Development and Characterization of Lyophilized Liposomal Garlic Extract Nanoparticles as Natural Antimicrobial Dusting Powder

Because garlic (Allium sativum) contains special organosulfur compounds including thiosulfinates and allicin, it has long been known for its exceptional antibacterial, antifungal, and therapeutic qualities. However, the medicinal chemicals in garlic have a strong odor and are highly volatile and unstable. Their sensitivity to chemical and environmental changes also limits their practical use. By encasing these delicate molecules and improving their stability, bioavailability, and controlled release characteristics, nanotechnology-based delivery techniques—in particular, liposomes—offer a possible treatment (2).

Biocompatible phospholipids make up liposomes, which offer a flexible delivery system that may release and shield hydrophilic and lipophilic bioactive compounds throughout time. The resulting liposomal nanoparticles enhance their shelf life and storage durability when paired with lyophilization (freeze-drying), which qualifies them for usage in powder-based compositions. Lyophilized liposomal garlic extract nanoparticles have the potential to be a novel, natural antibacterial dusting powder that can safely and effectively replace topical therapies in wound care, personal hygiene, and other biomedical areas, according to recent research (3).

In order to create, improve, and characterize such nanoparticles, this study will assess important factors like shape, antibacterial efficiency, encapsulation effectiveness, and particle size. This work is significant because it combines green chemistry, nanotechnology, and traditional medical knowledge to create new treatments for microbial contamination and resistance (4).

Due to the negative effects of synthetic medications and the developing global problem of antibiotic resistance, garlic extract is gaining popularity as a natural antibacterial. For topical, dental, and food safety applications in particular, garlic presents a promising, safe, and affordable alternative to treat infections with little disruption of commensal bacteria or resistance-inducing effects. In conclusion, garlic extract's many, scientifically proven anti-infection qualities, as well as its value as a substitute or addition to traditional antimicrobials, account for its history and importance as a natural antibacterial (5).

Using complex nano-formulations known as liposomal garlic extract nanoparticles, the therapeutic components of garlic, including allicin and related organosulfur compounds, are enclosed in spherical lipid-based vesicles known as liposomes. The sensitive garlic ingredients are shielded from environmental deterioration and odor problems by this encapsulation, which improves stability and controlled release and makes it perfect for antibacterial and therapeutic uses (6).

Liposomes are small, spherical vesicles that have one or more phospholipid bilayers surrounding an aqueous core. They are widely used as carriers for the delivery of drugs, bioactive molecules, and nutrients due to their biocompatibility, ability to encapsulate both hydrophilic and hydrophobic substances, and ability to protect active chemicals from degradation (7). Because of their capacity to replicate cell membranes, encapsulate a variety of compounds, and effectively transport them to target locations, liposomes are fundamental to nanomedicine and biotechnology.

Liposomes are small, spherical vesicles that have one or more phospholipid bilayers surrounding an aqueous core. Their structure resembles real cell membranes quite a bit. With hydrophilic (water-attracting) heads and hydrophobic (water-repelling) tails, amphiphilic phospholipids self-organize in water to create the vesicle structure that makes up the lipid bilayer. Cholesterol is often present in the bilayer to regulate the membrane's fluidity and stability. Depending on their size and lamellarity, liposomes can have one bilayer (unilamellar) or several concentric bilayers (multilamellar). Because they are biocompatible  and can encapsulate both water-soluble molecules inside their watery core and fat-soluble compounds within the lipid bilayer, they are perfect carriers for medication administration and other biological uses (8).

