PHYTOCONSTITUTENTS OF MEDICINALLY IMPORTANT PLANT ANDROGRAPHIS PANICULATA NEES
HTML Full TextPHYTOCONSTITUTENTS OF MEDICINALLY IMPORTANT PLANT ANDROGRAPHIS PANICULATA NEES
Sanjay R. Biradar, D. S. Birajdar * and Bhagyashri D. Rachetti
Tissue Culture Research Center, Department of Botany, Department of Physics Shri Chhatrapati Shivaji College Omerga, Osmanabad - 413606, Maharashtra, India.
ABSTRACT: Andrographis paniculata Nees is a plant that has been effectively used in traditional Asian medicines for centuries. It’s perceived “blood purifying” property and also for medicinal properties. The present paper deals to study the phytochemical screening of Andrographis paniculata for various medicinally important compounds. In the present investigation, it was found that phenols, alkaloids, tannins, flavonoids are present in leaves, stem, and root of the plant. Saponin is absent.
Keywords: |
Phytochemicals, Andrographis paniculata, secondary metabolites
INTRODUCTION: Over the past twenty years, interest in medicinal plants has grown enormously from the use of herbal products as natural cosmetics and for self-medication by the general public to the scientific investigations of plants for their biological effects in human beings. Beyond this pharmaceutical approach to plants, there is a wide tendency to utilize the herbal product to supplement the diet, mainly with the intention of improving the quality of life and preventing the diseases of elderly people 1. India has been identified as a major resourceful area in traditional and alternative medicines globally. Medicinal plants constitute an important component of flora and are wildly distributed in India.
Andrographis paniculata Nees is one of the wildly distributed medicinal plants in India and using since ancient times in traditional ayurvedic systems of medicines. Andrographis paniculata Nees is a medicinal plant belonging to the family of Acanthaceae. Diterpenoids and flavonoids are the main chemical constituents of A. paniculata, and these compounds are believed to be responsible for the biological activities of the plant 2, 3.
It is widely used in Chinese and Ayurvedic medicine for the treatment of gastric disorders, infectious diseases, and common colds. It has multiple pharmacological properties such as antiprotozoal, hepatoprotective, anti-HIV, anti-inflammatory 4, antipyretic, anticancer 5, antitumor, hypoglycaemic 6, hypotensive activities and has been used for the treatment of snake bites.
The primary modern use of A. paniculata is for the prevention and treatment of the common cold. It appears to have antithrombotic actions, suggesting a possible benefit in cardiovascular disease 7.
Pharmacological and clinical studies suggest the potential for beneficial effects in diseases like Cancer 8-12 and HIV infections 13. A. paniculata has been reported as having antibacterial, antifungal, antiviral, choleretic, hypoglycemic, hypocholes-terolemic, and adaptogenic effects 14.
MATERIALS AND METHODS:
Collection of Plant Material: The fresh parts of Andrographis paniculata Nees. were collected in the flowering period from Amrutkund Tq. Basavkalyan, Dist. Bidar near Maharashtra-Karnataka border. The plant material was properly washed with tap water and then rinsed with distilled water.
Extraction:
Preparation of Ethanolic Extracts Callus: Fresh leaves, stem, and roots of Andrographis paniculata were washed thoroughly under running tap water, shade dried and used for extraction. Dried leaves stem and roots were homogenized to a fine powder and stored in airtight bottles. 10 gm of leaves, stem, and seeds powders were extracted with 100 ml of different solvents (absolute alcohol, methanol, and chloroform) for 72 h. After 72 h of extraction, each extract was filtered through Whatman’s filter paper no. 1. The filtrate was evaporated to dryness at room temperature & store at 5 ºC in the refrigerator. Extracts were used for different tests.
Qualitative Analysis: Extracts were tested for the presence of active principles. Following standard procedures were used 15 16.
Test for Alkaloids: Ethanolic extract was warmed with 2% H2SO4 for two minutes. It is filtered, and a few drops of reagents were added and indicated the presence of alkaloids.
