Aegle Marmelos - Excipient2

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JOURNAL OF NATURAL REMEDIES

DOI: 10.18311/jnr/2023/31704 RESEARCH ARTICLE

Formulation and Evaluation of Sustained Release


Linezolid Tablet using Natural Antibacterial
Polymer - Aegle marmelos
Shrey Patel and Rupal Jani*
Department of Pharmaceutics, Parul Institute of Pharmacy and Research, Parul University,
Waghodia, Vadodara - 391760, Gujarat, India; [email protected]

Abstract
The objective of the current research study was to formulate and evaluate a sustained-release linezolid tablet using
the natural antibacterial polymer Aegle marmelos. The natural gum of Aegle marmelos is becoming increasingly
used in pharmaceutical formulations as a beneficial medication with excipients. Natural-based plant materials
are biocompatible, free of side effects, biodegradable, and economic. Therefore, in order to maintain the drug
releases from the matrix system, Aegle marmelos fruit gum as a natural polymer and HPMC grade (K100M) as a
synthetic polymer were used in the formulation of the linezolid matrix tablet. The formulation of sustained-release
matrix tablets included the wet granulation technique. The formulated matrix tablets were evaluated in terms of
weight variation, hardness, diameter, physical appearance, friability, thickness, and in vitro drug release. Each
formulation’s matrix tablet passed the required physical assessment tests. The formulation analyses of the tablets’
dissolution showed sustained releases of drugs for up to 10–12 hours. Additionally, several polymer combinations
and fillers were used to improve drug release factors using the 32 factorial design approach, drug release kinetics
were optimized, and the antibacterial study was evaluated.

Keywords: Aegle marmelos, Fruit Gum, HPMC K100M, Linezolid, Sustained Release

1. Introduction plasma drug concentration at a therapeutically acceptable


level2. Sustained-release tablets offer patients comfort
One of the most widely used methods of medication while also increasing compliance with cost-effective illness
administration is the oral route of drug delivery due to treatment. The two primary categories of sustained release
its convenience for patient compliance, administration, tablets - active drug dissolution and dissolved drugs - that
cost-effectiveness, self-medication, and flexible design are used to categorise and control drug release are further
of the dosage form. More than ninety percent (90%) of clarified by the processes. These mechanisms could
today’s formulations are taken orally1. It demonstrates function singly or in succession3. Linezolid (Figure 1)
that the formulation’s greatest popularity, availability in is regarded as the first member of the oxazolidinone
all countries, simplicity of production, dosage accuracy, antibiotic family. Its treatment involves simple infections
ease of administration, and improved stability have drawn of the skin and skin structure. Impetiginous lesions, simple
the majority of the researcher’s focus in this area. These abscesses and cellulitis furuncles are bacterial infections
medications act as a reservoir, releasing medication of the skin and adjacent tissues that are often caused by
continuously over a sufficient period of time to keep the Streptococcus pyogenes and Staphylococcus aureus4.

Article Received on: 21.10.2022 Revised on: 18.01.2023 Accepted on: 03.02.2023
192 Formulation and Evaluation of Sustained Release Linezolid Tablet using Natural Antibacterial Polymer...