1. INTRODUCTION TO DUSTING POWDER

• Finely crushed antimicrobial dusting powders are used topically to the skin to treat and prevent minor infections caused by burns, scrapes, small wounds, or skin conditions. These powders typically combine antibacterial agents derived from natural extracts or synthetic compounds with absorbents such as talc, starch, or bentonite to reduce friction and absorb moisture. This aids in skin preservation and healing (9).
• Herbal extracts (such as carrot seed, neem, or turmeric), antibiotics, or antiseptics are examples of active antimicrobial substances.
• On the skin, inert bases like silicon dioxide, talc, and starch enhance spreadability and flowability.
• Other ingredients that improve efficacy and shelf life include salicylic acid and benzoic acid, which have anti-inflammatory and preservation properties. Applications and advantages
• By preventing bacterial and fungal growth on afflicted skin areas, antimicrobial dusting powders lessen itching and inflammation and guard against secondary infections.
• They are frequently used to treat small burns, abrasions, ringworm, eczema, and athlete's foot.
• By absorbing perspiration and moisture, the powders aid in maintaining dryness and stop additional irritation or microbial growth.

This study examined the effects of sonication period (10–30 min), cholesterol-to-lecithin ratio (CLR) (0–1), and garlic extract (GE) (0%–3%) on the physicochemical and antibacterial properties of phytosomal nanocarriers (PNCs). Fourier-transform infrared (FT-IR) analysis was done to see if there were any interactions between the GE and other component constituents. Particle size, zeta potential, encapsulation efficiency, turbidity, and stability were all significantly impacted by the duration of sonication, and a strong correlation between these factors was discovered. The CLR and GE concentrations were strongly correlated with the electrical conductivity and pH levels. The tiny particle size and excellent penetration efficiency of GE-containing PNCs likely contributed to their antibacterial and antioxidant properties. The CLR and the sonication time were shown to be important parameters affecting the stability of the synthesized PNCs throughout a 4-week storage period. A CLR of 0.52, a sonication duration of 23.59 minutes, and a concentration of 3% GE were determined to be the ideal parameters during the optimization phase. Lastly, it is crucial to emphasize that the FT-IR analysis verified that GE and other PNC components did not interact (10).

One of the ingredients of garlic extract (Allium sativum L.) is allicin, which breaks down readily. A garlic extract was created in a phytosome system to increase the chemical stability of allicin. Ostwald ripening can lead to an increase in the distribution of particle sizes in phytosomes, which are colloidal systems. The system is physically unstable if the size distribution changes. The study's objective was to evaluate the garlic extract phytosome's physical stability after four weeks of storage at three distinct temperatures (11).

In this study, garlic essential oil (GEO) was used to embed lecithin (LT) and β-sitosterol (β-S). Ethanol was then injected into liposomes (L-S-G) to boost the bioavailability of GEO (51%). Next, L-S-G's stability, encapsulation efficiency, and particle size were investigated. The stability of L-S-G samples under different storage conditions, such as light and dark, was also evaluated. The average particle size of the implanted L-S-G was 375.8 nm, and the encapsulation effectiveness of the liposomes was 81%, according to the study's results. Particle diameters of 529 nm and 473 nm were found in liposomes that were stored for 15 days in both light and dark conditions. Additionally, the gastrointestinal trial results in this study showed that L-S-G had a 51% bioavailability. The long-term bacteriostatic tests in this study showed that garlic essence liposomes effectively inhibited the growth of Listeria monocytogenes in beef, poultry, and pork when compared to garlic essential oil (12).

High dose carrying capacity, stability, patent protection, propellant-free nature, controlled drug release, reduced local and systemic toxicities, patient compliance, and selective localization of the drug within the lung are just a few of the promising features that liposomal drug dry powder formulations have shown for pulmonary drug administration. A critical analysis of recent developments will provide a fair appraisal of the benefits of liposomal encapsulation when developing dry powder formulations. It will also help researchers stay up to date and focus their work on more relevant areas. In liposomal dry powder formulations (LDPF), drug-encapsulated liposomes are homogenized, dispersed throughout the carrier, and dried into powder form using freeze drying, spray drying, and spray freeze drying. Alternatively, LDPF may be produced using supercritical fluid technology. Drug-encapsulated liposomes rehydrate in the lung and release the medication gradually when breathed with the proper inhalation equipment. The generated LDPF is evaluated for lung deposition behavior and medication distribution in the lung both in vitro and in vivo using a suitable inhaler device. The most often used liposomes are composed of lung surfactants and synthetic lipids. Anticancer drugs for lung cancer, corticosteroids for asthma, immunosuppressants to prevent lung transplant rejection, antifungal drugs for lung fungal infections, antibiotics for cystic fibrosis and local pulmonary infections, and opioid analgesics for pain relief are a few examples. Many liposomal formulations have reached the clinical trial stage for the treatment of pulmonary pain, cystic fibrosis, lung cancer, and lung fungal infections. These formulations have produced very promising results in both in vitro and in vivo studies. However, alterations to innovative medications for respiratory illnesses and systemic distribution will make it more challenging to conduct well-designed inhalation toxicity studies to support these treatments, especially for chronic ailments (13).

