Formulation and valuation of E thanolic extract of e ... · International Research Journal of Pharmaceutical and Biosciences (IRJPBS) 4 (4) 13 constant mixing at 800 rpm. Mixing was

Ayesha Siddiqua and Sirisha Mittapally, 2017/ Formulation and evaluation of ethanolic extract

International Research Journal of Pharmaceutical and Biosciences (IRJPBS) 4 (4) 9

RESEARCH ARTICLE

International Research Journal of Pharmaceutical and Biosciences

Pri -ISSN: 2394 - 5826 http://www.irjpbs.com e-ISSN: 2394 - 5834

Formulation and Evaluation of ethanolic extract of Cissus quadrangularis herbal gel Ayesha Siddiqua* and Sirisha Mittapally

Deccan School of Pharmacy, Dar-Us-Salam, Aghapura, Hyderabad, Telangana-500001.

Article info Abstract

Article history: Received 29 July 2017 Accepted 04 AUG 2017

*Corresponding author: [email protected]

Copyright 2017 irjpbs

In this study topical as well as ethosomal gels of Ethanolic extract of Cissus quadrangularis was formulated and is made for better patient compliance. The topical gels F1-F8 were formulated using different concentrations of polymer carbopol (1-2.5%). The ethosomal gels E1-E8 were formulated with ethanol (10%) and phospholipid (2-3%). The prepared topical and ethosomal gels were subjected to physical evaluations, in vitro diffusion studies, PH determination, viscosity, spreadability, entrapment efficiency, zeta potential and SEM. The diffusion studies have shown that the percentage drug release of optimized formulations F4,F8 and E2,E6 after 6 hours were found to be 78.21%, 83.78 % of 20 and 40% topical gels of Cissus quadrangularis and 97.68, 99.76 of 20 and 40% ethosomal gels of Cissus quadrangularis respectively. All the formulations were found to be easily washable, spreadable and free from grittiness. The in vitro drug release studies, evaluation, stability and accelerated stability studies of the topical and ethosomal gels shown that the entrapment efficiency of ethosomes containing 10% w/w ethanol has highest value 96.45% with respect to all other formulations. Hence, the formulation E6 containing 40% w/w of Ethanolic extract of Cissus quadrangularis has shown the promising results with 99.76% drug release after 6 hours with PH 6.8, viscosity 40,300 cps, Zeta potential -60.9 (mV), with specific vesicular size and shape was observed by SEM. Key words:-Cissus quadrangularis gel, Herbal gel, Ethosomes, Phosholipids, Entrapment efficiency.

INTRODUCTION A transparent gel has been in use both in cosmetics and in pharmaceutical preparations to provide a faster release of drug substance. Gels for dermatological use have several advantageous properties such as thixotropic, greaseless, easily spreadable, easily removed, emollient, non-staining and compatible with many excipients and water soluble or miscible.



Transdermal route offers several potential advantages over conventional routes like avoidance of first pass metabolism, predictable and extended duration of activity, minimizing undesirable side effects, utility of short half-life drugs, improving physiological and pharmacological response, avoiding the fluctuation in drug levels.

Ethosomes has permeation of drug molecules (relatively small vesicle size) to and through the skin to the systemic circulation. In ethosomes, ethanol interacts with lipid molecules in the polar head group region resulting in a reducing the rigidity of the stratum corneum lipids, increasing their fluidity. In addition to the effect of ethanol on stratum corneum structure, the ethosome itself may interact with the stratum corneum barrier. Better stability, solubility and patient compliance. Though, Ethosomes are advantageous over other vesicular system these have poor yield.

Cissus quadrangularis is a succulant herbal plant belonging to family, Vitaceae. It is fleshy, cactus in nature. It is also known as Vitis quadrangularis, Lycopodium imbricatum or Heliotropium indicum. In Ayurveda, it is used as Pachana (digestive aid), Sara (relieves constipation), Athiyuk (strengthening bones), Vrushya (Aphrodisiac), etc. In Unani, it is used to treat gastritis. The whole plant is used in treatment of asthma, Powdered root is specifically used in treatment of bone fractures.

