CHARACTERIZATION AND DEVELOPMENT OF EMBELIA RIBES …

www.wjpps.com Vol 9, Issue 8, 2020.

2570

Jain et al. World Journal of Pharmacy and Pharmaceutical Sciences

CHARACTERIZATION AND DEVELOPMENT OF EMBELIA RIBES

PHYTOSOMES

Chanchal Jain*, Sonal Gupta and Dr. A. K. Singhai

M. Pharm, Department of Pharmaceutics, Lakshmi Narain College of Pharmacy, Bhopal

(M.P.).

ABSTRACT

Embelia ribes, in particular embelin isolated from dried berries of

embelia ribes has a wide spectrum of biological activities. Belong to

the family Myrsinacae It is commonly known as false black pepper or

Vidanga it is widely distributed throughout india.in ayurveda and

siddha it is considered widely beneficial in variety of disease. E. Ribes

Seeds are rich in alkaloids, flavonoids and phenolic compounds

Hence,in present work we have analyzed The DPPH free radical

scavenging activity of Hydroalcoholic extract of Embelia ribes were

21.64%, 27.43%, 32.39%, 37.36%, 41.25%, 49.96% at different

concentration of 10 µg/ml, 20 µg/ml, 40 µg/ml, 60 µg/ml, 80 µg/ml,

100 µg/ml respectively. The Embelia ribes seed extract has been showed the robust

scavenging activity at 100µg/ml, which was 49.96% as compared to Ascorbic acid 84.13%.

This activity was lower than ascorbic acid, Embelia ribes hydroalcoholic extract exhibited

potent scavenging activity (IC50-103.93). Reference standard ascorbic acid showed IC50 -

17.681. In this study, we prepared the, Embelia ribes -phospholipids complex to improve the

lipophilic properties of Embelia ribes. We prepared the complex with different quantity ratios

of phospholipids: cholesterol and Embelia ribes such as 1:1:1, 1:2:1, 2:1:1, 2:3:1.The

phytosomes formulation having 2:1 ration of phospholipid and cholesterol and 1.0% w/v

extract of embelia ribes (F10) showing the greatest entrapment efficiency,(73.32+0.25) with

small particle size (313.25) was further evaluated for drug release study. The phytosomal

formulation of Embelia ribes (a complex of E.ribes with phosphatidylcholine) has been

shown to improve E.ribes bioavailability.

WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

SJIF Impact Factor 7.632

Volume 9, Issue 8, 2570-2591 Research Article ISSN 2278 – 4357

*Corresponding Author

Chanchal Jain

M. Pharm, Department of

Pharmaceutics, Lakshmi

Narain College of Pharmacy,

Bhopal (M.P.).

Article Received on

24 June 2020,

Revised on 14 July 2020,

Accepted on 03 August 2020

DOI: 10.20959/wjpps20208-16987


2571


KEYWORDS: embelia ribes, phytosomes, antioxidant activity antidiabetic, hydro alcoholic

extraction.

INTRODUCTION

The nano-carrier used in novel drug delivery system of herbal drugs has a potential future,

improving the activity and overcoming the problem associated with herbal constituents. Any

type of medicament could be delivered to its specific site of action with the help of novel

drug delivery system. Researchers are currently underway to develop an ideal drug delivery

system which satisfies the need of targeted site of action. Biological membrane presents a

barrier through which a drug must pass before it gets absorbed or excreted. Lipid solubility

and molecular size of drug molecule pose two major limiting factors to pass the biological

membrane by which drug can be absorbed systematically following oral or topical

administration. During the last century chemical and pharmacological studies have been

performed on a lot of plant extracts in order to know their chemical composition and confirm

the indications of traditional medicine. Preparations of Phyto medicine has been used for

health maintenance since ancient times. The Phytomedicines posses a lot of therapeutic uses.

Phytosome is vesicular drug delivery system in which phytoconstituents of herb extract

surround and bound by lipid (one phyto-constituent molecule linked with at least one

phospholipid molecule). Phytosome protect valuable component of herbal extract from

destruction by digestive secretion and gut bacteria and because of which they shows better

absorption which produces better bioavailability and improved pharmacological and

pharmacokinetic parameters than conventional herbal extract. Phytosomes is the combination

of two words, the term “PHYTO” means plant while “SOME” means cell-like. The

formulation is developed by encapsulating the plant material or plant extract within the

spherical cell like structure, which is an advanced nano-sphere or cell forms of herbal

products that are better absorbed. Phytosomes produces better pharmacokinetic and

pharmacodynamic profile of drug than conventional herbal formulations. It‟s a novel

emerging technique that is applied to phytopharmaceuticals for the enhancement of

bioavailability of natural plant extract for medicinal applications. Phytomedicines, complex

chemical mixture prepared from plants, have been used in India and worldwide from the very

beginning of human civilization and continue to have widespread popular use. Phytosomes

are more bioavailability as compared to herbal extract owing to their enhancedcapacity to

cross the lipid rich biomembranes and finally reaching the blood. Phosphatidylcholine is


2572


phospholipids. It is a key component of phytosomes process. Phospholipids are employed as

natural digestive aids and carriers for water soluble and lipid soluble nutrients.

