Formulation Development and In-Vitro Evaluation of Tetrazosin Loaded Chitosan Nanoparticles For The Treatment of Hypertension

World Journal of Pharmaceutical Research
Marabathuni et al. World Journal of Pharmaceutical Research

SJIF Impact Factor 8.084
Volume 12, Issue 17, 939-955. Research Article ISSN 2277– 7105
FORMULATION DEVELOPMENT AND IN-VITRO EVALUATION OF

TETRAZOSIN LOADED CHITOSAN NANOPARTICLES FOR THE
TREATMENT OF HYPERTENSION
Daggumati Pravallika1, *V. Jhansi Priya Marabathuni and Naidu Narapusetty
1
Department of Pharmaceutics, Bellamkonda Institute of Technology and Science, Podili. A.
P-523240.
ABSTRACT
Article Received on
12 August 2023, The purpose of this research was to prepare Tetrazosin loaded Chitosan
Revised on 01 Sept. 2023, Nanoparticles for controlled release of drug, to improve the solubility,
Accepted on 22 Sept. 2023
DOI: 10. 20959/wjpr202317-29825
reduce the dosing frequency, thereby increasing patient compliance to
the therapy. Tetrazosin is formulated as Nanoparticles by ionic-
gelation method using Chitosan as polymer, Sodium tripolyphosphate
*Corresponding Author
as a polyanionic agent (cross linking agent) and the lyophilized
V. Jhansi Priya
Marabathuni nanoparticles filled in hard gelatin capsules. The particle size analysis
Department of was done by Horriba scientific Nano SZ 100 particle size analyzer
Pharmaceutics, showed that mean particle size 188.3 nm and Z- Average 229.0 nm
Bellamkonda Institute of
respectively. The Zeta potential study was done by Horriba scientific
Technology and Science,
Nano SZ 100. The Zeta potential for the optimized formulations F5
Podili. A. P-523240.
was found to be 25.8mV and shows that the formulation is stable. Post
formulation parameters (uniformity of weight, disintegration test, drug content, and in vitro
drug release) for nano particulate capsules were evaluated. The results were found to be
complying with official specifications. The dissolution data of the optimized formulation was
fitted to various kinetic models and the formulation F5 was best fitted to Zero order kinetics.
The slope of the Korsmeyer Peppas plot indicating the diffusion was Anomalous diffusion
(Non-Fickian diffusion). From the overall results, it is clear that the formulations F5
containing 0.3% polymer concentration (Chitosan) is the optimal formulation, as it produces
controlled drug release.
KEYWORDS: Chitosan Nanoparticles, particle size Zeta potential, kinetic study, Non-
Fickian diffusion.
www.wjpr.net │ Vol 12, Issue 17, 2023. │ ISO 9001:2015 Certified Journal │ 939
INTRODUCTION
A drug delivery system (DDS) is defined as a formulation or a device that enables the
introduction of a therapeutic substance in the body and improves its efficacy and safety
by controlling the rate, time and place of release of drugs in the body. This process
includes the administration of the therapeutic product, the release of the active
ingredients by the product, and the subsequent transport of the active ingredients across
biological membrane.[1]
The objective of any drug delivery system is to deliver a therapeutic amount of drug to
the sites of action and to maintain the desired amount of drug level in the tissue or the
body that can elicit a desired pharmacological effect without causing any serious adverse
reactions.
A perfect drug delivery system possesses two elements: the capacity to target and to
control the release of drug. Targeting will ensure high efficiency of the drug and reduce
the side effects, especially when dealing with drugs that are presumed to kill cancer cells
but can also kill healthy cells when delivered to them. The side effects can be reduced or
prevented by the control of drug release.[2]
Drawbacks associated with conventional dosage forms[3]