This study aims to optimize the preparation factors, such as sonication time (5–20 min), cholesterol to lecetin ratio (CHLR) (0.2–0.8), and essential oil content (0.1–0.3 g/100 g) in the solvent evaporation method for formulation of liposomal nanocarriers containing garlic essential oil (GEO), in order to find the highest encapsulation efficiency and stability with the strongest antioxidant and antimicrobial activity. Droplet size, zeta potential, encapsulation efficiency, turbidity, changes in turbidity after storage (as a measure of instability), antioxidant capacity, and antibacterial activity were measured for each produced nanoliposome sample. Sonication length is known to have the biggest effect on droplet size, zeta potential, encapsulation efficiency, turbidity, and instability, however CHLR was the most effective factor on zeta potential and instability. The GEO content had a significant effect on the antioxidant and antibacterial action, particularly against gram-negative bacteria (Escherichia coli). The FTIR results based on the identification of functional groups confirmed the presence of GEO in the created nanoliposome's spectra; also, no interaction between the components of the nanoliposome was seen. Response surface methodology (RSM) was employed to identify the overall ideal conditions based on attaining the most stability and efficiency as well as the strongest antioxidant and antibacterial activities. The factors under investigation (sonication time: 18.99 min, CHLR: 0.59, and GEO content: 0.3 g/100 g) were anticipated to have these values (14).

• Size and size distribution: Determined using methods such as dynamic light scattering (DLS), size is important because it affects cellular absorption and circulation time. The regularity of the size distribution is indicated by the polydispersity index (PDI).
• Zeta potential, or surface charge, influences biological system interaction and stability. obtained via electrophoretic light scattering.
• Drug entrapment efficiency: The proportion of drug used that is entrapped in liposomes.
• Morphology and shape: Examined by microscopy techniques like transmission electron microscopy (TEM) and scanning electron microscopy (SEM).
• Stability: Assessed by tracking changes in drug retention, size, and charge under different storage circumstances (pH, temperature, etc.).
• In vitro drug release: To forecast in vivo behavior, drug release kinetics under various conditions are studied (15).
• Flow properties: Powder flow is critical for application and measured by angle of repose, bulk density, tapped density, Carr’s index, and Hausner’s ratio.
• pH: Ensures compatibility with skin, typically measured in a 1% powder solution (16).

In order to create a natural antibacterial dusting powder, this study focuses on creating and assessing lyophilized liposomal nanoparticles loaded with garlic extract. To improve stability, bioavailability, and controlled release of its active chemicals (mostly allicin), garlic, which has potent antibacterial and antifungal qualities, is encapsulated in liposomes composed of phospholipids and cholesterol(17).

To create a stable, free-flowing powder that can be applied topically, the formulation will be made using nanotechnology-based methods like thin-film hydration, sonication, thin-film hydration, reverse phase evaporation, ethanol and solvent injection, and lyophilization.

Applications include biomedical coatings, cosmeceuticals, wound care, and personal hygiene. Benefits including increased stability, less odor, better penetration, biocompatibility, and sustainability are highlighted in the study(18).
Future studies will focus on multi-herbal formulations, clinical validation, large-scale production, and intelligent delivery systems for broader application in the food packaging, veterinary, and medicinal industries(19).

