World Journal of Pharmaceutical Research

Rishi et al. SJIF Impact

World Journal of Pharmaceutical Factor 8.074
Research
Volume 8, Issue 9, 812-825. Research Article ISSN 2277– 7105

PURIFICATION CHARACTERIZATION AND ANALYTICAL

EVALUATION OF SUSPENDING PROPERTIES OF AZADIRACHTA
INDICIA AND ACACIA NILOTICA GUM

Punet Kumar1, Rishi Kumar*3, Vidhan Chand Bala2, Sangam1 and Harjeet Singh3

1
Department of Pharmaceutical Chemistry, Faculty of Oxford College of Pharmacy, Hapur
U.P. India.
2
Department of Pharmacology, Faculty of Oxford College of Pharmacy, Hapur U.P. India.
3
Department of Pharmaceutics, Faculty of Oxford College of Pharmacy, Hapur U.P. India.

ABSTRACT
Article Received on
24 May 2019, Aim: The present research work was aimed at purification
Revised on 14 June 2019,
characterization and analytical evaluation of the suspending properties
Accepted on 04 July 2019,
DOI: 10.20959/wjpr20199-15399 of Azadirachta indicia and Acacia nilotica gum. Methods: The present
study aims to extract the gum from Acacia nilotica and Azadirachta

*Corresponding Author indicia and then characterized to be used as an excipient. This study
Rishi Kumar also aimed to search the cheap and effective excipients and evaluate
Department of the suspending properties of the different natural polymer (Acacia
Pharmaceutics, Faculty of
nilotica gum, Azadirachta indicia) on paracetamol suspension and
Oxford College of
compared with each other Acacia nilotica gum and Azadirachta indicia
Pharmacy, Hapur U.P. India.
at the deliberation of 1-4%w/v be used to practice the suspension in
which paracetamol is used as a model drug. Prepared suspensions were evaluated for
sedimentation volume, flow rate, viscosity, and redispersibility number. The values
performed are used as the basis for evaluating the suspended agents studied. Results: The
results of the physiochemical investigation confirm that polysaccharides were absent in both
polymers. Results obtained from the micromeritic characterization and flow behaviour
showed that Acacia nilotica gum is a good candidate as compared to Azadirachta indicia to
be used as a pharmaceutical excipient. Particles suspended with Azadirachta indicia gum at
concentration 4%w/v re-dispersed easily than those formulated with the Acacia nilotica gum.
Conclusions: Evaluation of suspending properties of both the gums suggests that Neem gum
has good suspending properties than that of Babool gum. Therefore it is confirmed that both
the gums possess good properties to use them as a pharmaceutical excipient.

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KEYWORDS: Polymer, suspending, sedimentation, Excipient, Acacia nilotica, Azadirachta

indicia.

INTRODUCTION
Polymers are widely found in nature. In recent year, natural polymers play an important role
in the pharmacy field and are widely used as a pharmaceutical excipient. When two or more
than two smaller molecules combined together they form a large molecule. Direct attachment
of the monomer unit to the small molecules helps in the formation of the large molecule unit;
this process is known as polymer synthesis. Many no glucose monomer joins together to form
starch molecules. A natural polymer such as proteins and nucleic acid is also found in the
human body.[1-2] Natural herbal polymers have many important applications in various forms
of pharmaceutical dosage such as embarrassed matrix systems, film coating agents, buccal
films, microspheres, nanoparticles, viscose liquid formulations, such as ophthalmic solutions,
suspensions, implants and also used as viscosity stimulators, stabilizers, Stabilizer,
Solubilizers, emulsifiers, suspension agents, gelling, Bioadhesives and binders in the above
dosage forms.[3-4] They are non-toxic, biodegradable, accessible and cost-effective for use as
pharmaceutical excipients.[5] Acacia, Neem gum, Babool gum, gum karaya, tragacanth, gum
ghatti are some examples of plant gums. Natural gums extracted from plants are high
molecular weight of hydrogenated carbohydrate polymers. They are generally composed by
joining the monosaccharide unit through glycosidic bond. They are insoluble in organic
solvents such as hydrocarbons, ethers or alcohols and are also insoluble in oils.[6]

