Standard Operating Procedure for Use of Cryoprotectants in Nanoparticle Formulations

1) Purpose

This SOP describes the use of cryoprotectants to prevent aggregation and maintain the stability of nanoparticle formulations during lyophilization and subsequent storage. Cryoprotectants help protect the structural integrity of nanoparticles when frozen and dried.

2) Scope

This SOP applies to personnel involved in the preparation of nanoparticle formulations that require lyophilization and need the addition of cryoprotectants to maintain stability.

3) Responsibilities

• Operators: Responsible for accurately adding cryoprotectants to the nanoparticle formulations as outlined in this SOP.
• QA: Verifies that the correct concentrations of cryoprotectants are used and ensures product stability after lyophilization.

4) Procedure

4.1 Selection of Cryoprotectants

4.1.1 Cryoprotectant Types

• 4.1.1.1 Select a cryoprotectant based on the type of nanoparticle and its sensitivity to freezing and drying. Common cryoprotectants include sucrose, trehalose, and mannitol.

4.2 Preparation of Cryoprotectant Solution

4.2.1 Solution Preparation

• 4.2.1.1 Prepare a cryoprotectant solution by dissolving the selected cryoprotectant in distilled water or a suitable buffer at the required concentration (e.g., 5–10% w/v).

4.3 Addition to Nanoparticle Formulation

4.3.1 Incorporation of Cryoprotectant

• 4.3.1.1 Add the cryoprotectant solution to the nanoparticle suspension before freezing. Stir gently to ensure the cryoprotectant is uniformly distributed.

4.4 Lyophilization

4.4.1 Freeze-Drying Process

• 4.4.1.1 Proceed with the lyophilization process as per the lyophilization SOP, ensuring that the cryoprotectant remains effective in protecting the nanoparticles during freezing and drying.

4.5 Storage of Lyophilized Nanoparticles

4.5.1 Storage Conditions

• 4.5.1.1 Store the lyophilized nanoparticles at 4°C or room temperature, depending on the stability of the formulation and the cryoprotectant used.

5) Abbreviations, if any

• QA: Quality Assurance

6) Documents, if any

• Cryoprotectant Usage Logbook

7) References, if any

• Guidelines for using cryoprotectants in nanoparticle formulations

8) SOP Version

Version 1.0

Annexure

Cryoprotectant Usage Logbook Template

  

    
Date
    
Batch Number
    
Cryoprotectant Type
    
Concentration (%)
    
Nanoparticle Stability
    
Operator Initials
    
QA Initials
  
  

    
DD/MM/YYYY
    
Batch Number
    
Cryoprotectant Used
    
Concentration %
    
Pass/Fail
    
Operator Name
    
QA Name
  
  

    
 
    
 
    
 
    
 
    
 
    
 
  

Standard Operating Procedure for Lyophilization of Nanoparticle-Based Formulations

1) Purpose

This SOP outlines the process for lyophilizing nanoparticle-based formulations, which helps to preserve their stability, increase shelf life, and facilitate easier transportation. The purpose is to standardize the procedure for freeze-drying nanoparticle formulations.

2) Scope

This SOP applies to laboratory personnel responsible for lyophilizing nanoparticle formulations used in drug delivery systems and other applications.

3) Responsibilities

• Operators: Responsible for accurately following the lyophilization procedure and ensuring the nanoparticles retain their stability after the process.
• QA: Ensures that the lyophilized products meet the required quality and stability standards.

4) Procedure

4.1 Preparation for Lyophilization

4.1.1 Pre-Freeze Preparation

• 4.1.1.1 Ensure the nanoparticle suspension is prepared and filtered to remove any aggregates before freeze-drying.
• 4.1.1.2 Dispense the nanoparticle formulation into vials, leaving sufficient headspace for sublimation during the lyophilization process.

4.2 Freezing

4.2.1 Initial Freezing

• 4.2.1.1 Place the vials in the lyophilizer’s pre-freezing chamber and freeze the formulation at temperatures between -40°C and -80°C, depending on the material’s characteristics.

4.3 Primary Drying

4.3.1 Sublimation

• 4.3.1.1 Set the lyophilizer to initiate the primary drying process by gradually reducing the pressure and maintaining low temperatures to allow sublimation of the frozen solvent.
• 4.3.1.2 Monitor the pressure and temperature to ensure sublimation occurs efficiently and completely.