Principle of Phytosome Technology

The phytochemical constituents (flavonoids and terpenoides) of the extracts provide them for

the direct complexation with Phosphatidylcholine. Phytosome results from the reaction of a

stoichiometric amount of the phospholipid with the standardized extract or polyphenolic

constituents in a non-polar solvent. The Phosphatidylcholine is a bi-functional compound

composed of lipophilic phosphatidyl moiety and the hydrophilic choline moiety. The choline

head of phosphatidylcholine molecule binds to phytocomponent while the lipid soluble

phosphatidyl portion comprises the body and tail which then envelops the choline bound

material. Hence, the Phytoconstituents build up a lipid compatible molecular complex with

phospholipid also called as phyto-phospholipid complex.

Biological properties

Phytosomes are novel complex utilized; hence they produce more bioavailability and better

result thanes which are better absorbed and the conventional herbal extract or non-complex

extracts, which has been demonstrated by pharmacokinetic studies or by pharmacodynamic

tests in experimental animals and in human subjects.

Method for preparations of phytosomes

Phospholipids

Dissolved in organic solvent containing Drug/Extract

Solution of phospholipids in organic solvent with drug/extract

Drying

Formation of thin film

Formation of phytosomal suspension


2573


MATERIAL AND METHODS

Collection of Plant material

Seeds of Embelia Ribes were collected from local area of Bhopal in the month of January,

2020 The plant material was identified and authenticated by depositing the herbarium

specimens in Botany Department, Dr.javinder Mehta career college, Bhopal Madhya

Pradesh,india. under the voucher no. Career/Herb/2020/010 Accession no.

1 Defatting of plant material

Seeds of Embelia ribes were shade dried at room temperature. The shade dried plant material

was coarsely powdered and subjected to extraction with petroleum ether by maceration. The

extraction was continued till the defatting of the material had taken place.

2 Extraction by maceration process

100 gm of dried powdered seeds of Embelia ribes has been extracted with hydroalcoholic

solvent (ethanol: water: 70:30) using maceration method for 48 hrs, filtered and dried using

vacuum evaporator at 400C.

Table 1: Phytochemical screening of extract of Embelia ribes.

S. No. Constituents Hydroalcoholic extract

1.

Alkaloids

Wagner‟s Test

Hager‟s test

-ve

-ve

2. Glycosides

Legal‟s test

-ve

3.

Flavonoids

Lead acetate

Alkaline test

+ve

-ve

4. Phenolics

Ferric Chloride Test

+ve

5. Proteins

Xanthoproteic test

+ve

6. Carbohydrates

Fehling‟s test

+ve

7.

Saponins

Froth Test

Foam test

+ve

+ve

8. Diterpins

Copper acetate test

-ve

9. Tannins

Gelatin Test

+ve


2574


Antioxidant activity of extract using DPPH method

DPPH scavenging activity was measured by the spectrophotometer. Stock solution (6 mg in

100ml methanol) was prepared such that 1.5 ml of it in 1.5 ml of methanol gave an initial

absorbance. Decrease in the absorbance in presence of sample extract at different

concentration (10- 100 µg/ml) was noted after 15 minutes. 1.5 ml of DPPH solution was

taken and volume made till 3 ml with methanol, absorbance was taken immediately at 517

nm for control reading. 1.5 ml of DPPH and 1.5 ml of the test sample of different

concentration were put in a series of volumetric flasks and final volume was adjusted to 3 ml

with methanol. Three test samples were taken and each processed similarly. Finally the mean

was taken. Absorbance at zero time was taken for each concentration. Final decrease in

absorbance was noted of DPPH with the sample at different concentration after 15 minutes at

517 nm.

Figure 1: Photographs during examination.

Results of antioxidant activity using DPPH method

Table 2: % Inhibition of ascorbic acid and Hydroalcoholic extract of Embelia ribes

using DPPH method.

S. No. Concentration

(µg/ml)

% Inhibition

Ascorbic acid Hydroalcoholic

extract

1 10 44.65 21.64

2 20 48.62 27.43


2575


The outcome of DPPH (1,1-Diphenyl-2-picrylhydrazyl radical) radical scavenging activity of

Hydroalcoholic extract of Embelia ribes was showed and The DPPH free radical scavenging

activity of Hydroalcoholic extract of Embelia ribes were 21.64%, 27.43%, 32.39%, 37.36%,

41.25%, 49.96% at different concentration of 10 µg/ml, 20 µg/ml, 40 µg/ml, 60 µg/ml, 80

µg/ml, 100 µg/ml respectively. The Embelia ribes seed extract has been showed the robust




17.681.

