Phytochemical and pharmacological actions of Ixora coccinea

G. Shiny*, Dr. M. Nagulu, P. Mounika, G. Radha Krishna, K. Aravind, D. Priyanka

Swami Ramananda Tirtha Institute of Pharmaceutical Sciences, Nalgonda

ABSTRACT:

The present work is aimed at investigating the phyto chemical constituents, physico chemical studies, anti microbial and anti oxidant activities of Ixora coccinea. Petroleum ether, chloroform, methanol and water are used for extraction. Cup plate method is used for assaying antimicrobial activity using both bacterial and fungal strains and phosphomolybdenum method is followed for assessing total antioxidant capacity. The extracts have shown considerable bacterial activity but not have shown antifungal activity. It has good antioxidant activity comparable to ascorbic acid.

KEYWORDS: Ixora, phosphomolybdenum method, antimicrobial, cup plate method, phyto chemical screening.

INTRODUCTION:

The use of medicinal plants as a source for relief from illness can be treated back over 5 million years ago. Plants are used medicinally in different countries and are a source of many potent and powerful drugs¹. They have been important to mankind, both socially and economically for thousands of years. They contribute to be important to people that do not have access to modern medicines and moreover pharmaceuticals rely heavily on the same active principles, be they natural or synthetic². Most of the antibiotics are becoming less effective against certain illness with much more toxicity, hence it is essential to investigate newer drugs with lesser resistance. According to WHO – 2001, 80% of world population use medicinal plants in treatment of diseases.

The increased prevalence of antibiotics resistant bacteria due to extensive use of antibiotics may render current antimicrobial agents insufficient to control some bacterial diseases and hence research for identifying novel substances that are active against human pathogens is an urgent need³. Oxidative stress is related to many of the major diseases. Plants contain a wide range of secondary metabolites and so, the present study aimed at screening the selected plants for presence of secondary metabolites in them with view of bioprospecting medicinal plants and to examine anti microbial and in vitro antioxidant activities.

Ixora coccinea belongs to the family of Rubiaceae, commonly known as flame of woods, Jungle geranium⁴. Traditionally it is used as hepatoprotective, chemo protective, antimicrobial, antioxidant, antiulcer, anti inflammatory. Roots are used in hiccups, nausea. Flowers are used on reddened eyes and eruptions in children⁵.

MATERIALS AND METHODS:

Collection of plant material::

Fresh, healthy plants of Ixora coccinea free from microbial attack were collected from domestic places of Nalgonda and was authenticated by Botanist P. Suresh, N.G. College, Nalgonda.

All the chemicals used for research work were of analytical grade and were purchased from S. D Fine Chem Ltd., Mumbai.

Test organisms like bacterial and fungal isolates namely Staphylococcus aureus, Escherishia coli and Candida albicans were procured from Department of Microbiology, N.G. College, Nalgonda.

Physico chemical studies::

Ash values, extractive values were performed according to the official methods prescribed in Indian Pharmacopoeia⁶. Flourescence analysis of drug powder was carried out according to the method of Chase and Pratt⁷.

Extraction⁸

Fresh leaves of the plant were macerated by petroleum ether (PEE), chloroform (CEE), methanol (MEE) and water (AEE) for 5 days at room temperature separately and extracts were collected and weighed. Colour and consistency were recorded. Preliminary phyto chemical screening is done to all the extracts according to the standard procedures described by Kokate and Harborne⁹ .

Antimicrobial activity:

Antimicrobial activity for all the extracts was done by cup plate method. Nutrient agar media ¹⁰was used for bacterial studies and Sabouraud’s dextrose agar¹⁰ was used for fungal studies.

Test solutions of all the four extracts are prepared in concentration of 500µg/ml, 1000µg/ml and 1500µg/ml using DMSO as solvent. Standard solutions are prepared using streptomycin (100µg/ml) for bacterial studies and flucanozole (100µg/ml) for fungal studies as positive control and DMSO is negative control. After inoculation, all the petriplates were incubated at 37^oC for 24 hrs and zone of inhibitions are recorded after 24 hrs for assaying the activity.

In vitro antioxidant activity:

Total antioxidant capacity was determined by Phosphomolybdenum method¹¹ using ascorbic acid as standard.

RESULTS AND DISCUSSION:

Physico chemical studies are mainly used in judging purity and quality of the drug. Ash values give an idea of the earthy matter or inorganic composition or other impurities present along with drug. Extractive values give an idea about chemical constituents present in the drug as well as useful in determination of adulterants. All the results are displayed in table I.

Flourescence analysis of drug powder is observed both in UV light and day light. Powder showed green colour indicating the presence of chromophore. Results of which are displayed in table II.

Extraction is done by maceration for 5 days using four solvents, petroleum ether, chloroform, methanol and water and percentage yields and nature of extracts is tabulated in table III.

Preliminary phytochemical screening for all the four extracts is carried out and results are displayed in table IV. Various extracts showed the presence of alkaloids, carbohydrates, glycosides, flavonoids and phytosterols.

Antimicrobial activity: Zone of inhibitions are measured for all the extracts and methanolic and chloroform extracts showed considerable antimicrobial activity in comparision with standard and also with other extracts. Results are shown in table V.

In vitro antioxidant capacity: Methanolic and chloroform extracts are studied for antioxidant activity and both showed good activity comparable to ascorbic acid, results are shown in table VI.