4.4 Secondary Drying

4.4.1 Desorption of Bound Water

• 4.4.1.1 Raise the temperature slowly to between 20°C and 30°C while maintaining a low pressure to remove any residual water bound to the nanoparticles.

4.5 Storage of Lyophilized Nanoparticles

4.5.1 Sealing and Storage

• 4.5.1.1 Seal the vials under sterile conditions and store them at 4°C or room temperature, depending on the stability of the formulation.

5) Abbreviations, if any

• QA: Quality Assurance

6) Documents, if any

• Lyophilization Process Logbook

7) References, if any

• Manufacturer guidelines for lyophilizers

8) SOP Version

Version 1.0

Annexure

Lyophilization Process Logbook Template

  

    
Date
    
Batch Number
    
Freezing Temperature
    
Drying Pressure
    
Final Product Stability
    
Operator Initials
    
QA Initials
  
  

    
DD/MM/YYYY
    
Batch Number
    
Freezing Temp (°C)
    
Pressure (mbar)
    
Pass/Fail
    
Operator Name
    
QA Name
  
  

    
 
    
 
    
 
    
 
    
 
    
 
  

Standard Operating Procedure for Preparation of Nanoparticles for Targeted Drug Delivery

1) Purpose

The purpose of this SOP is to outline the procedure for preparing nanoparticles designed for targeted drug delivery. These nanoparticles are engineered to deliver therapeutic agents directly to specific cells or tissues, thereby enhancing drug efficacy and minimizing off-target effects.

2) Scope

This SOP applies to personnel involved in the formulation and characterization of nanoparticles for targeted drug delivery, particularly in pharmaceutical development and clinical research settings.

3) Responsibilities

• Operators: Responsible for following the outlined procedures for the preparation and characterization of the nanoparticles.
• QA: Ensures that the nanoparticle formulations meet the required specifications for particle size, drug loading, targeting efficacy, and sterility.

4) Procedure

4.1 Selection of Materials

4.1.1 Nanoparticle Material

• 4.1.1.1 Select appropriate materials such as PLGA (poly(lactic-co-glycolic acid)), PEGylated lipids, or other biodegradable polymers that provide stability and facilitate targeted delivery.

4.1.2 Drug and Targeting Ligand

• 4.1.2.1 Select the therapeutic agent based on its suitability for targeted delivery (e.g., chemotherapeutic agents, peptides).
• 4.1.2.2 Choose targeting ligands (e.g., antibodies, peptides) specific to receptors on the target cells or tissues.

4.2 Nanoparticle Preparation

4.2.1 Nanoparticle Synthesis

• 4.2.1.1 Use solvent evaporation, nanoprecipitation, or emulsification methods to prepare nanoparticles. Optimize the process to ensure a particle size between 100-200 nm for effective targeting.

4.2.2 Surface Functionalization

• 4.2.2.1 Conjugate targeting ligands (e.g., antibodies, aptamers) to the surface of the nanoparticles using techniques such as carbodiimide cross-linking (EDC/NHS) or thiol-maleimide reactions.

4.3 Characterization and Testing

4.3.1 Particle Size and Surface Charge

• 4.3.1.1 Measure the particle size using dynamic light scattering (DLS) and surface charge (zeta potential) to ensure the nanoparticles are within the desired range for targeting.

4.3.2 Drug Loading Efficiency

• 4.3.2.1 Quantify the amount of drug loaded into the nanoparticles using techniques such as UV-Vis spectrophotometry or high-performance liquid chromatography (HPLC).

4.3.3 In Vitro Targeting Efficacy

• 4.3.3.1 Perform in vitro studies on target cells (e.g., cancer cells) expressing the desired receptors to confirm the targeting capability of the nanoparticles.

4.4 Sterility and Storage

4.4.1 Sterilization

• 4.4.1.1 Sterilize the nanoparticle formulation by passing it through a 0.22 µm filter to remove contaminants.

4.4.2 Storage Conditions

• 4.4.2.1 Store the nanoparticles in sealed, sterile containers at 4°C to maintain stability and functionality of the targeting ligands.