MATERIALS AND METHODS

Preparation of Aqueous Garlic Extract Maceration Method (Warm Water Extraction Method)

• Maceration Method

1. Principle

The aqueous extraction of garlic is based on the solubility of water-soluble phytoconstituents present in Allium sativum. When garlic powder is mixed with warm water (40–50°C), bioactive compounds such as allicin, flavonoids, and other polar constituents are released into the aqueous medium. Mild heating facilitates extraction without causing significant degradation of thermolabile compounds.

2. Materials Required

• Garlic powder (Allium sativum)
• Distilled water
• Magnetic stirrer or mechanical stirrer
• Beaker or conical flask
• Whatman filter paper
• Funnel

3. Methodology

Step 1: Preparation of Extraction Mixture

A measured quantity of garlic powder is accurately weighed and transferred into a clean beaker. A suitable volume of distilled water, preheated to 40–50°C, is added to the powder.

Step 2: Extraction by Stirring

The mixture is subjected to continuous stirring using a magnetic or mechanical stirrer for a duration of 5–10 minutes. Stirring ensures proper dispersion of the powder and enhances the contact between solvent and plant material, thereby improving the extraction efficiency of active constituents.

Step 3: Filtration

After stirring, the mixture is allowed to settle briefly and then filtered using Whatman filter paper. Filtration removes insoluble plant residues and yields a clear aqueous extract.

Step 4: Collection of Extract

The filtrate obtained is collected in a clean container. This filtrate represents the aqueous garlic extract, which can be used for further formulation or analysis.

4. Mechanism of Extraction

During the extraction process, the enzyme alliinase present in garlic becomes activated when the garlic cells are disrupted (powdered form). This enzyme converts alliin into allicin, the principal bioactive compound. The warm aqueous environment enhances this enzymatic reaction and facilitates the diffusion of allicin and other soluble constituents into the solvent.

5. Advantages of the Method

• Simple and cost-effective
• Does not require organic solvents
• Suitable for extraction of polar compounds
• Mild temperature preserves heat-sensitive constituents
• Safe and environmentally friendly

6. Limitations

• Limited extraction of non-polar compounds
• Allicin is unstable and may degrade over time
• Short shelf-life of aqueous extract
• Possibility of microbial growth if not stored properly

7. Applications

The obtained aqueous garlic extract can be used in:

• Herbal formulations
• Wound healing preparations
• Antimicrobial studies
• Gel and cream formulations (e.g., ethosomal gel research)

Preparation of Liposomes by Thin Film Hydration Method

Liposomes are spherical vesicles composed of one or more phospholipid bilayers enclosing an aqueous core. They are widely used as drug delivery systems due to their ability to encapsulate both hydrophilic and lipophilic drugs. Among various preparation techniques, the thin film hydration method is one of the most commonly employed methods due to its simplicity and reproducibility.

The thin film hydration method is based on the formation of a dry lipid film followed by its hydration to produce multilamellar vesicles (MLVs). Phospholipids and cholesterol are first dissolved in an organic solvent such as chloroform or a chloroform–methanol mixture. Upon evaporation of the solvent under reduced pressure, a thin, uniform lipid film is formed on the inner surface of a round-bottom flask.

When this lipid film is hydrated with an aqueous phase under controlled temperature and agitation, the lipid layers swell and peel off to form vesicles. The hydrophilic drug gets entrapped in the aqueous core, whereas lipophilic drugs are incorporated within the lipid bilayer.

The temperature during hydration is maintained above the phase transition temperature (Tc) of the lipid to ensure proper vesicle formation and stability.

