School of Pharmacy & Research, People’s University, Bhopal, Madhya Pradesh, India.
Qualitative phyto chemical screening of Nyctanthes arbor tristis showed the presence of active metabolites such as Alkaloids, carbohydrates, glycosides, tannin and phenols is present. Quantitative phytochemical assay was performed by calculating total phenolic content (TPC) and total flavonoid content (TFC). Microspheres were prepared by the chemical denaturation method using glutaraldehyde as a cross-linking agent. The microspheres were characterized by particle size analysis, zeta potential, scanning electron microscopy (SEM) and stability study. Several factors such as stirring rate, temperature, and viscosity show an effect on the size. The sizes of the microspheres were found to be 156.5 nm. The current study involved the formulation and characterization of extract-loaded microspheres. SEM photographs confirmed the shape and formation of the microspheres. These microspheres are employed in the delivery of drugs and extracts; the drugs may be encapsulated or entrapped. In future it can also be formulated in various dosage forms.
Novel drug delivery system:
Compared to traditional multi-dose therapy, novel drug delivery systems have many advantages. Micro particulate drug delivery systems, according to recent trends, are particularly well suited to achieving controlled release and delayed-release oral formulations with low risk of dose dumping, blending flexibility to achieve different release patterns and reproducible and short gastric residence time.
Microspheres:
Microspheres are small spherical particles, with diameters in the micrometer range (typically 1 μm to 1000 μm). Microspheres are sometimes referred to as microparticles. Microspheres can be manufactured from various natural and synthetic materials. Microspheres as drug carriers are one of the most cutting-edge methods for sustaining and controlling pharmacological action in a specific location. Microspheres made of degradable materials are used to provide transient embolization. They should, in theory, be expelled from the body once they have achieved their clinical aim without interfering with the operation of other organs.
Plant Profile
Nyctanthes arbor-tristis
Taxonomical Classification
Kingdom: Plantae
Division: Magnoliophyta
Class: Magnoliopsida
Order: Lamiales
Family: Oleaceae
Genus: Nyctanthes
Species: arbor-tristis
Uses:
Experimental Work
Plant collection
The medicinal plant Nyctanthes arbor tristis (300 gm) was collected. After cleaning, plant parts (leaves) were dried under shade at room temperature for 3 days and then in oven dried at 45°C till complete dryness. Dried plant parts were stored in air tight glass containers in dry and cool place to avoid contamination and deterioration. Authentication of selected traditional plant - Medicinal plant Nyctanthes arbor tristis was authenticated by a plant taxonomist in order to confirm its identity and purity.
Extraction
In the current investigation, plant material was extracted utilizing the Soxhlet apparatus and a continuous hot percolation method. Nyctanthes arbor tristis powder was added to a soxhlet apparatus thimble.At 60°C, soxhlation was carried out with petroleum ether acting as a nonpolar solvent. After being dried, the exhausted plant material (marc) was extracted again using an ethanol solvent. For every solvent, the extraction process was carried out until no visual color shift was seen in the siphon tube, and the absence of any residual solvent upon evaporation confirmed that the extraction was complete. The obtained extracts were evaporated at 40°C in a Buchi-type rotating vacuum evaporator. Weighing the dried extract allowed us to calculate the % yield for each extract using a formula:
Prepared extracts was observed for organoleptic characters (percentage yield, colour and odour) and was packed in air tight container and labelled till further use
Design Of Experiment
Design of experiment for the formulation of Microsphere was performed by Design Expert (Version 12.0.1.0) software. The quadratic response surfaces were represented by the secondorder polynomial model.
Evaluation Parameters of Microspheres Formulation
The zeta potential was measured for the determination of the movement velocity of the particles in an electric field and the particle charge.
The size of microspheres was measured using Malvern Zeta sizer (Malvern Instruments). The dispersions were diluted with Millipore filtered water to an appropriate scattering intensity at 25°C and sample was placed in disposable sizing cuvette.
The electron beam from a scanning electron microscope was used to attain the morphological features of the optimized extract loaded microspheres were coated with a thin layer (2–20 nm) of metal(s) such as gold, palladium, or platinum using a sputter coater under vaccum.
Formulation Of Microsphere Loaded Gel
Initially carbopol-934 was immersed in 50 mL of warm water (A) for 2 hr and was homogeneously dispersed using magnetic stirrer at 600 rpm. In separate container carboxymethyl cellulose and methyl paraben was added into 50 ml warm water (B) and stirred continuously to make stiff gel. Both the mixtures A and B were mixed with the continuous stirring. Then tri-ethanol amine (Drop wise) was added to neutralize the pH and microspheres of optimized formulation were incorporated into the dispersion to obtained Gel. At this stage, permeation enhancer (Propylene glycol) was added. The final dispersion was agitated until smooth gel was formed without lumps
Characterization Of Microsphere Loaded Gel
RESULT AND DISCUSSION
In phytochemical extraction the percentage yield is very crucial in order to determine the standard efficiency of extraction for a specific plant, various sections of the same plant or different solvents used. The yield of extracts received from the Nyctanthes arbor tristisis 8.3 Quantitative Analysis Preliminary phytochemical testing of crude extracts confirmed the presence of phenolics and flavonoids in plant material. To estimate their amount total phenolic (TPC) and total flavonoid content (TFC) assays were performed.
