Formulation and In-Vitro
Characterization of Oral Nanocrystals of Meloxicam
Anuja P. Patil, Harshal D. Mahajan*, Tanvirahmad J. Shaikh,
Vinod S. Ahire Rajendra D. Wagh
DCS’s A.R.A. College of Pharmacy,
Nagaon, Dhule, Maharashtra, India.
*Correspondence: h.d.mahajan@gmail.com;
DOI: https://doi.org/10.71431/IJRPAS.2025.41108
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Article
Information
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Abstract
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Review Article
Received: 03/11/2025
Accepted: 08/11/2025
Published:30/11/2025
Keywords
Meloxicam;
Bioavailability; Stability;
Solubility.
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The assessed the safety,
tolerability, and efficacy of the tablet formulation of meloxicam (10 mg), a
new, nonsteroidal anti-inflammatory drug (NSAID), in patients with confirmed
osteoarthritis (OA) of the hip or knee.
Poor water solubility and a slow dissolution rate are problems for the
majority of new and existing biologically active compounds, solubility was
improved in this work in a novel way. Through the use of Nanocrystal
technology, the current study aimed to improve the oral bioavailability of the
drug Meloxicam, a poorly water soluble drug, by speeding up its rate of
dissolution. In the present work Nanocrystal is made by nanoprecipitation
technique in the presence of Sodium lauryl sulfate as a surfactant, Poloxamer
188 as a wetting agent and HPMC E-50 as a stabilizer and PVP K-30. Particle
size, DSC, FTIR, XRD, SEM and Redispersibility study were used to analyse and
characterize the prepared Nanocrystal. FTIR spectrum revealed that there are
no interactions between drug and carriers. Drug content, entrapment
effectiveness and in-vitro dissolution were all assessed for the
formulations. SEM results showed the particle size of the formulated
Nanocrystal in nanosize. Finally it was concluded that formulating poorly
soluble drugs in the form of Nanocrystal would be a promising approach in
delivery of poor water soluble drugs by oral route in a simple and effective
way. It optimized Nanocrystal formulation containing Meloxicam showed higher
in vitro drug release, as compared to marketed formulation. The stability
study of Nanocrystal has shown suitable results. It revolves that there’s no
change in drug content, redispersibility study and particle size.
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INTRODUCTION
The
physical, chemical, and biological properties of all drug substances and
pharmaceutical ingredients to be used in the preparation of a dosage form must
be considered during its design and formulation. Solubility, particularly
aqueous system solubility, is an important property of a drug substance.1 One
of the most serious issues with poorly soluble drugs is their low
bioavailability and erratic absorption due to their slow dissolution rates.2The
oral bioavailability of a drug is significantly influenced by its
solubility-dissolution behavior. The issues with these conventional approaches
for solubility dissolution and bioavailability improvement can be resolved
using nanotechnology. In Nanocrystal technology, the drug is kept in the
necessary crystalline state with smaller particles, which increases surface
area and speeds up dissolution, improving bioavailability.3Because of their
increased surface area and saturation solubility, drug particles reduced to the
nanometer range have a faster rate of dissolution.
Meloxicam
(MLX) is a non-steroidal anti-inflammatory drug that is prescribed in the
treatment of rheumatoid arthritis and osteoarthritis with low gastrointestinal
complications. A rapid onset of action is important for patients, particularly
under the pain condition of rheumatism and osteoarthritis. Consequently, a
suitable oral dosage form of MLX, with enhanced aqueous solubility, could
facilitate its oral absorption and reduce its onset of action for the treatment
of acute pain.
In this situation,
increasing aqueous solubility is a worthwhile objective to boost therapeutic
effectiveness. The solubility and surface area of the drug are two factors that affect how quickly the
drug dissolves; as a result, the dissolution rate will rise if the solubility
and surface area of the drug both rise.4 & 5
In the current study, the drug solution is combined with an
aqueous solution that contains a surfactant using the nanoprecipitation
technique. The supersaturated solution causes the nucleation and growth of drug
particles during mixing, which surfactants can stabilise.6
The purpose of this work is to improve the dissolution rate
and formulate the Meloxicam Nanocrystal by nanoprecipitation method. Scanning
electron microscopy was used to further characterise the optimised formulation (SEM). In distilled water, a dissolution study of Nanocrystal
formulations was conducted.
MATERIALS AND METHODS
Meloxicam was
supplied by Umedica Laboratories Private Limited vapi, Gujrat as a gift sample.
