In vitro Antioxidant, Antimicrobial, Anti-inflammatory, Anthelmintic Activity and Phytochemical Analysis of Indian Medicinal Spices

 

V. Manju*, R. Revathi and M. Murugesan.

Department of Biochemistry, Periyar University, Salem-636 011, Tamil Nadu, India.

*Corresponding Author E-mail: vaisell@gmail.com

 

ABSTRACT:

This study reports the in vitro antioxidants, antimicrobial, anti-inflammatory, anthelmintic activity and phytochemical analysis of Indian medicinal spices. The use of medicinal spices or their crude extract provides the unlimited better opportunities to develop the new drugs against infectious diseases. Fresh spices of Quercus infectoria and Papaver somniferum were dried in the shade at room temperature (25º±2ºC) and pulverized to powder using an electric blender. The powder (10g) was percolated in 80mL of absolute ethanol for 48 h after which it was filtered. The ethanol in the filtrate was allowed to evaporate at room temperature to yield the extract concentrate, giving a 10 % yield, were subjected to phytochemical screening for the presence of secondary metabolites by the method of Fadeyi and Harbone. Antimicrobial activity was investigated using the method of agar dish diffusion technique using five clinical strain of human pathogenic organism and anti-inflammatory activity was investigated by measuring the percentage of haemolysis using the method of HRBC and antihelmintic activity was investigated using earthworm test. In addition to this antioxidant activity was determined by measuring both enzymatic and non-enzymatic antioxidants such as catalase, glutathione peroxidase, glutathione -S- transferase, vitamin C and E. Our results revealed that both spices have biologically active compounds such as flavonoids, alkaloids, glycosides, phenols and resins and which exhibit the significant levels of antioxidant, antimicrobial, anti-inflammatory and anthelmintic properties. The results obtained from this study suggested that Quercus infectoria and Papaver somniferum can be used in treatment of infectious diseases caused by various microbes.

 

KEYWORDS: Quercus infectoria, Papaver somniferum, antioxidant, antimicrobial

 


 

INTRODUCTION:

Infectious diseases the leading cause of death worldwide. Antibiotic resistance has become a global concern1. The use of spices has evoked interest as source of natural products for their potential uses as alternative remedies to heal many infectious diseases2. Synthetic drugs are not only expensive and inadequate for the treatment of diseases but also often with adulterations and side effects3. Therefore, researchers are increasingly turning their attention to folk medicine, looking for new infection-fighting strategies to control microbial infections4. Natural antimicrobial agents are non-toxic and non-allergenic and do not cause the problems of microbial resistance5.

 

Spices are frequently used as an active ingredient in certain medicines and reported to possess a number of pharmacological effects to treat different human alignments6. Moreover several scientific reports have described the inhibitory effect of spices on a variety of microorganisms7.

 

Traditional medicinal systems, like the ayurveda and siddha mention the use of plants and spices in treatment of various human aliments. Spices have a diverse array of natural phytochemicals that have complementary and overlapping actions, including antioxidants effects, modulation of steroid metabolism and antimicrobial effects8.

 

Quercus infectoria is a small tree native of Greece, Asia and Iran. The galls arise on young branches of this tree as a result of attack by the gall wasp Adleria gallae tinctoria9. The galls are locally known as manjakani in Malaysia, and are used in combination with other herbs as drinking remedy by women after childbirth to restore the elasticity of the uterine wall10. Galls of Quercus infectoria also used to treat haemorrhages, chronic diarrhea and dysentery11.

 

Papaver somniferum is one of the oldest cultivated medicinal plants commonly known as opium poppy12,13 is most important renewable resources for alkaloids and produce more than a 100 different alkaloids14 mainly morphine, codeine, thebaine, papaverine and norcotine15 in which morphine and codetine is only source for analgesic and antitissue drugs16.

 

Nowadays natural products have long been recognized as an important source of therapeutically effective medicines. Therefore the present study was carried out to evaluate the phytochemical composition and in vitro antioxidant, antimicrobial, anti-inflammatory and anthelmintic properties of Quercus Infectoria, Papaver Somniferum.

 

MATERIALS AND METHODS:

Collection and identification of spices:

The spices used in the study Quercus Infectoria, Papaver Somniferum was collected from nearby gardens of Salem town at 2009. The voucher of these plant and spices was deposited at the herbarium of Tamil Nadu Agricultural University, Coimbatore.