Mayer’s Reagent: A creamy- white colored precipitation positive.
Wagner’s Reagent: A reddish-brown precipitation positive.
Picric Acid (1%): A yellow precipitation positive.
Test for Steroids Terpenoid and Triterpenoids:
Liebermann Burchard Test: The crude extract was mixed with few drops of acetic anhydride, boiled and cooled. Concentrated sulphuric acid was then added from the sides of the test tube and observed for the formation of a brown ring at the junction of two layers. Green coloration of the upper layer and the formation of deep red color in the lower layer would indicate a positive test for steroids terpenoid and triterpenoids respectively.
Salkowski Test: The extract was mixed with 2 ml of chloroform and concentrated H2SO4 (3 ml) is carefully added to form a layer. A reddish-brown coloration of the interface is formed to show the positive result of the presence of steroids terpenoid and triterpenoids respectively.
Test for Saponins: The crude extract was mixed with 5ml of distilled water in a test tube and it was shaken vigorously. The formation of stable foam was taken as an indication for the presence of saponins.
Test for Phenols and Tannins: The crude extract was mixed with 2 ml of 2% solution of FeCl3. A blue-green or black coloration indicated the presence of phenols and tannins.
Test for Flavonoids: A small quantity of the extracts is heated with 10 ml of ethyl acetate in boiling water for 3 min. The mixture is filtered differently, and the filtrates are used for the following test.
Ammonium Test: The filtrate was shaken with 1 ml of dilute ammonia solution (1%). The layers were allowed to separate. A yellow coloration was observed at ammonia layer. This indicates the presence of the flavonoid.
Aluminum Chloride Test: The filtrates were shaken with 1 ml of 1% aluminum chloride solution and observed for light yellow color. It indicated the presence of flavonoid and diluted NaOH and HCl was added. A yellow solution that turns colorless indicated positive.
Test for Carbohydrate:
Benedict's test: The test solution was mixed with few drops of Benedict's reagent (alkaline solution containing cupric citrate complex) and boiled in a water bath, observed for the formation of reddish-brown precipitate to show a positive result for the presence of carbohydrate.
Test for Glycosides:
Fehling’s Test: An equal volume of Fehling A and Fehling B reagents were mixed, and 2 ml of it was added to the crude extract and gently boiled. A red brick precipitate appeared at the bottom of the test tube indicated the presence of reducing sugars.
Test for Proteins:
Millon’s Test: Crude extract when mixed with 2 ml of Millon’s reagent, a white precipitate appeared which turned red upon gentle heating that confirmed the presence of protein.
Test for Free Amino Acids: 19
Ninhydrin Test: Test solution when boiled with 0.2% solution of Ninhydrin, would result in the formation of purple color suggesting the presence of free amino acids.
Test for Vitamin C: 18
DNPH Test: The test solution was treated with Dinitrophenylhydrazine dissolved in concentrated sulphuric acid. The formation of yellow precipitate would suggest the presence of Vitamin C. For Carboxylic acid, test for NH2, Nitrogen, Sulphur, Halogen, Amides, test for unsaturation, test for Aromaticity 17.
RESULT AND DISCUSSION: The preliminary phytochemical screening tests may be useful in the detection of the bioactive principles and subsequently may lead to drug discovery and development. The phytochemical analysis of Andrographis paniculata Nees tested was summarized in Table 1, which revealed that the presence of a medicinally active compound in plant Leaf, stem and roots.