is sweet, the odour is aromatic and minute oil glands dot,


hard woody shell of the fruit surface, as well as colour, is
grey-green until completely ripe when they turn yellow10.
In one study, the release retardant properties of Aegle
marmelos fruit gum were investigated in manufactured
sustained-release formulations. The sustained-release
method of the matrix tablets was shown to be enhanced
Figure 1. Structure of linezolid. for up to 10–12 hours11. According to release rate
The substance is a synthetic antibiotic that binds to characteristics, swelling behavior, and an in vitro
rRNA to prevent bacteria from synthesizing proteins5. dissolution investigation, to create sustained release
Additionally, it stops the initiation complex during protein matrix tablets, the fruit gum from Aegle marmelos is an
synthesis, which can shorten the formed peptide chains appropriate matrix-forming ingredient. According to
and slow down the pace at which translation elongation the kinetics of the chosen formulation, which followed
occurs6. Linezolid promotes diabetic foot infections, limb- zero order, Aegle marmelos gum is an outstanding
threatening infections, pneumonia, and bloodstream matrix-forming polymer that has been shown to delay
infections, and is effective for the symptomatic treatment the release of pharmaceuticals from the formulations12.
of skin structure bacterial infection. Rapid heartbeat and Wet granulation, which gives improved formulation
extreme sickness. Backaches, burning or soreness, frequent efficacy, was employed in this research to formulate
urination, trouble peeing, and all of the above7. Due to the matrix tablets. The composition of linezolid matrix
its rapid plasma elimination and high water solubility, tablets includes the addition of a natural polymer, Aegle
linezolid is a useful model medication for creating marmelos fruit gum, as well as synthetic hydroxypropyl
controlled-release dosage forms. Linezolid is classified as methylcellulose, a hydrophilic polymer, in grades with
a Class I drug by the BCS and exhibits high solubility and varying viscosities and concentrations13. The study’s goal
permeability8 as well as 100% oral bioavailability. Linezolid was to develop a novel formulation of prolonged-release
mostly bounds to serum albumin, accounting for 31% of linezolid tablets and assess their effectiveness using Aegle
its total binding. A hydroxyethyl glycine metabolite, and marmelos, a naturally occurring anti-bacterial polymer.
an aminoethoxyacetic acid metabolite, both of which are
formed by morpholine ring oxidation, are the two inactive 2. Material and Method
metabolites that linezolid mostly breaks down into. Most
likely formed by non-enzymatic processes, hydroxyethyl 2.1 Materials
glycine is the more abundant of the two metabolites6. The fresh Aegle marmelos fruit was collected from
Linezolid does not appear to be susceptible to metabolism Gujarat, India. Linezolid, Hydroxypropyl Methylcellulose
via the CYP450 enzyme system, nor does it appear to (HPMC), calcium phosphate dibasic, talc, and magnesium
substantially inhibit or stimulate these enzymes, even stearate were obtained from Alembic Pharmaceuticals
though the precise enzymes involved in linezolid’s bio- Pvt. Ltd., Vadodara, as gift samples. Acetone (99.5%) and
transformation are unknown. However, the reversible, AR-grade chemicals, respectively. The experiment also
non-selective inhibitor of monoamine oxidase enzymes is included the use of distilled water.
linezolid5.
Natural gum is now being used more often in
2.2 Methodology
pharmaceutical formulations as a useful medicinal
excipient. Natural plant-based substances are more cost- 2.2.1 Aegle marmelos Fruit Extraction
effective, free of side effects, biodegradable, renewable, Ripe bael fruits were taken from the A. marmelos tree and
biocompatible, processed in an environmentally used to make bael fruit gum (Figure 2). The hard-wooded,
responsible way, as well as better tolerated by patients. spherical fruits were cut into two halves. The pulp and
Aegle marmelos fruit gum (Rutaceae family) is frequently seeds separating the fruit’s exterior wall, coupled with
known as bael9. Its plant part is used as a fruit, which is the amber-coloured viscous, extremely sticky, gummy
oval, or oblong in shape, 50-20 cm in diameter, the taste material, transparent, were designated as the desirable

Journal of Natural Remedies | ISSN: 2320-3358 http://www.informaticsjournals.com/index.php/jnr | Vol 23 (1) | January 2023
Shrey Patel and Rupal Jani 193

2.2.3.1.3 Detection of Alkaloids


Wagner’s test: Take 1-3 ml of filtrate sample with a few
drops of Wagner’s reagent to give a reddish brown ppt that
is formed.
Dragendroff ’s test: Take 2-3 ml of the filtrate sample
and add a few drops of Dragendroff ’s reagent. An orange-
brown precipitate is formed.

