Phytochemical studies and in vitro Anti-inflammatory activity of  Melia azedarach (L) flower

 

S. Sumathi1*, M. Sinthanai Selvi2

1Guest Lecturer, Department of Biochemistry, Dharamapuram Gnanambigai Govt. Arts College, Mayiladuthurai India-609001

2Department of Biochemistry, Dharamapuram Gnanambigai Govt. Arts College,

Mayiladuthurai India-609001

*Corresponding Author E-mail: ssumibio@yahoo.co.in

 

ABSTRACT:

Physiochemical parameters such as colour, taste, odour, ash values, moisture content were determined. Carbohydrate  and protein  content were also determined. Various colour change were observed when treated with different chemical reagents and Preliminary phytochemical screening showed the presence of alkaloids, flavonoids, terpenoids, anthraquinonens, tannins, saponins and cardic glycosides. The aqueous extracts of the flower of Melia azedarach (L) were studied for in vitro anti-inflammatory activity by HRBC membrane stabilization method. The flower extract exhibited membrane stabilization effect by inhibiting hypotonicity induced lysis of erythrocyte membrane. The aqueous extract shows significant anti inflammatory activity at the concentration of 800 µg /ml which is comparable to the standard drug (Diclofenac sodium 200µg/ml)The anti inflammatory activity of the extracts were concentration dependent, with the increasing concentration the activity is also increased.

 

KEYWORDS: Phytochemicals, Melia azedarach, anti inflammatory.

 

 

 

INTRODUCTION:

Nature is an immense source of pharmacologically active molecules which has been used for the treatment of the incurable diseases without any adverse effects. Ayurvedic plants are more potent and known to have human medicinal properties. The products obtained from the plant source are clinically used for health benefits without any chemical modification (Kokate et al., 2000)

 

Plants have contributed lot of medicinal compounds being used today to treat diseases like cancer, hormonal imbalances, jaundice, diabetes, inflammation etc. Medicinal plants are very commonly available in abundance especially in the tropics. They are the vital sources of wide variety of chemicals from which novel anti-inflammatory agents can be discovered (Padmanabhan., 2012) Inflammation is a normal protective response to tissue injury and it involves a complex array of enzyme activation. Mediators release, fluid extravasations, cell migration, tissue breakdown and repair. It is a complex process, which is frequently associated with pain and involves occurrences such as the increase in vascular permeability, increase of protein denaturation and membrane alterations. (Umapathy et al., 2010)

 

MATERIALS AND METHODS:

Collection of plant material:

Fresh flower of Melia azedarach (L) were collected from in and around Mayiladuthurai. Mayiladuthurai belongs to Nagapattinam district of Tamil Nadu State, India.

 

Preparation of flower extract:

Fresh flowers was washed thoroughly under running tap water and shade dried. Dried flowers were homogenized to a fine powder and stored in air tight bottles. Powder characteristics analysis was done. 10gm of flower powder was extracted with 100ml of solvents (water) for 24 hours by soaking method. This extract filtered through a filter paper and further used for different test (Manimegalai and Rakkimuthu, 2012).

 

Methods:

Fluorescence analysis (Xavier, 2012), preliminary Phytochemical  test (Harbone, 1973), Determination of total carbohydrates (Mahadevan and Sridhar,1982), Determination of total protein (Lowry et al., 1951), Determination of ash content (AOAC 2000), Determination of moisture content (Sadasivam and Manikam 1996), Calculation of solid content (Hossain et al., 2007), in vitro anti –inflammatory activity (Azeem et al., 2010).

 

RESULT:

Table 1 shows the powder characteristics such as colour, odour and taste of flower of Melia azedarach (L) were observed. The physiochemical parameters such as Total Ash, Moisture and Solid content were measured. The total carbohydrate content in Melia azedarach (L) has been found to be 177g/100ml Biochemical studies on the flower extract of Melia Azedarach (L) revealed that the protein content has been found to be 105 g /100 ml .

 

TABLE: 1. PHYSIOCHEMICAL AND BIOCHEMICAL ANALYSIS OF FLOWER EXTRACT OF Melia azedarach (L)

S.NO

CHARACTERS/

PARAMETERS

PROPERTIES

1

Colour

Brown colour

2

Taste

Bitter taste, Acceptable

3

Odour

Acceptable

4

Moisture

6.8g

5

Ash

1.15g

6

Solid content

0.15

7

Carbohydrate

177g/100ml

8

Protein

105g/100ml

 

Table 2, From the results it was proved that the powder of the flower were treated with chemicals like NaOH, Conc. H2SO4, Acetic acid,HCl,conc.HNO3 and Iodine various colours were obtained. The powder as such expressed brown colour and when it was dissolved in water, no colour changes was observed. Various colour change were observed when treated with different chemical reagents.