An ideal dosage regimen in the drug therapy of any disease is one which immediately attains
the desired therapeutic concentration of drug in plasma and maintains it constant for the
entire duration of treatment. This is possible through the administration of drug delivery
system in a particular dose and at particular frequency. The frequency of administration or
dose interval of any drugs depends upon its life or mean residence time and its therapeutic
index. In most cases, dosing interval is much shorter than the half-life of the drug, resulting in
number of limitations associated with such a conventional dosage form which are, A drug
with short biological half-life which needs a close succession.
The uncontrolled fluctuation of drug level may leads to either below effective range or over
the effective range.
Plasma concentration verses time profile of dosage form and it is difficult to achieve the
steady state active drug level. The rise and fall of drug levels may give to accumulation of
adverse effects, especially for a drug having less therapeutic index. To overcome the above
drawbacks, drug delivery system capable of controlling the rate of drug delivery, sustain the
duration off therapeutics action or targeting the drug to a particular tissue was developed.
Terazosin is a member of quinazolines, a member of piperazines, a member of furans and a

primary amino compound. It has a role as an antineoplastic agent, an antihypertensive agent
and an alpha-adrenergic antagonist. It is chemically known as [4-(4-amino-6,7-
dimethoxyquinazolin-2-yl)piperazin-1-yl]-(oxolan-2-yl)methanone.
Fig. 1: Structure of Terazosin.
MATERIALS AND METHODS

Materials Used In Formulation
Tetrazosin collected from Active pharmaceutical, Bangalore; Chitosan 50k, Acetic acid,
Sodium tripolyphosphate, Ethanol, Sodium Hydroxide and Distilled Water from Lab
Chemicals, Chennai, and Potassium dihydrogen phosphate were collected from Merck
specialities Pvt. Ltd, Mumbai.
Determination of Melting point[68]

The melting point of Tetrazosin was determined by the capillary tube method as per USP. A
Sufficient quantity of Tetrazosin powder was filled into the capillary tube to give a compact
column of 4-6 mm in height. The tube was introduced in electrical melting point apparatus
and the temperature was raised. The temperature at which the last solid particle of Tetrazosin
in the tube passed into liquid phase was noted as melting point.
Preparation of 6.8 pH Phosphate buffer[69]

0.2M solution of potassium dihydrogen phosphate was prepared by dissolving 27.218gm of
substance in 1000ml of distilled water.0.2M solution of sodium hydroxide solution was
prepared by dissolving 8gm of substance in 1000 ml of distilled water. 250 ml above
prepared potassium dihydrogen phosphate solution & 112 ml of sodium hydroxide solution
were mixed together and made up to 1000ml and pH was adjusted to 6.8.
Determination of lambda max (λmax)[32]

100 mg of Tetrazosin was weighed and transferred to 100ml of volumetric flask. The drug
was dissolved in 10 ml of ethanol and volume was made up to 100ml using phosphate buffer
pH 6.8 to obtain a stock solution of 1000µg/ml (stock solution I). 10ml of this stock solution
was again diluted with phosphate buffer pH 6.8 up to 100ml to obtain a solution of 100µg/ml
(stock solution II). From the stock solution-II, 10 ml was pipette out in 100ml volumetric
flask. The volume was made up to 100 ml using phosphate buffer pH 6.8 get a concentration
of 10µg/ml. this solution was then scanned at 200-400nm in UV-Visible spectrophotometer
to attain the absorption maxima (λmax).
Standard curve for Tetrazosin[33]

100 mg of Tetrazosin was weighed and transferred to 100ml of volumetric flask. The drug
was dissolved in 10 ml of ethanol and volume was made up to 100ml using phosphate buffer
pH 6.8 to obtain a stock solution of 1000µg/ml (stock solution I). 10ml of this stock solution
was again diluted with phosphate buffer pH 6.8 up to 100ml to obtain a solution of 100µg/ml
(stock solution II). From the stock solution-II 2, 4,6,8,10,12 ml were transferred to series of
100 ml volumetric flasks. The volume was made up with phosphate buffer pH 6.8. The
absorbance of these solutions was measured at 268nm against the blank.
Solubility studies of pure Tetrazosin[70]