HPMC E-50 and Sodium Lauryl Sulphate were obtained from Anmol Chem. Ltd,
Mumbai. Poloxamer 188 was obtained from Anmol Chem. Ltd, Mumbai. Ethanol was obtained from SD Fine-Chem. Limited,
Gujarat.
Preparation of drug loaded Nanocrystal using Nanoprecipitation technique
The nanoprecipitation technique was used for the preparation
of Meloxicam Nanocrystal. Nine
formulation were prepared by Nanoprecipitation method. The drug is dissolved in
suitable organic solvent ethanol in which the drug is soluble. This was poured
into different amount of water containing different amount of Poloxamer 188,
HPMC E-50 and SLS at maintained at room temperature and subsequently stirred
magnetically to allow volatile solvents to evaporate. After 30 min of stirring
the volume of Nanocrystal dispersion was concentrated to 10 mL under reduced
pressure using a Rota evaporator with vacuum (KNF, vaccum pumps & system).
The aggregates were removed by filtration through a 0.45µm syringe filter.
Separation of non-encapsulated drug was performed by ultracentrifugation
(Beckman Coulter) at 50,000rpm at 40C for 30 min.7
Table 1: Composition of Meloxicam Loaded Nanocrystal
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Sr.No
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Ingredients
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F1
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F2
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F3
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F4
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F5
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F6
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F7
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F8
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F9
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1
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Meloxicam (mg)
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10
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10
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10
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10
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10
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10
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10
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10
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10
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2
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Poloxamer
188 (mg)
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1.5
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1.5
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2
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2.5
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1.5
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1.5
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2
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2.5
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2.5
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3
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HPMC E-50 (mg)
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-
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20
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-
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10
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12.5
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5
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5
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15
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5
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4
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PVP K30 (mg)
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20
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-
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20
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10
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12.5
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15
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15
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5
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15
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5
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Dimethyl
Formamide (ml)
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1
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1
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1
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1
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1
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1
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1
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1
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1
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6
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Water (d/w)
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15
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15
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15
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15
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15
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15
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15
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15
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15
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Characterization of Nanocrystal
Determination of the Particle Size
By using Zeta-particle size, Model Nano ZS, photon
correlation spectroscopy was used to determine size distribution, average
particle size, and PDI. Measurement and dilution with distilled water were applied
to the separated Nanocrystal. At a scattering angle of 900 degrees
and a temperature of 25 degrees, the particle size and PDI measurements were
performed. Experiments were conducted in triplicate.8
Entrapment Efficiency (EE)
Using a cooling ultracentrifuge, the freshly made Nanocrystal
was centrifuged for 20 minutes at 20,000 rpm at 5°C temperature. By comparing
the absorbance of the appropriately diluted 25 ml of supernatant solution at
350 nm with a blank/control Nanocrystal, the amount of unincorporated drug was
determined. EE was calculated by deducting the initial dose of the drug from
the amount of free drug in the supernatant. For each batch, the experiment was
run in triplicate, and the average was computed. The following equation could result in the entrapment efficiency
(EE %).9
EE of the drug= (amount
of encapsulated drug) /
(total amount of the drug) X
100
………Equation No.1
Redispersibility of
Nanocrystal
The chosen formulation was freeze-dried to produce a dry
powder for additional research. Additionally, the effect of the cryoprotectant
on the freeze-drying and dispersibility of the prepared Nanocrystal was
investigated. In the formulation, mannitol was used as a cryoprotectant at a
concentration of five times the total solid contents. Each flask contained two
samples of Nanocrystal. The required amount of mannitol was added to one sample
and shaken to dissolve it, while the second sample was left without cryoprotectants.
For primary freezing, these flasks were placed in a deep freezer and kept there
for 12 hours at - 20°C. The lyophilizer's hoover adapter was then connected to
the container. For 48–72 hours, the solvent sublimated at a pressure of 80 mmHg.10
Differential Scanning
Calorimetry
A differential scanning calorimeter (DSC DA 60 Shimadzu,
Japan) with a liquid nitrogen subambient accessory was used to perform the
differential scanning calorimetric measurements. The DSC was run on the
Poloxamer407, drug-loaded Nanocrystal formulation, and pure Meloxicam. In a
flat-bottomed aluminium pan, sample 2 mg were loaded and heated at a rate of 100
C/min over a range of 40 to 4000 C. The rate of heating and cooling
was managed by a stream of nitrogen gas. Purified indium was used as the
calibration standard for the instrument's temperature and energy scales.11
Fourier Transform Infrared Spectrophotometry
(FT-IR)
The pressed pellet technique was used to conduct the FTIR
spectral analysis. Any substance's IR spectrum can reveal the groups that are present
in that substance. (KBr potassium bromide) pellets were used to administer a drug with an
IR spectrum. Small amounts of the drug sample were combined with oil, and a drop
was then evenly spread between two KBr pellets. An infrared spectrum was taken
after the pellets had been placed in the holder. The scanning range was
400–4000 cm1. Various peaks in the infrared spectrum were
interpreted to indicate the presence of various groups in the drug's structural
makeup.12
X-ray diffraction
The drug's crystallinity in the Nanocrystal formulation was
determined using X-ray diffraction analysis, which was carried out on a Philips
PW 3710 x-ray diffractometer(XRD) with a copper target and nickel filter.