5) Abbreviations, if any

• PLGA: Poly(lactic-co-glycolic acid)
• DLS: Dynamic Light Scattering
• EDC: 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide
• NHS: N-Hydroxysuccinimide

6) Documents, if any

• Nanoparticle Preparation Logbook
• Targeting Ligand Conjugation Logbook

7) References, if any

• Guidelines for nanoparticle preparation for drug delivery
• FDA Guidance on Nanotechnology

8) SOP Version

Version 1.0

Annexure

Nanoparticle Preparation Logbook Template

  

    
Date
    
Batch Number
    
Nanoparticle Material
    
Targeting Ligand
    
Particle Size
    
Drug Loading Efficiency
    
Operator Initials
    
QA Initials
  
  

    
DD/MM/YYYY
    
Batch Number
    
Material Name
    
Ligand Name
    
Size in nm
    
Efficiency (%)
    
Operator Name
    
QA Name
  
  

    
 
    
 
    
 
    
 
    
 
    
 
  

Targeting Ligand Conjugation Logbook Template

  

    
Date
    
Batch Number
    
Ligand Conjugated
    
Conjugation Method
    
Ligand Density
    
Binding Efficiency
    
Operator Initials
    
QA Initials
  
  

    
DD/MM/YYYY
    
Batch Number
    
Ligand Name
    
Method Used
    
Density (mol/mol)
    
Efficiency (%)
    
Operator Name
    
QA Name
  
  

    
 
    
 
    
 
    
 
    
 
    
 
  

Standard Operating Procedure for Formation of Solid Lipid Nanoparticles (SLNs)

1) Purpose

This SOP provides the steps for preparing solid lipid nanoparticles (SLNs), which are used in drug delivery systems for improving the bioavailability of poorly soluble drugs.

2) Scope

This SOP applies to laboratory personnel involved in the preparation of SLNs using the hot and cold homogenization methods.

3) Responsibilities

• Operators: Responsible for accurately preparing SLNs according to this SOP.
• QA: Ensures SLNs meet the required specifications for particle size, stability, and drug encapsulation.

4) Procedure

4.1 Preparation of Lipid Phase

4.1.1 Lipid Selection

• 4.1.1.1 Select a solid lipid suitable for the drug to be encapsulated (e.g., glyceryl monostearate, Compritol).

4.1.2 Melting the Lipid

• 4.1.2.1 Melt the solid lipid at a temperature 5–10°C above its melting point.

4.2 Drug Incorporation

4.2.1 Drug Loading

• 4.2.1.1 Dissolve or suspend the drug in the molten lipid phase and stir until the drug is uniformly dispersed.

4.3 Formation of SLNs

4.3.1 Emulsification

• 4.3.1.1 Add the molten lipid-drug mixture to an aqueous phase containing a surfactant (e.g., Poloxamer 188) under high-speed stirring to form an emulsion.
• 4.3.1.2 Use a high-pressure homogenizer to reduce the particle size.

4.3.2 Cooling

• 4.3.2.1 Cool the emulsion to room temperature, allowing the solid lipid to recrystallize, forming SLNs.

4.4 Washing and Collection of SLNs

4.4.1 Centrifugation

• 4.4.1.1 Centrifuge the SLN suspension at 10,000 g to collect the nanoparticles and wash with distilled water.

4.5 Particle Size Measurement

4.5.1 DLS Analysis

• 4.5.1.1 Use dynamic light scattering (DLS) to measure particle size and ensure that SLNs meet the required size specifications (typically 50–300 nm).

4.6 Storage

4.6.1 Storage Conditions

• 4.6.1.1 Store the SLN suspension in sterile, sealed containers at 4°C to maintain stability.

5) Abbreviations, if any

• DLS: Dynamic Light Scattering

6) Documents, if any

• SLN Formation Logbook

7) References, if any

• SLN formulation protocols for drug delivery

8) SOP Version

Version 1.0

Annexure

SLN Formation Logbook Template

  

    
Date
    
Batch Number
    
Lipid Type
    
Particle Size
    
Encapsulation Efficiency
    
Operator Initials
    
QA Initials
  
  

    
DD/MM/YYYY
    
Batch Number
    
Lipid Type
    
Size in nm
    
% Encapsulation
    
Operator Name
    
QA Name
  
  

    
 
    
 
    
 
    
 
    
 
    
 
  

Standard Operating Procedure for Solvent Evaporation Method for Nanoparticles

1) Purpose

This SOP provides the detailed steps for the solvent evaporation method used in nanoparticle preparation. The purpose is to ensure consistency and control in the formation of nanoparticles for drug delivery applications.