Solubility of Tetrazosin pure drug was tested in distilled water and phosphate buffer pH 6.8.
An excess amount of Tetrazosin pure drug was added in the pertinent media. The mixtures
were stirred in a mechanical shaker at speed 50 rpm for 24 hours and the temperature was
maintained at 37±0.5ºC. Visual inspection was carefully made to ensure there were excess
Tetrazosin solids in the mixture, indicating saturation had been reached. Then the mixtures
were filtered using 0.45µm filter and filtrates were suitably diluted with same media. The
absorbance of the solution was measured at 268nm in UV-Visible spectrophotometer.
Formulation Development
Preparation of Chitosan Nanoparticles – Ionic gelation Method[4,5,6]
The preparation of Chitosan nanoparticles was based on ionic interaction between positively
charged Chitosan solution and negatively charged STPP solution, with and without drug and
it was prepared in the presence of Tween 80 as a re-suspending agent to prevent particle
aggregation, at ambient temperature while stirring and Chitosan solution were raised to pH
4.6 to 4.7. Seven formulations (F1,F2,F3,F4,F5,F6,F7) of Tetrazosin loaded Chitosan
nanoparticles were prepared by dissolving Tetrazosin in 30ml of Chitosan with varying

concentrations (0.1,0.15,0.2,0.25,0.3,0.4,0.5%w/v) containing 0.5% w/v tween 80,
TPP(0.1%w/v) was added drop wise under magnetic agitation(1000 rpm).The formed
nanoparticle suspensions were lyophilized at -40oC for 24hrs.
Table No.1: Composition of Nanoparticles.

Chitosan 1% Aqueous DRUG TWEEN 80 TPP Distilled TPP
Formulation Chitosan
(%w/v) acetic acid (ml) (mg) (%w/v) (%w/v) water (mg)
F1 0.10% 30ml 30 20 0.5 0.1 30 30
F2 0.15% 30 ml 45 20 0.5 0.1 30 30
F3 0.20% 30 ml 60 20 0.5 0.1 30 30
F4 0.25% 30 ml 75 20 0.5 0.1 30 30
F5 0.30% 30 ml 90 20 0.5 0.1 30 30
F6 0.40% 30 ml 120 20 0.5 0.1 30 30
F7 0.50% 30 ml 150 20 0.5 0.1 30 30
Characterization of tetrazosin loaded chitosan nanoparticle

Percentage yield[43]
The nanoaparticle yield was calculated according to the equation given below.
Determination of Drug Content

Equivalent to 60 mg of the prepared formulation were weighed and dissolved in minimum
quantity of ethanol mixture and made up to 100ml with phosphate buffer pH 6.8. The
solution kept for 24 hours and filtered to separate fragments. Drug content was analyzed after
suitable dilution by UV- Visible spectrophotometer at a wave length 268nm against
phosphate buffer pH 6.8 as blank. From the absorbance the drug content in the batches were
calculated.
Drug entrapment efficiency

For the determination of drug entrapment, the nanosuspension with known amount of drug
was centrifuged at 4000 rpm for 15 minutes. The supernatant solution was separated. 5ml of
supernatant was distributed with 100ml of phosphate buffer solution pH 6.8 and the
absorbance was measured using UV-Visible spectrophotometer at 268nm using of phosphate
buffer solution pH 6.8 as blank. The amount of drug unentrapped was calculated. The
percentage of entrapment efficiency was determined according to the following equation

given below.
amount of unbound
total amount of drug - drug
% Drug Entrapment = ×100
total amount of drug
Solubility Studies of Tetrazosin loaded Chitosan Nanoparticle

The solubility of the Tetrazosin loaded Chitosan nanoparticles formulations were tested in
various medium (distilled water and phosphate buffer pH 6.8) by adding an excess amount of
formulations. The mixtures were stirred in a mechanical shaker at speed 50 rpm for 24 hours
at room temperature. Visual inspection was carefully made to ensure there were excess
Tetrazosin solids in the mixture, indicating saturation had been reached. Then the mixtures
were filtered using 0.45µm filter and filtrates were suitably diluted with same media. The
absorbance of the solution was measured at 268nm in UV-Visible spectrophotometer.
In vitro drug release studies