In vitro anti diabetic activity of Hydroalcoholic extract of Embelia ribes

1 Inhibition of alpha amylase enzyme

Preparation of standard: 10 mg acarbose was dissolved in 10 ml methanol, and various

aliquots of 100- 1000μg/ml were prepared in methanol.

Preparation of sample: 100 mg of dried extract was extracted with 100 ml methanol, filter,

and make up the volume up to 100 ml. 500 µl of this extract was for the estimation of enzyme

inhibition.

METHOD: A total of 500 µl of test samples and standard drug (100-500µg/ml) were added

to 500 µl of 0.20 mM phosphate buffer (pH 6.9) containing α-amylase (0.5mg/ml) solution

and were incubated at 25°C for 10 min. After these, 500 µl of a 1% starch solution in 0.02 M

sodium phosphate buffer (pH 6.9) was added to each tube. The reaction mixtures were then

incubated at 25°C for 10 min. The reaction was stopped with 1.0 ml of 3, 5 dinitrosalicylic

acid colour reagent. The test tubes were then incubated in a boiling water bath for 5 min,

cooled to room temperature.

3 40 65.34 32.39

4 60 69.65 37.36

5 80 77.41 41.25

6 100 84.13 49.96

IC 50 17.681 103.93


2576


The reaction mixture was then diluted after adding 10 ml distilled water and absorbance was

measured at 540 nm. Control represent 100% enzyme activity and were conducted in similar

way by replacing extract with vehicle.

A 0 - A t

% Inhibition = ________ X 100

A 0

Where A 0 is the absorbance of the control and A t is the absorbance of the sample.

Results of in vitro anti diabetic activity of Hydroalcoholic extract of Embelia ribes.

Table 4: Absorbances of Hydroalcoholic extract of Embelia ribes.

S. No Acarbose Embelia ribes

Conc. Absorbance Absorbance

1. 100 0.369 0.458

2. 200 0.226 0.321

3. 300 0.145 0.225

4. 400 0.098 0.125

5. 500 0.065 0.098

Control Absorbance = 0.698

Table 4.1: % Inhibition of Hydroalcoholic extract of Embelia ribes.

S. No. Acarbose Embelia ribes

Conc. % Inhibition % Inhibition

1. 100 47.13467 40.974

2. 200 67.62178 50.573

3. 300 79.22636 55.301

4. 400 85.95989 62.034

5. 500 90.68768 65.473

IC 50 value 71.52 221.16

Formulation development of phytosomes

Preparation of phytosomes

The complex was prepared with phospholipids: Cholesterol and Embelia ribes in the ratio of

1:1:1, 1:2:1, 2:1:1, 2:3:1 respectively. Weight amount of extract and phospholipids and

cholesterol were placed in a 100ml round-bottom flask and 25ml of dichloromethane was

added as reaction medium. The mixture was refluxed and the reaction temperature of the

complex was controlled to 50°C for 3 h. The resultant clear mixture was evaporated and 20

ml of n-hexane was added to it with stirring. The precipitated was filtered and dried under

vacuum to remove the traces amount of solvents. The dried residues were gathered and


2577


placed in desiccators overnight and stored at room temperature in an amber colored glass

bottle.

Figure 2: Microscopic observation of optimized batch F10.

Table 5: Different formulations of phytosomes.

Formulation

Ratio of

Phospholipids and

Cholesterol

Extract

Concentration

(%)

Dichloromethane

Concentration

Optimization of Phospholipids and Cholesterol

F1 1:1 1 25

F2 1:2 1 25

F3 2:1 1 25

F4 2:3 1 25

Optimization of Drug Concentration

F5 2:1 0.5 25

F6 2:1 1.0 25

F7 2:1 1.5 25

F8 2:1 2.0 25

Optimization of solvent concentration

F9 2:1 1.0 10

F10 2:1 1.0 25

F11 2:1 1.0 50

F12 2:1 1.0 75

Evaluation of phytosomes

Microscopic observation of prepared phytosomes

An optical microscope (cippon, Japan) with a camera attachment (Minolta) was used to

observe the shape of the optimized Phytosome formulation.


2578


Drug Excipient compatibility study by FT-IR

IR spectra of physical mixture of drug and excipients were recorded by ATR (Attenuated

total reflection) techniques using Fourier transform infrared spectrophotometer. A base line

correction was made and the sample was directely mounted in IR compartment and scanned

at wavelengths 4000 cm-1

to 400 cm-1

.

Entrapment efficiency

Phytosome preparation was taken and subjected to centrifugation using cooling centrifuge

(Remi) at 12000 rpm for an hour at 4.

The clear supernatant was siphoned off carefully to separate the non entrapped flavonoids

and the absorbance of supernatant for non entrapped Embelia ribes was recorded at λmax

420.0 nm using UV/visible spectrophotometer (Labindia 3000+). Sediment was treated with

1ml of 0.1 % Triton x 100 to lyse the vesicles and diluted to 100 ml with 0.1 N HCl and

absorbance taken at 420.0 nm. Amount of quercetin in supernatant and sediment gave a total

amount of Embelia ribes in 1 ml dispersion. The percent entrapment was calculated by

following formula.