The usual dose of the powder is 30-40 grains. Leaves and young shoots are frequently taken with curry in Southern India. In Chennai, young shoots of the plant are dried, powdered, burnt to ashes in a closed vessel. These ashes are administered in dyspepsia, indigestion and certain bowel complaints. Leaves and young shoots are also considered as powerful alternatives in the gastro intestinal treatments. Juice of stem is dropped into the ear in otorrhoea and into the nose in epistaxis. The plant has many therapeutic uses [13].

Phytochemical studies on ethanolic extract revealed the presence of triterpenes including β-sitosterol, α- and β- amyrins, ketosteroids, phenols, tannins, carotene and vitamin C. Seven alicyclic lipids constituents have also been reported from Cissus quadrangularis. unsymmetric tetracyclic triterpenoids such as d-amyrin, onocer-7-ene-3a, 21b-diol, d-amyrone and 3,3',4,4'-tetra hydroxy biphenyl, 3,3',4,4'-tetrahydroxybiphenyl have been isolated from plant and were quantitatively determined by HPTLC method.Several other constituents such as flavonoids quercetin and kaempferol, and stilbene derivatives, quadrangularins A,B,C and many others e.g. resveratrol, piceatanon, pallidol, perthenocissi and phyto-sterols have been isolated from plant.Stem extract contains a high percentage of calcium ions and phosphorus, both essential for bone growth [1] .



Quadrangularin-A MATERIALS AND METHODS

Table no.1: Table showing list of materials with manufacturer S.NO

NAME OF THE MATERIAL

CATEGORY

MANUFACTURER

1. ETHANOLIC EXTRACT OF CISSUS QUANDRANGULARIS

HERBAL PLANT EXTRACT

HIMALAYA DRUG COMPANY, BANGAL ORE, KARNATAKA.

2. LECITHIN CHOLESTEROL

PHOSPHOLIPID S.D FINE CHEMICALS, MUMBAI, INDIA.

3. POLYETHYLENE GLYCOL AS A SKIN PENETRATION ENHANCER

FISCHER SCIENTIFIC, MUMBAI, INDIA.

4. CARBOPOL 934

GELLING AGENT RESEARCH LAB FINE CHEM. INDUSTRIES, MUMBAI, INDIA.

5. ETHANOL VOLATILE SOLVENT SIGMA-ALDRICH CORPORATION.

6. SODIUM BENZOATE BACTERIOSTATIC AGENT

FISCHER CHEMICALS, MUMBAI,INDIA.



PHYTO CHEMICAL EVALUATION OF ETHANOLIC EXTRACT OF CISSUS QUADRANGULARIS The qualitative tests for various phytoconstituents were carried out by using Ethanolic extract of Cissus quadrangularis.

Table no.2: Qualitative Chemical Examination of Cissus quadrangularis

S.NO Test Test for Phyto-

constituent Ethanolic extract

1. Libermann-Burchard test

Sterols +

2. Gelatin test, Ferric chloride test Tannins + 3. Vanillin hydrochloride test Phenols + 4. Shinoda test, Zn-HCL reduction test Flavonoids + 5. Salkowski reaction. Triterpenoids + 6. Millon’s test, Nin-hydrin test Proteins and

