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IN-VITRO INVESTIGATION OF BIOLOGICAL ACTIVITY OF AERIAL PART OF SYZYGIUM CUMINI

 


Azima Sultana Julie, TanjinaSultana, Farhina Rahman Laboni, Samira Karim and Zubair Khalid Labu*
 Department of Pharmacy, World University of Bangladesh, 51/8, Green road, Dhaka, Bangladesh

 

ABSTRACT

 Plants parts and plant products of all the descriptions, particularly those with medicinal properties, are invariably used as principal components or ingredients of various traditional medicines. The ancient time of modern age the race successfully used plants and plant products as therapeutics tools for fitting against disease and various other health hazards. In this study the ethanolic extract and its different partitionates of leaves of Syzygiumcumini were observed to different phytochemical investigations such as antimicrobial activity, thrombolytic activity and membrane stabilizing activity. The different extractives of Syzygiumcumini also significantly confirm that the plant has moderate membrane stabilizing activity 59.33 %( hypotonic) and 29.46 %( heat induced) where standard drug acetyl salicylic acid showed 72.57 %( hypotonic) and 49.25% (heat induced) membrane stabilizing activity.  However, the standard drug Streptokinase showed thrombolytic activity 65% and the ethanolic soluble fraction of plant showed (ESF-45.33%). In antimicrobial screening the plant showed satisfactory activity with the comparison of standard drug Ciprofloxacin. The crude extract of Syzygiumcuminigave 15mm and standard drug 24mm of diameter zone of inhibition against Vibrio parahimolyticus.

Key words- Syzygiumcumini, Ciprofloxacin, membrane stabilizing, thrombolytic, antimicrobial activity

INTRODUCTION

Research on the medicinal plants can also contribute immensely to the development of modern pharmacology. The discovery of a novel chemical compound from a medicinal plant or a plant medicinal may form the basis for the development of important therapeutic agent with better pharmacological properties. Over the year medicinal plants have been found useful in the treatment and management of the various health problems and natural products compounds discovered from medicinal plants have provided numerous clinically useful drugs [1]. Even with recent advances in modern medicine practice is gaining more follower ship. An enumeration of the WHO from the late 1970s listed 21000 medicinal species. However in chaina alone 4941 to 26092 native species are used as drug in Chinese traditional medicine an astonishing 18.9 percent [1]. If this proportion is calculated for other well-known medicinal floras and then applied to the global total of 422,000 flowering plant speceies [2] it can be estimate that the number of plant species used for medicinal purposes is more than 50,000.

The genus Syzygium is one of the genera of the myrtle family Myrtaceae which is native to the tropics, particularly to tropical America and Australia. It has a worldwide, although highly uneven, distribution in tropical and subtropical regions [3].

Syzygiumcumini is a large evergreen and densely foliaceous tree with greyish-brown thick bark, exfoliating in woody scales. The leaves are leathery, oblong-ovate to elliptic or obovate-elliptic with 6 to 12 centimeters long. Flowers are scented, greenish-white, in clusters of just a few or 10 to 40 and are round or oblong in shape, including some improved races bearing purple to violet or white colored flesh and seedless fruits have been developed. The fruits are berries and are often obviously oblong, 1.5 to 3.5 centimeters long, dark-purple or nearly black, luscious, fleshy, and edible; it contains a single large seed. According to Ayurveda, its bark is acrid, sweet, digestive, and astringent to the bowels, anthelmintic and in good for sore throat, bronchitis, asthma, thirst, biliousness, dysentery, blood impurities and to cure ulcers [4]. The fruits are acrid and sweet, cooling, dry and astringent to bowels. They increase “Vata” and remove bad smell from the mouth. As per Unani system of medicine they acts as liver tonic, enriches blood, strengthens teeth and gums and forms good lotion for removing ringworm infection of the head.  This study was designed to isolate ethanolic extract as well as to observe biological activities phytochemical screening, antimicrobial, thrombolytic, membrane stabilizing activity of the Syzygiumcumini (Family: Myrtaceae).

 MATERIALS AND METHOD

 Syzygiumcumini leaves were collected in the month of April 2016 from Comilla during rainy season when weeds were in their maximum densities. The plant was identified by the experts of Bangladesh National Herbarium, Mirpur, Dhaka and a voucher specimen was also deposited there.

At first the leaves were thoroughly washed to remove dust, soil and then dried under sunlight. The dried samples were grounded to coarse powder with a mechanical grinder (Grinding Mill) and powdered samples were kept in clean closed glass containers. The fine powder of leaves (250g) was soaked in 1.5 Litre ethanol for 8 days. After the extraction process the plant extracts was filtered with sterilized cotton filter and Whatman filter paper number 1. The rotary evaporator was used for the evaporation of filtrates extract under vacuum. Solvent-solvent partitioning of crude extract (5gm) of Syzygiumcumini was done using the protocol designed by modified Kupchan [5]. It was extracted with Petroleum Ether, carbon tetrachloride, Chloroform and fractions were evaporated to dryness. Then finally obtained petroleum ether (PESF) soluble fraction, carbon tetrachloride (CTSF) soluble fraction, chloroform (CLSF) soluble fraction and aqueous (AQSF) soluble fraction (Table 1)

Table 1: Fractions from Syzygiumcumini extract by modified Kupchan 

Crude extract or Fractions Syzygiumcumini(g)
ESF 5.00
PESF 0.55
CTSF 0.50
CLSF 0.40
AQSF 2.50

Membrane stabilizing activity: The erythrocyte membrane resembles to lysosomal membrane and as such, the effect of drugs on the stabilization of erythrocyte could be extrapolated to the stabilization of lysosomal membrane [6]. In heat induced and hypnotic solution induced hemolysis, analiquots (2mg/ml of each) containing different extractive of Syzygiumcumini were taken to investigate the anti-inflammatory activity.