Particle size and size distribution

The particle size, size distribution and zeta potential of optimized phytosomes formulation

were determined by dynamic light scattering (DLS) using a computerized inspection system

(Malvern Zetamaster ZEM 5002, Malvern, UK). The electric potential of the phytosomes,

including its Stern layer (zeta potential) was determined by injecting the diluted system into a

zeta potential measurement cell.

Transmission Electron Microscopy: Surface morphology was determined by TEM, for

TEM a drop of the sample was placed on a carbon-coated copper grid and after 15 min it was

negatively stained with 1% aqueous solution of phosphotungustic acid. The grid was allowed

to air dry thoroughly and samples were viewed on a transmission electron microscopy (TEM

Hitachi, H-7500 Tokyo, Japan).


2579


In Vitro dissolution rate studies

In vitro drug release of the sample was carried out using USP- type I dissolution apparatus

(Basket type). The dissolution medium, 900 ml 0.1N HCl was placed into the dissolution

flask maintaining the temperature of 37±0.50C and 75 rpm. 10 mg of prepared phytosomes

was placed in each basket of dissolution apparatus. The apparatus was allowed to run for 8

hours. Sample measuring 3 ml were withdrawn after every interval (30 min, 1 hrs, 2 hrs, 4

hrs, 6 hrs, 8 hrs, and 12 hrs.) up to 12 hours using 10 ml pipette. The fresh dissolution

medium (370C) was replaced every time with the same quantity of the sample and takes the

absorbance at 256.0 nm using spectroscopy.

Mathematical treatment of in-vitro release data: The quantitative analysis of the values

obtained in dissolution/release tests is easier when mathematical formulas that express the

dissolution results as a function of some of the dosage forms characteristics are used.

1. Zero Order kinetics: The pharmaceutical dosage forms following this profile release the

same amount of drug by unit of time and it is the ideal method of drug release in order to

achieve a pharmacological prolonged action. The following relation can, in a simple way,

express this model.

Qt = Qo + Ko t

where Qt is the amount of drug dissolved in time t, Qo is the initial amount of drug in the

solution (most times, Qo=0) and Ko is the zero order release constant58-62

.

2. First Order kinetics: The following relation expresses this model.

where Qt is the amount of drug dissolved in time t, Qo is the initial amount of drug in the

solution and K1 is the zero order release constant.

In this way a graphic of the decimal logarithm of the released amount of drug versus time

will be linear. The pharmaceutical dosage forms following this dissolution profile, such as

those containing water-soluble drugs in porous matrices, release drug in a way that is

proportional to the amount of drug remaining in its interior, in such way, that the amount of

drug released by unit of time diminish.


2580


3. Higuchi Model: Higuchi developed several theoretical models to study the release of

water-soluble and low soluble drugs in semi-solid and/or solid matrixes. Mathematical

expressions were obtained for drug particles dispersed in a uniform matrix behaving as the

diffusion media. The simplified Higuchi model is expressed as.

Where Q is the amount of drug released in time t and KH is the Higuchi dissolution constant.

Higuchi model describes drug release as a diffusion process based in the Fick‟s law, square

root time dependent. This relation can be used to describe the drug dissolution from several

types of modified release pharmaceutical dosage forms such as transdermal systems and

matrix tablets with water-soluble drugs.

4. Korsmeyer Peppas Model: Korsmeyer et al. used a simple empirical equation to describe

general solute release behaviour from controlled release polymer matrices:

where Mt/M is fraction of drug released, a is kinetic constant, t is release time and n is the

diffusional exponent for drug release. ‟n‟ is the slope value of log Mt/M versus log time

curve. Peppas stated that the above equation could adequately describe the release of solutes

from slabs, spheres, cylinders and discs, regardless of the release mechanism.. Peppas used

this n value in order to characterize different release mechanisms, concluding for values for a

slab, of n =0.5 for fickian diffusion and higher values of n, between 0.5 and 1.0, or n =1.0, for

mass transfer following a non-fickian model. In case of a cylinder n =0.45 instead of 0.5, and

0.89 instead of 1.0. This equation can only be used in systems with a drug diffusion

coefficient fairly concentration independent. To the determination of the exponent n the

portion of the release curve where Mt/M < 0.6 should only be used. To use this equation it is

also necessary that release occurs in a one-dimensional way and that the system width-

thickness or length-thickness relation be at least 10. A modified form of this equation was

developed to accommodate the lag time (l) in the beginning of the drug release from the

pharmaceutical dosage form.

When there is the possibility of a burst effect, b, this equation becomes.


2581


In the absence of lag time or burst effect, l and b value would be zero and only atn

is used.

This mathematical model, also known as Power Law, has been used very frequently to

describe release from several different pharmaceutical modified release dosage forms.

Stability studies

Stability studies of optimize phytosomes formulation

The prepared phytosomes subjected to stability studies at 40±2°C/75±5% RH and

30±2°C/60±5% RH as per ICH guidelines for a period of 3 months. Samples were withdrawn

at 1 month time intervals and evaluated for physical appearance and drug content.

