8

Views:
 
Category: Entertainment
     
 

Presentation Description

No description available.

Comments

Presentation Transcript

slide 1:

Gunji Venkateswarlu et al IJCTPR 2015 34: 997–1003 ISSN: 2321-3760 International Journal of Current Trends in Pharmaceutical Research 997 International Journal of Current Trends in Pharmaceutical Research Journal Home Page: www.pharmaresearchlibrary.com/ijctpr Research Article Open Access Preparation and Evaluation of Antimicrobial and Antioxidant Activity of Polyherbal Ointment G. Swarupa Rani 1 Raghavendra Kumar Gunda 2 Y. Mansa 3 CH. Khushi Vardhan 3 V. Dileep Kumar 3 Gunji Venkateswarlu 4 1 Asst Professor Department of Pharmacology Narasaraopeta Institute of Pharmaceutical Sciences Narasaraopet Guntur Dt Andhra Pradesh. 2 Asst Professor Department of Pharmaceutics Narasaraopeta Institute of Pharmaceutical Sciences Narasaraopet Guntur Dt Andhra Pradesh. 3 Narasaraopeta Institute of Pharmaceutical Sciences Narasaraopet Guntur Dt Andhra Pradesh. 4 Asst Professor Department of Pharmacognosy Narasaraopeta Institute of Pharmaceutical Sciences Narasaraopet Guntur Dt Andhra Pradesh. A B S T R A C T The main objective of the present study is to formulate and evaluate a poly herbal ointment with antiseptic activity. Ointments were formulated using methanolic extracts of Eclipta alba Ocimum sanctum Azadiracta indica and Achyranthes aspera which were evaluated for its physicochemical property antibacterial and antioxidant activity. Ointments were prepared using different concentrations of the extracts such as 2 4 6 w/w by fusion method using emulsifying ointment as base. Formulations were then tested for its physicochemical properties which gave satisfactory results. The prepared formulations were also stable at 4ºC 25ºC and 37ºC. Further Polyherbal formulations were evaluated for its anti- bacterial activity against Betadine 5w/w as the standard. All the formulations showed Predominant activity against selected species. Formulations were also evaluated for anti-oxidant activity through reducing power assay nitric oxide and hydrogen peroxide scavenging method. The results showed that the scavenging activity of the formulations increased with increase in concentration and this is due to the presence of flavanoids and tannins. The presence of both antibacterial and antioxidant activity reveals that the prepared ointment can also be used for wound healing. Hence an attempt was made to formulate a Polyherbal ointment and to evaluate for its physical parameter in-vitro anti-oxidant activity and to compare its antibacterial activity with a marketed formulation 5 w/w Betadine.Overall result of this study reveals that this is an effective Polyherbal antiseptic ointment. Keywords: Eclipta alba Ocimum sanctum Azadiracta indica Achyranthes aspera Formulations Spread ability Extrudability A R T I C L E I N F O CONTENTS 1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 998 2. Materials and Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .998 3. Results and discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1000 4. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . .1002 5. Acknowledgement. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . 1002 6. References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1003 Article History: Received 28 May 2015 Accepted 30 June 2015 Available Online 15 July 2015 Citation: Gunji Venkateswarlu et al. Effect of Daily consumption of green tea and hibiscus on Some Biochemical Parameters in a Sample of Healthy Volunteers Preliminary Study. Int. J. Curnt. Tren. Pharm Res. 2015 34: 997-1003.

slide 2:

Gunji Venkateswarlu et al IJCTPR 2015 34: 997–1003 ISSN: 2321-3760 International Journal of Current Trends in Pharmaceutical Research 998 PAPER-QR CODE Copyright© 2015 Gunji Venkateswarlu et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License which permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited. 1. Introduction Preparation of medium and nutrient broth: Weighed about 0.4g of nutrient broth and dissolved in 30ml of water. Then the broth was suspended in each of test tube. The Muller Hintonagar medium was prepared which contain 9.7g of MHA was suspended in 250ml of water. Then both the medium and broth were for sterilization. After sterilization the nutrient broth was allowed to cool and then the organism were inoculated and incubated for 4hours.The MHA medium were poured in the Petri dish before cooling and allowed to solidify for about 3-4 hours. Ayurvedic medicine is a time-tested system of medicine which has been in clinical use for centuries in India. Being a time-tested system it has an edge over other existing systems of health management 1. When two or more herbs are used in formulations they are known as Polyherbal formulations. Ayurveda and herbal medicine has roots in medicinal herbs and they have been practiced for centuries. Herbal medicine is making dramatic comeback and increasing number of patients are visiting alternative medicine clinics. Side effects of synthetic medicine are alarming and recent time has seen risk of herbal and herbal- synthetic drug interactions 2. In India from ancient times different parts of medicinal plants have been used to cure specific ailments. Today there is widespread interest in drugs derived from plants. This interest primarily stems from the belief that green medicine is safe and dependable compared with costly synthetic drugs that have adverse effects. Natural antimicrobials can be derived from plants animal tissues or microorganisms. The shortcomings of the drugs available today propel the discovery of new Pharmacotherapeutic agents in medicinal plants. To determine the potential and promote the use of herbal medicine it is essential to intensify the study of medicinal plants that find place in folklore 3. The herbal drugs are boon to our society. These herbal drugs are considered as a therapeutic weapon to fight against various diseases in birds humans and animals without having any side effects Under the prevailing circumstances further investigations into the concept of Polyherbal formulations should be undertaken .so in the present work we formulated a Polyherbal ointment with better antimicrobial as well as anti-oxidant activity can be used for skin infections .In recent years there has been a great demand for plant derived products in developed countries. The literatures have reported that the usage of the traditional medicines brought a great benefit in skin related diseases. Hence the plant entities derived from the natural source need to be identified and formulated in to suitable dosage form for the management and treatment of various antimicrobial diseases. Azadirachta indica known as Neem is well known for its medicinal properties 4. Its leaves possess broad spectrum of activity against Gram+ve and Gram–ve bacteria including M.tuberculosis Vibrio cholera4.Ocimum snactum is known as scared holybsil possess anti-inflammatory used in treating leprosy leucoderma etc 5. Eclipta alba known as bringeraj antifungal as well as anti microbial activities antioxident6. Achyranthes aspera as known as uttareni and the methanolic extract of the plant have properties like anti bacterial antifungal activity anti oxidant activity and anti-inflammatory 6. The astringent properties of the leaf extracts of Achyranthes aspera on the blood vessel has made it a popular plant in the prevention of blood loss from wounds also its anti-microbial properties has made it a popular choice in disinfecting and treating open wounds 7. Hence an effort has been made to establish the scientific validity to investigate the possible antimicrobial and antioxidant activity of the formulated ointments made from the methanolic extracts of the above four herbs. From this investigaton and the results this polyherbal ointment posses significant antimicrobial as well as anti-oxidant activity can be used for the treatement of burns wounds rashes etc. 2. Materials and Methods Collection of plants: The Leaves of Eclipta alba ocimum sanctum azadiracta indica achyranthes aspera species was collected in the month of November at kotappakonda which is near to Narasaraopet Guntur Dt A.P and the collected plants were authentified by Prof. Jayaraman plant anatomist at plant anatomy and research center thambram Chennai. Chemicals and reagents: Emulsifying wax I. P Lobe Chem. Liquid paraffin I.P Nice Chemicals White soft paraffin I.P Lobe Corresponding Author Gunji Venkateswarlu Asst Professor Department of Pharmacognosy Narasaraopeta Institute of Pharmaceutical Sciences Narasaraopet Guntur A. P India Manuscript ID: IJCTPR2583 