Pharmacognostic and Preliminary Phytochemical Investigation of Argyreia nervosa Roots (Convolvulaceae)

 

Atar Mujum, Wasimuzzama Khan, Tausif Shaikh and Rukhsana A. Rub*.

 

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

 

 

ABSTRACT:

Argyreia nervosa Linn. (Argyreia speciosa) belonging to family Convolvulaceae, commonly known as elephant creeper, has been reported to have antimicrobial, hepatoprotective, nootropic, anticonvulsant, hypoglycemic, anti-oxidant, antiviral, nematicidal, aphordiasic, immunomodulatory, analgesic and anti-inflammatory activity.

 

Pharmacognostical parameters for the roots were studied with the aim of contributing towards establishing monograph for this species. In pharmacognostical investigations, examination of macroscopical, microscopical characters, ash values, extractive values, foreign matter and LOD were carried out. Phytochemical screening included qualitative chemical examination and thin layer chromatography of the detected phytoconstituents.

 

Microscopical study showed well defined tissues like thick walled cork, cortex and phloem with resin canals, drum shaped xylem vessels etc. Preliminary phytochemical investigation of aqueous and methanolic extract of roots of Argyreia nervosa revealed the presence of alkaloids, glycosides, amino acid, tannins and flavonoids which were further confirmed by thin layer chromatography. However flavonoids were found to be absent in chloroform extract and ethereal extract showed presence of only tannins. Thus the present work justifies our aim of detailed pharmacognostic and phytochemical investigation and chromatographic studies of the plant.

 

KEYWORDS: Argyreia nervosa, Phytochemical, convolvulaceae flavonoids

 

1. INTRODUCTION:

Argyreia nervosa (Elephant creeper), is an ornamental plant of Convulaceae family, having various synonyms such as Argyreia speciosa, Samudrasoka, vridarukand, hawian baby wood rose etc. It is a very large climbing shrub with woody, White tomemtose stem, leaves with acute apex and cordate. Flowers are large purple, silky pubscent, corolla tubular-infundiulifom. Fruits are dry globes, apiculate. Seeds are tringular, brown coloured, hard, two side slightly concave and third sight convex, two folded cotyledons. It is distributed throughout India in areas up to 300 m elevation13. As per literature survey, the aqueous and various organic extracts of the plant are reported to have antimicrobial1,2, hepatoprotective3, nootropic4, anticonvulsant5, hypoglycemic6, anti- oxidant7,antiviral8, nematicidal9, aphordiasic10, immunomodulatory11, analgesic and anti-inflammatory activity12.

 


Hallucinogenic property of plant discovered recently. Seeds contain the highest concentration of psychoactive compounds in the entire family. 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.

 

2. MATERIAL AND METHODS:

Sample of Argyreia nervosa was collected in the month of July- Agust from Jeur Pune district-and was identified by P. G. Diwakar, Botanical Survey of India, Pune.

(BSI/WC/Tech/2009/464)

 

2.1 Pharmacognostic study:

The fresh roots plant of Argyreia nervosa was studied for external visible characters such as size, shape, diameter and organoleptic properties of roots were recorded. The characters were matched with the morphological characters reported in the literature.

 

2.2 Microscopical study:

A thin transverse section and powder characteristic of the root were studied.14,15

 

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. Since these structures have birefringent property, under polarized light they appear bright against background. Magnifications of the figures are indicated by scale-bars.

 

2.3 Proximate analysis:

Physicochemical constants such as Extractive values, Ash values (Total, Acid insoluble and water soluble ash values), L.O.D. were studied using standard methods16.

 

2.4 Qualitative Phytochemical Screening:

Crude dried powder was extracted by using Soxhlet apparatus  different organic solvent and Aqueous extract was prepared by decoction method and extract was concentrated and subjected to phytochemical screening, for presence of various phytoconstituents like alkaloids, glycosides, flavonoids, tannins, terpenes, steroids etc17.

 

2.5 Thin Layer Chromatography:

Thin layer chromatography of aqueous and organic extracts of Argyreia nervosa roots were carried out. Various solvent systems along with visual detectors with respect to various phytoconstituents and/or UV light were used18.

