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 10 was 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 37oC 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

 

-

-

-

 

 

-

10

-

 

 

-

-

-

 

chloroform

500µg/ml

1000µg/ml

1500µg/ml

 

 

-

-

-

 

 

05

10

15

 

 

-

-

-

 

Petroleum ether

500µg/ml

1000µg/ml

1500µg/ml

 

 

-

-

10

 

 

-

-

-

 

 

-

-

-

 

 

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.

4.        Yasmeen et. al,  “Evaluation of the anti diarrhoeal activity of the leaves of I. coccinea Linn, in rats” Pharmaceutical Biology, 43(2), 2005, 147 – 152.

5.        Neelapu Neelima et. Al, “Hypolipidemic activity and HPTLC analysis of Ixora coccinea L leaves”, Journal of Applied Pharmaceutical Sciences 01 (10),2011,172 – 175.

6.        Kokate C.K, Purohit, Gokhale S.B, Pharmacognosy, 36 th edition, Nirali Prakashan, Pune, 2006

7.        Vadivi R et. Al, “Pharmacognostic standardization of leaves of Ixora coccinea:, Journal of Pharmaceutical Sciences and Research 1(4), 2009, 151 – 157.

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

© A&V Publication all right reserved

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