Suspending agent is defined as a formulation for reducing the sedimentation rate in the
suspension of the particles. These formulations are natural polysaccharides, semi-synthetic or
synthetic materials.[7] like other distributed systems, the pharmaceutical suspensions are also
thermodynamic and unstable, and if necessary, suspensions or stabilizers are added to the
dosage form.[22] the stabilizer reduces the rate of sedimentation and makes any sedimentary
particle material easy to redistribute by improving the consistency of the suspended medium.
The gum consists mainly of calcium, potassium salt and other basic acids and magnesium.[8-9]
Gums are primarily used to increase the suspension of insoluble substance in liquid drug
preparations.[10] In this work, Azadirachta indicia and Acacia nilotica gums regulations that
are as natural polymers and paracetamol taken as a research model for drug requires a
suspending agent in preparing the liquid dosage form.[11]

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MATERIAL AND METHODS

Collection, Identification and purification of plant material
Crude plant materials (Acacia nilotica gum and Azadirachta indicia) were purchased from
the local shop of Ghaziabad, India. Collected plant materials were identified by Department
of Biotechnology, Gautam Buddha University (State Govt. University) Greater Noida.
Furthermore, the plant material was purified by the following procedure.

As the author describes elsewhere, plant raw materials were soaked in hot water for 4 hours,
boiled for 2 hours and held aside for 2 hours for the release of gums into the water.
Afterwards, the material is pressed into a muslin bag to remove the mark from the strain. For
the isolation of gums, the same amount of ethyl alcohol is added to the flat to separate the
gums. After separation, the gums are dried in the oven at 45˚C, powder and filtered in sieve
no. 80. Gum powder has been stored in the desicator until further use.[12]

Physicochemical Properties Characterization of Isolated Gums

Tests for the identification of carbohydrates, proteins and tannins: As identified by the
author elsewhere, aqueous solution (1%) of extracting gums was utilized for chemical
characterization. Carbohydrate, protein and tannin tests are carried out using standard
procedures.

Organoleptic Evaluation of Isolated gum: As studied elsewhere, isolated gums are

characterized by biological properties such as colour, smell, taste, fracture and texture.

pH of gum: As reported by the author elsewhere, the pH of a 1% w/v gum solution

was evaluated using a digital pH meter.[13]

Swelling Index of Isolated gum: As studied elsewhere, the swelling index is the volume (in
ml) absorbed by the swelling of 1 g of test material under specified conditions. The gum
swelling index was determined by accurately weighing 1 g of gum, which was further
introduced into a 25 ml glass-Stoppard measuring cylinder. 25 ml of water has been added
and the mixture is thoroughly displaced every 10 minutes for 1 hour. It was then allowed to
stand for room temperature for 3 hours. Then the volume inhabited by the gum was
measured. The same procedure was repeated three times and the average value was
calculated.[14]

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Bulk density and bulkiness: As the author describes, the volume is the opposite of bulk
density. For the determination of bulk density, the volume of 4g was correctly introduced in a
graduated measurement glass and the cylinder was attached to the mass density apparatus.
The amount of dust was noted.[12]

Tapped density: As described by the author elsewhere, the density of pressure was calculated
by pressing the powder in a bulk density device until the fixed volume was reached. The final
amount has been noted.[15]

Powder flow property: As the author describes elsewhere, the property powders flow was
calculated by measuring the angle of repose. The use of the resting angle of the type was
calculated three times.

Powder compressibility: As the authors describe elsewhere, the compressibility of the

powder is measured by the Carr Index. This finely powder, gum (4g) is transferred to the
measuring cylinder and is calculated using a density unit.[13]

Viscosity of gum: As identified by the author elsewhere, the viscosity was measured 1% w/v
solution of the gums by using an Ostwald Viscometer.