2.2.3.1.4 Detection of Tannins


Ferric chloride test: The sample should first be dissolved
in water and ethanol, and then drops of a diluted ferric
Figure 2. Aegle marmelos fruit. chloride solution (FeCl3) should be added. Phenols are
present when the sample develops a red, green, purple, or
section. This gum and the seeds were collected in a beaker blue coloration.
filled with a glacial acetic acid solution that was 2% (v/v). 2.2.3.1.5 Detection of Gums
The solution (slurry) was heated in a water bath for 40-45 Ruthenium test: Observe a little volume of dried gum
minutes with stirring, then it was allowed to cool. To powder under a microscope after mounting it on the slide
avoid particles, the slurry was filtered through muslin with ruthenium red solution. If a pink coloration appears,
fabric. After adding acetone, the filtering slurry produced mucilage is present15.
the gum, which then precipitated. The precipitates were
crushed into a light brown, fine powder after being baked 2.2.3.1.6 Detection of Chloride
in an oven at 50°C14. Add a few drops of the 10% AGNO3 solution to the 3 ml
of HNO3-prepared test solution. AgCl2 is noticed as white
2.2.2 Characterization of Aegle marmelos Gum dilutable ppts in nitrate solution16.
2.2.2.1 Taxonomical Classification 2.2.3.1.7 Detection of Sulphate
The collected Aegle marmelos fruit was classified for its Fill the 5 ml filter with a few drops of the 5% BACl2
family, genus, class, kingdom, species, and order. solution. BASO4 appears as a white crystalline ppt. that is
insoluble in HCL. Lead acetate with the reagent results in
2.2.3 Physical Characterization a white ppt that is soluble in NAOH15.
The collected gum was assessed for its physical properties
2.2.3.2 Flow Properties
including solubility, odour, appearance, smell, percentage
The flow parameters of the dried Aegle marmelos fruit
yield, the percentage loss of drying, and moisture content.
gum, including the angle of repose, tapped density, bulk
All values were assessed.
density, Carr’s index, and Hausner’s ratio, were assessed.
2.2.3.1 Preliminary Phytochemical Characterization 2.2.3.2.1 Determination of Swelling Index
2.2.3.1.1 Detection of Carbohydrates The volume in milliliters filled by 1 gm of medication,
Molisch’s test: 1 ml of the extract, 2 drops of Molisch’s including any adherent mucilage, is known as the swelling
reagent, and 2 ml of pure (H2SO4) sulfuric acid were index, after 4 hours of swelling in an aqueous liquid. 1 gm
carefully added along with the side of the test tube. At of gum from each sample was allowed to swell for 4 hours
the intersection, a violet ring that formed suggested the with a small quantity of water, and then 1 ml of ethanol was
presence of carbohydrates. added to each cylinder. After a few minutes, water was added
to the measurement cylinders to bring the volume up to 25
2.2.3.1.2 Detection of Protein ml. This was vigorously shaken once every ten minutes for
Millon’s test: To mix 1-2 ml of sample, add 1 ml of an hour and then left standing for 24 hours. An occupied
Million’s reagent, if required, heat the solution till it boils volume was calculated. The average of 3 measurements was
and observe the pink to brick red precipitate. used to determine of the swelling index17.

Journal of Natural Remedies | ISSN: 2320-3358 http://www.informaticsjournals.com/index.php/jnr | Vol 23 (1) | January 2023
194 Formulation and Evaluation of Sustained Release Linezolid Tablet using Natural Antibacterial Polymer...

Swelling Index % (SI) = V2/V1 2.2.3.2.5 Carr’s Index


V1 = Initial Volume in ml Compressibility, which is used to determine how smoothly
V2 = Final Volume in ml a powder flows, may be calculated using the formula
below,
2.2.3.2.2 Angle of Repose Carr’s index = Dt – Db/Dt × 100
It is a useful tool for calculating frictional force in loose Where, Dt = Tapped density of the powder,
powder. This is the greatest angle that may be formed Db = Bulk density of the powder.
between a pile of powder or grains’ surface and the
horizontal plane18. 2.2.3.2.6 Hausner Ratio
tanθ = h/r The Hausner ratio is a proximate indicator of powder flow
θ = tan-1 (h/r) ease. The formula used to compute it is as follows18.
Where, θ is the angle of repose, Hausner ratio = Dt/Db
h is the height of the pile, Where Dt = Tapped density.
r is the radius of the base of the pile. Db = Bulk density.
Each sample’s 5 g of powder was poured onto the
graph paper using a funnel at a set height. Measurements 2.2.4 Method of Preparing the Calibration Curve
were taken of the mounds’ heights. A pencil was used to In a dry container, 100mg of pure medication
outline the piles’ perimeters. The large and small squares (Linezolid) that had been precisely weighed was
that were present inside the produced circles were used to consumed. The medication was put in a pristine
compute the areas of the circles, and the parameter ‘r’ that 100 mL volumetric flask. Using the solvent 0.1N HCl
was determined from the area of the circle was then used and the phosphate buffer solution pH 6.8, the volume is
to calculate the angle of repose. properly made up to 100 ml, and after that, it receives
a few slow knocks. Let the 0.1N HCl and pH 6.8 buffer
2.2.3.2.3 Bulk Density (Db) solution thoroughly dissolve the tablet. This solution
Each 50 g of gum was weighed precisely before being contains 1000 g/mL of solutions. The stock solution
placed into a graduated measuring cylinder. The cylinders was formed by pipetting 10 ml of the aforementioned
were fastened to the bulk density device, and the powder’s solution into 100 ml of the flask and filling the remaining
volume was recorded. The initial volume is the bulk volume with 0.1N HCl and buffer solution pH 6.8. This
volume. The following formula is used to calculate the formed the stock solution, which had a concentration of
bulk density using this information. 100 mg/ml20.
Db = M/Vb
Where M = Mass of powder, 2.2.4.1 Working Standard Solution
In each case, transfer 0.2, 0.4, 0.6, 0.8, 1.0, 1.2, 1.4, 1.6,
Vb = Bulk volume of the powder.
and 1.8 ml of the solution to a separate 10 ml volumetric
flask. The volume is precisely made up of 10 ml. Different
2.2.3.2.4 Tapped Density (TD)
concentrations of the stock solution were collected,
Following the bulk volume notation, the same number of
including 2 g/ml, 4 g/ml, 6 g/ml, 8 g/ml, 10 g/ml, 12 g/ml,
samples (each weighing 50 g) were placed into graduated
16 g/ml, and 18 g/ml.
measurement cylinders. The powder was tapped 100
times in a bulk-density device after that. The completed 2.2.4.2 Procedure of Standard Plot
volumes were recorded. It is computed using the formula
Using a UV-visible spectrophotometer, different
below19.
concentrations were created by properly diluting working
Dt = M/Vt standard solutions. The data was then plotted using the
Where M = Mass of powder, typical absorbance vs. concentration chart.
Vt = Volume of the powder.