 

Preparation of extracts:

Fresh spices of the plant were dried in the shade at room temperature (25º±2ºC) and pulverized to powder using an electric blender. The powder (10g) was percolated in 80mL of absolute ethanol for 48 h after which it was filtered. The ethanol in the filtrate was allowed to evaporate at room temperature to yield the extract concentrate, giving a 10 % yield17.

 

Phytochemical screening:

Both qualitative and quantitative analysis of the phytochemicals present was carried out using the methods of 18, 19.

 

Microorganism:

Staphylococcus aureus, Streptococcus pyogenes, E.coli, Pseudomonas aeruginosa, Candida albicans were procured from the Department of Microbiology, Tamil Nadu Agricultural University, Coimbatore.

 

Antimicrobial activity:

The antimicrobial activity was determined by modified agar well diffusion method of20, 21.

 

Antioxidant activity:

The activity of catalase (CAT) was determined by the method of22. Reduced glutathione (GSH) was determined by the method of23. The activity of glutathione-S-transferase (GST) was assayed by the method of24. Glutathione peroxidase (GPx) was assayed by the method of25. Vitamin C was measured according to the method of26. Vitamin E was determined by the method of 27.

 

Anti-inflammatory activity:

Anti-inflammatory activity was determined by the method of slightly modified28.

 

 

Anthelmintic activity:

the anthelmintic assay was carried by the method of 29.

 

RESULTS:

Qualitative phytochemical analysis confirms the presence of flavonoids, glycosides, phenols, resins and alkaloids were indicated to various extends in the Quercus infectoria and Papaver somniferum for secondary metabolites. Steroids and saponins were not detected in the present studies. Quantitative analysis confirms that flavonoids and glycosides were found to be present in high amount in Quercus infectoria than Papaver Somniferum. In Papaver Somniferum the levels of alkaloids, resins and phenols were found in high amount than the Quercus infectoria (Table 1).

 

The maximum antimicrobial activity was showed in Quercus infectoria, Papaver Somniferum. Quercus infectoria shows in maximum activity against all the organisms Staphylococcus aureus, Streptococcus pyogenes, E.coli, Pseudomonas aeruginosa and Candida albicans. Papaver somniferum shows in minimum effect against all the organisms, but there is no antimicrobial activity against Cadida albicans (Table 2).

 

The levels of catalase and glutathione-S-transferase are significantly increased in Papaver somniferum compared to Quercus infectoria. But the level of glutathione peroxidase, reduced glutathione, vitamin E and vitamin C significantly increased in Quercus infectoria compared to Papaver somniferum (Table 3). The anti-inflammatory activity of Quercus infectoria is (47.3%) and this is found to be higher than the other spices Papaver somniferum. It is quite apparent that the ethanolic extracts possess significant anti-inflammatory activity (Table 4). Quercus infectoria extract exhibited more potent activity at 10% concentration against the pheretima posthuma earth worm. Papaver somniferum extract at high concentration is more effective as compared to standard 50 mg/mL (Table 5).

 

DISCUSSION:

Results from our phytochemical analysis revealed the ethanol fractions of the spices Quercus infectoria and Papaver somniferum showed the presence of flavonoids, glycosides, phenols, resins and alkaloids which could be responsible for beneficial efficacies of these spices on human health. The ethanolic extract of the spices of Quercus infectoria and Papaver somniferum were subjected to preliminary screening for antimicrobial activity against four standard bacteria Staphylococcus aureus, Streptococcus pyogenes, Escherichia coli, Pseudomonas aeruginosa and one fungal strain Candida albicans.


Table 1: Qualitative and Quantitative Phytochemical Analysis of Quercus infectoria and Papaver somniferum

Phytochemical constituents

Quercus infectoria

Papaver somniferum

Quercus infectoria

Papaver somniferum

Flavonoids

+

+

65 ± 7.07

52 ± 6.15

Glycosides

+

+

95.2 ± 3.42

74.8 ± 7.41

Resins

+

+

16.6 ± 5.42

29 ± 2.82

Phenols

+

+

184 ± 5.65

275.6 ± 3.72

Alkaloids

+

+

137.6 ± 3.44

155.4 ± 6.31

Saponins

-

-

-

-

Steroids

-

-

-

-

+ (Present), - (Absent)


Table 2: Screening of Antimicrobial Activity of Quercus infectoria, Papaver somniferum

Strains

Control

Quercus infectoria

Papaver somniferum

Staphylococcus aureus

30 mm

26 mm

8 mm

Streptococcus pyogenes

29 mm

20 mm

11 mm

E.Coli

35 mm

22 mm

9 mm

Pseudomonas aeruginosa

35 mm

25 mm

8 mm

Candida albicans

45 mm

36 mm

Absent

 

