ANTI TUBERULAR AND ANTHELMINTHIC ACTIVITIES OF AQUEOUS METHANOLIC EXTRACT OF CARALLUAMA ATTENUATAHTML Full Text
ANTI TUBERULAR AND ANTHELMINTHIC ACTIVITIES OF AQUEOUS METHANOLIC EXTRACT OF CARALLUAMA ATTENUATA
- Mounika 2, M. Vijaya Jyothi *1, P. Ramalingam 1, Vinusha Reddy J. 2 and K. Anusha 2
Department of Pharmaceutical chemistry 1, Department of Pharmacology 2, Raghavendra Institute of Pharmaceutical Education &Research, KR Palli cross, Ananthapuramu, Andhra Pradesh, India.
ABSTRACT: Caralluma attenuata is a perennial small, leafless succulent plant which grows in tropical and subtropical areas. It belongs to the family Apocyanaceae. The folkal information reveals that the tribes generally chew the whole plant to treat peptic ulcer, diabetes, in weight reduction to suppress hunger and also to produce endurance. It has been reported for the presence of steroids, flavanoids, saponins, triterpenes and pregnane glycosides etc., several biological activities like anti-diabetic and anti ulcerogenic were reported. In the present study an attempt is made to evaluate Anti tubercular and Anthelminthic activities for aqueous methanolic (50:50) extract after carrying out the phytochemical investigations. Anti tubercular activity was evaluated by Micro plate Alamar blue assay method against streptomycin, ciprofloxacin and pyrazinamide as standard drugs. Anthelminthic activity was evaluated on Perithima posthuma using Piperazine citrate as standard drug. The results obtained for biological activities were shown to posses considerable Anti tubercular activity at 50 µg/ml and Anthelminthic activity at 300mg/ml.
Anti Tubercular Activity,
Micro Plate Alamar Blue Assay Method, Anthelminthic Activity and Piperazine Citrate
INTRODUCTION: Tuberculosis is the second major global infectious disease. As per WHO survey.8.7 million cases were reported in 2012 and 1.4 million people died. In low income and middle income countries over 95%of TB deaths were occurred due to malnutrition which leads to impaired immunity. It is one of important reason for prognosis of TB infection.
Intestinal parasitic infections are the most common form of public health problem even in the developed countries. Chronic helminthiasis causes severe anaemia which leads to death because of vital organ infections. According to WHO survey the children of age group 7-11 years are more prone to the helminth infections because of poor sanitation. However, some of the foods which contain bitter glycosides, tannins etc; act as natural Anthelminthics.
Since the literature survey revealed that tribal have been chewed the stem parts of Caralluma attenuate 2 as the best remedy to cure several diseases like obesity 3-5, diabetes 6-7, athereosclerosis 8 etc because of their medicinal properties, the researcher aimed to evaluate Anti tubercular and Anthelminthic activities for aqueous methanolic extract of whole stem part of Caralluma attenuata belongs to family Apocynaceae1. The researcher has used advanced methods to evaluate the activities. The methods and results were explained as following.
MATERIALS AND METHODS:
Collection of plant material:
The whole stem parts of Caralluma attenuata was collected in Ananthapuramu district, Andhra Pradesh. The plant was authenticated by Taxonomist Professor. J. Sreeramulu, Sri Krishna Devaraya University, Anantapuramu.
Preparation of extract:
The dried and powdered stem part of Caralluma attenuata was passed through a sieve no.22 and each kilo gram was extracted successively by cold maceration 9 with 2.5 litres of aqueous methanol (50:50). The extract was concentrated to dryness under reduced pressure using rotary vacuum evaporator. The extract was subjected for the phytochemical investigations 10. The identified chemical constituents were reported in Table 1. The extract was utilised to evaluate Anti tubercular and Anthelminthic activities. The results obtained were tabulated in Table 2 and 3 respectively.
Anti tubercular activity:
The aqueous methanolic extract was dissolved in chloroform (RP1), ethanol (RP2) and ethyl acetate (RP3) then these solutions was subjected for anti TB activity.