Journal of Natural Remedies | ISSN: 2320-3358 http://www.informaticsjournals.com/index.php/jnr | Vol 23 (1) | January 2023
Shrey Patel and Rupal Jani 195

2.2.5 Fourier Transforms Infrared Spectroscopic 2.2.7.2 Tablet Thickness


Studies (FTIR) The thickness of the tablets was measured using the
To explore the interaction and compatibility between Vernier calliper. The average value was determined, and
excipients and drugs, FTIR spectra for both the drug (5) five pills are needed. Tablet variations might lead to
and the excipients will be recorded using an FTIR issues with counting and packing. Tablet thickness needs
spectrophotometer and KBr pellets. 4000-400 cm-1 on to be kept within 5% of a defined value22.
the spectrometer. The KBr will be taken and maintained
in a hot-air oven for 2 hours to remove moisture content 2.2.7.3 Hardness
before being used in the KBr press to press pellets for Using a Monsanto hardness tester, the tablets’ hardness
the FTIR research. The medication pellets and chosen was measured. The unit of measurement is kg/cm2. 10 pills
formulation excipients will next be prepared using the are chosen at random, and their hardness is measured.
aforementioned dried KBr. It should be documented how The tablet passed the test for hardness test and was found
the infrared experiments for the medicine and powder to be resistant to mechanical shocks23.
mixture turned out21.
2.2.7.4 Weight Variation
2.2.6 Preparation of Linezolid Tablet Choosing 20 tablets at random and weighing them
The Linezolid sustained-release matrix tablets were individually to check for weight variation24.
created using the wet granulation method. Both natural
2.2.7.5 Friability
polymers and artificial polymers were employed in
Twenty pills were randomly chosen and weighed from
varying percentages of 20%, 30%, 40%, and 50%. A
each batch. The Friabilator is used to test the friability
total of 9 formulations were created, utilizing linezolid
of tablets for 100 rotations. For 4 minutes, the friability
at a constant dose of 300 mg together with various
is run at 25 (revolution per minute) rpm. The tablets are
excipient doses. Aegle marmelos gum and Hydroxypropyl
thrown from a height of six inches on each revolution
Methylcellulose (HPMC) are the polymers utilized in
while being subjected to a plastic chamber that combines
formulations (Table 1).
the effects of abrasion and shock. The pills were taken
out, cleaned, and weighed once more. It is stated as a
2.2.7 Post-compression Evaluation
(%) percentage. Tablets with less than 1% friability are
2.2.7.1 Physical Appearance deemed acceptable25.
The tablets are visually observed for changes in color,
(initial weight)-(final weight)
capping, lamination and chipping. %Friability= × 100
(initial weight)

Table 1. Formulation of linezolid sustained release matrix tablets


Components F1 F2 F3 F4 F5 F6 F7 F8 F9
Linezolid 300 300 300 300 300 300 300 300 300
Aegle marmelos fruit gum 70 80 70 80 70 80 70 75 75
HPMC K100M 10 10 20 20 15 15 10 20 15
PVP K30 25 15 15 5 20 10 20 10 15
Dicalcium phosphate 10 10 10 10 10 10 10 10 10
Magnesium stearate 10 10 10 10 10 10 10 10 10
Talc 5 5 5 5 5 5 5 5 5
Isopropyl alcohol q.s q.s q.s q.s q.s q.s q.s q.s q.s
Total 430 430 430 430 430 430 430 430 430

Journal of Natural Remedies | ISSN: 2320-3358 http://www.informaticsjournals.com/index.php/jnr | Vol 23 (1) | January 2023
196 Formulation and Evaluation of Sustained Release Linezolid Tablet using Natural Antibacterial Polymer...