RESULTS AND DISCUSSION

The Hydroalcoholic extract of seeds of Embelia ribes had revealed the presence of

flavonoids, Phenols, Protein, carbohydrates, tannins and saponins. Alkaloids, glycosides,

diterpenes, were found to be absent. the results of total phenolic and flavonoid contents in the

Hydroalcoholic extract of seeds of Embelia ribes. The total phenolic and flavonoid contents

of seeds of Embelia ribes (Hydroalcoholic extract) showed the content value 0.247 mg

GAE/100mg extract and 0.838 mg QAE/100mg extract respectively. The Embelia ribes

(Hydroalcoholic extract) showed the highest concentration of flavonoid followed by phenols

indicate plant may hold better therapeutic application. The DPPH free radical scavenging

activity of Hydroalcoholic extract of Embelia ribes were 21.64%, 27.43%, 32.39%, 37.36%,

41.25%, 49.96% at different concentration of 10 µg/ml, 20 µg/ml, 40 µg/ml, 60 µg/ml, 80

µg/ml, 100 µg/ml respectively. The Embelia ribes seed extract has been showed the robust




17.681.

Many herbal extracts have been reported to have antidiabetic activities and are used in

Ayurveda for the treatment of diabetes. Herbal extracts have been used directly or indirectly

for the preparation of many modern medicines. In this study, an in vitro inhibitory effect of

hydroalcoholic extracts of Embelia ribes on alpha amylase was evaluated. In our study, the

hydroalcoholic extract (at a concentration 500 μg/mL) showed 65.47% of α-amylase


2582


inhibitory activity with IC50 value 221.16μg/mL. At the same time, when compared with

standard acarbose IC50 value was found to be 71.52μg/ml.

Different formulation of Phytosomes were prepared using different amount of phospholipids:

cholesterol and extract and were evaluated for Drug Excipient compatibility study,

Entrapment efficiency and particle size analysis, High performance liquid chromatography,

Transmission Electron Microscopy (TEM), In vitro drug release study of prepared

Phytosomes formulation and Stability study. The appearance or disappearance of peaks

and/or the shift of their positions are often indications of interactions such as hydrogen

bonding. The IR spectra of extract, Fig. 5.7-5.8, shows stretching vibrations at 1600.9880 cm-

1 attributed predominantly to the overlapping stretching vibrations of alkenes (C=C) and

carbonyl (C=O) character. Infrared of extract show stretching vibration at 2941.0572cm-1

due

to O-H groups, C=C aromatic stretching vibration at 1436.7392cm-1

. When the data obtained

from FTIR spectra is compared with the spectra studied it was observed that there are similar

peaks for functional groups in Phytosomes.

From the FTIR data of the physical mixture it is clear that functionalities of drug have

remained unchanged including intensities of the peak. This suggests that during the process

drug, Phospholipids and Cholesterol has not reacted with the drug to give rise to reactant

products. So there is no interaction between them which is in favor to proceed for formulation

of Phytosomes as drug delivery system. The entrapment efficiency of all the prepared

formulations is shown in Table 5.8. The entrapment efficiency of the phytosomes was found

in the range of 50.23±0.36 to 73.32±0.25%.

Particle size of all formulations found within range 313.25-465.32nm. Concentration of lipid

has shows significant impact on size of phytosomes. Formulation F10 was found best one

which is further evaluated for drug release study, transmission electron microscopy (TEM),

and stability studies.chromatographic analysis of phytosomes, performed by using

acetonitrile and methanol 50:50 (v/v) solutions as mobile phase and used a flow rate of 1ml

per min and absorbance at 256nm. It gave good separation of quercetin at RT 3.965 min. The

HPLC results of the Hydroalcoholic extract loaded Phytosomes showed major peaks at the

RT 3.962 (min) at a wavelength of 256nm The retention time of peaks was found to similar

retention time of standard 3.965, which confirm the presence of quercetin in Phytosomes. The

Percentage of Quercetin was found in Phytosomes (0.014%).


2583


TEMs are capable of imaging at a significantly higher resolution than light microscopes,

owing to the small de Broglie wavelength of electrons. This enables the instrument's user to

examine fine detail even as small as a single column of atoms, which is thousands of times

smaller than the smallest resolvable object in a light microscope. TEM forms a major analysis

method in a range of scientific fields, in both physical and biological sciences. TEM

characterization revealed that the Phytosomes are spherical in shape. However, some

variation in size distribution was observed in the TEM image, which might be attributed to an

uncontrolled charge neutralization process involved between oppositely charged chains

occurring during the formation of phytosomes. When the regression coefficient values of

were compared, it was observed that „r‟ values of Korsmeyer Peppas was maximum i.e. 0.992

hence indicating drug release from formulations was found to follow Korsmeyer Peppas

kinetics. Results of stability studies clearly indicates that optimized batches of phytosomes

were stable over the chosen temperature and humidity conditions up to 3 months as were

found no significant variation in physical appearance and % drug content.