TABLE 1: PHYTOCHEMICAL SCREENING FOR DIFFERENT EXTRACTS OF ANDROGRAPHIS PANICULATA
S. no. | Compound | Test | Plant part | ||||||||
Leaf extract | Stem extract | Root extract | |||||||||
E | M | C | E | M | C | E | M | C | |||
1 | Alkaloid | Mayer’s reagent | ++ | + | − | ++ | + | ++ | ++ | + | + |
Wagner’s Reagent | + | + | − | + | + | + | + | + | − | ||
Picric acid | ++ | + | − | + | + | − | + | + | − | ||
2 | Amides | Hydrolysis with alkali | + | + | + | + | + | + | + | + | + |
3 | Amines | Amines test | + | + | + | + | + | + | + | + | + |
4 | Ascorbic acid | DNPH test | + | ++ | + | + | +++ | + | + | +++ | +++ |
5 | Carbohydrates | Benedict test | + | + | + | + | + | + | + | + | + |
6 | Carboxylic acid | Sodium bicarbonate test | + | + | + | + | + | + | + | + | + |
7 | Flavonoids | Ammonium Test | + | +++ | + | + + | +++ | ++ | + | ++ | + |
Aluminum chloride test | + | + | + | + | + | + | + | + | + | ||
8 | Glycosides | Fehling solution | ++ | +++ | ++ | ++ | +++ | ++ | + | +++ | ++ |
9 | Phenol | Ferric chloride test | + | + | + | + | + | + | + | + | + |
10 | Proteins | Millons Reagent test | + | + | − | + | + | − | + | + | − |
11 | Reducing Sugar | Fehling solution test | − | − | + | − | − | +++ | − | − | +++ |
12 | Saponin | Frothing test | − | − | − | − | − | − | − | − | − |
13 | Starch | Starch test | + | + | + | + | + | + | + | + | + |
14 | Steroids | Liebermann - Burchard’s test | ++ | +++ | ++ | ++ | +++ | + | + | +++ | + |
Salkowski’s Test: | + | +++ | ++ | + | +++ | ++ | + | ++ | + | ||
15 | Tannin | Ferric chloride test | ++ | +++ | ++ | + + | +++ | + | +++ | ++ | + |
16 | Terpenoides | Liebermann - Burchard’s test | ++ | ++ | ++ | + | +++ | ++ | + | ++ | ++ |
Salkowski’s Test: | + | +++ | ++ | + | +++ | ++ | + | ++ | ++ | ||
17 | Amino acid | Ninhydrin Reagent test | + | + | + | + | + | − | + | + | − |
18 | Aromaticity | Flame test (Ignition test) | + | + | + | + | + | + | + | + | + |
19 | Unsaturation | Test for unsaturation | − | − | − | − | − | − | − | − | − |
- =absent, + =Presence, ++ = Moderate, +++ = Maximum
For extraction of phytochemicals, ethanolic, methanolic and chloroform extracts were used. Preliminary phytochemical analysis of Andrographis paniculata compounds shows various types of chemical compounds which provide the baseline for the occurrence of the medicinally active constituents like alkaloids, flavonoids, glycosides, tannins. The A. paniculata were rich in alkaloid, ascorbic acid, Flavonoids, glycosides, steroids, tannins, terpenoids, etc. It lacks saponin. From the table, it is revealed that the methanolic extract of leaf stem, the root of A. paniculata shows maximum Phytoconstituents.
CONCLUSION: In the present study callus extract showed the presence of bioactive compound such as alkaloids, flavonoids, terpenoids, ascorbic acid, tannin, glycosides, proteins, triterpenoids, starch, phenol, etc. This study also leads to further research in the way of isolation and identification of the active compound from the A. paniculata using extraction, chromatographic and spectroscopic techniques.
ACKNOWLEDGEMENT: Author Dr. Sanjay R. Biradar is thanks to Principal of Shri Chhatrapati Shivaji College, Omerga, Dist. Osmanabad, (M.S.), India for providing all necessary facilities and encouragement for the present research work.
CONFLICT OF INTEREST: Nil
REFERENCES:
- Taylor, Francis and Maffei M: Dietary supplements of plant origin-nutrition and health approach. E-Library 2003; 18.
- Tang W and Eisenbrand G: Chinese drugs of plant origin, chemistry, pharmacology and use in traditional and modern medicine. Springer Verlag, Berlin 1992: 97-103.