2.2.7.6 In Vitro Dissolution Study Table 3. Independent and dependent variables of


In vitro dissolution of tablet formulations was studied formulation
using a rotating paddle (USP technique II). The linezolid Amount of Amount of
dissolving medium would comprise phosphate buffers polymer polymer
Level
with a pH of 1.2 and 6.8. Given that the medicine is only Aegle HPMC
moderately soluble at low pH levels, this pH value was marmelos (X1) K100M (X2)
selected to produce the highest volume of drug release Independent
Low 70 10
Variable
from the tablets. The dissolving medium temperature Medium 75 15
was kept at 37± 0.5 °C, with the spinning speed set at High 80 20
75 rpm. Using filter paper, 5 ml samples are collected at 2
Dependent Hardness (kg/cm )
intervals of 0.5, 1.0, 1.5, 2.0, 3.0, 4.0, 5.0, 6.0, 8.0, 10.0,
Variable % Cumulative Drug Release at 10-12 Hrs
12.0, 18.0, and 24 h. Then, using a spectrophotometric
instrument with a 251 nm wavelength, all samples were
examined26.
2.2.9 Kinetic Modelling of in vitro Drug Release
2.2.9.1 Zero Order Kinetics
2.2.8 Factorial Design Drug dissolution from dosage forms that do not
32 Full Factorial Design was employed, using the amount disaggregate and release the drug slowly can be represented
of Aegle marmelos gum in mg and the amount of HPMC by the equation (1):
K100M in mg as independent variables. Hardness Qt  Q0  K 0t    (1)
(kg/cm2) and % cumulative drug release at 10-12 hrs were
dependent variables. The responses were assessed using Where Qt is the amount of drug dissolved in time t,
an interactive and polynomial statistical model (Tables 2 Q0 is the initial amount of drug in the solution (most
and 3). times, Q0 = 0) and
K0 is the zero-order release constant expressed in units
Y = 0 +1X 1 + 2 X 2 +12 X 1X 2 +11X 12 + 22 X 22 + of concentration/time.
Where Y is the dependent variable, X1 and X2 are the To study the release kinetics, data obtained from in
independent variables26. vitro drug release studies was plotted as the cumulative
amount of drug released versus time.

Table 2. 32 Full factorial design layout 2.2.9.2 First-Order Kinetics


32 Full Factorial Design Layout This model describes the absorption and/or elimination of
some drugs, although it is difficult to this mechanism on a
Independent Variable
theoretical basis on the release of the drug which followed
Amount first-order kinetics can be expressed by the equation (2):
Amount of
Batch No. Coded Coded of HPMC
Aegle marmelos Kt
value value K100M Log C  log C0     (2)
gum (mg) 2.303
(mg)
F1 -1 70 -1 10 Where C0 is the initial concentration of the drug,
F2 -1 70 0 15 k is the first-order rate constant, and t is the time.
F3 -1 70 1 20 The data obtained are plotted as a log cumulative
percentage of drug remaining vs. time, which would yield
F4 0 75 -1 10
a straight line with a slope of -K/2.30327.
F5 0 75 0 15
F6 0 75 1 20 2.2.9.3 Higuchi Model
F7 1 80 -1 10 Accordingly, the model expression is given by the
F8 1 80 0 15 equation (3):
F9 1 80 1 20 ft  Q  A D (2C  Cs ) Cst    (3)

Journal of Natural Remedies | ISSN: 2320-3358 http://www.informaticsjournals.com/index.php/jnr | Vol 23 (1) | January 2023
Shrey Patel and Rupal Jani 197