The Antitubercular activity 11-14 of extract was assessed against M. tuberculosis using Micro plate Alamar Blue Assay (MABA)15.This methodology is non-toxic, uses a thermally stable reagent and shows good correlation with proportional and BACTEC radiometric method. Briefly, 200µl of sterile deionised water was added to all outer perimeter wells of sterile 96 wells plate to minimized evaporation of medium in the test wells during incubation. The 96 wells plate received 100 µl of the Middle brook 7H9 broth and serial dilutions of compounds were made directly on plate. The final drug concentrations tested was 100 to 0.2 µg/ml. Plates was covered and sealed with Para film and incubated at 37ºC for five days. After this time, 25µl of freshly prepared 1:1 mixture of Alamar Blue reagent and 10% tween 80 was added to the plate and incubated for 24 hrs. A blue colour in the well was interpreted as no bacterial growth and pink colour was scored as growth. The MIC was defined as lowest drug concentration which prevented the colour change from blue to pink as shown in the Fig. 1 and Fig. 2.
FIG.1: ANTI TB ACTIVITY OF SAMPLES
FIG. 2: ANTI TB ACTIVITY OF STANDARD DRUGS
The Anthelminthic activity 16-18 was carried out using adult earthworms (pheretima posthuma), belongs to the group of Annelida. 20ml of aqueous methanolic(50:50) extract of caralluma attenuata (test solutions) containing three different concentrations (100mg/ml, 200mg/ml, and 300mg/ml in distilled water) were prepared and taken in three different petridishes and 5 earthworms were placed in each petridish. 20 ml of Piperazine citrate 19 (10mg/ml concentration) was used as reference standard while distilled water was used as the control. Time taken for paralysis of worms was noted for the sample and standard solution; where there was no movement of any sort could be observed except the worms was shaken vigorously. Time of death of worms was recorded after ascertaining that the worms neither moved when shaken vigorously nor when dipped in warm water at 50degrees centigrade. The observations were explained in results Table 3.
RESULTS AND DISUSSION:
TABLE 1: PHYTOCHEMICAL STUDIES
|Compound||Name of chemical test||Result|
|Cardiac glycosides||Killer killani test||Positive|
|Anthraquinone glycosides||Borntragers test||Positive|
|Cyanogenetic glycosides||Guignaurd reagent test||Negative|
|Guaiacum resin and copper sulphate test||Negative|
|Caumarin glycosides||Aromatic odour test||Negative|
|Filter paper test||Negative|
|Tannins and phenolic compounds||5%FeCl3 solution||Positive|
Anti tubercular activity:
The Micro plate alamar blue assay method was used for evaluation of Anti tubercular activity. The Anti tubercular activity was Sobserved at 50 µg/ml solution for all the three solvents.
TABLE 2: ANTI TUBERCULAR ACTIVITY
|12.5 μg/ml||6.25 μg/ml||3.12 μg/ml||1.6
S: Sensitive R: Resistance
The results obtained for Anthelminthic activity was represented as follows.
TABLE 3: ANTHELMINTHIC ACTIVITY USING AQUEOUS METHANOLIC EXTRACT
|Paralysis time||Death time|
(Aqueous methanolic extract)
|Distilled water (control)|
|Piperazine citrate||10mg/ml||25 min||64min|
The aqueous methanolic extract of Caralluama attenuata showed significant Anthelminthic activity at all concentrations, but the maximum activity was observed at 300mg/ml.
DISCUSSION: The aqueous methanolic extract was found to posses the considerable Anti tubercular and Anthelminthic activities. Hence the researcher has evaluated the activities by in vitro methods; the present study becomes one of the positive evidence to explore the research for further investigations.
CONCLUSION: The aqueous methanolic extract of Caralluma attenuata may be considered as preventive natural herb for helminthiasis since it has shown the better anthelminthic activity within 16 minutes than the standard drug. Further the principle active constituents which are responsible for antitubercular activity can be isolated. The mechanism for both of these activities would be explained by carrying out the in-vivo methods in future research.
ACKNOWLEDGEMENT: The researchers are thankful to Dr. Y. Padmanabha Reddy, principal of RIPER and S. Nagarjuna Department of pharmacology, RIPER for their support.
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How to cite this article:
Mounika P, Jyothi MV, Ramalingam P, Vinusha Reddy J and Anusha K: Anti tuberular and anthelminthic activities of aqueous methanolic extract of Caralluama attenuata. Int J Pharm Sci Res 2016; 7(11): 4561-64.doi: 10.13040/IJPSR.0975-8232.7(11).4561-64.
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P. Mounika, M. Vijaya Jyothi *, P. Ramalingam, Vinusha Reddy J. and K. Anusha
Department of Pharmaceutical chemistry 1, Raghavendra Institute of Pharmaceutical Education &Research, Ananthapuramu, Andhra Pradesh, India
02 June, 2016
21 July, 2016
02 August, 2016
01 November, 2016