The in vitro release rate studies of Tetrazosin loaded Nanoparticles formulations were carried
out by dissolution test apparatus USP Type-I (Basket). Tetrazosin loaded Chitosan
Nanoparticles were filled in capsule and placed in a dissolution medium and rotated at
100rpm. 10 ml of samples were withdrawn predetermined intervals up to 12 h and replaced
with equal amount of phosphate buffer pH 6.8 for further dissolution testing the absorbance
determined by spectrophotometrically at 268nm.
FTIR study of optimized formulations

FT-IR spectra of optimized formulations of Chitosan Nanoparticle were recorded by grinding
and dispersing the samples with micronized IR grade Potassium bromide powder and
subjected to FT-IR measurement over the range of 4000- 400cm-1.
Surface Morphology by SEM analysis

The Surface Morphology of the Chitosan nanoparticle can be measured by SEM at Anna
University (Model Vega3 – Tescan, USA). Scanning Electron Microscopy was used to
analyse particle size, shape and surface morphology of Nanoparticles. The sample was
mounted directly onto the SEM sample holder using double sided sticking tape and image
were recorded at different magnification at acceleration voltage of 10 kv using scanning
electron microscope.
Particle size characterization

The samples of the optimized formulations were analyzed for their particle size using Horiba
Scientific SZ-100 particle size analyzer, Particle size (Z-average diameter), Polydispersity
index (as a measure of the width of the particle size distribution) of Tetrazosin loaded
Chitosan Nanoparticles dispersion is performed by dynamic light scattering also known as
photon correlation spectroscopy (PCS) using a Horiba Scientific Nano SZ-100 at 25°C.
Prior to measurements all samples were diluted using ultra – purified water to yield a suitable
scattering intensity. The diluted nanoparticles dispersion was poured into the disposable
sizing cuvette which is then placed in the cuvette holder of the instrument and analyzed. Air
bubbles were removed from the capillary before measurement.
Polydispersity Index (PDI)

PDI indicates the width of the particle size distribution, which ranges from 0 to1.
Monodisperse samples have a lower PDI value, whereas higher PDI value indicates a wider
particle size distribution and the polydisperse nature of the samples can be calculated by
following equation:
PDI = d/d avg
Where,
d is the width of distribution donated by SD
d avg is the average particle size denoted by MV (nm) in particle size data sheet.
Zeta potential
Zeta Potential is a crucial factor to evaluate the stability of colloidal dispersion surface
charge on the Tetrazosin loaded chitosan NPs were determined using Horiba Scientific Nano
ZS100. 1 ml of sample of Tetrazosin suspension was filled in clear disposable zeta cell
ensured there was no air bubble within the sample and the system was set at 25°C
temperature and the test can be carried.
RELEASE KINETICS OF THE OPTIMIZED FORMULATIONS

Different kinetic models such as zero order (cumulative amount of drug released vs. time),
first order (log cumulative percentage of drug remaining vs. time), Higuchi model
(cumulative percentage of drug released vs. square root of time), Korsmeyer-Peppas model
and Hixson Crowell model were applied to interpret the drug release kinetics from the
formulations. Based on the highest regression values for correlation coefficients for
formulations, the best‐ fit model was decided.
To study the in vitro release kinetics of the optimized formulation, data obtained from
dissolution study were plotted in various kinetics models.
 Zero-order
 First-order
 Higuchi
 Hixson-Crowell cube root law
 Korsmeyer-Peppas model
RESULTS AND DISCUSSION

Pre-Formulation Studies
The optimization of a formulation can be done only after a thorough investigation of its
physicochemical properties of the drug and excipients. The drug and the polymer must be
compatible for a successful formulation.
COMPATIBILITY STUDIES
Physical Compatibility study
Inference
The Physical compatibility study was performed for 3 months. There was no change of
color therefore the drug and excipients are physically compatible with each other.
Chemical Compatibility Study