Gunji Venkateswarlu et al IJCTPR 2015 34: 997–1003 ISSN: 2321-3760 International Journal of Current Trends in Pharmaceutical Research 998 PAPER-QR CODE Copyright© 2015 Gunji Venkateswarlu et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License which permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited. 1. Introduction Preparation of medium and nutrient broth: Weighed about 0.4g of nutrient broth and dissolved in 30ml of water. Then the broth was suspended in each of test tube. The Muller Hintonagar medium was prepared which contain 9.7g of MHA was suspended in 250ml of water. Then both the medium and broth were for sterilization. After sterilization the nutrient broth was allowed to cool and then the organism were inoculated and incubated for 4hours.The MHA medium were poured in the Petri dish before cooling and allowed to solidify for about 3-4 hours. Ayurvedic medicine is a time-tested system of medicine which has been in clinical use for centuries in India. Being a time-tested system it has an edge over other existing systems of health management 1. When two or more herbs are used in formulations they are known as Polyherbal formulations. Ayurveda and herbal medicine has roots in medicinal herbs and they have been practiced for centuries. Herbal medicine is making dramatic comeback and increasing number of patients are visiting alternative medicine clinics. Side effects of synthetic medicine are alarming and recent time has seen risk of herbal and herbal- synthetic drug interactions 2. In India from ancient times different parts of medicinal plants have been used to cure specific ailments. Today there is widespread interest in drugs derived from plants. This interest primarily stems from the belief that green medicine is safe and dependable compared with costly synthetic drugs that have adverse effects. Natural antimicrobials can be derived from plants animal tissues or microorganisms. The shortcomings of the drugs available today propel the discovery of new Pharmacotherapeutic agents in medicinal plants. To determine the potential and promote the use of herbal medicine it is essential to intensify the study of medicinal plants that find place in folklore 3. The herbal drugs are boon to our society. These herbal drugs are considered as a therapeutic weapon to fight against various diseases in birds humans and animals without having any side effects Under the prevailing circumstances further investigations into the concept of Polyherbal formulations should be undertaken .so in the present work we formulated a Polyherbal ointment with better antimicrobial as well as anti-oxidant activity can be used for skin infections .In recent years there has been a great demand for plant derived products in developed countries. The literatures have reported that the usage of the traditional medicines brought a great benefit in skin related diseases. Hence the plant entities derived from the natural source need to be identified and formulated in to suitable dosage form for the management and treatment of various antimicrobial diseases. Azadirachta indica known as Neem is well known for its medicinal properties 4. Its leaves possess broad spectrum of activity against Gram+ve and Gram–ve bacteria including M.tuberculosis Vibrio cholera4.Ocimum snactum is known as scared holybsil possess anti-inflammatory used in treating leprosy leucoderma etc 5. Eclipta alba known as bringeraj antifungal as well as anti microbial activities antioxident6. Achyranthes aspera as known as uttareni and the methanolic extract of the plant have properties like anti bacterial antifungal activity anti oxidant activity and anti-inflammatory 6. The astringent properties of the leaf extracts of Achyranthes aspera on the blood vessel has made it a popular plant in the prevention of blood loss from wounds also its anti-microbial properties has made it a popular choice in disinfecting and treating open wounds 7. Hence an effort has been made to establish the scientific validity to investigate the possible antimicrobial and antioxidant activity of the formulated ointments made from the methanolic extracts of the above four herbs. From this investigaton and the results this polyherbal ointment posses significant antimicrobial as well as anti-oxidant activity can be used for the treatement of burns wounds rashes etc. 2. Materials and Methods Collection of plants: The Leaves of Eclipta alba ocimum sanctum azadiracta indica achyranthes aspera species was collected in the month of November at kotappakonda which is near to Narasaraopet Guntur Dt A.P and the collected plants were authentified by Prof. Jayaraman plant anatomist at plant anatomy and research center thambram Chennai. Chemicals and reagents: Emulsifying wax I. P Lobe Chem. Liquid paraffin I.P Nice Chemicals White soft paraffin I.P Lobe Corresponding Author Gunji Venkateswarlu Asst Professor Department of Pharmacognosy Narasaraopeta Institute of Pharmaceutical Sciences Narasaraopet Guntur A. P India Manuscript ID: IJCTPR2583 