Surface tension of gum: As the authors describe elsewhere, the surface tension of the
solution 1% w / v gum was tested by Stalagmometer.[13]

Particle size analysis: As described elsewhere, the particle size was determined using
microscopy.[15]

Ash value: As described by the author elsewhere, ash value was calculated by weighing
2,2gms of Acacia nilotica and Azadirachta indicia powder in a tarred silica crucible. These
were then incinerated in a muffle furnace up to 450 °C till the powder completely changes to
ash. The crucibles were then kept in desicator after complete incineration. Weight of ash was
noted and total ash was calculated in terms of percentage.[16-17]

IR: As described elsewhere, Acacia nilotica and Azadirachta indicia powders were dried in
oven at 70-80 °C for 4 hr and desiccated over night prior to FTIR analysis. FTIR spectra
were recorded at the absorbance mode from 400 to 4000 cm-1.[18]

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Preparation of suspension: Paracetamol suspensions were prepared by adding natural

polymer (0.5g) in 3ml distilled water, triturated with mortar pastel. Add paracetamol (1g),
triturated well and transferred in to the 10ml measuring cylinder. Prepared
with distilled water, and stirred for 2 minutes, thus 1% w/v gum is prepared. The procedure
was repeated in natural gums, in 2%, 3% and 4%.[17]

Evaluation of Suspension Properties

Sedimentation volume: As described by the author elsewhere, 10 ml of each suspension was
stored in a 10 ml volumetric cylinder for 7 days at 35oC. Observations are performed every
24 hours. For 7 days. Sedimentation volume F (%) was calculated on the basis of the
following equation1.

Where it is,
Vu is the total amount of the sediment (funnel volume) and Vo is the original amount of
suspension.

Re-dispersion number: As the author described elsewhere, after recording the sedimentation
volume for a week, the suspensions were re-dispersed and the re-dispensable number was
determined for each of them. This was done by reversing the cylinders and observing the
number of times the cylinders were reversed until the bottom of the cylinder was free of
particles.[19]

Rheology: As described by the author elsewhere, the time required for each flow of sample
suspension through a 5 ml, determined pipette and the actual viscosity (1 gm per cm per Sec)
is calculated using Equation 2.

The viscosity (in prose) of the samples was determined using the Ostwald Viscometer. All
determinations have been made at least three times and the results are expressed as the
average rates.[20-21]

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RESULT AND DISCUSSION

After isolation of gum by ethyl alcohol, the percentage yield of gum was found to be 48%.
The phytochemical examination of isolated gum showed the presence of carbohydrates,
lacking the reduction of sugar, glucose, tannins, proteins, polysaccharides, fats and volatile
oils. Organoleptic property of gum was found to be acceptable which is shown in table1 and
phytochemical tests are summarized in table 2.

Table 1: Organoleptic properties of Azadirachta indicia and Acacia nilotica gum.

Polymer Colour Odour Taste Texture Fracture
Azadirecta indica Ambered /light cream Odourless Not bitter Irregular Rough
Acacia nilotica Off white Odourless Tasteless Irregular Smooth

Table 2: The chemical properties of the isolated Azadirachta indicia gum and Acacia
nilotica.
Test Azadirachta indicia Acacia nilotica
Carbohydrate + +
Reducing sugar + -
Glucose + -
Tannins - -
Proteins - -
Polysaccharide - -
+ Present, -Absent tests.

The dissolution profile of the gums is shown in table 3. As a result, Azadirachta

indicia and Acacia nilotica gum are shown to swell to form a gel when dissolved in cold
water. Both gums are insoluble in organic solvents.

Table 3: Solubility profile of Azadirachta indicia and Acacia nilotica gum.

Solubility
Solvents
Azadirachta indicia Acacia nilotica
Cold water Swell to form gel Swell to form gel
warm water Soluble Soluble
Methanol Insoluble Insoluble
Ethanol Insoluble insoluble
Diethyl ether Insoluble Insoluble
Petroleum ether Insoluble Insoluble
Acetone Insoluble Insoluble

The pH of the solution of 1% of Acacia nilotica gum and the Azadirachta indicia was found
to be 6.3 ± 0.057 and 6.80±0.06 indicating that the Mucilages were less irritating in GIT. Ash
value of isolated gums was found 3.5 ± 0.144% and 2.50±0.10 respectively. Swelling index
of gums was found to be 23.07 ± 0.0115% and 14.28±0.02 for isolated gums. Bulk density