Materials Required

• Phospholipids (e.g., lecithin)
• Cholesterol
• Organic solvents (chloroform, methanol)
• Drug (hydrophilic or lipophilic)
• Distilled water or buffer solution
• Round-bottom flask
• Rotary evaporator
• Water bath
• Vacuum pump
• Sonicator or extruder

Preparation of Garlic-Loaded Liposomes by Thin Film Hydration Method

• 1. Garlic Extract
• Biological Source: Obtained from bulbs of Allium sativum (Family: Amaryllidaceae)
• Chemical Constituents: Contains sulfur compounds such as allicin, alliin, ajoene, diallyl disulfide
• Molecular Formula (Allicin): C?H??OS?
• Molecular Weight: 162.27 g/mol
• Description: Pale yellow liquid with a characteristic pungent odor
• Solubility: Soluble in ethanol and partially soluble in water
• Mechanism of Action: Acts by reacting with thiol (-SH) groups of microbial enzymes, inhibiting metabolism and leading to cell death
• Uses: Antibacterial, antifungal, antioxidant
• Stability Issues: Highly unstable; degrades rapidly when exposed to heat, light, and oxygen

The molecular weight of the principal bioactive compound of garlic extract, allicin, is 162.27 g/mol. Molecular weight represents the sum of the atomic masses of all atoms present in a molecule and is an important physicochemical parameter influencing the behavior of a compound in biological systems. Allicin has the molecular formula C?H??OS?, which consists of carbon, hydrogen, oxygen, and sulfur atoms. The relatively low molecular weight of allicin facilitates its rapid diffusion across biological membranes, contributing to its effective antimicrobial activity. Due to its small size, allicin can easily penetrate microbial cell walls and interact with intracellular targets. Furthermore, molecular weight plays a significant role in drug delivery and formulation development, particularly in liposomal systems. Compounds with lower molecular weight are more efficiently encapsulated and can be released in a controlled manner from lipid vesicles. In the case of garlic extract, the molecular weight of allicin supports its incorporation into liposomal carriers, enhancing its stability and bioavailability. Thus, the molecular weight of allicin is a critical parameter that contributes to its pharmacokinetic properties, permeability, and overall therapeutic effectiveness in antimicrobial applications (42).

1. 2. Soya lecithin

Soya lecithin is a naturally occurring phospholipid mixture extracted from soybean oil during degumming, primarily comprising phosphatidylcholine (~20-40%) phosphatidylethanolamine, and phosphatidylinositol. This amphiphilic compound serves as an exceptional emulsifier, stabilizer, and wetting agent due to its hydrophilic head and hydrophobic fatty acid tails, enabling oil-water phase integration in formulations. In pharmaceutical sciences, particularly drug delivery systems, soya lecithin forms the structural backbone of liposomes and niosomes for encapsulating hydrophobic actives like thymol, enhancing bioavailability, controlled release, and membrane stability in wound healing applications. Its GRAS status, biocompatibility, and cost-effectiveness make it ideal for nanocarrier development targeting antimicrobial and anti-inflammatory therapies. Soya lecithin, derived from soybean oil, is a powerful natural emulsifier and functional ingredient widely used across the food, pharmaceutical, and animal feed industries. Known for its excellent emulsifying, stabilizing, and wetting properties, non-GMO soya lecithin is increasingly in demand as brands shift towards clean-label, plant-based, and sustainable solutions. In this guide, we explore everything you need to know about soya lecithin, including its benefits, applications, types, and why choosing non-GMO soy lecithin can give your products a market advantage.

Synonyms: Phosphatidylcholine, Soy phospholipid.

Methodology

Step 1: Preparation of Lipid Phase

Accurately weighed quantities of lecithin (phospholipid) and cholesterol are dissolved in a suitable organic solvent such as chloroform or a chloroform–methanol mixture in a clean, dry round-bottom flask. The lipid components are completely solubilized to form a clear lipid solution, ensuring uniform distribution of all constituents.

Step 2: Formation of Thin Lipid Film

The organic solvent is removed using a rotary evaporator under reduced pressure at a controlled temperature (typically 40–60°C). As the solvent evaporates, a thin, uniform lipid film is deposited on the inner wall of the flask. This step is critical for achieving proper vesicle formation. The film is further dried under vacuum to remove any residual solvent.