Result of Drug-Excipient compatibility study

Figure 3: FT-IR spectra of hydroalcoholic extract.

http://en.wikipedia.org/wiki/Optical_resolution

http://en.wikipedia.org/wiki/Optical_microscope

http://en.wikipedia.org/wiki/De_Broglie_wavelength


2584


Table 6: Interpretation of FT-IR spectrum.

S. No. Functional group Peak wave number(cm

-1)

Experimental Theoretical

1. C-C str. and C=O str. 1600.9880 1550-1650

3. O-H str. 2941.0572 3200-2800

5. C=C aromatic str. 1436.7392 1400-1450

Figure 3.1: FT-IR spectra of prepared phytosomes formulation.

Particle size and entrapment efficiency of drug loaded phytosomes

Formulation

Code

Particle size

(nm)

Entrapment Efficiency

(%)

F1 435.65 50.23±0.36

F2 422.47 53.65±0.25

F3 465.32 58.89±0.65

F4 390.56 65.45±0.32

F5 365.58 63.12±0.47

F6 430.14 69.98±0.56

F7 452.32 63.32±0.47

F8 436.65 65.48±0.62

F9 346.65 62.32±0.14

F10 313.25 73.32±0.25

F11 415.65 65.45±0.36

F12 412.32 69.98±0.32


2585


Average of three determinations (n=3)

Figure 3.2: Graph of Particle size and entrapment efficiency.

Figure 3.3: Particle size of optimized batch F10.


2586


Dissolution rate studies

In vitro drug release study of prepared Phytosomes formulation.

Table 7: In-vitro drug release data for optimized formulation F10.

Time

(h)

Square

Root of

Time(h)1/2

Log

Time Cumulative*%

Drug Release

Log

Cumulative

% Drug

Release

Cumulative

% Drug

Remaining

Log

Cumulative

% Drug

Remaining

0.5 0.707 -0.301 22.32 1.349 77.68 1.890

1 1.000 0.000 30.35 1.482 69.65 1.843

2 1.414 0.301 42.32 1.627 57.68 1.761

4 2.000 0.602 65.56 1.817 34.44 1.537

6 2.449 0.778 75.65 1.879 24.35 1.386

8 2.828 0.903 82.32 1.916 17.68 1.247

12 3.464 1.079 95.65 1.981 4.35 0.638

Figure 7.1: Cumulative % drug released Vs Time (Zero Order Kinetics).


2587


Figure 7.2: Log cumulative % drug remaining Vs Time (First Order Kinetics).

Figure 7.3: Cumulative % drug release Vs Root time (Higuchi Release Kinetics).


2588


Figure 7.4: Log Cumulative % drug release Vs Log time (Korsmeyer Peppas Model).

Table 8: Regression analysis data of optimized formulation F10.

Batch Zero Order First Order Higuchi Korsmeyer Peppas

R² R² R² R²

F10 0.910 0.980 0.984 0.992

CONCLUSION

In this study, the combined hydroalcoholic extract of Embelia ribes in ratio of 2:1:1 found to

exhibit significant results. Phytosomes has better physical characteristics than that of extract. In-

vitro studies revealed that phytosomes showed control release of phytoconstituents. Hence,

phytosomal formulation of this herbal drug combination can be used for clinical application to

enhance the therapeutic effect.

REFERENCE

1. Rana Mahendra: Drug Delivery through Targeted Approach with Special References to

Phytosomes. https://www.intechopen.com/books/role-of-novel-drug-delivery-vehicles-in-

nanobiomedicine/drug-delivery-through-targeted-approach-with-special-references-to-

phytosomes.


2589


2. Pande S. D: Preparation and Evaluation of Phytosomes of Pomegrane Peels. Research

Journal of Pharmacy and Technology, 2015; 8.

3. Sharma D, Bhujbale AA; Phytosomes is a Novel Drug Delivery System based herbal

formulation: An Review; Pharma Tutor, 2018; 6(3): 23-26.

4. Solmaz Rasaie: Nano Phytosomes of Quercetin: A Promising Formulation for Fortification

of Food Products with Antioxidants, Pharmaceutical sciences, 2014; 20: 96-101.

5. American Diabetes Association. Standards of medical care in diabetes. Diabetes Care, 2009;

32(suppl 1): s15.

6. Modak, M., Dixit, P., Londhe, J. Devasagayam. Indian herbs and herbal drugs used for the

treatment of diabetes. J Clin Biochem Nutr, 2007; 40: 163-73.

7. Rathore P. Planterosomes: potential phyto-phospholipid carriers for the bioavailability

enhancement of herbal extracts. International journal of pharmaceutical science and research,

2012; 3: 737-755.

8. Valenzuela A, Aspillaga M, Vial S, Guerra R. Selectivity of silymarin on the increase of the

glutathione content in different tissues of the rat. Planta Med, 1989; 55: 420-422.