Gunji Venkateswarlu et al IJCTPR 2015 34: 997–1003 ISSN: 2321-3760 International Journal of Current Trends in Pharmaceutical Research 998 PAPER-QR CODE Copyright© 2015 Gunji Venkateswarlu et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License which permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited. 1. Introduction Preparation of medium and nutrient broth: Weighed about 0.4g of nutrient broth and dissolved in 30ml of water. Then the broth was suspended in each of test tube. The Muller Hintonagar medium was prepared which contain 9.7g of MHA was suspended in 250ml of water. Then both the medium and broth were for sterilization. After sterilization the nutrient broth was allowed to cool and then the organism were inoculated and incubated for 4hours.The MHA medium were poured in the Petri dish before cooling and allowed to solidify for about 3-4 hours. Ayurvedic medicine is a time-tested system of medicine which has been in clinical use for centuries in India. Being a time-tested system it has an edge over other existing systems of health management 1. When two or more herbs are used in formulations they are known as Polyherbal formulations. Ayurveda and herbal medicine has roots in medicinal herbs and they have been practiced for centuries. Herbal medicine is making dramatic comeback and increasing number of patients are visiting alternative medicine clinics. Side effects of synthetic medicine are alarming and recent time has seen risk of herbal and herbal- synthetic drug interactions 2. In India from ancient times different parts of medicinal plants have been used to cure specific ailments. Today there is widespread interest in drugs derived from plants. This interest primarily stems from the belief that green medicine is safe and dependable compared with costly synthetic drugs that have adverse effects. Natural antimicrobials can be derived from plants animal tissues or microorganisms. The shortcomings of the drugs available today propel the discovery of new Pharmacotherapeutic agents in medicinal plants. To determine the potential and promote the use of herbal medicine it is essential to intensify the study of medicinal plants that find place in folklore 3. The herbal drugs are boon to our society. These herbal drugs are considered as a therapeutic weapon to fight against various diseases in birds humans and animals without having any side effects Under the prevailing circumstances further investigations into the concept of Polyherbal formulations should be undertaken .so in the present work we formulated a Polyherbal ointment with better antimicrobial as well as anti-oxidant activity can be used for skin infections .In recent years there has been a great demand for plant derived products in developed countries. The literatures have reported that the usage of the traditional medicines brought a great benefit in skin related diseases. Hence the plant entities derived from the natural source need to be identified and formulated in to suitable dosage form for the management and treatment of various antimicrobial diseases. Azadirachta indica known as Neem is well known for its medicinal properties 4. Its leaves possess broad spectrum of activity against Gram+ve and Gram–ve bacteria including M.tuberculosis Vibrio cholera4.Ocimum snactum is known as scared holybsil possess anti-inflammatory used in treating leprosy leucoderma etc 5. Eclipta alba known as bringeraj antifungal as well as anti microbial activities antioxident6. Achyranthes aspera as known as uttareni and the methanolic extract of the plant have properties like anti bacterial antifungal activity anti oxidant activity and anti-inflammatory 6. The astringent properties of the leaf extracts of Achyranthes aspera on the blood vessel has made it a popular plant in the prevention of blood loss from wounds also its anti-microbial properties has made it a popular choice in disinfecting and treating open wounds 7. Hence an effort has been made to establish the scientific validity to investigate the possible antimicrobial and antioxidant activity of the formulated ointments made from the methanolic extracts of the above four herbs. From this investigaton and the results this polyherbal ointment posses significant antimicrobial as well as anti-oxidant activity can be used for the treatement of burns wounds rashes etc. 2. Materials and Methods Collection of plants: The Leaves of Eclipta alba ocimum sanctum azadiracta indica achyranthes aspera species was collected in the month of November at kotappakonda which is near to Narasaraopet Guntur Dt A.P and the collected plants were authentified by Prof. Jayaraman plant anatomist at plant anatomy and research center thambram Chennai. Chemicals and reagents: Emulsifying wax I. P Lobe Chem. Liquid paraffin I.P Nice Chemicals White soft paraffin I.P Lobe Corresponding Author Gunji Venkateswarlu Asst Professor Department of Pharmacognosy Narasaraopeta Institute of Pharmaceutical Sciences Narasaraopet Guntur A. P India Manuscript ID: IJCTPR2583