Step 3: Hydration with Garlic Extract

The dried lipid film is hydrated with the previously prepared aqueous garlic extract. Hydration is carried out at a temperature above the phase transition temperature of the lipid with continuous gentle agitation. During this process, the lipid film swells, detaches from the surface, and forms multilamellar vesicles (MLVs). The bioactive constituents of garlic are encapsulated within the aqueous core and partially within the lipid bilayer depending on their solubility.

Step 4: Sonication

The resulting liposomal suspension is subjected to sonication using a probe or bath sonicator for a specified duration. Sonication reduces the size of the vesicles and converts multilamellar vesicles into small unilamellar vesicles (SUVs), thereby improving uniformity, stability, and drug encapsulation efficiency. Care is taken to avoid excessive heat generation during this process.

Preparation of Liposomal Dusting Powder

1. Introduction

Liposomal dusting powder is a novel topical formulation in which drug-loaded liposomes are incorporated into a dry powder base. This system combines the advantages of liposomes (enhanced drug delivery, controlled release, and improved penetration) with the convenience and stability of a dusting powder. It is particularly useful for topical applications such as wound healing, antifungal, and antimicrobial treatments.

2. Principle

The preparation of liposomal dusting powder involves two major steps:

1. Preparation of drug-loaded liposomes (e.g., garlic extract-loaded liposomes)
2. Conversion of the liposomal dispersion into a dry powder form by adsorption or drying techniques

The liposomal suspension is adsorbed onto suitable carriers like talc or starch, or converted into dry powder by methods such as lyophilization (freeze-drying). This ensures better stability, ease of application, and prolonged shelf-life.

3. Materials Required

• Prepared liposomal suspension (garlic-loaded liposomes)
• Adsorbent base (talc, starch, kaolin)
• Glidant (optional, e.g., magnesium stearate)
• Preservatives (if required)
• Mortar and pestle
• Sieve (e.g., #80 mesh)
• Tray dryer or lyophilizer (optional)

4. Procedure

Step 1: Preparation of Liposomes

Liposomes are prepared using the thin film hydration method. Lecithin and cholesterol are dissolved in an organic solvent, followed by evaporation to form a thin film. The film is then hydrated with garlic extract and subjected to sonication to obtain a uniform liposomal suspension.

Step 2: Selection and Preparation of Powder Base

A suitable adsorbent powder base such as talc or starch is selected. The powder is passed through a sieve (e.g., #80 mesh) to ensure uniform particle size and to remove any impurities or aggregates.

Step 3: Adsorption of Liposomal Suspension

The prepared liposomal suspension is gradually added to the powder base in a mortar. The mixture is triturated continuously to ensure uniform adsorption of the liposomal dispersion onto the powder particles. Care is taken to avoid lump formation.

Step 4: Drying

The wet mass obtained after adsorption is subjected to drying. Drying can be performed by:

• Air drying at room temperature, or
• Tray drying at controlled temperature (not exceeding 40–50°C) to prevent degradation of liposomes

Alternatively, freeze-drying (lyophilization) may be used for better stability and preservation of vesicle structure.

Step 5: Pulverization and Sieving

The dried mass is gently pulverized using a mortar and pestle to obtain a fine powder. The powder is then passed through a sieve (e.g., #80 mesh) to ensure uniform particle size and smooth texture.

Step 6: Addition of Excipients (Optional)

If required, excipients such as glidants (e.g., magnesium stearate) or preservatives may be added and mixed uniformly.

Step 7: Packaging

The final liposomal dusting powder is packed in airtight containers or sifter-top containers to protect it from moisture and contamination.