Where Q is the amount of drug released in time t per Table 4. T axonomical classification of Aegle
unit area A, marmelos gum
C is the drug initial concentration,
Kingdom Plantae
Cs is the drug solubility in the matrix media and
Subfamily Aurantioideae
D is the diffusivity of the drug molecules (diffusion
coefficient) in the matrix substance. The data obtained Order Sapindales
were plotted as cumulative percentage drug release versus Family Rutaceae
square root of time28. Genus A. marmelos
Species Aegle species
2.2.9.4 Korsmeyer-Peppas Model Tribe Clauseneae
To find out the mechanism of drug release, the first 60%
of drug release data were fitted in the Korsmeyer-Peppas Table 5. P
 hysical characterization of Aegle marmelos
mode of the equation (4): fruit gum
Mt Parameters Observation of gum
 Kt n    (4)
M Color Yellowish brown
Where Mt/M∞ is a fraction of the drug released at Odour Characteristic
time t, Taste Characteristic
k is the release rate constant and Nature Amorphous
n is the release exponent.
The n value is used to characterize different releases Table 6. P
hytochemical characterization of Aegle
for cylindrical-shaped matrices28. marmelos fruit gum
Test Observed gum
2.2.10 Antibacterial Study (Zone of Inhibition) Carbohydrates Present
Mueller–Hinton agar plates were used for the test. Glycoside Absent
After pouring approximately 20 ml of molten agar Tannins Absent
media into sterile Petri dishes, the agar plates were
Saponins Absent
allowed to solidify under a laminar flow hood. It had
Protein Present
been demonstrated with 105 organisms/ml suspensions
of each strain of Methicillin-Resistant Staphylococcus Gum Present
aureus (MRSA). The inoculum of these cultures was Polysaccharide’s Present
spread on the surface of the agar plates. Plates were
allowed to air dry for 10 minutes. After creating a zone Table 7. Flow properties of Aegle marmelos fruit gum
in each plate, pouring the drug on it, and placing it Parameter Observed gum
in the incubator at 37.8°C for 24 hours, the zones of
pH (1% w/v solution) 6.4
inhibition (diameter in mm) are measured on the agar
Swelling index 16.8%
surface with a scale29.
Bluk density 0.47 ± 0.01 (g/cm3)
Tapped density 0.56 ± 0. 01
3. Results and Discussion Hausner’s ratio 1.19 ± 0. 006
3.1 Gum Characterization Carr’s index 16.23 ± 0. 44

3.1.1 Taxonomical Classification Angle of repose 28.09 ± 0.70

It is based on the taxonomy of Aegle marmelos, which The physicochemical properties and micromeritics
is classified as belonging to the Kingdom of Plantae, properties of gums were performed as per the procedure.
Order of Sapindales, and family Rutaceae. The detailed It was observed that gum was acidic in nature but they
classifications were shown in Table 4. Characterization are near neutral. Information on the pH of polymers is an
and flow properties are shown in Tables 5, 6 and 7. important parameter in determining their appropriateness

Journal of Natural Remedies | ISSN: 2320-3358 http://www.informaticsjournals.com/index.php/jnr | Vol 23 (1) | January 2023
198 Formulation and Evaluation of Sustained Release Linezolid Tablet using Natural Antibacterial Polymer...

in formulations, while the stability and physiological Sr. No. Concentration (ug/ml) Absorbance
action of most preparations are influenced by pH. The pH 6 12 0.693±0.0012
of the 1% w/v test solution of gum in water was 6.4. The
7 14 0.792±0.0012
neutral pH suggested that when it was used in uncoated
8 16 0.923±0.0012
tablets, it may be less irritating to the GI tract. It may find
beneficial application in the formulation of acidic, basic Calibration curve,
and neutral drugs. Aegle marmelos (16.8)% gum has a Correlation Coefficient (R²) = 0.999
good swelling index. The gum had a high swelling index, Where y = Absorbance   X = Concentration (ug/ml)
suggesting that the gums may perform well as matrixing
agent. Tapped density and bulk density of gum were
found to be 0.47 ± 0.01 and 0.56 ± 0.01, respectively, and
results of Hausner’s ratio, angle of repose, and Carr’s index
showed passable flow properties of gum powder.

3.1.2 FTIR Spectroscopy Study (Figure 3, Table 8)

Figure 4. Calibration curve of linezolid in 0.1 N HCL.