2) Scope

This SOP applies to laboratory staff involved in the formulation of nanoparticles using the solvent evaporation method.

3) Responsibilities

• Operators: Responsible for carrying out the solvent evaporation technique as per this SOP.
• QA: Ensures quality control by verifying particle size and encapsulation efficiency.

4) Procedure

4.1 Polymer Dissolution

4.1.1 Solvent Selection

• 4.1.1.1 Select a suitable organic solvent (e.g., dichloromethane) and dissolve the polymer (e.g., PLGA) under gentle stirring.

4.1.2 Drug Loading

• 4.1.2.1 Dissolve or suspend the drug in the polymer solution based on its solubility profile.

4.2 Formation of Emulsion

4.2.1 Emulsification

• 4.2.1.1 Add the polymer-drug solution into an aqueous phase containing a surfactant (e.g., PVA) under continuous stirring to form an emulsion.
• 4.2.1.2 Use a high-speed homogenizer to reduce the size of emulsion droplets.

4.3 Solvent Evaporation

4.3.1 Solvent Removal

• 4.3.1.1 Evaporate the solvent under reduced pressure or stirring at room temperature, leading to nanoparticle formation as the polymer solidifies.
• 4.3.1.2 Continue until no solvent remains in the suspension.

4.4 Washing and Collection of Nanoparticles

4.4.1 Centrifugation

• 4.4.1.1 Centrifuge the nanoparticle suspension to collect the solid particles and discard the supernatant.
• 4.4.1.2 Wash the particles with distilled water and repeat the centrifugation process.

4.5 Particle Size Measurement

4.5.1 Dynamic Light Scattering (DLS)

• 4.5.1.1 Use DLS or another method to measure the particle size of the nanoparticles and ensure they meet the desired specifications (e.g., 100–200 nm).

4.6 Storage

4.6.1 Storage Conditions

• 4.6.1.1 Store the nanoparticles in sterile, sealed containers at 4°C or freeze-dry them for long-term stability.

5) Abbreviations, if any

• PLGA: Poly(lactic-co-glycolic acid)
• PVA: Polyvinyl Alcohol
• DLS: Dynamic Light Scattering

6) Documents, if any

• Solvent Evaporation Method Logbook

7) References, if any

• Formulation protocols for nanoparticles using solvent evaporation

8) SOP Version

Version 1.0

Annexure

Solvent Evaporation Method Logbook Template

  

    
Date
    
Batch Number
    
Polymer Used
    
Particle Size
    
Encapsulation Efficiency
    
Operator Initials
    
QA Initials
  
  

    
DD/MM/YYYY
    
Batch Number
    
Polymer Type
    
Size in nm
    
% Encapsulation
    
Operator Name
    
QA Name
  
  

    
 
    
 
    
 
    
 
    
 
    
 
  

Standard Operating Procedure for Preparation of Nanoparticle Suspensions

1) Purpose

This SOP outlines the procedure for preparing stable nanoparticle suspensions used in drug delivery, diagnostics, and other applications. The goal is to ensure that nanoparticles are uniformly dispersed in the suspension and maintain stability over time.

2) Scope

This SOP applies to laboratory personnel responsible for the preparation of nanoparticle suspensions in a sterile and controlled environment.

3) Responsibilities

• Operators: Responsible for following the SOP to prepare stable nanoparticle suspensions.
• QA: Ensures that nanoparticle suspensions meet the required size, stability, and quality parameters.

4) Procedure

4.1 Preparation of Nanoparticles

4.1.1 Nanoparticle Selection

• 4.1.1.1 Select the desired type of nanoparticles based on the specific application (e.g., polymeric, metallic, or lipid-based nanoparticles).

4.2 Preparation of Suspension Medium

4.2.1 Solvent Selection

• 4.2.1.1 Prepare a suspension medium using distilled water, saline solution, or a buffer depending on the application.

4.2.2 Stabilizers

• 4.2.2.1 Add a stabilizer (e.g., PVA, Tween 80) to the suspension medium to prevent nanoparticle aggregation.

4.3 Suspending the Nanoparticles

4.3.1 Dispersion

• 4.3.1.1 Add the nanoparticles to the suspension medium under gentle stirring.
• 4.3.1.2 Use an ultrasonic bath or homogenizer to ensure uniform dispersion of the nanoparticles in the suspension.