- Saxena S, Jain DC, Bhakuni RS and Sharma RP: Indian Drugs 1998; 35: 458-467.
- Sheeza K, Shihab PK and Kuttan G: Antioxidant and anti-inflammatory activities of the plant Andrographis paniculata Nees, Immunopharmacol Immunotoxicol 2006; 28: 129-140.
- Li J, Cheung HY, Zhang Z, Chan GKL and Fong WF: Andrographolide induces cell cycle arrest at G2/M Phase and cell death in HepG2 cells via alteration of reactive oxygen species, J. Pharmacol 2007; 568: 31-44.
- Borhanuddin M, Shamsuzzoha M and Hussain AH: Hypoglycaemic effects of Andrographis paniculata Nees on nondiabetic rabbits. Bangladesh Med Res Counc Bull 1994; 20: 24-26.
- Amroyan E, Gabrielian E and Panossian A: Inhibitory effect of andrographolide from Andrographis paniculata on PAF-induced platelet aggregation. Phytomedicine 1999; 6: 27-31.
- See D, Mason S and Roshan R: Increased tumor necrosis factor alpha (TNF-alpha) and natural killer cell (NK) function using an integrative approach in late stage cancers. Immunol Invest 2002; 31: 137-153.
- Sheeja K, Guruvayoorappan C and Kuttan G: Antiangiogenic activity of Andrographis paniculata extract and andrographolide. Int Immunopharmacol 2007; 7: 211-221.
- Shi MD, Lin HH and Lee YC: Inhibition of cell-cycle progression in human colorectal carcinoma Lovo cells by andrographolide. Chem Biol Interact 2008; 174: 201-210.
- Yang L, Wu D and Luo K: Andrographolide enhances 5-fluorouracil-induced apoptosis via caspase-8-dependent mitochondrial pathway involving p53 participation in hepatocellular carcinoma (SMMC-7721) cells. Cancer Lett 2009; 276: 180-188.
- Zhao F, He EQ, Wang L and Liu K: Anti-tumor activities of andrographolide, a diterpene from Andrographis paniculata, by inducing apoptosis and inhibiting VEGF level. J Asian Nat Prod Res 2008; 10: 467-473.
- Calabrese C, Berman SH and Babish JG: A phase I trial of andrographolide in HIV positive patients and normal volunteers. Phytother Res 2000; 14: 333-338.
- Bhatnagar SS, Santapau H and Desa JD: Biological activity of Indian medicinal plants. I. Antibacterial, antitubercular and antifungal action. Indian J Med Res 1961; 49: 799-813.
- Raman N: Phytochemical Technique. New Indian Publishing Agencies: New Delhi 2006: 19.
- Harborne JB: (Reprint. Edn.). Phytochemical Methods. New Delhi: Springer (India) Pvt. Ltd., 2005: 17.
- Devi P: Principles and methods of plant molecular biology. Biochemistry and Genetics. Agrobios (India) 2003.
- Sethi A: Lab experiments in organic chemistry, New Age International (P) Limited, Publisher 2003; ISBN 81- 224-1491-5.
- Thacher CH: A handbook of organic analysis, IV Edn., CBS Publishers, New Delhi 2007.
How to cite this article:
Biradar SR, Birajdar DS and Rachetti BD: Phytoconstitutents of medicinally important plant Andrographis paniculata Nees. Int J Pharmacognosy 2015; 2(1): 55-58. doi: 10.13040/IJPSR.0975-8232.2(1).55-58.
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Article Information
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55-58
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English
IJP
S. R. Biradar, D. S. Birajdar * and B. D. Rachetti
Tissue Culture Research Center, Department of Botany, Department of Physics Shri Chhatrapati Shivaji College Omerga, Osmanabad, Maharashtra, India.
sanjaybiradar2006@rediffmail.com
12 September 2014
27 December 2014
30 December 2014
http://dx.doi.org/10.13040/IJPSR.0975-8232.IJP.2(1).55-58
01 January 2015