Table 10. Absorption of the drug in phosphate buffer 6.8

Figure 3. FTIR spectra of Aegle marmelos gum. Sr. No. Concentration (ug/ml) Absorbance
1 2 0.309±0.0012
Table 8. Wave number of Aegle marmelos gum
2 4 0.385±0.0016
Sr. No. Functional group present Wave numbers (cm–1) 3 6 0.466±0.0012
of gum
4 8 0.537±0.0008
1 OH – Stretching 3362.05
5 10 0.616±0.0016
2 C-H Stretching 2931.58
6 12 0.686±0.0016
3 C=O Stretching 1746.86
7 14 0.777±0.0012
4 C-O Stretching 1673.69
8 16 0.905±0.0012
5 C-C Deformation 1421.25
9 18 0.984±0.0012
6 Secondary OH 1022.75

3.1.3 Calibration curve of Linezolid (Figure 4,


Table 9)

Table 9. Absorption of the drug in 0.1 N HCL


Sr. No. Concentration (ug/ml) Absorbance
1 2 0.108±0.0012
2 4 0.235±0.0012
3 6 0.336±0.0012
4 8 0.465±0.0012
5 10 0.567±0.0008
Figure 5. Linezolid in phosphate buffer 6.8.

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Shrey Patel and Rupal Jani 199

The regression coefficient value was found to be 0.999 3.1.4 Factorial Design Formulations of Linezolid
and the line equation was y = 0.057x + 0.00 for HCL and y Sustained Release Matrix Tablet (Figure 6,
= 0.041x + 0.208, R² = 0.993 for a buffer, R2 value of drug Tables 11, 12 and 13)
showed linearity of the standard curve (Figure 5, Table 10).

Table 11. Results of powder blends of factorial design tablet


Batch No. Angle of Repose (°) Bulk Density (g/ml) Tapped Density (g/ml) Carr’s Index (%) Hausner Ratio
F1 28.15±0.04 0.394±0.02 0.465±0.03 15.26±0.03 1.18±0.02
F2 29.23±0.02 0.395±0.04 0.471±0.03 16.13±0.02 1.19±0.02
F3 30.65±0.03 0.356±0.03 0.468±0.02 23.93±0.03 1.31±0.03
F4 30.44±0.04 0.384±0.02 0.472±0.03 18.64±.0.02 1.22±0.02
F5 34.9±0.03 0.378±0.03 0.571±0.02 33.80±0.03 1.51±0.03
F6 36.75±0.02 0.394±0.02 0.471±0.02 16.34±0.02 1.19±0.04
F7 29.85±0.04 0.395±0.03 0.471±0.03 16.34±0.03 1.19±0.03
F8 30.65±0.03 0.384±0.02 0.472±0.02 18.64±0.02 1.22±0.03
F9 30.86±0.02 0.355±0.04 0.448±0.03 20.75±0.03 1.26±0.02
(Where n = 3, Mean ± SD)

Table 12. Results of post-compression of factorial design tablet


Batch no. Weight variation Friability (%) Thickness (mm) Hardness (kg/cm3) Drug content
F1 429±0.01 0.464±0.04 3.5±0.04 5.3±0.02 91.02
F2 427±0.05 0.465±0.05 3.6±0.03 7.2±0.03 96.65
F3 429±0.02 0.466±0.06 3.4±0.02 5.8±0.02 93.21
F4 429±0.02 0.464±0.05 3.7±0.03 8.0±0.03 98.52
F5 422±0.01 0.463±0.06 3.6±0.04 5.5±0.02 92.86
F6 429±0.02 0.465±0.04 3.5±0.06 7.7±0.03 97.86
F7 431±0.01 0.465±0.05 3.6±0.04 6.0±0.03 95.01
F8 428±0.02 0.463±0.05 3.8±0.05 6.9±002 96.11
F9 431±0.04 0.464±0.04 3.6±0.03 6.2±0.02 95.65

Table 13. Results of in vitro dissolution study of factorial design batch F1-F9
Time (Hours) F1 F2 F3 F4 F5 F6 F7 F8 F9
0 0 0 0 0 0 0 0 0 0
1 7.95 8.06 8.18 7.92 7.96 7.59 8.03 9.45 8.02
2 19.82 18.75 20.95 19.82 17.78 20.36 18.75 22.75 31.75
3 22.97 22.09 27.45 22.97 31.32 32.02 35.39 26.54 39.48
4 33.56 36.55 35.69 33.58 39.49 42.45 36.52 36.25 48.36
5 40.35 39.54 39.46 40.58 48.35 49.88 45.62 46.15 56.45
6 47.45 46.48 46.83 47.45 56.45 58.45 55.38 50.66 67.78
7 53.65 52.68 58.45 53.65 67.88 69.55 56.55 58.23 73.26
8 65.26 66.49 63.68 65.26 73.28 75.95 67.96 68.36 78.76
(Continued )

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200 Formulation and Evaluation of Sustained Release Linezolid Tablet using Natural Antibacterial Polymer...