5. Evaluation Parameters

• Particle size distribution
• Flow properties (angle of repose, bulk density)
• Drug content uniformity
• Moisture content
• In-vitro drug release
• Stability studies

6. Advantages

• Improved stability of liposomes in dry form
• Easy application and better patient compliance
• Controlled drug release
• Enhanced penetration through skin
• Reduced risk of microbial contamination

7. Limitations

• Possible aggregation of liposomes during drying
• Loss of vesicle integrity
• Additional processing steps required
• Optimization needed for adsorption efficiency

8. Applications

• Wound healing formulations
• Antifungal and antibacterial powders
• Dermatological treatments

Herbal topical drug delivery systems

1. 1. Solubility study

The solubility of Garlic extract in different solvents was determined. It was clear from the results in Table 7.4.1 that the solubility of Garlic extract in Ethanol is higher as compare to other solvents.

Table 1 Solubility of Garlic extract in different vehicle

1. 2. Determination of Calibration Curve

Standard calibration curve of Garlic extract was estimated by UV-spectrophotometer in Ethanol. Garlic extract was analysed using double beam ultraviolet spectrophotometer. The values of absorbance are mentioned in following tables. The plot of absorbance vs concentration was plotted and given in fig. Data in this range was further subjected to linear regression analysis. Regression coefficient was found 0.99 with linear graph hence obey Beer Lamberts Law in the concentration range of 2-10µg/ml at 275 nm against ethanol as blank for Garlic extract.

Fig.no. 20 Calibration curve of Garlic extract

Table 2 Calibration curve of Garlic extract

1. 3. Determination of λ-max

The λ-max of Garlic extract was taken on UV-visible Spectrophotometer (Shimadzu, UV- 1800 Japan) by scanning a diluted solution that was prepared in concentration range of 10 mg/ml and analyzed the solution and absorbance was taken. The λ-max of Garlic extract was found to be 275 nm. The wavelength in triplicate study shown in the table.

Table 3 Maximum wave length of garlic extract

1. 4. Drug Entrapment Efficiency

The entrapment efficiency (EE) of garlic extract in the prepared liposomes was determined using the ultracentrifugation method. The liposomal formulation, prepared with 300 mg of soya lecithin, 3 ml of ethanol, 1 ml of propylene glycol, 20 mg of tween 80,  6 ml of distilled water, exhibited high entrapment efficiencies for both compounds. The calculated entrapment efficiencies were found to be 77% for Garlic extract. The high entrapment efficiency can be attributed to the effective encapsulation of garlic extract within the liposomal vesicles. The soya lecithin provided a stable bilayer structure, which enhanced the drug loading capacity. Ethanol, a crucial component of the liposomal system, likely contributed to the fluidization of the lipid bilayer, thereby facilitating drug entrapment.

1. 5. In Vitro Diffusion Study

The in vitro drug release study was conducted to evaluate the diffusion profile of Garlic extract from the formulated Liposomes over a period of 24 hours. The cumulative drug release data is summarized in Table and graphically represented in figure.

Table 4 (a) Cumulative Release of garlic extract Over Time (suspension)

Figure: Graph showing diffusion study garlic extract (suspension).

A quick initial release of 3.33% garlic extract was seen at one hour.Drug release rose to 8.56% after two hours, marking the beginning of a more regulated release phase.Diffusion-controlled release was demonstrated by the drug's 18.84% release at 4 hours and its subsequent 30% release at 6 hours.44.06% were released by 8 hours, demonstrating the liposomal system's capacity to deliver prolonged drug release. The findings demonstrate that liposomal formulations have the potential to improve anti-microbial and wound healing efficacy through longer drug retention and absorption by effectively sustaining the release of Garlic extract over an extended period of time.

RESULT AND INTERPRETATION:

Particle Size Analysis of Liposomes

1. Result

The particle size of the prepared liposomal formulation was determined using the Dynamic Light Scattering (DLS) technique. The analysis revealed that the liposomes' average particle size (Z-average) was in the nanometer range.

• • Average Particle Size: 152.76 nm
  • Polydispersity Index (PDI): 0.257

Results

Cars index of powder

Particle size

Zeta potential