Table 3: Screening of Antioxidant Activity of Quercus infectoria, Papaver somniferum

Antioxidants

Quercus infectoria

Papaver somniferum

Catalase

0.31 ± 0.042

0.87 ± 0.014

Glutathione peroxidase

0.518 ± 0.036

0.22 ± 0.080

Glutathione-S- transferase

0.482 ± 0.089

0.672 ± 0.050

Reduced glutathione

0.646 ± 0.1

0.63 ± 0.014

Vitamin E

0.614 ± 0.085

0.694 ± 0.089

Vitamin C

0.26 ± 0.102

0.216 ± 0.049

 

 

Table 4: Screening of Anti-Inflammatory Activity of Quercus infectoria, Papaver somniferum

Ethanolic extract of Plants

Anti-inflammatory activity

Quercus infectoria

47.3%

Papaver somniferum

10.8%

 

Table 5: Screening of Antihelmintic Activity of Quercus infectoria, Papaver somniferum

Ethanolic extract of Plants

Treatment vehicle

Concentration used

Time taken for paralysis (min)

Time taken for death (min)

Control

Piperazine Citrate

15 mg/ml

10

12

Quercus infectoria

Alcoholic Extract

50 mg/ml

15

19

Papaver somniferum

Alcoholic Extract

50 mg/ml

14

17

 

 

 


Quercus infectoria shows high levels of phenols, alkaloids and flavonoids and shows antimicrobial activity. Our results are in line with previous findings30. Papaver somniferum shows high levels of phenols, alkaloids and moderate levels of flavonoids but it shows considerable levels of antimicrobial activity against bacterial strain. Our results are in line with previous findings31Papaver somniferum shows no activity against fungal strain of Candida albicans. It was clear from Table 2 and it is not surprising that there are defenses in the antimicrobial effects of spices, due to phytochemical properties and difference among spices. It has been suggested that antimicrobial activity is mainly due to the presence of essential oils, flavanoids and triterpenoids and other natural polyphenolic compounds or free hydroxyl groups and alkaloids also highly correlated with antimicrobial activity32.

 

Quercus infectoria had rich source of GPx, GSH and vitamin C. In the previous study, Nutagalls of Quercus infectoria shows potent antioxidant activity33, 34. Papaver somniferum had rich source of CAT, GST and vitamin E. But in the previous findings shows Papaver somniferum plant extract had moderate range of antioxidant activity35. Finally our results concluded that both spices show good antioxidant activity. This suggests that the physiochemical nature of the individual phenolic in the extract may be important to contributing the antioxidant activity36. Polyphenols are the major plant compounds with antioxidant activity. Although are not the only once. The antioxidant activity of the phenolic compounds is reported to be mainly due to their redox properties, which can play an important role in observing and neutralizing free radicals, quenching single and triplet oxygen or decomposing peroxides37, 38.

 

Ethanolic extract of Quercus infectoria and Papaver somniferum were subjected to preliminary screening for anti-inflammatory activity. Quercus infectoria it shows 47.3% of anti-inflammatory activity. This results correlate with the previous finding shows that substantial anti-inflammatory activity39. But Papaver somniferum shows 10.8% of anti-inflammatory activity and Quercus infectoria and Papaver somniferum also give maximum membrane stability due to the presence of biologically active compounds such as alkaloids and flavonoids which have been reported to be promising the anti-inflammatory activity in animal model40.

 

As shown in Table 4 ethanolic extract of Quercus infectoria and Papaver somniferum in dose dependent manner showed anthelminitic activity. Results were comparable with standard drug piperzine citrate. Phytochemical analysis of the crude extract revealed that presence of biologically active compounds in Quercus infectoria and Papaver somniferum. The biologically active compounds are joinly or independently involved in anthelminthic activity against parasites41. Quercus infectoria exhibit more potent activity at 10% concentration against the earthworm. Papaver somniferum shows at 50% concentration is more effective as compared to standard 50 mg/ml.

 

Based on the results, we may concluded that Quercus infectoria and Papaver somniferum contained many biologically active compounds with important medicinal properties. Hence the Quercus infectoria and Papaver somniferum can be further subjected to isolation of the therapeutic value and carry out further pharmacological evolution.

 

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Received on 02.12.2010          Modified on 20.12.2010

Accepted on 30.12.2010         © RJPT All right reserved

Research J. Pharm. and Tech. 4(4): April 2011; Page 596-599