 

TABLE: 2 FLUORESCENCE ANALYSIS OF FLOWER EXTRACT OF Melia azedarach (L)

S.NO

CHEMICAL REAGENTS

APPEARANCE OF COLOUR

1

Powder colour

Brown/colour

2

5%  NaOH

Reddish yellow

3

10%  NaOH

Reddish yellow

4

Conc.H2SO4

Purple

5

5% Acetic acid

Light pink

6

5% HCl

Light red Yellow

7

Con. HNO3

Yellow

8

10%  HCl

Light Red

9

Iodine

Light yellow

10

10% Acetic acid

Yellow

 

Table 3 shows the qualitative analysis of phytochemical evaluation revealed the presence of alkaloids, flavonoids, terpenoids, anthraquinonens, tannins, saponins and cardic glycosides. So the presence of this compound may be responsible for anti inflammatory activity

 

Table 4 depicts the flower extract exhibited membrane stabilization effect by inhibiting hypotonicity induced lysis of erythrocyte membrane. The lysosomal enzymes released during inflammation produce a variety of disorders. Since HRBC membrane are similar to lysosomal membrane components the prevention of hypotonicity induced HRBC membrane lysis is taken as a measure of anti inflammatory activity of drugs. The results were reported in table 4. It was observed from the table 4 and figure 1 that the aqueous extract shows significant anti inflammatory activity at the concentration of 800 µg /ml which is comparable to the standard drug (Diclofenac sodium 200µg/ml) The anti inflammatory activity of the extracts were concentration dependent, with the increasing concentration the activity is also increased.

 

DISCUSSION:

Vijaya Kumar et al.,(2012) reported the physiochemical parameters such as foreign matter, total ash, acid insoluble ash, water soluble ash, loss on drying and swelling index was measured in bark extract of Melia azedarach (L) Naseerbanu et al., (2003) estimated carbohydrate contents in Amaranthus viridis and Spinacea oleracea. In which, A viridis showed higher carbohydrate (3.562 mg /100 ml) than Spinacea oleracea.

 

Babu et al. ,(2007) reported that the presence of carbohydrates and proteins in both Origanum vulgare and Althea officinalis based on the preliminary phytochemical analysis

 

During a pharmacognostic study carried out on the flower of Pterospermum cicerifolium (L) by Shome and Mehrotra (1990) greenish purple colour was noted on treatment with 1N HCl and nitro cellulose.

 

Vijaya Kumar et al.,(2012) reported the phytochemical profiling of the bark extract of Melia azedarach (L) revealed the presence of carbohydrates, steroid, glycosides, triterpenoids and flavonoids. The therapeutic benefits of secondary Metabolite of plant origin have been researched in several recent studies (Nayak., 2010).

 

 

TABLE :3 PHYTOCHEMICAL SCREENING OF AQUEOUS EXTRACT OF Melia azedarach (L)

S.No

TEST

NATUREOF COLOUR CHANGE

INFERENCE

PHYTOCHEMICAL CHANGES

1

Alkaloids

White

Presence

Presence Of Alkaloids

2

Glycosides

Yellow

Absence

Absence Of Glycosides

3

Flavanoids

Reddish Pink

Presence

Presence Of Flavanoids

4

Reducing Sugar

Orange Red

Presence

Presence Of Sugar

5

Tannins

White

Presence

Presence Of Tannins

6

Anthroquinones

Rose Pink

Presence

Presence Of Anthroquinones

7

Saponins

Foam

Presence

Presence Of Saponins

8

Cardiac Glycosides

Blue

Presence

Presence Of Cardiac Glycosides

 

TABLE :4 In vitro ANTI-INFLAMMATORY ACTIVTY OF FLOWER EXTRACT OF Melia azedarach (L)

S.NO

GROUPS

CONCENTRA-TION (µg/ml)

 % PROTECTION

 MEAN ± S.E.M

1

Aqueous

Extract

100

18.31 ± 0.30

 

 

200

24.62 ± 0.22

 

 

400

31.05 ± 0.62

 

 

800

55.09 ± 0.09

2

Standard

100

49.32 ± 0.18

 

 

200

60.15 ± 0.35

 

The earlier studies reported that the presence of tannins (Mota, 1985) flavanoids (Huang, 2009) in extracts are responsible for inflammatory activity. So the membrane stabilizing activity of the extract may be due to the presence of flavonoids, alkaloids, tannins and saponins present in aqueous extract of Melia azedarach (L).

 

The lysosomal enzymes released during inflammation produce a variety of disorders. The extra cellular activity of these enzymes is said to be related to acute or chronic inflammation. The non steroidal drugs act either by inhibiting these lysosomal enzymes or by stabilizing the lysosomal membrane. (Rajendran , 2008).

 

From the above study it was concluded that the aqueous extract Melia azadrach (L) has significant membrane stabilization property compared to the standard drug Diclofenac sodium. The present study suggests that the Melia azedarach (L) would serve as a source for the discovery of novel anti-inflammatory agents.

 

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Received on 23.11.2013       Modified on 11.12.2013

Accepted on 14.12.2013      ©A&V Publications All right reserved

Res.  J. Pharmacognosy & Phytochem. 6(1): Jan.-Mar. 2014; Page 19-21