Pharmacognostic and Preliminary Phytochemical investigation of Salvadora persica Linn (Salvadoraceae)

 

Wasimuzzama Khan, Atar Mujum, Tausif Shaikh,       S.M. Katekar, Rashmi Tambe and Rukhsana A. Rub*

 

Dept. of Pharmacognosy, M.C.E. Society’s Allana College of Pharmacy, Pune-411001 (M.S) India

 

 

1. ABSTRACT:

Salvadora persica linn. (Salvadoraceae), a desert plant, also known as toothbrush tree or Miswak is used as tooth cleaning stick in many third world countries. The aqueous and alcoholic extracts of roots, leaves, bark and pulp of Salvadora persica linn have been found to have antimicrobial, anti-plaque, analgesic, antipyretic, antioxidant and anti-ulcer activity. Since the plant is not much explored pharmacognistically and phytochemically, the present work is planned to study the pharmacognostic, phytochemical and chromatographic behavior and to establish the monograph of the plant.

 

Microscopy of stem showed well defined single layered quadrangular celled epidermis, 7 to 8 layers of cellular cortex embedded into which are bunches of lignified pericyclic fibers and cocentric vascular bundle, Medullary rays and pith.  Powder showed presence of epidermis, bordered and pitted xylem vessels, reticulate xylem parenchyma and lignified pericyclic fibers etc

 

The proximate analysis showed satisfactory results with respect to foreign matter, moisture content and ash values. The water soluble extractive value was found to be the highest (25.4%), whereas alcohol soluble extractive, chloroform soluble extractive and ether soluble extractive values were found as 7.6%, 6% and 3.6% respectively.

 

The phytochemical investigation of the stem of Salvadora persica, showed presence of Alkaloids, Glycosides, Flavonoids, Carobohydrates, Tannins and Saponins, in the aqueous extract. The ethanolic extract of the stem showed presence of Glycosides, Steroids and Flavonoids whereas Acetone extract showed presence of Steroids and Flavnoids which were confirmed by the Thin Layer Chromatography. Chloroform and ethereal extract failed to show the presence of any phytoconstituents.

 

2. KEYWORDS:  Salvadora persica, Miswak, Pharmacognostic study, TLC

 

3.  INTRODUCTION:

Miswak (Salvadora persica) is a desert plant of Salvadoraceae family and is commonly known as toothbrush tree, Mustard tree, Arak tree, Peelu tree etc. Its roots and branches are used as tooth cleaning stick in many third world countries.

 

As per literature survey the aqueous and alcoholic extracts of the plant are reported to have antimicrobial1-3, anti-plaque4-7, analgesic8, antipyretic9, antioxidant10 and anti-ulcer activity11,12. 


However the plant is not much explored pharmacognistically and no extensive work is seen with respect to its phytochemical account.  Therefore the present work is planned to explore the plant for its pharmacognostic, phytochemical and chromatographic behavior and to establish the monograph of the plant.

 

4. MATERIAL and METHODS:

Authentification of plant was done by P.G. Diwakar, Botanical Survey of India, Pune. BSI/WRC/Tech./2010/959

 

4.1 Pharmacognostic study:

The fresh plant of Salvadora persica was collected from Malkapur, Buldana district, (Maharashtra). External visible characters such as size, shape, diameter and organoleptic properties of stem were studied and recorded. The characters were matched with the morphological characters reported in the literature.

 

4.2 Microscopical study:

A thin transverse section and powder characteristic of the stem were studied.

Microscopic descriptions of tissues were supplemented with micrographs.  Photographs of different magnifications were taken with Motic Image Plus 2.0 Microscopic Unit.  For normal observations bright field was set. For the studies of crystals, starch grains and lignified cells, polarized light was employed. Magnifications of the figures are indicated by scale-bars.

 

4.3 Proximate analysis:

Physicochemical constants such as Extractive values, Ash values (Total, Acid insoluble and water soluble ash values), L.O.D. were studied as per Pharmacopeial procedures.

 

4.4 Qualitative Phytochemical Screening:

The Crude dried powder was extracted by cold maceration process and subjected to phytochemical screening, for presence of various phytoconstituents like alkaloids, glycosides, flavonoids, tannins, terpenes, steroids etc.

 

4.5 Thin Layer Chromatography:

TLC of aqueous and organic extracts of stem of Salvadora persica were carried out, the various solvent systems along with various visual detectors with respect to various phytoconstituents and/or UV light were used.

 

5. RESULTS:

5.1 Pharmacognostic study, of the stem of Salvadora persica showed presence of well defined tissues like single layer of quadrangular celled epidermis, 7 to 8 layer of cellular cortex embedded into which are bunches of lignified pericyclic fibers and cocentric vascular bundle, Medullary rays and pith (Fig. I and II).  Powder showed presence of epidermis, bordered and pitted xylem vessels, reticulate xylem parenchyma and lignified pericyclic fibers etc (Fig. III to VI).

 

5.2 Physicochemical constants:

Table I: Table representing the physicochemical constants

Sr. No.