amino acids -

7. Extract+ Fehling’s reagents (A and B) Glycosides - 8. Mayer’s test, Dragendroff’s test Alkaloids -

HPTLC HPTLC systems equipped with Linomat-5 applicator with WinCATS-4 software were used. All the solvents used for HPTLC analysis were of selective grade. Cissus quadrangularis ethanolic extract was dissolved in methanol and used for HPTLC analysis. The samples (5 μl) spotted in the bands of width 8 mm with a Camag microlitre syringe on pre-coated silica gel glass plate. The sample loaded plate was kept in TLC twin trough developing chamber (after saturated with Solvent vapour) with respective mobile phase and the plate was developed up to optimum level. The Toluene: Ethyl acetate (80:20) and Hexane:Ethyl acetate (70:30) were employed as mobile phase for separation. Linear ascending development was carried out in glass chamber saturated with the mobile phase. The developed plate was dried by hot air to evaporate solvents from the plate. The plate was scanned by UV at 254 and 366 nm. Densitometric scanning was performed on Camag scanner and operated by CATS software. Preparation of Ethanolic Extract of Cissus quadrangularis Ethosomal gels A gift sample of ethanolic extract of the Plant Cissus quadrangularis was given by Himalaya Drug Company, Bangalore, Karnataka.

Ethosomal gels containing Ethanolic Extract of Cissus quadrangularis were prepared by using the method suggested by Touitou et al., with modification. The ethosomal formulation of Ethanolic extract of Cissus quadrangularis was formulated using different compositions of 2-3% phospholipid, 10-30% ethanol, 5% of polyethylene glycol (PEG) and 5g of cholesterol.

The ethanolic extract of Cissus quadrangularis was dissolved separately in a covered vessel at room temperature by vigorous stirring and polyethylene glycol was added slowly to this mixture and heated to 300C at 800 rpm. Lecithin and cholesterol dissolved in ethanol and added to the above mixture. Double distilled water was added slowly as a fine stream with



constant mixing at 800 rpm. Mixing was continued for additional 5 minutes. The size of the ethosomes vesicles can be decreased using sonication or extrusion method. Ethosomes formulation was stored under refrigeration.

Ethosomal vesicles suspension were incorporated into carbopol gel (1%, 1.5%, 2% and 2.5% w/w). the specified amount of carbopol 934 powder was slowly added to ultrapure water and kept at 1000C for 20min. tri ethanolamine was added to it drop wise. Appropriate amount of formulation of ethosomes containing Ethanolic extract of Cissus quadrangularis was then incorporated into gel-base. Water q.s was added with other formulation ingredients with continuous stirring until homogenous formulation were achieved [16].

Table no.3: Composition of different ethosomal formulations

NAME OF THE INGREDIENTS

Quantities in w/w %(100 gm)

E1 E2 E3 E4 E5 E6 E7 E8 Ethanolic extract of Cissus quadrangularis

20 20 20 20 40 40 40 40

Lecithin (w/v)

2 2 2 2 2 2 2 2

Cholesterol 5 5 5 5 5 5 5 5 Ethanol (V/V) 10 10 10 10 10 10 10 10 PEG 400 (V/V) 5 5 5 5 5 5 5 5 Carbapol 934 gel base (% W/V)

1 1.5 2 2.5 1 1.5 2 2.5

Sodium benzoate (W/W)

1 1 1 1 1 1 1 1

Preparation of Ethanolic extracts of Cissus quadrangularis topical gels Topical Gels containing 20 and 40% of ethanolic extract of Cissus quadrangularis were prepared by using different concentrations of polymer such as carbopol 934 1% to 2.5%. The specified amount of carbopol 934 powder was slowly added to ultrapure water and kept for 12 hours for the polymer to swell. Appropriate amount of ethanolic extract of Cissus quadrangularis was dissolved, polyethylene glycol and sodium benzoate was added to it, this mixture is incorporated to the above mixture. Water q.s was added with other formulation ingredients with continuous stirring at 800rpm after complete addition the mixture is stirred till the homogeneous gels was obtained.(23) These formulations were then stored in the wide mouthed bottles for stability studies and all the samples were allowed to equilibrate at room temperature [19].