FT-IR spectroscopic study
FT-IR spectroscopy gives the possible information about the interaction between the drug
and polymer. The results are follows
Figure 2: FT-IR spectrum of Tetrazosin.
MELTING POINT- The melting point of Tetrazosin was measured using capillary tube
method in the range 157-158ºC.
SOLUBILITY STUDY
The solubility study of Tetrazosin in different dissolution medium is performed by
saturation solubility method.
Table 2: Saturation solubility of Tetrazosin in phosphate buffer pH 6.8 and Distilled

water.
Medium pH Solubility (mg/ml)
Phosphate buffer pH 6.8 6.8 0.004093
Distilled water 7.0 0.000216
Inference
The solubility of the drug at pH 6.8 was significantly higher than in that of distilled water.
Pure drug of Tetrazosin in distilled water and phosphate buffer pH 6.8 was found be
insoluble.
DETERMINATION OF LAMBDA MAX (λmax) FOR TETRAZOSIN

The maximum absorbance of the Tetrazosin was studied. The maximum absorbance of the
Tetrazosin was found to be 268nm. Hence the wavelength of 268nm was selected for
analysis of drug in dissolution media.
Calibration Curve For Tetrazosin

Table 3: Data for standard curve of Tetrazosin in Phosphate buffer pH 6.8.
S.No Concentration (µg/ml) Absorbance
1 0 0
2 2 0.115
3 4 0.2203
4 6 0.3445
5 8 0.4686
6 10 0.6035
7 12 0.7089
Mean ± SD (n=3)
Figure 3: Standard curve of Tetrazosin in Phosphate buffer pH 6.8.
Inference
The constructed calibration curve of Tetrazosin in Phosphate buffer pH 6.8 is shown figure
9.5. It was found that the solutions show linearity (R2 = 0.9995) in absorbance at a
concentration of 2-12 µg/ml and obeys Beer-Lambert’s law.
SOLUBILITY STUDIES OF TETRAZOSIN LOADED NANOPARTICLES

The solubility study of Chitosan Nanoparticles formulations in distilled water and phosphate
buffer pH 6.8 were studied by saturation solubility method. The Nanoparticles formulations
compared with pure and tabulated below.
Table 4: Solubility of Nanoparticles and Tetrazosin in various medium.

Solubility Medium
S. No Formulation code Distilled water Phosphate buffer
pH 7 (mg/ml) pH 6.8 (mg/ml)
1 Pure drug 0.000216 0.004093
2 F1 6.0998 6.9676
3 F2 6.7923 7.4355
4 F3 7.5888 8.7722
5 F4 7.9351 9.6748
6 F5 9.4933 13.251
7 F6 7.5190 8.1373
8 F7 6.4802 7.6360
Inference
The solubility of Tetrazosin in distilled water and phosphate buffer pH 6.8 were found to be
0.000216 mg/ml and 0.004093 mg/ml respectively. The solubility of all formulations was
improved (from insoluble to slightly soluble) compared to pure drug of Tetrazosin. Among
all the formulations F5 show higher solubility in distilled water and phosphate buffer pH 6.8.
Inference
Thus the solubility of formulation F5 in Distilled water and Phosphate buffer pH 6.8 were
improved and compared with pure drug.
COMPARATIVE IN VITRO DRUG RELEASE FOR ALL FORMULATION

The formulated Nanoparticles preparation containing drug and polymer were evaluated for
drug release and results were tabulated below
Table 5: In vitro drug release for all formulations
Time
In Vitro Drug Release For Nanoparticles Formulations
(h)
F1 F2 F3 F4 F5 F6 F7
0.5 17.25 17.45 16.04 15.09 13.19 9.71 8.32
1 20.12 19.89 19.2 18.99 16.17 11.14 10.14
2 36.4 29.13 28.83 27.02 24.02 17.35 14.14
3 44.8 38.03 37.92 36.21 34.52 31.3 29.27
4 60.2 50.08 47.22 44.1 41.23 34.02 31.73
5 72.27 63.07 59.12 53.8 50.14 36.37 32.13
6 82.29 79.02 69.03 61.78 57.71 42.13 39.92
7 97.04 89.9 78.97 73.2 64.21 50.12 43.78
8 - 96.02 89.17 86.51 73.22 62.79 59.11
9 - - 94.18 91.52 78.01 71.23 69.24
10 - - - 94.47 86.22 79.13 75.17
11 - - - - 91.89 83.2 81.2
12 - - - - 95.03 86.03 83.04
In-vitro Drug Release