9. Saraf S, Kaur CD. Phytoconstituents as photoprotective novel cosmetic formulations.

Pharmacogn Rev, 2010; 4: 1-11.

10. Pandey S. Phytosome: Technical Revolution in Phytomedicine. International Journal of

PharmTech Research, 2010; 2: 627-631.

11. Amin T, Bhat S. A Review on Phytosome Technology as a Novel Approach to Improve the

Bioavailability of Neutraceuticals. International Journal of Advancements in Research and

Technology, 2012; 1: 1-15.

12. Saha S, Sarma A, Saikia P, Chakrabarty T. Phytosome: A Brief Overview. Scholars

Academic Journal of Pharmacy, 2013; 2: 12-20.

13. Battacharya S. Phytosome: Emerging strategy in delivery of herbal drugs and nutraceuticals.

PharmTimes, 2009; 41: 3.

14. Gupta A, Ashawal MS, Saraf S. Phytosome: a novel approach towards functional cosmetics.

J Plant Science, 2007; 644-649.

15. Kareparamban J, Nikam P, Jadhav A, Kadam V. Phytosome: a novel revolution in herbal

drugs. International journal of research in pharmacy and chemistry, 2012; 2: 299-310.

https://rjptonline.org/search.aspx?key=Pande%20S.%20D.


2590


16. Tawheed Amin, Suman Vikas Bhat, A Review on Phytosome Technology as a Novel

Approach to Improve The Bioavailability of Nutraceuticals, International Journal of

Advancements in Research & Technology, 2012; 1(3): 1-15.

17. A. Vidyalakshmi, S. Esaki Selvi. Protease Activity of Floral Extracts of Jasminum

grandiflorum L., a Wound Healing Herb. Journal of Medicinal Plants Studies, 2013; 1(4):

11-15.

18. Hossan MS, Fatima A, Rahmatullah M, Khoo TJ, Nissapatorn V, Galochkina AV, Slita AV,

Shtro AA, Nikolaeva Y, Zarubaev VV, Wiart C. Antiviral activity of Embelia ribes Burm. f.

against influenza virus in vitro. Arch Virol, 2018; 163(8): 2121-2131.

19. Durg S, B NK, Vandal R, Dhadde SB, Thippeswamy BS, Veerapur VP, Badami S.

Antipsychotic activity of embelin isolated from Embelia ribes: A preliminary study. Biomed

Pharmacother, 2017; 90: 328-331.

20. Meenu Bist and Shyam Baboo Prasad. Embelia ribes: A valuable medicinal plant. Journal of

Chemical and Pharmaceutical Research, 2016; 8(4): 1229-1233.

21. Sudhakaran MV. Botanical Pharmacognosy of the Fruit of Embelia ribes Burm. F.

Sudhakaran, J Pharmacogn Nat Prod, 2015; 1: 1.

22. K. Souravi and P. E. Rajasekharan. Ethnopharmacological Uses of Embelia ribes Burm. F. -

A Review. IOSR Journal of Pharmacy and Biological Sciences, 2014; 9(3): 23-30.

23. M. Nazam Ansari & U. Bhandari. Antihyperhomocysteinemic Activity of an Ethanol Extract

from Embelia ribes. in Albino Rats. Journal Pharmaceutical Biology, 2008; 46(4): 11-18.

24. Amudha S., Prabal Kumar Manna, Jeganathan N. S. Formulation and evaluation of capsules

of Syzygium Cumini phytosomes. J. Pharm. Sci. Innov, 2018; 7(3): 70-78.

25. Arun Kumar, Bimlesh Kumar, Sachin Kumar Singh, Barinder Kaur, Saurabh Singh, A

review on phytosomes: novel approach for herbal phytochemicals, Asian J Pharm Clin Res,

2017; 10(10): 41-47.

26. Ashwini S Dhase, Shweta S Saboo, Preparation and Evaluation of Phytosomes Containing

Methanolic Extract of Leaves of Aegle Marmelos (Bael), International Journal of PharmTech

Research, 2015; 8(6): 231-240.

27. Solmaz Rasaie, Saeed Ghanbarzadeh, Maryam Mohammadi, Hamed Hamishehkar, Nano

Phytosomes of Quercetin: A Promising Formulation for Fortification of Food Products with

Antioxidants, Pharmaceutical sciences, 2014; 20: 96-101.