Figure 1: Ethosomal Formulation

Table.4: Composition of different Topical formulations

NAME OF THE INGREDIENTS

Quantities in W/W % (100gm) F1 F2 F3 F4 F5 F6 F7 F8

Ethanolic extract of Cissus quadrangularis

20 20 20 20 40 40 40 40

Carbapol 934 gel base (% w/w)

1 1.5 2 2.5 1 1.5 2 2.5

PEG 4000 (w/w)

5 5 5 5 5 5 5 5

Sodium benzoate 1 1 1 1 1 1 1 1

Figure.2: Topical Formulation

Evaluation of Ethanolic extract of Cissus quadrangularis ethosomal and topical gels Visualization The surface morphology of ethosomal and topical preparations was determined using scanning electron microscopy (SEM).



Entrapment Efficiency Separation of unentrapped drug and evaluation of entrapment efficiency can be measured by ultracentrifugation. Zeta Potential Measurement Zeta potential was used to measure the electrostatic or charge repulsion or attraction between particles of ethosomal formulation. Zeta potential was determined using a computerized inspection system. In-Vitro Drug Release Studies Using Franz Diffusion Apparatus In vitro absorption studies are generally carried out in vertical Franz diffusion apparatus. It has a receptor and a donor chamber, which is filled with phosphate buffer (PH 6.8). The dialysis membrane (Hi media 60) is sandwiched between the two chambers and clamped in place tightly.

Figure.3: Diffusion release of gels by using Franz-diffusion apparatus

PH measurements The PH of topical and ethosomal gels was measured by using electrode based digital PH meter.

Drug Content The drug content of topical and ethosomal gel formulation was determined by using phosphate buffer PH 6.8. Drug entrapment efficiency Drug entrapment efficiency of Ethanolic extract of Cissus quadrangularis ethosomal gel.



Spreadability and density of topical and ethosomal gels The spread ability of gel formulations was determined by measuring the spreading diameter of 1g of gel between two horizontal plates (20 cm × 20 cm) and the density of gels is determined by using densitometer. Viscosity Viscosities (in cps) of the prepared topical gels and ethosomal gels formulations of Ethanolic extract of Cissus quadrangularis were determined with LV Brookfield, DV-E viscometer using spindle no 64. The viscosities of the formulations were found to be 38,200 of cps for topical F8 gel formulation and 40,300 of cps for ethosomal E6 formulation were found to be satisfactory. Stability study The stability study was studied by using ICH guidelines. Ethosomal formulations were observed for any change in appearance or color for a period of 8 weeks. Loss in percentage of drug release during stability studies of the optimized batch (4±2°C & 30±2° C). The prepared formulations were tested for accelerated stability. They were kept in stability chambers at 30° C ± 2° C and 60% ± 5 % Relative humidity for 6 months and observed for the physical appearance.

RESULTS AND DISCUSSIONS FTIR Drug-Excipient Compatibility Studies FTIR of the formulation of topical and ethosomal gels of ethanolic extract of Cissus quadrangularis have shown that there is no change in the functional groups. Hence, the activity of active constituent i.e, β-sitosterol is protected in the formulations showing compatibility with the formulations of gels. FTIR of β-sitosterol

Figure.4: FTIR of β-sitosterol



FTIR of Ethanolic extract of Cissus quadrangularis

Figure.5: FTIR of Ethanolic Extract of Cissus quadrangularis

FTIR of 20% Ethanolic extract of Cissus quadrangularis topical gel

Figure.6: FTIR of 20% Ethanolic extract of Cissus quadrangularis topical gel



FTIR of 40% Ethanolic extract of Cissus quadrangularis topical gel

Figure.7: FTIR of 40% Ethanolic extract of Cissus quadrangularis topical gel

• FTIR of 20% Ethanolic extract of Cissus quadrangularis ethosomal gel:

Figure.8: FTIR of 40% Ethanolic extract of Cissus quadrangularis ethosomal gel

CALIBRATION CURVE OF β-SITOSTEROL Preparation of Standard curve of β-sitosterol in Phosphate buffer PH 6.8