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and tapped density were calculated as 0.67 ± 0.0057 g/cm3 and 0.87 ± 0.023 g/cm3 for
Acacia nilotica gum and 0.59±0.02 and 0.79±0.02 for Azadirachta indicia. Bulkiness was
established to be 1.47 ± 0.0115 cm3/g and 1.70±0.02. Hausner’s ratio and Carr’s index
measured be 1.28 ± 0.0305 and 22.48 ± 1.788% for Acacia nilotica gum and 1.38±0.01 and
1.38±0.01 for Azadirachta indicia. Angle of repose of isolated gums was found to be 2.84 ±
0.0709 and 3.08±0.11 respectively suggested that the powdered gums possess excellent flow
property. The average size of 50 particles calculated were 62.30 ± 0.05 for Acacia nilotica
gum and 31.65±0.07for Azadirachta indicia. Surface tension of isolated gums calculated
were 71.22 ± 3.083dyne/cm and 88.38±0.05dyne/cm and viscosity were 5.04 ± 0.01poise and
7.76±1.47 poise respectively. Evaluation of the different gum parameters and shown in Table
4. Figure 1, Figure 2 shows the IR spectrum of purified gums shown in Table 5, 6.

Table 4: Different parameter of Acacia nilotica gum and Azadirichta indicia gum.
Observations
Parameters
Acacia nilotica gum Azadirichta indica gum
pH 6.30±0.05 6.80±0.06
Total Ash (%) 3.50±0.14 2.50±0.10
Bulk density (g/cm3) 0.67±0.01 0.59±0.02
tap density (g/cm3) 0.87±0.02 0.79±0.02
Bulkiness (cm3/g) 1.47±0.01 1.70±0.02
Hausner’s ratio 1.28±0.03 1.38±0.01
Carr’s index (%) 22.48±1.79 27.85±1.52
Angle of repose (°) 2.84±0.07 3.08±0.11
Particle size (µm) 62.30±0.05 31.65±0.07
Surface tension (dyne/cm) 71.22±3.08 88.38±0.05
Viscosity (poise) 5.04±0.01 7.76±1.47
Swelling Index (%) 23.07±0.01 14.28±0.02

Figure 1: IR spectra of Acacia nilotica gum.

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Acacia nilotica gum IR spectra show wave numbers (cm-1) 1034.47, 1397.55, 1455.88,
1514.96, 1645.14, 1740.63 etc. Confirming the presence of an Amines, Keaton, anhydride,
carboxylic acid, and aromatic rings.

Table 5: IR study of Acacia nilotica gum.

S. No. Wavenumber (cm-1) Functional Group
1. 1034.47 -C-O(alcohol)
2. 1397.55 -C-NO2
3. 1455.88 -C=C (aromatic)
4. 1514.96 -C-C (aromatic)
5. 1559.73 -C-O (stretching)
6. 1645.14 -C=C
7. 1706.13 -C=O
8. 1740.63 -C=O (anhydride)
9. 1796.16 -C=O (Ketone)
10. 2361.92 -COOH
11. 3735.62 -NH2

Figure 2: IR spectra of Azadirichta indicia.

Azadirichta indicia gum IR spectra shows the wave numbers (cm-1)

1338.76,1387,73,795.29.1425.26,1454.98 etc, which confirms the presence of Carboxylic
acid, Amine, Alcohol, Ketone, Alcohol, Aromatic ring etc.

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Table 6: IR study of Azadirichta indicia gum.

S. No. Wavenumber (cm-1) Functional Group
1. 1387.73 -C-NO2
2. 1454.98 -C=C(Aromatic)
3. 1514.57 -C-C(Aromatic)
4. 1645.18 -C=C
5. 1792.34 -C=O(Ketone)
6. 1740.09 -C=O(Anhydride)
7. 2360.42 -COOH
8. 3596.20 -OH
9. 3754.29 -NH2

Suspensions of Babool and Neem gum in the concentration range of 1, 2, 3, 4% w/v were
made in batches where paracetamol was used as a model drug. Based on their sedimentation
volume, flow volume, viscosity and re-dispersion number, the following preparations were
evaluated.

Sedimentation and flow volume: Table 7 shows the results of suspension sedimentation rate
experiments developed with Acacia nilotica and Azadirichta indicia at the same
concentration. It was observed that the suspension formulated with Azadirichta indicia 1, 2, 3
and 4%w/v have high sedimentation rate than Acacia nilotica at same concentrations. The
flow rate at the highest concentration of each suspension agent demonstrates the fact that
Azadirichta indicates that the gum is a slightly weak flow rate compared to Acacia nilotica
gum when performing a suspension using paracetamol as a exemplary drug. Suspension
using different concentrations of (a) Babool gum and (b) Neem gum in fig. 3.