The formulations F1 TO F7 were prepared using Chitosan as a polymer (0.1%, 0.15%,
0.2%, 0.25%, 0.3%, 0.4% and 0.5%) and the Sodium tripolyphosphate as a cross linking
agent.
Figure 4: In vitro release study of Chitosan Nanoparticles (F1-F7).
Inference
 The in vitro drug release profile for formulated Tetrazosin loaded Chitosan Nanoparticles
obtained from F1-F7 formulations were shown in figure 4.
 Among all the formulations F5 formulations shows 95.03% of drug release at the end of
12h in controlled manner. Thus F5 was selected as the optimized formulation.
CHARACTERIZATION OF OPTIMIZED TETRAZOSIN LOADED CHITOSAN

NANOPARTICLES
FTIR Study of Optimized NPs
FTIR of optimized formulations of Tetrazosin loaded Chitosan Nanoparticles (F5)
Figure 5: FT-IR spectral of Tetrazosin + Chitosan + Sodium tripolyphosphate.
SCANNING ELECTRON MICROSCOPE (SEM) ANALYSIS
Figure 6: SEM Image of optimized formulation F5.
Inference
 The shape and surface morphology of optimized formulations F5 was observed in
scanning electron microscope.
PREFORMULATION STUDIES OF OPTIMIZED NPs

After lyophilisation the preformulation studies of optimized Chitosan Nanoparticles
formulations were carried out to check the flow property. The optimized formulation F5
are evaluated for flow property and the results are shown in table 6.
Table 6: Flow property measurements of optimized NPs.

Formulation Bulk density Tapped density Carr’s Index Hausner’s Angle of
code (g/ml) (g/ml) (%) ratio repose (ϴ)
Pure drug 0.2013±0.000531 0.29393±0.001247 32.75±0.3657 1.49±0.008165 51.14±1.3609
F5 0.3356±0.000245 0.3885±0.000327 13.62±0.00474 1.16±0.00471 33.20±0.1087
Mean ± SD (n=3)
Inference
The above results reveal that the optimized formulation F5 shows good flow property
compared with Tetrazosin pure drug.
RELEASE KINETIC OF OPTIMIZED FORMULATION

RELEASE KINETIC OF C-NPs. F5
Table No. 7: Release Kinetics of C-NPs.F5.
% Cum. % Cum. Log % Cum. Square cubic root
Time Log % Cum.
Drug Drug Drug root of Log time of % drug
(h) Drug Release
Release Remaining Remaining time Remaining
0 0 100 2 0 ∞ ∞ 4.6416
0.5 13.19 86.81 1.9386 0.70711 -0.30103 1.1202 4.4278
1 16.17 83.83 1.9234 1 0 1.2087 4.3766
2 24.02 75.98 1.8807 1.4142 0.30103 1.3805 4.2355
3 34.52 65.48 1.8161 1.73205 0.4771 1.53807 4.0306
4 3.6806 58.77 1.7692 2 0.60205 1.6152 3.8879
5 50.14 49.86 1.6978 2.23607 0.69897 1.70018 3.6806
6 57.71 42.29 1.6262 2.44949 0.7782 1.76125 3.4840
7 64.21 35.79 1.5538 2.64575 0.8451 1.8076 3.2955
8 73.22 26.78 1.4278 2.82843 0.9031 1.8646 2.9918
9 78.02 21.99 1.3422 3 0.9542 1.89215 2.8016
10 86.22 13.78 1.1393 3.162278 1 1.9356 2.3975
11 91.89 8.11 0.9090 3.316625 1.04139 1.96327 2.0091
12 95.03 4.97 0.6964 3.46410 1.07918 1.97786 1.7066
Kinetic study
The coefficient of determination (R2) was taken as criteria for choosing the most appropriate
model. The R2 values of various models are given in table 9.
Table R2 Values of F5 in various kinetic models.