Table 13. (Continued)


Time (Hours) F1 F2 F3 F4 F5 F6 F7 F8 F9
9 72.36 70.05 71.45 72.36 78.85 81.16 74.56 75.06 83.85
10 79.55 73.65 76.69 79.55 83.85 85.16 83.25 80.23 87.48
11 86.45 85.36 83.68 86.45 87.48 89.26 87.45 87.65 91.82
12 90.22 97.12 92.54 98.29 91.25 92.76 93.56 96.25 94.23

Figure 6. Drug release profile of batches F1-F9.

3.1.5 32-factorial Design (Figures 7 and 8)


• Response 1: Hardness

  
Figure 7. C
 ontour plot for the effect of concentration of gum (X1) and concentration of polymer (X2) on the hardness.

Journal of Natural Remedies | ISSN: 2320-3358 http://www.informaticsjournals.com/index.php/jnr | Vol 23 (1) | January 2023
Shrey Patel and Rupal Jani 201

• Response 2 - % CDR

Figure 8. Contour plot for effect of gum (X1) and amount of HPMC K100M (X2) on % cumulative drug release at 10-12 hrs.

Figure 9. Overlay plot showing the combined effect of Aegle marmelos gum (X1) and polymer (X2).

Discussion: The overlay plot revealed that the area of


interest is in the “yellow zone” of Figure 9. Formulation show the validity of the produced model. Batch F4 was
having a concentration of Aegle marmelos gum (X1) (80) selected as the optimized formulation (Figure 10).
and an amount of polymer (X2) (20 mg) with a hardness Table 14. Results of checkpoint batch OF1 and OF2
of 8.0 kg/cm2, and in the experimental area of the overlay
plot, the %CDR is 97.29% showed the greater acceptability Batch Predicted Experimental
Parameters
than other checkpoint batches. It was therefore chosen as Code Value Value
batches OF1 and OF2 (Table 14). Hardness 7.92 8.01±0.03
Discussion: The checkpoint batch findings showed
OF1 % cumulative drug
no significant difference between the two results. As a 98.63 98.12±0.87
release up to 12 hrs.
result, in certain variable ranges, the equation obtained
Hardness 7.56 7.35±0.52
for specific responses is validated. The similarity between
OF2 % cumulative drug
observed and expected response values was used to 97.58 96.98±0.42
release up to 12 hrs
measure the robustness of prediction and these statistics

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202 Formulation and Evaluation of Sustained Release Linezolid Tablet using Natural Antibacterial Polymer...

Figure 10. Tablets of Optimized formulation batch F4.

3.1.6 Kinetic Modelling of In Vitro Drug Release (Table 15)


Table 15. Release kinetics of linezolid from sustained release matrix tablets
Zero-order kinetics First-order kinetics Higuchi kinetics Korsmeyer–Peppas kinetics
Formulation
R2 R2 R2 R2 n
F1 0.9961 0.7975 0.9782 0.8363 1.30
F2 0.9921 0.8323 0.9663 0.8366 1.13
F3 0.9939 0.8065 0.9739 0.7835 1.18
F4 0.9972 0.7960 0.9696 0.8425 1.15
F5 0.9948 0.7787 0.9819 0.6835 1.21
F6 0.9705 0.8978 0.9949 0.7635 1.23
F7 0.9861 0.7888 0.9875 0.7585 1.29
F8 0.9949 0.7877 0.9838 0.5862 1.35
F9 0.9412 0.7960 0.9874 0.8030 1.37

The mechanism of drug release from the sustained- the release of drug from matrix tablets of Aegle marmelos
release matrix tablet, formulations were subjected to gum contained tablets fitted best into zero-order kinetics
modelling and drug release kinetic studies such as with maximum values of R2 = 0.9972 and a n value of 1.15.
zero-order, first-order, the Higuchi model, and the It was revealed that the mechanism of drug release was
Korsmeyer–Peppas model. The value of the Regression a super case-II transport, indicating the drug release rate
coefficient (R2) from the drug release kinetic model was does not change over time and the release is attributed to
tabulated. The result of the modelling study revealed that zero-order.

3.1.7 Antibacterial Study (Figure 11, Table 16)

     
(a) (b) (C)
Figure 11. Zone of inhibition (a) Linezolid, (b) AMG, (c) Formulation.

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Shrey Patel and Rupal Jani 203

Table 16. Zone of inhibition of different sample 2. Bangale GS, Rathinaraj SB, Shinde GV, Umalkar DG,
Rajesh KS. Formulation and evaluation of natural
Sr. No. Sample Zone of inhibition gum based matrix tablets for oral controlled delivery
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