Table.5: Calibration curve data of β-sitosterol CONCENTRATION (µg/ml) ABSORBANCE

50 0.101 100 0.221 150 0.325 200 0.489 300 0.655 400 0.882 500 0.991



Figure.9: Calibration curve of β-sitosterol

HPTLC Of Ethanolic extract of Cissus quadrangularis:-

Figure.10: HPTLC of Cissus quadrangularis

Mobile phase: Toluene : Ethyl acetate:- 80:20 Spray: Vanillin/H2SO4 Sample preparation: 0.5 gm of sample in 10ml methanol Volume of injection: 5ul

Figure.11: HPTLC of Cissus quadrangularis by employing different mobile phase

y = 0.0021x + 0.0207 R² = 0.9897

0

0.2

0.4

0.6

0.8

1

1.2

0 200 400 600

A

bsor

banc

e

Concentration (µg/ml)



Mobile phase: Hexane : Ethyl acetate:- 70:30 Spray: Vanillin/H2SO4 Sample preparation: 0.5 gm of sample in 10ml methanol Volume of injection: 5ul

Evaluation of topical gel Color: Dark brown Physical Appearance: The gel containing 1.5 and 2.0%

of carbapol was too liquidy in appearance. Greasiness: Non greasy Grittiness: Free from grittiness Ease of application: Easily/smoothly applied Skin irritation: No skin irritation Washability: Easily washable Size And Shape Analysis: Microscopic analysis was performed under different

magnification to visualize the vesicular structure, lamellarity and to determine the size of topical preparations.

Scanning Electron microscopy of Topical formulations

Figure.12 and 13: SEM of 20% and 40% of Ethanolic extract of Cissus quandrangularis topical gels.

Evaluation of Ethosomal gels Colour: Dark brown Physical Appearance: The gel containing 2.0 and 2.5%

of carbapol was too tough in nature. Greasiness: Non greasy Grittiness: Free from grittiness Ease of application: Easily/smoothly applied Skin irritation: No skin irritation Washability: Easily washable Size And Shape Analysis: Microscopic analysis was performed under different

magnification to visualize the vesicular structure, lamellarity and to determine the size of ethosomal preparations.



Scanning Electron microscopy of Ethosomal formulations

Figure.14 and 15: SEM of 20% and 40% of Ethanolic extract of Cissus quandrangularis Ethosomal gels.

Zeta Potential Zeta potential of Ethanolic extract of Cissus quadrangularis Topical gel.

Figure.16: Zeta potential of Cissus quadrangularis Topical gel



Zeta potential of Ethanolic extract of Cissusquadrangularis Ethosomal gel

Figure.17: Zeta potential of Cissus quadrangularis Ethosomal gel PH measurements The PH of topical and ethosomal gels was measured by using electrode based digital PH meter.

Table.6: PH Values of gels. Formulation PH Value Formulation PH Value Ethanolic extract of Cissus quadrangularis 20% Topical gel (F4)

6.9 Ethanolic extract of Cissus quadrangularis 20% Ethosomal gel (E2)

7.2

Ethanolic extract of Cissus quadrangularis 40% Topical gel (F8)

7.1 Ethanolic extract of Cissus quadrangularis 40% Ethosomal gel (E6)

7.3

Drug Content The drug content of topical and ethosomal gel formulations of the extract was also evaluated. Table.7: Drug content in gel formulations Formulation Drug content (%) Formulation Drug content (%) Ethanolic extract of Cissus quadrangularis 20% Topical gel (F4)

88.6 ±1.3

Ethanolic extract of Cissus quadrangularis 20% Ethosomal gel (E2)

98.3 ±1.1


89.8 ±1.8

Ethanolic extract of Cissus quadrangularis 40% Etosomal gel (E6)

99.6±0.4



Spreadability and density of topical and ethosomal gels The Spreadability and density of gel formulations was also evaluated.