Evaluation parameters

Value (%w/w)

1

Foreign matter

1.3

2

Moisture content

0.25

3

Total ash value

9.4

4

Water-soluble ash value

2

5

Acid-insoluble ash value

3

6

Water soluble extractive value

25.4

7

Alcohol soluble extractive value

7.6

8

Chloroform soluble extractive value

6

9

Petroleum ether soluble extractive value

3.6

 

6. DISCUSSION:

Microscopical examination of the stem of Salvadora persica was conducted with the aim to study and understand the characteristic features of the plant. The transverse section of the stem showed a unique and well defined cellular structure.  Transverse section showed presence of single layered quadrangular celled epidermis, 7-8 layers of cortical parenchyma with lignified pericyclic fibers, and concentric vascular bundles. Supporting vascular tissues like Phloem fibers, Xylem fibers and xylem vessels were also seen prominently (Fig. I and II).

 

Microscopical examination of powder showed presence of well defined bordered and pitted xylem vessels, lignified fibers, reticulate parenchyma etc. These unique and characteristic microscopical features of the plant can be considered as an identifying tool for the plant (Fig. III to VI).

5.2 Phytochemical investigation

Table II: Phytochemical Investigation of various extracts of Salvadora persica

Sr. No.

Chemical Test

Aqueous Extract

Ethanolic Extract

Chloroform Extract

Ether Extract

Acetone Extract

1

Carbohydrate

1. Molish’s Test

2. Fehlings Test

3. Benedict’s Test

4. Barfoed’s Test

 

+

+

-

-

 

-

-

-

-

 

-

-

-

-

 

-

-

-

-

 

-

-

-

-

2

Protein

1. Biuret Test

2. Million’s Test

 

+

-

 

-

-

 

-

-

 

-

-

 

-

-

3

Amino Acid

1. Ninhydrin Test

 

-

 

-

 

-

 

-

 

-

4

Steroids

1. Salkowski Test

2. Lieberman Buchard Reaction

 

+

+

 

+

+

 

-

-

 

-

-

 

+

+

5

Glycoside

1. Keller Killani’s Test

 

+

 

+

 

-

 

-

 

-

6

Saponins

1. Foam Test

 

+

 

-

 

-

 

-

 

-

7

Alkaloids

1. Dragendroff’s reagent

2. Mayer’s reagent

3. Wagner’s reagent

4. Hager’s reagent

 

+

+

+

+

 

-

-

-

-

 

-

-

-

-

 

-

-

-

-

 

-

-

-

-

8

Flavonoids

1. Shinoda Test

2. Lead Acetate

3. NaOH

4. FeCl3

 

+

+

+

+

 

+

+

+

+

 

-

-

-

-

 

-

-

-

-

 

+

+

+

+

9

Tannins

1. Fecl3

2. Lead Acetate

3. Bromine water

4. Gelatin

 

+

+

+

+

 

-

-

-

-

 

-

-

-

-

 

-

-

-

-

 

-

-

-

-

 

5.3 Thin Layer Chromatography

Table III: TLC profile of extracts of Salvadora persica

Sr. No.

Chemical constituents

Mobile phase

Detecting agent

1

Alkaloids

Chloroform: Methanol: Diethyl amine (80:20:10)

Iodine Chamber

2

Glycoside

Methanol: Water: Chloroform (65:25:4)

UV at  254 nm

3

Flavonoid

Ethyl acetate: Formic acid: Glacial acetic acid: water  (100:11:11:26)

Anisaldehyde-sulphuric acid

4

 

Sugars

 

Benzene : GAA : Methanol

(20:20:60)

Anisaldehyde-sulphuric acid

5

Tannins

Ethyl acetate: GAA: water (90:10:10)

5% Fecl3

 

6

Steroids

Benzene : Ethyl Acetate (5 :95)

Anisaldehyde-sulphuric acid

 

7

Saponin

Chloroform: GAA : Methanol : Water

(64 : 32 : 12 : 8 )

Anisaldehyde-sulphuric acid

 

 

 

The proximate analysis of the stem powder showed satisfactory results with respect to foreign matters, moisture content and ash values. The water soluble extractive value was found to be the highest (25.4%), whereas alcohol soluble extractive, chloroform soluble extractive and ether soluble extractive values were found as 7.6%, 6% and 3.6% respectively (Table I).

 

In the phytochemical investigation of the stem of Salvadora persica, the aqueous extract showed presence of Alkaloids, Glycosides, Flavonoids, Carbohydrates, Tannins and Saponins which were further confirmed by Thin Layer Chromatography with the Rf values as, Alkaloid (0.32), Glycoside (0.95), Flavonoids (Spot1- 0.38, Spot2- 0.45 and Spot3- 0.8, Carbohydrate (0.94), Tannins (0.96) and Saponins (0.98). (Fig. VII).

 

Fig VII: Thin Layer Chromatography of aqueous extract

Ethanolic extract of the stem of Salvadora persica showed presence of Glycosides, Steroids and Flavonoids which were also further confirmed by TLC with Rf values for Glycosides, Steroids and Flavonoids as 0.97, 0.78 and 0.57 respectively (Fig. VIII), whereas Acetone extract showed presence of Steroids and Flavnoids with the Rf values 0.84 and 0.97 respectively (Fig. IX).

 

                   

Chloroform extract and ethereal extract failed to show the presence of any phytoconstituents (Table II).

 

This significant data pertaining to morphological, microscopical and phytochemical nature of the plant, obtained in the present research work can effectively be utilized in establishing the monograph of the plant.

 

7. ACKNOWLEDGEMENTS:

We are grateful to our Principal, Dr. Kiran Bhise, for providing us the necessary infrastructure for the research work.

 

8. REFERENCES:

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Received on 19.05.2010

Accepted on 14.06.2010        

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Research Journal of Pharmacognosy  and Phytochemistry. 2(4): July-Aug. 2010, 319-323