 

 

3. RESULTS:

3.1. Pharmacognostic study

 

Table I: Morphological characteristics of the bark of Argyreia nervosa

Morphological Characteristics

Observation

Color

Brown

Odour

Odourless

Taste

Sweet

 

3.2. Microscopy:

T.S. of the roots of Argyreia nervosa showed presence of well defined tissues like, 6 to 9 layers of thick walled cork cells; thin walled tangentially elongated cortical cells containing circular starch grains, rosette crystals of calcium oxalate and few resin canals; a wide zone of secondary phloem consisting of sieve tubes, companion cells and numerous resin canals; phloem parenchyma traversed by Medullary rays containing circular starch grains; secondary xylem comprising of xylem vessels, tracheids, and fibres.

 

Microscopy of root powder showed cortical cells filled with circular starch grain measuring between 3 to 16 μ in diameter; brown colouring matter and rosette crystals of calcium oxalate, xylem parenchyma, fibres, drum shaped vessels, pitted with large end perforations; tracheids, much longer with bordered pits and fibres having pointed ends.

 

 


3.3. Physicochemical constants:

Table II: Table representing the physicochemical constants:

Sr. No.

Evaluation parameters

Value (%w/w)

1

Foreign organic matter

2.0

2

Moisture content

8.0

3

Total ash value

5.8

4

Water-soluble ash value

2.5

5

Acid-insoluble ash value

1.1

6

Water soluble extractive value

13.8

7

Alcohol soluble extractive value

8.84

8

Ethyl acetate soluble extractive value

2.22

9

Chloroform soluble extractive value

1.71

10

Petroleum ether soluble extractive value

1.23

 

3.4. Preliminary Phytochemical Investigation:

Table III: Phytochemical investigations of various aqueous and organic solvent extracts:

Sr. No.

Chemical Test

Aqueous Extract

Methanolic Extract

Chloroform Extract

Ether 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. Gelatin

 

+

+

+

 

+

+

+

 

+

+

+

 

+

+

+

 

 

 

3.5. Thin Layer Chromatography of Argyreia nervosa:

Table IV: TLC profile of various extract of Argyreia nervosa:

Sr. No.

Chemical constituents

Mobile phase

1

Flavonoid

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

2

Sugars

Benzene : GAA : Methanol (20:20:60)

3

Glycoside

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

5

Alkaloid

Chloroform: Methanol: Diethyl amine

6

Steroids

Benzene : Ethyl acetate (95:5)

7

Amino acid

N – Butanol : GAA : Water (10:1:3)


 

4. DISCUSSION:

Microscopical examination of roots of Argyreia nervosa showed presence of well defined cellular structure. Transverse section showed presence of 6-9 layers of thick walled cork cells, cortex containing starch grains, calcium oxalate crystals and resin canals, A well developed phloem with parenchyma, medullary rays and numerous resin canals scattered throughout the tissue, and xylem with xylem vessels and tracheids were also seen (Fig.I).

 

Microscopical examination of powder also showed well identifiable tissues like thick walled cork cells, xylem vessels, xylem parenchyma and fibres having pointed ends (Fig. II to V). Thus these unique microscopical observations can be considered as an identifying tool for the plant.

 

The proximate analysis with respect to foreign matter, moisture content and ash values etc was found within limit as depicted in Ayurvedic pharmacopeia. The water soluble extractive value was found to be the highest (13.8%), whereas alcohol soluble extractive value, ethyl acetate soluble extractive value  chloroform soluble extractive value and  pet ether soluble  extractive values were found as 13.8%, 8.84%, 2.22%., 1.7% and 1.23% respectively (Table II).

 

In the phytochemical investigation of the roots of Argyreia nervosa, the aqueous extract showed presence of Alkaloids, Glycosides, Amino acids Flavonoids and Tannins which were confirmed by thin layer chromatography with the Rf values as 0.82,0.84,0.86,0.92, and 0.96 respectively (Fig.VI).