Powders were mounted on glass-bottomed aluminium stages with a level surface.
Each sample's XRD pattern was evaluated from 10 to 50 degrees. 2-theta with a
0.1 2-theta degree step increment and a 1 second dwell time between each step.13
Scanning electron
microscopy
SEM (JEOL Model JSM - 6390LV) was used to characterise the
surface morphology of Nanocrystal surfaces. The Nanocrystal was scanned in a
high vacuum chamber with a focused electron beam after being mounted directly
on the SEM stub with double-sided, sticking
tape. Secondary electrons
emitted by the samples were detected and used to create the image. 14-16.
In vitro release studies
Using a membrane diffusion technique, the release study of
the Nanocrystal formulation for the release of Meloxicam from polymeric
Nanocrystal was conducted. A semi- permeable membrane called the dialysis
membrane was used to create an in vitro diffusion cell. A dialysis membrane with a molecular weight cut-off of 12,000–14,000 was used to both retain the Nanocrystal and
allow the free drug to diffuse freely into the release media. 1 mg of prepared
drug Nanocrystal were dissolved in 1 mL of isotonic, phosphate-buffered saline
at pH 7.4. The dialysis membrane was filled with the Nanocrystal dispersions
and clamped at both ends. The dialysis bag was submerged in tightly-capped
glass vials (7 x2.8 cm) containing 10 mL of a 0.5% (w/v) sodium lauryl sulphate
solution in distilled water in order to maintain sink condition. The glass
vials were put through the release test in a thermostatically-controlled
shaking water bath that was set to 37±0.5°C and was shaking at a constant speed
of 100 rpm. The entire release medium was removed and replaced with new release
medium at predetermined time intervals. UV was used to determine the drug's release's concentration. The majority of
the experiments were done in triplicate. 17
Stability Studies
On an improved formulation, stability tests were performed
for 6 months at 30 °C in a stability
chamber (Thermolab). The glass bottle with a tight seal contains the optimized
formulation. Stability Studies on drug content, particle size, and
redispersibility were conducted after six months.18
RESULTS AND DISCUSSION
Preparation of drug loaded Nanocrystal:
Meloxicam Nanocrystal were successfully prepared by using
nanoprecipitation technique as it is
rapid and easy to perform. Nanocrystal formation is an instantaneous, one-step
process. After adding the polymer solution to the non-solvent, there is rapid
desolation and then nanoprecipitation. The polymer precipitates, resulting in
immediate drug entrapment, as soon as the solvent containing the polymer has
diffused into the dispersing medium. Additionally, this method consistently
yields carriers with sizes in the nanometer range and uses low-toxic
ingredients that are suitable for oral administration.
Characterization of
Nanocrystal
Determination of particle size
The size of the particle plays a significant role in the
design of an oral drug delivery system when considering irritation and comfort.
Figure 2 displays
the average particle
size of the
prepared Nanocrystal formulation. The range of particle sizes is 130.1–409.1
nm. The effect of various formulation factors, particularly the polymer
concentration, had a big impact on particle
size. The particle sizes obtained with lower polymer
concentration were noticeably
smaller than those obtained with higher polymer concentration, as shown in Fig.
1
Figure 1: Particle size of
the optimized Meloxicam Nanocrystal formulations (F6)
All formulations of Nanocrystal had their polydispersity
index (PDI) representing the particle size distribution measured. The range of
the mean PDI values is 0.402 to 1.0. The small PDI values indicate a narrow
distribution, which represents the uniformity of particle size in the
formulation of the Nanocrystal.
Entrapment Efficiency (EE)
The percentage of effective Nanocrystal entrapment ranged
from 81.8 to 94.9%.Each and every tested variable significantly affects the
EE%. It was found that the EE% of the Nanocrystal formulae decreased as the
polymer content was increased. This outcome in Fig. 3 can be attributed to the
organic phase's increasing viscosity as the polymer content increased.