slide 3:

Gunji Venkateswarlu et al IJCTPR 2015 34: 997–1003 ISSN: 2321-3760 International Journal of Current Trends in Pharmaceutical Research 999 Chem. Methanol n-hexane Sulphuric Acid Drangendroff’s reagent Molisch’s reagent Acetone. Equipments: Soxhlet apparatus Incubator Digital balance Bunsen burner pH meter Glass wares UV Spectrophotometer. Mediam: Nutrient Agar and Muller Hinton Agar Media Organisms: Mucus fungus Lactobacillus and Escherichia coli. Extraction: The collected plants Eclipta alba Ocimum sanctum Azadiracta indica Achyranthes aspera species were extracted by continuous hot percolation Soxhletation. 50g of powdered leaves of the above four plants were defatted using petroleum ether. The marc obtained from each of the powdered plant parts were successfully extracted separately with 250 ml of methanol by using Soxhlet apparatus. The extraction was carried out for 24 hours. After extraction the solvents were distilled out the concentrated residues were analyzed by chemical tests 8. Phytochemical analysis: The methanolic extract obtained after soxhletation was subjected to various phytochemical screening as per the standard procedure to reveals the presence of various active phytoconstituents.9 Formulation of ointment: Working formula emulsifying ointment base Emulsifying wax -300g White soft paraffin-500g Liquid paraffin-200g Procedure: Required quantities of emulsifying wax liquid paraffin and white soft paraffin were weighed and melted. To this adequate quantities of methanolic extract of the mentioned four plants were added and stirred well until a homogeneous mass were obtained 10. The compositions of different Polyherbal ointment are listed in Table II. Evaluation: Physicochemical parameters 11 12 Preliminary evaluation of formulations at different concentrations was carried out as follows Colour and odour - examined by visual examination. Loss on drying - determined by placing ointment in Petridish on water bath and dried for 105 0 C. The pH of various formulations was determined by using Digital pH meter. One gram of ointment was dissolved in 100 ml of distilled water and stored for two hours. The measurement of pH of each formulation was done in triplicate and average values were depicted in Table-III. Spreadability: Spreadability of the formulation was determined by an apparatus suggested by Muttimer et al. which was suitably modified in the laboratory and used for the study. It consists of a wooden block which was provided by a pulley at one end. A rectangular ground glass plate was fixed on this block. An excess of ointment about 3 g. under study was placed on this ground plate. The ointment was then sandwiched between this plate and another glass plate having the dimension of fixed ground plate and provided with the hook. A 1 Kg. weight was placed on the top of the two plates for 5 minutes to expel air and to provide a uniform film of the ointment between the plates. Excess of the ointment was scrapped off from the edges. The top plate was then subjected to pull of 80 g. with the help of string attached to the hook and the time in seconds required by the top plate to cover a distance of 10 cm. be noted. A shorter interval indicates better Spreadability. Spreadability is measured as S M × L /T M weight tide to upper slide L length of glass slides T Time Extrudability: A simple method was adopted for this study. The formulations were filled in the collapsible tubes after the ointments were set in the container. The extrudability of the different ointment formulations was determined in terms of weight in grams required to extrude a 0.5 cm of ribbon of ointment in 10 second. Diffusion study: The diffusion study was carried out by preparing agar nutrient medium of any Concentration. It was poured into petridish. A hole bored at the centre and ointment was placed in it. The time taken for the ointment to get diffused was noted. Stability studies: The stability studies were carried out for the prepared formulations at different temperature conditions 4o C 25o C and 37o C for 3 months. Evaluation of antimicrobial activity 12 Procedure: Microorganisms: Mucus fungus Lactobacillus Escherichia coli aerobic organism. Standard used: 5 w/w Betadine ointment. Sample preparation: About 10mg of ointments 2 4 6 w/w were weighed and dissolved in DMF dimethyl formamide and used for activity studies. Preparation of medium and nutrient broth: Weighed about 0.4g of nutrient broth and dissolved in 30ml of water. Then the broth was suspended in each of test tube. The Muller Hintonagar medium was prepared which contain 9.7g of MHA was suspended in 250ml of water. Then both the medium and broth were for sterilization. After sterilization the nutrient broth was allowed to cool and then the organism were inoculated and incubated for 4hours.The MHA medium were poured in the Petri dish before cooling and allowed to solidify for about 3-4 hours. Methodology: The bacterial culture was spread on the culture medium and a well was bored in the middle of the agar. Then different samples and standard solutions of 0.05ml was poured inside these wells and plates were incubated at 37oC overnight for observation. The presence of inhibition was noted and compared with the control. The susceptibility of the test organism to the tested plant extract was determined by observing the zone of inhibition around each well In-vitro antioxidant study: 13 14 Reducing power assay method Reagents: 0.1Ferric chloride 1Ferricyanide Phosphate buffer6.6 Trichloro acetic acid. Preparation of standard ascorbic acid solution:

slide 4:

Gunji Venkateswarlu et al IJCTPR 2015 34: 997–1003 ISSN: 2321-3760 International Journal of Current Trends in Pharmaceutical Research 1000 Weighed accurately 50mg of ascorbic acid and made up to 50ml with methanol. Preparation of sample solution: 50mg of ointments 2 4 6 w/w was dissolved in 50ml of methanol. From the above different concentrations 0.5 1 2 3 4 5ml were pipetted out and made up to 10ml with methanol. Added 2.5ml of phosphate buffer 6.6 and 2.5ml of potassium ferricyanide to each of the test tubes and was incubated at 40oC for 20min. After incubation 2.5ml of Tri chloro acetic acid was added and centrifuged the reaction mixture for 5min.To 2.5ml of this reaction mixture 0.5ml ferric chloride and 2.5ml water were added. The absorbance was measured using Double beam spectrophotometer at 700nm. Preparation of control: 10ml of methanol was taken. To it 2.5ml of potassium ferricyanide and 2.5ml of buffer 6.6 were added. The above reaction mixture was incubated at 40oC for 20min and centrifuged for 5min.To this add 0.5ml ferric chloride and 2.5ml of water. Nitric oxide scavenging method Reagents: Sodium nitroprusside phosphate buffer solution Griess reagent were procured. Fine chemicals and all the solvents used were of A R grade. Preparation of test sample: 50 mg of ointments was dissolved in 50ml of distilled methanol to obtain a solution of 1mg/ml. From this stock solution different working dilution were prepared to get concentration of 100 200 300 400 500 µg/ml. Methodology: Nitric Oxide was generated from sodium nitroprusside was measured by the Griess reagent. Sodium nitroprusside5mM in phosphate buffer of pH 7.4 saline was mixed with different concentrations of the ointments 100 200300400500 µg/ml dissolved in water and incubated at 25 ºC for 150 minutes. At different time interval sample 1.5 ml of the incubated solution were removed and diluted with 1.5 ml Griess reagent 1sulphanilamide 2 H3PO4 and 0.1naphthylethylenediamene dihydrochloride. Evaluation: The absorbance was read at 564nm. Ascorbic acid was used as the standard. The difference in the absorbance between test and control of Nitric Oxide were calculated and expressed as percent scavenging of Nitric Oxide radical. Capability to scavenge the nitric oxide radical was calculated by using equation. Hydrogen peroxide scavenging activity: Solution of hydrogen peroxide 40mM was prepared in phosphate buffer of the pH 7.4.The concentration of hydrogen peroxide was determined by absorption at 230nm using spectrophotometer. The ointments 1mg/ml in methanol were added to hydrogen peroxide solution 0.6ml 40mM.The absorbance at 230nm was determined after 10 minutes against a blank solution containing phosphate buffer without hydrogen peroxide. The of the hydrogen peroxide scavenging by the samples and the standard compounds were calculated. 3. Results and Discussion Literatures revealed that the selected four herbs Azadirachta indica Mimosa pudica Chromolaena odorata Samadera indica have antioxidant and antibacterial activity. Hence an attempt was made to formulate a polyherbal ointment and to evaluate for its physical parameter in vitro antioxidant activity and to compare its antibacterial activity with a marketed formulation 5 w/w Betadine.Extraction and the phytochemical screening was done using methanol as the solvent. Phytochemical screening confirmed the presence of various phytoconstituens like carbohydrate glycosides flavanoids and tannins. In the present study Polyherbal ointments were prepared by fusion method using emulsifying ointment as the base. The formulations were then evaluated for their physical parameters In-vitro antioxidant and compared with marketed 5w/w Betadine ointment for its antibacterial activity. These physical parameters were within the acceptable range. The stability studies were carried out and inferred that the formulations showed no signs of instability. The antibacterial activity of prepared ointments were compared with 5w/w Betadine ointment using selected species of microorganism such as Mucus fungus Lactobacillus Escherichia coli aerobic organism and it showed that formulations like F2 and F3 showed greater activity Staphylococcus aureus and Bacillus sp compared to 5 Betadine. So antimicrobial study shows that the prepared ointments has better activity Mucus fungus Lactobacillus Escherichia coli aerobic organism compared to standard 5 Betadine ointment. Anti-oxidant activity inferred that the formulated ointments showed similar activity as that of standard ascorbic acid and hence revealed that this activity is due to the presence of flavanoids and tannins. Hence the study concludes that an efficient antiseptic ointment with antimicrobial and antioxidant activities can be formulated from the methanolic plant extracts of Eclipta alba ocimum sanctum azadiracta indica achyranthes aspera which can also be used forwound healing and various skin infections. Table 1: Composition of Polyherbal Ointments Ingredients F12 F2 4 F3 6 Azadirachta indicam ethanolic extract 2g 4g 6g Ocimum sanctummethanolic extract 2g 4g 6g Eclipta alba methanolicextract 2g 4g 6g Achyranthes aspera methanolic extract 2g 4g 6g Emulsifying ointment q.s to 100g q.s to 100g q.s to 100g

slide 5:

Gunji Venkateswarlu et al IJCTPR 2015 34: 997–1003 ISSN: 2321-3760 International Journal of Current Trends in Pharmaceutical Research 1001 Table 2: Phytochemical Screening of the Methanolic Extract of Poly Herbal Extract Chemical Constituents Polyherbal extract Carbohydrates + Proteins - Alkaloids +++ Saponins - Tannins ++ Flavonoids ++ Steroids + Triterpenoids + Glycoside +++ Table 3: Physicochemical evaluation of formulated formulations Physicochemical parameters F1 2 F2 4 F3 6 Colour Dark green Dark green Characteristic Odour Characteristic Characteristic 41w/w Loss of drying 37w/w 40w/w 6.6 PH 7.02 6.88 13 Spread ability Seconds 12 12 182g Extrudability 180g 180g 0.7cm Diffusion study after 60 min 0.7cm 0.8 cm Stable Storage 4ºC 24ºC 37ºC Stable Stable Characteristic +present - absent Table 4: Antimicrobial activity of formulated ointment Ointment E. coli Mucus fungus Lactobacillus Herbal Ointment 1.25± 0.2 1.50±0.12 1.11±0.32 Polyherbal Ointment 1.85±0.42 1.93±0.31 1.71±0.13 StandardGentamycin 2.21±0.23 2.00±0.11 2.11±0.23 Control 0.166±0.11 0.41±0.23 0.08±22 Diameter of Zone of inhibition in cm Figure 1: Antimicrobial activity of formulated ointment Table 5: Antioxidant activity of formulated ointments by reducing power assay Concentration µg/ml Absorbance Standard F12 F24 F36 50 0.346 0.213 0.156 0.222 100 0.369 0.26 0.21 0.253 200 0.412 0.276 0.313 0.392 300 0.578 0.346 0.51 0.472 400 0.687 0.41 0.682 0.593 500 0.786 0.511 0.703 0.639 Control0.992

slide 6:

Gunji Venkateswarlu et al IJCTPR 2015 34: 997–1003 ISSN: 2321-3760 International Journal of Current Trends in Pharmaceutical Research 1002 Table 6: Antioxidant activity of formulated ointments by nitric oxide scavenging activity Table 7: Antioxidant activity of formulated ointments by hydrogen peroxide scavenging activity Figure 2: Plant Images Which Are Used For Poly Herbal Formulation 4. Conclusion A combinational therapy is the need of hour to treat eczema and pruritis. This can be achieved by Clotrimazole and Ichthammol an antifungal and antiseptic.In this study ointment was formulated with different bases like white soft paraffin cetostearyl alcohol hard paraffin and light liquid paraffin. By combining these drugs with appropriate ointment bases as polyherbal formulation a better therapy and patient compliance can be achieved. Both the formulations may shows the batter way to use these drugs with more significant way and more over WHO also emphasizes the herbal medicine for treatment. This study showed the convenient preparation and more effective use of both the herbs with modernized formulations with comparatively old form of medicine. This affords may leads a beginning of proper utilization and conservation of medicinally important herbs and cost effective treatment in future. 5. Acknowledgement The authors are thankful to Principal Dr. J N SURESH KUMAR principal Narasaraopeta College of pharmaceutical sciences providing the necessary facilities in the Colleges Sincerely thanks to Mr G. Ragavendra Kumar Assistant professor Department of Pharmacuetics Concentration µg/ml Absorbance Inhibition F1 F2 F3 STD F1 F2 F3 STD 100 0.637 0.641 0.637 0.613 60.67 60.43 60.67 62.16 200 0.624 0.628 0.626 0.603 61.48 61.23 61.35 62.39 300 0.619 0.624 0.613 0.586 61.79 61.48 62.16 63.83 400 0.598 0.615 0.605 0.562 63.08 62.03 62.65 65.31 500 0.548 0.589 0.569 0.525 66.17 63.64 64.87 67.59 Concentration µg/ml Absorbance Inhibition F1 F2 F3 STD F1 F2 F3 STD 100 1.179 0.61 0.515 0.495 45 71.5 76 76.95 200 0.77 0.456 0.451 0.394 64.13 78.76 78.99 81.64 300 0.551 0.427 0.441 0.295 74.33 80.11 79.45 86.25 400 0.455 0.369 0.399 0.195 78.8 82.81 81.41 90.91 500 0.372 0.366 0.376 0.134 82.67 82.95 82.48 93.75

slide 7:

Gunji Venkateswarlu et al IJCTPR 2015 34: 997–1003 ISSN: 2321-3760 International Journal of Current Trends in Pharmaceutical Research 1003 Narasaraopeta College of pharmaceutical sciences his valuable support and I very much thank full to my wife for her support and cooperation. 6. References 1. BHUx: A Patent Polyherbal Formulation to Prevent Atherosclerosis. 2. www.ayurlifestyle.com. 3. Nihal Singh Verma Sumeet Dwivedi Debadash Panigrahi and S.K. Gupta. Anti-bacterial activity of root bark of Nyctanthes arbor-tristis Linn”. International Journal of Drug Discovery and Herbal Research 2011 12April-June: 61-62. 4. Abhijeet Pandey Jui V. Jagtap Aditi A. Patil Richa N. Joshi and B. S. Kuchekar. Formulation and evaluation of anti-bacterial and anti-fungal activity of a herbal ointment containing Aloe-vera Azadirachta indica and Curcuma longa. Journal of Chemical and Pharmaceutical Research 2010 23: 182-186. 5. Rathnayaka RM. “Antibacterial Activity of Ocimum Sanctum Extracts against Fou Food- Borne Microbial Pathogens Sch. j. app. med. sci. 2013 16: 774-777. 6. Rajamurugan R. Deepa V. Sivashanmugam M. Raghavan C. M. Phytochemistry antioxidant and antibacterial activities of medicinal plants- a comparative study. Ijcrr. 2013 52: 08-19. 7. Abi Beaulah G Mohamed Sadiq A and Jaya Santhi R. “Antioxidant and antibacterial activity of Achyranthes aspera: An in vitro study.” Annals of Biological Research. 2011 2 5: 662-67. 8. Kokate C.K. 1997. Practical Pharmacognosy Vallabh Prakashan Delhi 4th Edition 107 - 111. 9. Abu Arra Basma Zuraini Zakaria Lacimanan Yoga Latha Sreenivasan Sasidharan “Antioxidant activity and phytochemical screening of the methanol extracts of Euphorbia hirta L” Asian Pacific Journal of Tropical Medicine. May 2011 45: 386–390. 10. Prabhudutta Panda. “Formulation and evaluation of topical dosage form of Pandanusfascicularis Lamk and their wound healingactivity” Drug Invention Today 2010 29: 417-420. 11. Tacey X Viegas Lise L Van Winkle a Paul A Lehman Sue F Franz Thomas J Franz “Evaluation of creams and ointments as suitable formulations for peldesine” International Journal of Pharmaceutics. May 2001 2191: 73–80. 12. Himal Paudel Chhetri Nisha Shrestha Yogol Jyoti Sherchan Anupa K.C S. Mansoor Panna Thapa “Formulation and evaluation of antimicrobial herbal ointment” Kathmandu University Journal of Science Engineering and Technology 2010 6:102-107. 13. Vidya Viswanad N. A. Aleykutty B. Jayakar Subin Mary Zacharia and Litha Thomas “Development and evaluation of antimicrobial herbal formulations containing the methanolic extract of Samadera indica for skin diseases” J Adv Pharm Technol Res. 2012 Apr-Jun 32: 106– 111. 14. Bhaskar G Arshia S Priyadarshini S. “Formulation and evaluation of topical polyherbal antiacne gels containing Garcinia mangostana and Aloe vera.” Phcog Mag. 2009 5 Suppl S2: 93-9. 15. Arun K. Mishra Amrita Mishra Pronobesh Chattopadhyay. Formulation and In-vitro evaluation of antioxidant activity of o/w sunscreen cream containing herbal oil as dispersed phase” International Journal of Biomedical Research 2010 15: 99-105.

authorStream Live Help