https://www.ncbi.nlm.nih.gov/pubmed/?term=Hossan%20MS%5BAuthor%5D&cauthor=true&cauthor_uid=29633078

https://www.ncbi.nlm.nih.gov/pubmed/?term=Fatima%20A%5BAuthor%5D&cauthor=true&cauthor_uid=29633078

https://www.ncbi.nlm.nih.gov/pubmed/?term=Rahmatullah%20M%5BAuthor%5D&cauthor=true&cauthor_uid=29633078

https://www.ncbi.nlm.nih.gov/pubmed/?term=Khoo%20TJ%5BAuthor%5D&cauthor=true&cauthor_uid=29633078

https://www.ncbi.nlm.nih.gov/pubmed/?term=Nissapatorn%20V%5BAuthor%5D&cauthor=true&cauthor_uid=29633078

https://www.ncbi.nlm.nih.gov/pubmed/?term=Galochkina%20AV%5BAuthor%5D&cauthor=true&cauthor_uid=29633078

https://www.ncbi.nlm.nih.gov/pubmed/?term=Slita%20AV%5BAuthor%5D&cauthor=true&cauthor_uid=29633078

https://www.ncbi.nlm.nih.gov/pubmed/?term=Shtro%20AA%5BAuthor%5D&cauthor=true&cauthor_uid=29633078

https://www.ncbi.nlm.nih.gov/pubmed/?term=Nikolaeva%20Y%5BAuthor%5D&cauthor=true&cauthor_uid=29633078

https://www.ncbi.nlm.nih.gov/pubmed/?term=Zarubaev%20VV%5BAuthor%5D&cauthor=true&cauthor_uid=29633078

https://www.ncbi.nlm.nih.gov/pubmed/?term=Wiart%20C%5BAuthor%5D&cauthor=true&cauthor_uid=29633078

https://www.ncbi.nlm.nih.gov/pubmed/29633078

https://www.ncbi.nlm.nih.gov/pubmed/?term=Durg%20S%5BAuthor%5D&cauthor=true&cauthor_uid=28376400

https://www.ncbi.nlm.nih.gov/pubmed/?term=B%20NK%5BAuthor%5D&cauthor=true&cauthor_uid=28376400

https://www.ncbi.nlm.nih.gov/pubmed/?term=Vandal%20R%5BAuthor%5D&cauthor=true&cauthor_uid=28376400

https://www.ncbi.nlm.nih.gov/pubmed/?term=Dhadde%20SB%5BAuthor%5D&cauthor=true&cauthor_uid=28376400

https://www.ncbi.nlm.nih.gov/pubmed/?term=Thippeswamy%20BS%5BAuthor%5D&cauthor=true&cauthor_uid=28376400

https://www.ncbi.nlm.nih.gov/pubmed/?term=Veerapur%20VP%5BAuthor%5D&cauthor=true&cauthor_uid=28376400

https://www.ncbi.nlm.nih.gov/pubmed/?term=Badami%20S%5BAuthor%5D&cauthor=true&cauthor_uid=28376400



https://www.tandfonline.com/author/Ansari%2C+M+Nazam

https://www.tandfonline.com/author/Bhandari%2C+U


2591


28. Sandeep Arora, Preparation and Characterization of Phytosomal-Phospholipid Complex of P.

Amarus and its Tablet Formulation, Journal of Pharmaceutical Technology, Research and

Management, Vol. 1 May 2013; 1–18.

29. Tawheed Amin, Suman Vikas Bhat, A Review on Phytosome Technology as a Novel

Approach to Improve The Bioavailability of Nutraceuticals, International Journal of

Advancements in Research & Technology, 2012; 1(3): 1-15.

30. Rajendra Awasthi, Giriraj T Kulkarni, Vivek K Pawar, Phytosomes: an approach to increase

the bioavailability of plant extracts, International Journal of Pharmacy and Pharmaceutical

Sciences, 2011; 3(2): 1-3.

31. Nilesh Jain Brahma P Gupta, Navneet Thakur, Ruchi Jain, Jitendra Banweer, Deepak Kumar

Jain, Surendra Jain, Phytosome: A Novel Drug Delivery System for Herbal Medicine,

International Journal of Pharmaceutical Sciences and Drug Research, 2010; 2(4): 224-228.

32. S Ambati; V Jyothi; A Jyothi, Int. J. Pharm. Tech, 2010; 2: 525-539.

33. B Lal, N Mishra; Int. J. Pharm. Sci. Res, 2013; 4: 3823-3838.

34. R Shelar R, C. Maurya, P. Tekale, K Katkar, V Naik, A Suthar, VS Chauhan; Int. J. Pharm.

Clin. Res, 2009; 1: 146-149.

35. S Asadulla; S Ramandang; Rajasekharan, Int. J. Res. Pharm. Chem, 2011; 1: 2236-2751.

36. MA Khan; A Naidu; M Akbar, Int. J. Pharm. Bio. Arch, 2010; 1: 267-270.

37. D Annapurna; A Srivastava; RT Singh, American J Plant Sci, 2013; 4: 28-35.

38. Vidya V. Kamble, and Nikhil B. Gaikwad. Fluorescence analysis, phytochemical, and

antioxidant activities in leaves and stem of embelia ribes burm. F. Asian Journal of

Pharmaceutical and Clinical Research, 2019; 12(4): 225-9.

CHARACTERIZATION AND DEVELOPMENT OF EMBELIA RIBES …

Documents

Transcript of CHARACTERIZATION AND DEVELOPMENT OF EMBELIA RIBES …