(a) Babool gum (b) Neem gum

Figure 3: Suspension using different concentration of (a) Babool gum and (b) Neem
gum.

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The result of flow rate of the formulated suspension with Babool gum and Neem gum at
different concentration are shown in Table 7.

Table 7: Values of Sedimentation volume (%) of the suspension using dissimilar

concentrations of Acacia nilotica and Azadirichta indicia gum.
Concentration Time (Days)
Polymers
(%w/v) 1 2 3 4 5 6 7
1 28 27 26 25 23 22 22
2 25 24 24 23 22 21 21
Babool gum
3 24 23 22 20 20 18 18
4 22 21 21 19 18 17 16
1 27 23 22 21 20 20 20
2 24 20 21 19 18 18 18
Neem gum
3 18 16 16 16 16 14 14
4 17 15 15 15 15 14 13

Table 8: The effects of the type and concentration of suspensions on the number of
suspensions of paracetamol.
Suspending agents Concentration(%w/v) Flow rate (ml s-1)
1 2.61
2 2.23
Babool gum
3 2.04
4 1.87
1 2.40
2 2.14
Neem gum
3 1.97
4 1.85

Redispersibility Number: The redispersed test indicates how easily a suspension is

redispersed after it is left to stand for a week. It was observed that particles suspended in
Acacia nilotica and Azadirichta indicia gum in concentrations 1, 2 and 3% w / v were easily
redispersed. Therefore suspension prepared with Acacia nilotica and Azadirichta indicia gum
at concentration 1%w/v was easily redispersed. The result of redispersibility number is
shown in table 9.

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Table 9: Redispersibility number of prepared suspension using Acacia nilotica and

Azadirichta indicia gum.
Suspending Concentratio Redispersibility
agents n (%w/v) number
1 3
2 5
Babool gum
3 12
4 16
1 3
2 5
Neem gum
3 12
4 10

Viscosity: The results of the viscosity of the suspension formulated in Acacia nilotica and
Azadirichta indicia gum are shown in table 10. It is observed that with the gum concentration
the viscosity of the suspension has increased respectively. The viscosity of Acacia niloticais
more as compare to Azadirichta indicia gum at different concentrations. Therefore, at
different concentrations, the viscosity of the gums decreases inversely proportional to the
sedimentation rate of the suspension, since the viscosity of the charge increases the
sedimentation rate of the suspension.

Table 10: Effect of different polymer suspension concentrations on viscosity.

Suspending agent Concentration(%w/v) Viscosity(Poise)
0.25 7.95
0.40 9.50
Babool gum
0.50 9.71
1.00 10.9
0.25 5.97
0.40 6.53
Neem gum
0.50 8.56
1.00 9.67

CONCLUSION
It was concluded from the research work that the organoleptic properties of both the extracted
gums were found to be acceptable. The phytochemical examination of both gums shows the
presence of carbohydrates, while the reducing of sugars, sugars, tannins, proteins,
polysaccharides, fats and volatile oils is absent. Different parameters are examined for both
rubbers, such as pH, swelling index, particle size, Ash Value, bulk density, tapped density,
powder flow properties, etc. The pH was found to be somewhat acidic for both gums. Total
ash was in limits. The flow property of Acacia nilotica (Babool gum) was found to be good in

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comparison with Azadirichta indicia (Neem gum). IR spectra of both the gums confirmed the
presence of Carboxylic acid, Amine, Ketone, anhydride and Aromatic ring. Evaluation of
suspending properties of both the gums suggests that Neem gum has good suspending
property than that of Babool gum. Therefore it is confirmed that both the gums possess good
properties to use them as pharmaceutical excipient.

ACKNOWLEDGEMENT
Authors are thankful to Dr. Arun Kumar Mishra, School of Pharmaceutical science, IFTM
University Moradabad U.P., INDIA and Dr. Ram Dayal Gupta, Oxford college of Pharmacy,
Ghaziabad. U.P., INDIA.

CONFLICT OF INTEREST
Authors declared no conflict of interest or otherwise.

FINANCIAL SUPPORT
Authors declared no financial support or otherwise.

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