 

    Alkaloid                    Amino acid    Flavonoid     Glycoside       Tannin

 

Methanolic extract of the roots of Argyreia nervosa showed presence of Alkaloids, Flavonoids, Glycosides, Amino acids, Steroids and tannins which were also confirmed by TLC with Rf values for Alkaloid-0.92, Flavonoid- spot10.90 and spot2 0.84, Glycoside-0.83, Amino acids- spot1 0.55 and spot2 0.60, Steroids- 0.23, and tannins- 0.93 (Fig. VII)

 

Alkaloid   Flavonoid Glycoside  Amino Acid  Steroid     Tannin

Fig VII: TLC Plates of methanolic extract:

Chloroform extract showed presence of Alkaloids (Rf values:spot1-0.75 and spot2-0.51), Glycosides (Rf value 0.82), Amino acids (Rf value 0.84) and steroids with, (Rf value 0.41) (Fig. VIII) However Pet ether extract showed presence of only Tannins with Rf value 0.96 (Fig. IX)

 

Thus significant data pertaining to morphological, microscopical and phytochemical nature of the plant could be obtained which can effectively be utilized in establishing the monograph of the plant successfully.

 

5. ACKNOWLEDGEMENT:

We are grateful to Dr. Kiran Bhise, Principal, Allana College of Pharmacy, for providing us the necessary infrastructure for the research work.

 

6. REferences:

1.        Shukla YN, Srivstava A, Sunil Kumar, Sushi Kumar (1999). Phytotoxic and antimicrobial constituents of Argyreia speciosa and Oenotherabiennis. J. Ethnopharmacol. 67: 241-245.

2.        Kelkar GM, Phalnikar NL, Bhinde BV (1947). Fatty oil from the seeds ofArgyreia speciosa Sweet. J. Indian Chem. Soc. 24: 83-86.

3.        Habbu PV, Shastry RA, Mahadevan KM, Hanumanthachar Joshi, Das SK (2008). Hepatoprotective effects of Argyreia speciosa in rats. Afr. J. Trad. CAM 5(2): 158-164

4.        Hanumanthachar J, Navneet K, Jyotibala C (2007). Evaluation of nootropic effect of Argyreia speciosa in mice, J. Health Sci. 53(4): 382-388.

5.        Vyavhare NS, Bodhankar SL (2009). Anticonvulsant activity of Argyreia speciosa in mice, Indian J. Pharm. Sci. 71(2): 131-134.

6.        Hemet LE, Satyanarayana T, Ramesh A, Durga Prasad, Routhu Y, Srinivas KV (2008). Hypoglycemic and antihyperglycemic effect of Argyreia speciosa Sweet. In normal and in alloxan induced diabetic rats.RLA press. J. Natl. Rem. 8(2): 203-208.

7.        Shreedhara C.S., Ashwata Ram,H.NSachin,B.Zanwar, Gajera P.Falguni, Antioxisant activity of Argyreia nervosa J.natural remedies vol 9/2 (2009) 216-223

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9.        Parveen N, Khan NU, Singhal KC (1990). Antifilarial activity of Argyreia speciosa against Setaria cervi in vitro. Phytother. Res. 4: 162

10.     Subramonium A, Madhavachandran V, Ravi K, Anuja VS (2007). Aphordiasic property of the elephant creeper Argyreia nervosa. J. Endocrinol. Rep. 11(2): 82-85.

11.     Gokhale AB, Damre AS, Saraf MN (2003). Investigation into the lmmuno- Modulatory activity of Argyreia speciosa. J. Ethanopharmacol. 84(1): 109-114.

12.     Shrivastava A, Shukla YN (1998). Aryl esters and a coumarin from Argyreia speciosa. Indian J. Chem. 37B: 192-194.

13.     Orient Longman.(2003)Indian medicinal plants, a compendium of 500 species, 4:264

14.     M.A.Iyenger, “Study of crude drug”, 12th Edition, Manipal press, 89.

15.     Khandelwal K.R, “Practical Pharmacognosy”, 9thEdition, Nirali prakashan, 149.

16.     Indian Pharmacopoeia 2007, vol-1, Indian pharmacopoeial commission, Gaziabad, 191.

17.     Pulok. K. Mukharjee, “Quality Control of Herbal drug”, 1st Edition, Business Horizone publication

18.     Wagner and Blad, “Plant drug analysis”, 5 th Edition, Springer publication. 196.

 

 

Received on 21.05.2010

Accepted on 04.06.2010        

© A&V Publication all right reserved

 Research Journal of Pharmacognosy and Phytochemistry. 2(5): Sept.-Oct. 2010, 359-363