Table.8: Spreadability, Density and Viscosity of Formulation gels. Formulation Spreadability [cm]

Density [gm/ml]

Viscosity (in cp)


8.3

2.15

37200


7.2

2.21 38200


7.0

1.20

39600


8.6 1.22

40300

Drug entrapment efficiency Drug entrapment efficiency of Ethanolic extract of Cissus quadrangularis ethosomal gel was evaluated. Table.9: Drug entrapment efficiency of Ethosomal gels Formulation β-sitosterol Entrapment efficiency(%) Ethanolic extract of Cissus quadrangularis 20% Ethosomal gel (E2)

94.34%


96.45%

In-vitro Diffusion studies

Table.10 showing % Drug Release Of Topical Gel containing Ethonolic extact of Cissus quadrangularis In Phosphate Buffer PH 6.8.

Time % CDR of 20% Ethanolic extract of Cissus quadrangularis topical gel (F4)

% CDR of 20% Ethanolic extract of Cissus quadrangularis topical gel (F8)

10 22.42 25.65 15 29.23 33.64 30 39.67 43.87 45 47.74 53.55 60 52.98 58.98 90 56.78 60.58

120 60.79 68.54 180 68.43 74.66 240 73.98 79.38 360 78.21 83.78



Figure.18: Diffusion Profile of Cissus quadrangularis Topical gels Table 11. Showing % Drug Release Of Ethosomal Gel containing Ethonolic extact of Cissus quadrangularis In Phosphate Buffer PH 6.8.

Time % CDR of 20% Ethanolic extract of Cissus quadrangularis ethosomal gel (E2)

% CDR of 40% Ethanolic extract of Cissus quadrangularis ethosomal gel (E6)

10 39.56 48.37 15 49.98 56.28 30 57.85 68.32 45 69.78 74.45 60 77.82 79.75 90 80.54 83.87

120 87.76 90.12 180 89.73 93.98 240 94.39 95.31 360 97.68 99.76

Figure. 19: Diffusion Profile of Cissus quadrangularis Ethosomal gels

0 10 20 30 40 50 60 70 80

0 50 100 150

% CDR F4 %CDR F8

%C

DR

Time in mins

0

20

40

60

80

100

120

0 100 200 300

%CDR OF E2 % CDR OF E6

%C

DR

Time in mins



Stability study Ethosomal formulations were observed for any change in appearance or color for a period of 8 weeks. There was no change in appearance in Ethosomal formulations throughout the period of study. The prepared formulations were tested for accelerated stability. They were kept in stability chambers at 30° C ± 2° C and 60% ±5 % Relative humidity for 6 months and observed for the physical appearance. It was found that there was no change in the formulations and were stable even after 6 months. Table.12 : Stability study of Ethanolic extract of Cissus quadrangularis Ethosomal gels Stability study period

% CDR of Ethanolic extract of Cissus quadrangularis 20% Ethosomal gel (E2)

% CDR of Ethanolic extract of Cissus quadrangularis 40% Ethosomal gel (E6)

Initial

4±2 ° C 99.5 99.7

30±2 ° C 98.6 99.8

After 2 weeks

4±2 ° C 98.4 99.6

30±2 ° C 98.4 99.4

After 4 weeks 4±2 ° C 98.3 99.1

30±2 ° C 98.1 98.9

After 6 weeks 4±2 ° C 97.9 98.8

30±2 ° C 97.8 98.7

After 8 weeks 4±2 ° C 97.5 98.5

30±2 ° C 97.3 98.3

RELEASE KINETIC PROFILE FOR ETHOSOMAL GEL (E6)

Table. 13: Release kinetic profile for Ethosomal gel (E6) ZERO ORDER FIRST ORDER HIGUCHI PEPPAS

%CDR Vs T Log % Remain Vs T

% CDR Vs √𝐓 Log C Vs Log T

Slope 1.5183393 -0.0179672 1.0476754 0.2819543 Intercept 15.720984 2.6221986 2.3677891 -0.0546732 Correlation 0.9678531 0.8954319 0.8795213 0.8943561 R2 0.9495612 0.9415982 0.9673241 0.9697525