Thrombolytic activity: Thrombolytic are used to dissolve the fibrin of blood clots which are potentially life-threatening. Significantly this method was expanded by Watson [7, 8] which was used to determine thrombolytic activity of ethanol extracts and its different fractions of Syzygiumcumini leaves by using Streptokinase as standard drug [9].

Antimicrobial screening: Antimicrobial activity of the different extracts of Syzygiumcuminileaves was determined by disc diffusion method. Disk diffusion technique is widely acceptable for the preliminary screening of antimicrobial activity. It is essentially a qualitative or semi-quantitative test indicating the sensitivity or resistance of microorganisms to the test materials. However, no distinction between bacteriostatic and bactericidal activity can be demonstrated by this method [10,11].

RESULTS AND DISCUSSION

 The different extracts of the leaf of concentration 2.0 mg/ml significantly protected the lysis of human erythrocyte membrane induced by hypotonic solution and heat induced (isotonic solution) as compared to the standard acetyl salicylic acid. The leaves of Syzugiumcumini were effective in the membrane stability activity as the extractive prevented the lysis of erythrocyte induced by hypnotic solution. The crude/ethanol extract (ESF) inhibited 59.33%, pet ether soluble fraction (PESF) inhibited 46.05%, Chloroform soluble fraction (CLSF) inhibited 40.44%, carbon tetra chloride soluble fraction (CTSF) inhibited38.16%, The aqueous soluble Fraction (AQSF) inhibited 33.54%, Acetyl salicylic acid (ASA) as standard drug that exhibited 72.57% inhibition of hemolysis of normal condition.

The crude/ethanol extract (ESF) inhibited 29.46%, pet ether soluble Fraction (PESF) inhibited 23.81%, Chloroform soluble fraction (CLSF) inhibited 20.41%, carbon tetra chloride soluble Fraction (CTSF) inhibited19.80%, the aqueous soluble Fraction (AQSF) inhibited 11.98%, and standard drug Acetyl salicylic acid (ASA) exhibited 49.25% inhibition of haemolysis of heating condition.

Table 2: Effects of different extraction of Syzugiumcumini hypnotic solution induced and heat induced haemolysis of erythrocyte membrane.

Sample % of inhibition  
  Syzugiumcumini  
  Hypotonic solution induced Heat induced
ESF 59.33 29.46
PESF 46.05 23.81
CHSF 40.44 20.41
CTSF 38.16 19.80
AQSF 33.54 11.98
STD(Acetyl salicylic acid) 72.57 49.25

In this study moderate thrombolytic activity was found in Ethanol soluble fraction (ESF –45.33%), Pet ether soluble fraction (PESF-30.33%), Carbon tetrachloride soluble fraction (CTSF-33.76%), Chloroform Soluble fraction (CLSF-22.85%) and aqueous soluble fraction (AQSF-17.46%) of Syzygiumcuminileaves.  The amounts of the thrombolytic activity in standard drug Streptokinase as positive control 62.66% and negative control (blank) 2.94% were found.

Table 3: Thrombolytic Activity (in terms of % of clot lysis) of the extractives of Syzygiumcumini leaves

Sample % of clot lysis
  Syzygiumcumini
Blank 2.94%
SK 62.66%
ESF 45.33%
PESF 30.33%
CLSF 22.85%
CTSF 33.76%
AQSF 17.46%

Standard (SK) = Streptokinase; Blank= negative control

The crude extract of Syzygiumcumini was tested for anti-microbial activity against a number (09) of both gram positive and gram-negative bacteria. Standard antibiotic discs of Ciprofloxacin (5µg/disc) were used for comparison purpose. The fractions gave zone of inhibition against these bacteria in the range of 4-14mm (diameter) and the standard drug Ciprofloxacin gave in the range of 18-22mm (diameter) zone of inhibition. The crude extract of Syzygiumcumini showed 15mm and standard drug 24mm of diameter zone of inhibition against Vibrio parahimolyticus.

 Table 4: In vitro antimicrobial screening of Syzygiumcumini leaf

Test organisms

 

Diameter of zone of inhibition(mm)
ESF PESF CLSF CTSF AQSF STD
Gram positive Bacteria
Bacillus subtilis 14 9 11 1 4 22
Sarcinalutea 10 7 8 8 6 19
Staphylococcus aureus 12 10 6 9 5 16
Bacillus cereus 13 8 10 11 7 19
Gram negative Bacteria
Escherichia coli 10 7 10 8 6 20
Shigelladysenteriae 13 8 6 9 7 21
Vibrio mimicus 12 6 7 9 6 16
vibrio parahimolyticus 15 9 8 6 7 24
salmonella typhi 12 8 12 8 8 18

CONCLUSION

The ethanolic extract and its different partitionates of leaves of Syzygiumcumini were subjected to different phytochemical investigations such as antimicrobial activity, thrombolytic activity and membrane stabilizing activity. The different extractives of Syzygiumcumini also significantly protect the lyses of human erythrocyte membrane induced by hypotonic solution and induced by heat which confirm that the plant also has high membrane stabilizing activity as it stabilized the membrane of RBCs. However, the plant has moderate thrombolytic activity. Although the plant has less antimicrobial activity to certain experimental organisms, the plant can be further screened against various diseases in order to find out its unexplored efficacy and can be a potential source of chemically interesting and biologically important drug candidates. Therefore there is huge potential to find active principles which could be beneficial for mankind for targeting various diseases and to rationalize their uses as traditional medicines.

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