Coefficient of determination (R2)
Kinetic Models
F5
Zero order 0.9878
First order 0.9216
Korsmeyer and Peppas 0.985
Higuchi 0.9717
Hixson crowell 0.9771
The in vitro release of optimized formulation F5 are fit into various kinetic models to find out
the mechanism of drug release from Tetrazosin loaded Chitosan Nanoparticles Thus, the
release kinetics of the optimized formulation was best fitted into Higuchi model that showed
zero order drug release with anomalous diffusion (Non Fickian diffusion) mechanism.
SUMMARY AND CONCLUSION

The purpose of this research was to prepare Tetrazosin loaded Chitosan Nanoparticles for
controlled release of drug, to improve the solubility, reduce the dosing frequency, thereby
increasing patient compliance to the therapy. Tetrazosin is formulated as Nanoparticles by

ionic-gelation method using Chitosan as polymer, Sodium tripolyphosphate as a polyanionic
agent (cross linking agent) and the lyophilized nanoparticles filled in hard gelatin capsules.
The preformulation studies like melting point, determination of absorption maximum
(268nm) were performed and the results evident that the drug and excipients are stable, safe
and effective within the range. Physical compatibility study showed that the drug and
excipients were physically compatible with each other. The chemical compatibility studies of
Tetrazosin with excipients were physically analyzed by using FTIR Spectrometer. The results
of the FTIR study proved that there was no interaction between the drug and polymer.
Standard graph was drawn for Tetrazosin and it was found that the solutions show linearity
(0.9995) and obeyed Beer – Lambert’s law. Tetrazosin loaded Chitosan Nanopaticles
prepared by ionic-gelation method using Chitosan as a polymer in different concentration
(0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.4%, and 0.5%). Sodium tripolyphosphate as a
polyanionic agent (cross linking agent), Tween 80 as a deaggregating agent. All seven
formulations characterized for percentage yield which found to be within the range of 78.84%
to 87.25% and the entrapment efficiency of the formulations was observed between 83.40%
to 93.15%. The results showed that the increase in polymer concentration, increase the
entrapment efficiency. The entrapment efficiency was found to be higher in F5- 93.15%
comparatively with other formulations. The Solubility analysis of Tetrazosin was carried out
before and after formulation in distilled water and phosphate buffer pH 6.8. The results show
that the solubility profile is improved after formulations (from insoluble to slightly soluble)
compared with pure drug. Thus the solubility of formulation F5 in distilled water and
phosphate buffer pH 6.8 were improved (9.4933 mg/ml and 13.251 mg/ml) respectively. The
in vitro release study was carried out for all seven formulations. The percentage of drug
release in formulation F5 was found to be 95.03% at the end of 12h and the release profile
was in controlled manner comparatively with other formulations.Based on the higher
entrapment efficiency, drug content and prolonged in vitro drug release F5 was selected as
optimized formulation.
The FTIR studies of optimized formulation F5 shows there was no change in the individual
peaks of the drug and the excipients. It concludes that there was no chemical interaction
between the drug and excipients. The optimized formulations F5 are characterized for SEM
analysis, particle size analysis and zeta potential. The SEM image showed that nanoparticles
were spherical with smooth surface. The particle size analysis was done by Horriba scientific
Nano SZ 100 particle size analyzer showed that mean particle size 188.3 nm and Z- Average
229.0 nm respectively. The Zeta potential study was done by Horriba scientific Nano SZ 100.
The Zeta potential for the optimized formulations F5 was found to be 25.8mV and shows that
the formulation is stable. Post formulation parameters (uniformity of weight, disintegration
test, drug content, and in vitro drug release) for nano particulate capsules were evaluated. The
results were found to be complying with official specifications. The dissolution data of the
optimized formulation was fitted to various kinetic models and the formulation F5 was best
fitted to Zero order kinetics. The slope of the Korsmeyer Peppas plot indicating the diffusion
was Anomalous diffusion (Non-Fickian diffusion). From the overall results, it is clear that the
formulations F5 containing 0.3% polymer concentration (Chitosan) is the optimal
formulation, as it produces controlled drug release.
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Formulation Development and In-Vitro Evaluation of Tetrazosin Loaded Chitosan Nanoparticles For The Treatment of Hypertension

