powder in a bulk density apparatus until constant volume was obtained. The final volume was noted. Tapped density is calculated by the formula given in equation 2 respectively. [10]

 

Tapped volume

Tapped density =----------------------------------------------------------------------------------------------------------- Equation 2

Mass

Powder flow property:

As described by the author elsewhere, flow property of powder was calculated by measuring angle of repose. Using the formula angle of repose was calculated thrice. Angle of repose was calculated by the formula given in equation 3. [11]

                                              h

Angle of repose (tan0) = --- ----   --------------------------------------------------------------Equation 3

                                             r

 

Where, h is the height of the pile and r is the radius of the pile.

 

Table No. 1 Effect of angle of repose  on flow property

Angle of repose

Type of flow

< 25

Excellent

25-30

Good

30-40

Passable

>40

Very poor

 

Powder compressibility:

As described by the author elsewhere, compressibility of powder is determined by Carr’s Index for this finely powdered mucilage (5g) was transferred into a measuring cylinder and using the bulk density apparatus calculations were done. Carr’s Index was calculated by the formula given in equation 4.[12]

 

Tapped density – Bulk density

Carr’s Index = --------------------------------------------------------------------------- Equation 4

Tapped density

 

Hausner's ratio:

It is another parameter for measuring flow ability of the microcapsules. It is calculated using the following formula given in equation 5.[12]

 

tapped density

Hausner’s ratio = -------------------------------------------------------------------------- Equation 5

Bulk density

 

Micromeritic Properties of neem and clove extracts are shown in table no 4 respectively.

 

 

 

FT-IR STUDY:

IR:

As described powdered extract were taken for FTIR studies. FTIR spectra were recorded at the absorbance mode from 4000 to 400 cm-1. [13]. IR functional groups are shown in table no 5 and 7, where as IR specta are shown in figure 1 and 2.

 

RESULT AND DISCUSSION:

Azadirachta indica is a very important in the world global context today because of it’s a fast-growing evergreen popular tree found commonly in India, Africa and America.

 

Syzygium aromaticum is commonly used as spices in households worldwide. Moreover, it has also been utilized as domestic medicine against various infections, a quality attributable to the existence of certain chemical moieties in it.

 


Table 2: Organoleptic properties of extracts

Extract

Color

Odor

Taste

Smell

Use

Neem

Yellowish-Green

Unpleasant

Acrid

Aroma (licorice\Soy sauce)

(Cardiovascular disease), fever, diabetes, gum disease (gingivitis), and liver problems

Clove

Brownish

Deep woody spicy

Pungent, strong and sweet with a bitter, astringent

Sweet-spicy, and hot

Antiseptic and pain-reliever

 

Identification test:

 

Table 3: Phytochemical components of neem leaf aqueous and methanol extract

Components

Tests

Scoring

Aqueous

Methanol

Alkaloids

Mayer’s reagent

+

++

Saponins

Frothing

+++

+

Tannins

Ferric choloride

++

++

Glycosides

Killer-Killiani test

++

+++

Flavonoids

Pew’s

+

++

Reducing sugars

Fehling’s

+

++

Note: + = low concentration, ++ = moderate concentration, +++ = high concentration

 

Table 4: Micromeritic Properties of Plant (Azadirachta indica)

Drug

Angle of repose (degree)

Bulk Density

Tapped Density

Compressibility %

Hausner’s Ratio

Neem

37.95±0.12

0.35±0.03

0.45±0.23

22.22±0.4

1.28±0.05

Clove

45±0.22

0.38±0.01

0.45±0.11

15.55±0.21

1.18±0.02

 


FT-IR Study of Azadirachta indica

 

Figure 1: FT-IR of Powder extract of Azadirachta indica

Table 5: Functional group of Azadirachta indica

Functional Group

Frequency cm-1

Alcohol C-O

1022

Alkane -C-H

1368

Alkeryl C=C

1647

Aldehyde C=O

1698

Aromatic C=C

1541

Amine C-N

1233

Ether C-O

1151


Table 6: Phytochemical Analyses of Clove

Components

Pet.Ether extract

Benzene Extract

Chloroform extract

Acetone extract

Alcohol extract

Water extract

Tannins

-

-

-

-

+

+

Saponins

-

-

-

-

+

+

Terpenoids

+

-

+

-

+

+

Cardiac Glycosides

-

-

-

-

-

-

Flavonoids

-

+

-

+

-

+

Alkaloids

1.Mayers reagent

-

-

-

-

-

-

Alkaloids

2.Dragendroff’s reagent

-

-

-

-

-

+

Note: -Absent and + Present

 


FT-IR Study of Syzygium aromaticum

 

Figure 2: FT-IR of Powder extract of Syzygium aromaticum

 

Table 7: Functional group of Syzygium aromaticum

Functional Group

Frequency cm-1

Alcohol C-O

1147

Alkane C-H

2920

Alkene =C-H

908

Alkyl Hylide C-F

1118

Aromatic C=C

1427

Amine C-N

1317

Ether C-O

1029

Nitro N-O

1364

Aldehyde =C-H

2846

 

CONCLUSION:

Neem has a long history as a medicinal plant with diverse therapeutic applications. The phytochemical and biochemical experiments performed during the current study confirm that the extracts of the plant are rich in chlorophyll and proline. Methanolic extracts showed the presence of some common phytochemicals like alkaloids, saponins, tannins, glycosides, flavonoids and reducing-sugars.

 

In this study we detected the presence of only flavonoids in the extracts of clove only water and acetone, cardiac glycosides were not identified while the rest of phyto-chemicals were present.

 

 

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Received on 14.11.2018          Accepted on 31.12.2018

© Asian Pharma Press All Right Reserved

Asian J. Pharm. Res. 2019; 9(1): 22-26.

DOI: 10.5958/2231-5691.2019.00005.4