Figure.20: Zero order Kinetics for E6 gel

Figure 21: First Order Kinetics for E6 gel

Figure 22: Higuchi’s model for E6 gel

y = -0.0172x + 2.6227 R² = 0.9417

0

0.5

1

1.5

2

2.5

3

0 20 40 60 80

LO

G %

DR

UG

RE

MA

INE

D

TIME

FIRST ORDER

y = 1.0474x + 2.3677 R² = 0.9672

0

20

40

60

80

100

120

0 50 100 150

% C D R

SQUARE ROOT OF TIME

HIGUCHI PLOT

y = 1.5181x + 15.721 R² = 0.9498

0

20

40

60

80

100

120

0 20 40 60 80

TIME IN HRS

% CDR

ZERO ORDER



Figure.23: Peppas model for E6 gel

R² of E6 formulation was found to be 0.949 of Zero order Kinetics therefore release pattern of E6 formulation follows zero order release. R² of E6 formulation was found to be 0.967 of Higuchi therefore release pattern of E6 formulation follows diffusion release mechanism. CONCLUSION

The Ethosomal system is a highly efficient drug delivery system. The Ethanolic extract of Cissus quadrangularis is used to prepare the gels. The method described by Touitou et al., (2000) was employed with modification for the preparation of various ethosomal formulations containing concentration of ethanol (10%) with sonication. The entrapment efficiency of ethosomes containing 40% w/w ethanolic extract of Cissus quadrangularis (E6) has shown highest value with respect to all other formulation The PH values for ethosomal gels formulations were in the range of 7.2 to 7.3. The PH values for topical gels formulations were in the range of 6.9 to 7.1. The viscosities of the formulations were found to be 38200 of cps for topical F8 gel formulation and 40300 of cps for ethosomal E6 Gel formulation. The maximum entrapment efficiency of ethosomal vesicles as determined by ultracentrifugation was 94.34 % for 20% Ethanolic extract of Cissus quadrangularis ethosomal gel (E2) and 96.45% for 40% Ethanolic extract of Cissus quadrangularis ethosomal gel (E6).Results of entrapment efficiency also suggest that 2% phospholipids is optimal concentration for entrapment efficiency. Ethosomal gel of Ethanolic extract of Cissus quadrangularis has shown 99.76% drug release, zeta potential of -60.9 (mV), viscosity was found to be 40,300 of cps, with specific vesicular size and shape observed by SEM Stability studies was carried out for a period of 8 weeks and showed no significance changes in the characteristics of ethosomes and further the loss of drug is not more than 2%. From the above observations it was concluded that ethosomal gel formulation E6 was found to be the best formulation among all the formulation of topical gels and ethosomal gels.

y = 0.2815x - 0.0544 R² = 0.9697

-0.2

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

0 2 4 6

LOG%CDR

PEPPAS

LOG TIME



ACKNOWLEDGEMENT

The Successful accomplishment of this Article would not have been possible but by the timely help and guidance rendered by many people. I would like to take this opportunity to place it in record. Though it’s not possible to name all of them, I would like to mention few of them. My first salutation goes to Almighty Allah and my Parents for being ever so kind and courteous. It gives me an immense pleasure to acknowledge a debt of gratitude to my guide Mrs. Sirisha Mittapally M.Pharm, PGDCA,(Ph.D) Associate Professor, Deccan School of Pharmacy for her constant encouragement, suggestions, supervision and support and also to Dr. Roshan.S M.Pharm, Ph.D Professor and Head of the Department of Pharmacognosy, Deccan School of Pharmacy for his strong support, guidance and co-operation. I would like to express profound gratitude to Dr. Syed Abdul Azeez Basha, honourable Principal of Deccan School of Pharmacy, Hyderabad, for guiding me as well. REFERENCES

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Formulation and valuation of E thanolic extract of e ... · International Research Journal of Pharmaceutical and Biosciences (IRJPBS) 4 (4) 13 constant mixing at 800 rpm. Mixing was