Preliminary phytochemical testing of crude extracts confirmed the presence of phenolics and flavonoids in plant material. To estimate their amount total phenolic (TPC) and total flavonoid content (TFC) assays were performed.
Total Phenolic Content (TPC) Estimation
Figure: Represent standard curve of Gallic acid
Total Flavonoids content (TFC) estimation
Figure: Represent Standard Curve of Rutin
Optimization of formulation by design of expert (DOE) software
The experimental nature based on this mixture of the component has resulted in 12 separate microsphere formulation batches. As indicated, numerous microsphere lots were prepared and then assessed for each of the responses. The responses observed were fit to 12 runs, and it has been noted that the best fit model was the linear model for the two dependent variables. The significance of the model with that of comparing with the other model for the analysis by analysis of variance (ANOVA). In All the responses recorded for 12 runs and the relation of independent and dependent variables recorded.
The particle size is one of the most important parameter for the characterization of
Microsphere. The average particle size of the prepared extract loaded microspherewas measured using Malvern zeta sizer. Particle size analysis showed that the average particle size of extract loaded Microsphere formulation was found to be 156.5 nm.
Table: Zeta potential
|
S. No |
Formulation |
Zeta potential (Predicted value) |
Zeta potential (Actual value) |
|
1. |
Microsphere |
0.6 mV |
0.50 mV |
Zeta potential analysis is carried out to find the surface charge of the particles to know its stability during storage. If the particles in Microsphere have a large positive zeta potential then they will tend to repel each other and there will be no tendency for the particles to come together. However, if the particles have low zeta potential values then there will be no force to prevent the particles coming together and flocculating for Microsphere. Zeta potential of Microsphere formulation was found to be range 0.5 mV with peak area of 100% intensity. These values indicate that the formulated Microsphere is stable.
Scanning electron microscope (SEM)
Discussion
SEM analysis was performed to determine their microscopic characters (shape & morphology) of prepared Microsphere. Microsphere were prepared and dried well to remove the moisture content and images were taken using scanning electron microscopy. Scanning electron micrograph of the prepared Microsphereat 39.78 kx magnification showed that the Microsphere were smooth surface morphology and spherical shape. The porous nature of Microsphere was clearly observed in the SEM images.
Characterization Of Microsphere Loaded Gel
Table: Physical appearance
|
S. No |
Parameter |
Result |
|
1. |
Colour |
Brown colour |
|
2. |
Odour |
Odourless |
|
3. |
Appearance |
Brown colour |
|
4. |
Homogeneity |
Homogeneous |
Discussion
An evaluation of the gel, including colour, odor, appearance and homogeneity, was conducted. Gel was discovered to have a Brown colour to it when tested. Gel does not have a distinctive odor and has a Brown colour appearance, according to research conducted on it.
Viscosity of Gel - 6830±0.32
Discussion
The viscosity was measured by the Brookfield viscometer spindle no. 61 at 100rpm. The viscosity of Gel was found to be 6830 centipoise respectively.
pH of Gel – 6.1
Discussion
The pH of the gel formulation was found to be 6.1, which lies in the
normal pH range of the skin and would not produce any skin irritation. There was no significant change in pH values as a function of time. The physicochemical properties of
prepared gel formulation were in good agreement.
Gel Formulation – 13.09
Discussion
One of the essential criteria for a Gel is that it should possess good spreadability. Spreadability depends on the viscosity of the formulation and physical characteristics of the polymers used in the formulation. A more viscous formulation would have poor spreadability. Spreadability is a term expressed to denote the extent of area on which the gel readily spreads on application to the skin. The therapeutic efficacy of a formulation also depends upon its spreading value. The spreadability of Gel formulation is found to be 13.09 g.cm/s.
Results of antimicrobial activity of microsphere loaded gel formulation
Figure: Antimicrobial activity
Table: Antimicrobial activity of microsphere loaded gel against E.coli
|
S. No. |
Sample Name |
Zone of Inhibition (mm) |
|
1. |
Gel (0.5mg/ml) |
8 mm |
|
2. |
Gel (1mg/ml) |
9 mm |
|
3. |
Gel (1.5mg/ml) |
11mm |
|
4. |
Gel (2 mg/ml) |
15mm |
Discussion: Gel (2 mg/ml)formulation had the highest antimicrobial efficacy against E. coli, with a zone of inhibition of 15mm. The higher effectiveness of microsphere loaded gel could be attributable to its modified composition, which may improve the antimicrobial agent's release and bioavailability, making it a good option for further development and possible therapeutic uses.
REFERENCES
Piyush Biloniya*, Eisha Ganju, Bhaskar Kumar Gupta, Design of Experiment Approach for Development of Microspheres Loaded Gel of Nyctanthes Arbor Tristis Leaves Extract for Enhances Stability as Well as Bioavailability of Phytoconstituents, Int. J. of Pharm. Sci., 2025, Vol 3, Issue 3, 2985-2995 https://doi.org/10.5281/zenodo.15105552
10.5281/zenodo.15105552