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Formulation Development and In-Vitro Evaluation of Tetrazosin Loaded Chitosan Nanoparticles For The Treatment of Hypertension

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World Journal of Pharmaceutical Research

Marabathuni et al. World Journal of Pharmaceutical Research

FORMULATION DEVELOPMENT AND IN-VITRO EVALUATION OF

Daggumati Pravallika1, *V. Jhansi Priya Marabathuni and Naidu Narapusetty

Drawbacks associated with conventional dosage forms[3]

Terazosin is a member of quinazolines, a member of piperazines, a member of furans and a

Fig. 1: Structure of Terazosin.

MATERIALS AND METHODS

Determination of Melting point[68]

Preparation of 6.8 pH Phosphate buffer[69]

Determination of lambda max (λmax)[32]

Standard curve for Tetrazosin[33]

Solubility studies of pure Tetrazosin[70]

nanoparticles were prepared by dissolving Tetrazosin in 30ml of Chitosan with varying

Table No.1: Composition of Nanoparticles.

Characterization of tetrazosin loaded chitosan nanoparticle

Determination of Drug Content

Drug entrapment efficiency

percentage of entrapment efficiency was determined according to the following equation

Solubility Studies of Tetrazosin loaded Chitosan Nanoparticle

In vitro drug release studies

FTIR study of optimized formulations

Surface Morphology by SEM analysis

Particle size characterization

Polydispersity Index (PDI)

RELEASE KINETICS OF THE OPTIMIZED FORMULATIONS

RESULTS AND DISCUSSION

Chemical Compatibility Study

Figure 2: FT-IR spectrum of Tetrazosin.

Table 2: Saturation solubility of Tetrazosin in phosphate buffer pH 6.8 and Distilled

DETERMINATION OF LAMBDA MAX (λmax) FOR TETRAZOSIN

Calibration Curve For Tetrazosin

Figure 3: Standard curve of Tetrazosin in Phosphate buffer pH 6.8.

SOLUBILITY STUDIES OF TETRAZOSIN LOADED NANOPARTICLES

Table 4: Solubility of Nanoparticles and Tetrazosin in various medium.

COMPARATIVE IN VITRO DRUG RELEASE FOR ALL FORMULATION

Table 5: In vitro drug release for all formulations

0.5 17.25 17.45 16.04 15.09 13.19 9.71 8.32

1 20.12 19.89 19.2 18.99 16.17 11.14 10.14

2 36.4 29.13 28.83 27.02 24.02 17.35 14.14

3 44.8 38.03 37.92 36.21 34.52 31.3 29.27

4 60.2 50.08 47.22 44.1 41.23 34.02 31.73

5 72.27 63.07 59.12 53.8 50.14 36.37 32.13

6 82.29 79.02 69.03 61.78 57.71 42.13 39.92

7 97.04 89.9 78.97 73.2 64.21 50.12 43.78

8 - 96.02 89.17 86.51 73.22 62.79 59.11

9 - - 94.18 91.52 78.01 71.23 69.24

10 - - - 94.47 86.22 79.13 75.17

11 - - - - 91.89 83.2 81.2

12 - - - - 95.03 86.03 83.04

In-vitro Drug Release

Figure 4: In vitro release study of Chitosan Nanoparticles (F1-F7).

CHARACTERIZATION OF OPTIMIZED TETRAZOSIN LOADED CHITOSAN

Figure 5: FT-IR spectral of Tetrazosin + Chitosan + Sodium tripolyphosphate.

SCANNING ELECTRON MICROSCOPE (SEM) ANALYSIS

Figure 6: SEM Image of optimized formulation F5.

PREFORMULATION STUDIES OF OPTIMIZED NPs

Table 6: Flow property measurements of optimized NPs.

RELEASE KINETIC OF OPTIMIZED FORMULATION