Figure 3: Entrapment Efficiency
of the Meloxicam in Nanocrystal
Redispersibility Test:
It was discovered that using mannitol as cryoprotectants
improved the dispersibility, and products spontaneously dispersed into primary
Nanocrystal within 1-3 minutes in both media (0.1 N HCl and phosphate buffer pH
6.8). It was implied that the presence of mannitol in the products would
enhance the hydrophobic drug's wetting and hasten the penetration of water into
them. On the other hand, as predicted by their agglomerated structure, the
products without cryoprotectants could not be dispersed well and instead
returned to the original Nanocrystal within 15 minutes.
Fourier Transform Infrared Spectrophotometry
(FT-IR)
The obtained sample's IR spectrum was performed and compared
to the IR spectrum of the Meloxicam reference standard. Similar characteristic
peaks can be seen in the sample drug's IR spectra and optimized Nanocrystal
formulation (F6). The interpretation is displayed in Table 2 for the IR spectra
of the sample drug and the standard drug Meloxicam, respectively, in Figures 4 and 5.
Figure 4:IR Spectra Analysis of Standard
Meloxicam
Figure 4:IR Spectra Analysis
of Poloxamer 188
DSC:
Each sample
was examined using
differential scanning calorimetry to confirm the existence of a physical interaction between
Meloxicam and excipients (DSC). Figure 6 displays a DSC thermogram of
Meloxicam, poloxamer-188, and Nanocrystal. The drug's distinctive endothermic
peaks a 1630C vanished from the thermogram of the drug-loaded Nanocrystal. This led to the conclusion
that Meloxicam was entrapped in the polymer matrix in an amorphous or molecular
dispersion state.
Figure 6:
A. DSC Thermogram of the
Meloxicam B. DSC Thermogram of the Polymer C. DSC Thermogram of the optimized Nanocrystal formulation (F6).
XRD:
By using powder x-ray diffraction, the crystallinity of the
in the Nanocrystal has been investigated. Figure 8 displays the optimised
formulation's powder x-ray diffraction patterns (F6). This supports the
findings of the DSC and suggests that the drug was present in the Nanocrystal
in an amorphous state.
Figure 7: XRD of the optimized Nanocrystal formulation (F6).
Scanning Electron
Microscopy (SEM):
SEM was used to access the morphology of drug-loaded
Nanocrystal (F6), which is depicted in figure 8. According to this
illustration, the Nanocrystal had a smooth surface that was crucial for the
delivery of oral drugs, were in the nanometer size range.
Figure 8: SEM of the optimized
Nanocrystal formulation (F6)
In vitro drug release from Nanocrystal. The drug formulation
made using the nanoprecipitation technique was tested for release in vitro.
Using a dialysis method, the amount of Meloxicam released from Nanocrystal was
measured. Figure 9 displays the release profiles of Meloxicam.
Figure 9: In vitro release study of the Meloxicam Nanocrystal formulation
The wetting-induced sustained release of the drug from
Meloxicam-based Nanocrystal is followed by
an immediate release as a result
of rapid diffusion
of the drug from biodegradable Nanocrystal.
Stability Studies:
On an improved formulation, stability tests were performed
for six months at 30±2°C in a stability chamber (Thermolab). The optimised mixture was kept in an aluminium foil bag that
was sealed. Studies on drug content, particle size, and redispersibility were conducted after six months.
Table 3: Stability of Meloxicam Nanocrystal during storage
(F6)
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Parameter
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0 Day
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1 Month
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3 Month
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6 Month
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Entrapment Efficiency
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94.9±0.5
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93.7 ± 0.3
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92.1 ± 0.4
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91.9 ± 1.0
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Particle Size
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130.1 ± 0.4
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132.7 ± 0.1
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133.2 ± 0.2
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134.3± 0.2
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Redispersibility Study
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Clear
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Clear
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Clear
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Clear
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CONCLUSION
Utilizing the nanoprecipitation technique, Meloxicam was
successfully suited within biodegradable nanosupension. The particle size and
encapsulation effectiveness of the Nanocrystal were significantly influenced by
the drug-polymer ratio. In terms of their particle size, drug loading capacity,
and redispersibility in vitro release characteristics, the formulated Meloxicam
Nanocrystal was found to be a suitable and potential natural carrier. The
stability analysis of ro from Nanocrystal has produced satisfactory findings.
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