Table I results showing the proximate analysis

Name of study	Yield of Ixora coccinea
Alcohol soluble extract	0.4g
Water soluble extract	0.17g
Total ash value	8%(w/w)
Acid insoluble ash value	49.905%(w/w)
Sulphated ash value	0.03mg(w/w)

Table II Fluorescence analysis of drug powder of leaves of I. coccinea

S.NO.	TREATMENT	DAYLIGHT	UV LIGHT
1.	Powder	Greenish brown	Pale green
2.	Powder + water	Yellowish green	Dark green
3.	Powder +1 N HCl	Yellowish green	Green
4.	Powder + 1 N HNO₃	Light brown	Green
5.	Powder + 1 N H₂SO₄	Pale brown	Green
6.	Powder + 1N NaOH	Yellowish brown	Blacksih green
7.	Powder + Alc.KOH	Dark brown	Dark green
8.	Powder +1 N KOH	Yellowish brown	Blakish green
9.	Powder + Alc. KOH	Light brown	Green
10.	Powder + ammonia	Yellowish brown	Dark green.

Table III showing percentage yield and appearance of various extracts of selected plants:

Name of the extract	State	Colour of the extract	% Yield. (w/w)
Pet ether	Semisolid	Yellowish green	12.17
Aqueous	Solid	Dark brown	7.85
Chloroform	Semisolid	Greenish yellow	8.34
Methanol	Semi solid	Reddish green	8.88

Table IV showing the nature of phyto constituents present in Ixora coccinea of different extracts

TEST

PEE

CEE

MEE

AQE

ALKALOIDS:

Dragendroff’S

Mayer’s

Wagner’s

Hager’s

AMINO ACIDS:

Million’s

Ninhydrine

CARBOHYDRATES:

Molish’s

Barfoed’s

Seliwinoff’s

Osazone formation

PROTEINS:

Warming’s

Biuret

Xanthoprotic

Hydrolysis

TANNINS:

Ferric chloride test

Gelatin

FLAVONOIDS:

Alkaline reagent

Zinc hydrochloride

FATS AND FIXED OILS:

Saponification

Copper sulphate

CELLULOSE:

Iodine+sulphuric acid

Iodine

STEROIDS AND TRITERPENOIDS:

Libermann buchard

Salkowiski

GLYCOSIDES:

Test A

Test B

ANTHROQUININE GLYCOSIEDS:

Brontragers

CARDIAC GLYCOSIDES:

Keller’s killani

Legal

Baljet

FROTH FORMATION

CONCLUSION:

The plant Ixora coccinea has shown good activity against bacterial strains and it also has good antioxidant capacity comparable to ascorbic acid. Hence further research can be done to achieve more benefits from this natural herb.

ACKNOWLEDGEMENTS:

The authors want to acknowledge the management and principal for giving support and encouragement for successful completion of this research work. We also want to thank Dept. of Microbiology, N.G. College, Nalgonda for supplying us the test organisms for fulfilling the research work.

Table V Showing zone of inhibition (in mm) of various extracts of Ixora coccinea

EXTRACT

E.COLI

S.AUREUS

C.ALBICANS

Aqueous

500µg/ml

1000µg/ml

1500µg/ml

Methanol

500µg/ml

1000µg/ml

1500µg/ml

chloroform

500µg/ml

1000µg/ml

1500µg/ml

Petroleum ether

500µg/ml

1000µg/ml

1500µg/ml

Table VI In-vitro antioxidant activity of Ixora coccinea.

Methanolic extract of I. coccinea:

Concentration	Absorbance
10µg/ml	0.244
30 µg/ml	0.246
50 µg/ml	0.260
70 µg/ml	0.248
100 µg/ml	0.275
200 µg/ml	0.217

Chloroform extract of I .coccinea:

Concentration	Absorbance
10 µg/ml	0.286
30 µg/ml	0.261
50 µg/ml	0.270
70 µg/ml	0.362
100 µg/ml	0.270
200 µg/ml	0.272

Table showing in-vitro anti oxidant activity of ascorbic acid (standard)

Concentration	Absorbance
3µg/ml	0.236
5µg/ml	0.238
10µg/ml	0.270
20µg/ml	0.272
30µg/ml	0.273
40µg/ml	0.278
50µg/ml	0.280

REFERENCES

1 . B. Mahesh et. Al, “Antimicrobial activity of some important medicinal plant against plant and human pathogens”, World Journal of Agricultural Sciences, 4(5): 839 – 843.

2. The Wealth of India, Dictionary of Indian Raw materials and Industrial products – Raw Materials, National Institute of Sciences communication, New Delhi, 2002

3. Mukesh Chandra Sharma et.al, “Preliminary phytochemical and antimicrobial investigation of aqueous extract of Ixora coccinea and Commelina benghalensis as gram +ve and gram –ve micro organisms”, Middle East Journal of Scientific Research 6(5):436 – 439.

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8. Pharmacognosy, C.K. Kokate, Purohit, Nirali prakashan, 2004

9. Pjytochemical methods, J.B. Harborne, Springer

10. Pelczar M.J and Reid J.D., Microbiology, Tata Mc graw Hill, New Delhi, 1974. Prieto P, Pineda M, Aguilar M, “Spectrophotometric quantitation of antioxidant formation of a phosphomolybdenum complex: capacity through the specific application to the determination of vitamin”, E. Anal Biochem, 1999:269: 337 – 41.

Received on 25.06.2012

Modified on 10.07.2012

Accepted on 14.07.2012

Research Journal of Pharmacognosy and Phytochemistry. 4(4): July- August 2012, 205-208