4.3.2 Monitoring Particle Size

• 4.3.2.1 Measure the particle size using dynamic light scattering (DLS) to ensure the nanoparticles remain in the desired size range (typically less than 200 nm).

4.4 Storage and Stability Testing

4.4.1 Storage

• 4.4.1.1 Store the nanoparticle suspension in sterile, sealed containers at 4°C.

4.4.2 Stability Testing

• 4.4.2.1 Periodically test the suspension for particle size, zeta potential, and any signs of aggregation or sedimentation.

5) Abbreviations, if any

• DLS: Dynamic Light Scattering
• PVA: Polyvinyl Alcohol

6) Documents, if any

• Nanoparticle Suspension Preparation Logbook

7) References, if any

• Relevant literature on the preparation and stabilization of nanoparticle suspensions

8) SOP Version

Version 1.0

Annexure

Nanoparticle Suspension Preparation Logbook Template

  

    
Date
    
Batch Number
    
Particle Type
    
Particle Size
    
Stabilizer Used
    
Operator Initials
    
QA Initials
  
  

    
DD/MM/YYYY
    
Batch Number
    
Type of Nanoparticles
    
Size in nm
    
Stabilizer Name
    
Operator Name
    
QA Name
  
  

    
 
    
 
    
 
    
 
    
 
    
 
  

Standard Operating Procedure for Preparation of Nanoemulsions

1) Purpose

This SOP outlines the procedure for preparing nanoemulsions, which are used as drug delivery systems to enhance the solubility of poorly soluble drugs. The aim is to ensure a consistent and controlled method for nanoemulsion preparation.

2) Scope

This SOP applies to all laboratory personnel involved in the formulation and preparation of nanoemulsions for drug delivery applications.

3) Responsibilities

• Operators: Responsible for following the SOP to accurately prepare nanoemulsions.
• QA: Ensures that the nanoemulsions meet quality and size specifications.

4) Procedure

4.1 Preparation of Oil Phase

4.1.1 Oil and Surfactant Selection

• 4.1.1.1 Select an appropriate oil phase based on the solubility of the drug and the desired application of the nanoemulsion.
• 4.1.1.2 Dissolve the drug in the oil phase under continuous stirring at room temperature.

4.2 Preparation of Aqueous Phase

4.2.1 Stabilizer Selection

• 4.2.1.1 Select a stabilizer or surfactant suitable for the formation of stable nanoemulsions.
• 4.2.1.2 Prepare the aqueous phase by dissolving the stabilizer in distilled water under gentle stirring.

4.3 Emulsification

4.3.1 Formation of Nanoemulsion

• 4.3.1.1 Add the oil phase to the aqueous phase under high-speed stirring to form an emulsion.
• 4.3.1.2 Continue stirring for 10–15 minutes to ensure proper emulsification.

4.3.2 Particle Size Reduction

• 4.3.2.1 Use a high-pressure homogenizer or ultrasonicator to reduce the particle size of the emulsion droplets to the nanoscale (typically less than 200 nm).
• 4.3.2.2 Monitor the particle size using dynamic light scattering (DLS).

4.4 Storage and Stability

4.4.1 Storage of Nanoemulsions

• 4.4.1.1 Store the prepared nanoemulsion in sterile, sealed containers at 4°C to maintain stability.

4.4.2 Stability Testing

• 4.4.2.1 Periodically test the nanoemulsion for particle size and physical stability (e.g., phase separation, creaming) to ensure the formulation remains stable over time.

5) Abbreviations, if any

• DLS: Dynamic Light Scattering

6) Documents, if any

• Nanoemulsion Preparation Logbook

7) References, if any

• Literature on nanoemulsion formulations and stability testing

8) SOP Version

Version 1.0

Annexure

Nanoemulsion Preparation Logbook Template

  

    
Date
    
Batch Number
    
Oil Phase
    
Particle Size
    
Stability Test
    
Operator Initials
    
QA Initials
  
  

    
DD/MM/YYYY
    
Batch Number
    
Oil Phase Used
    
Size in nm
    
Pass/Fail
    
Operator Name
    
QA Name
  
  

    
 
    
 
    
 
    
 
    
 
    
 
    
 
  

Standard Operating Procedure for High-Pressure Homogenization in Nanoparticle Production

1) Purpose

This SOP outlines the high-pressure homogenization method used for reducing particle size in nanoparticle production, a technique commonly employed to achieve nanoparticles for drug delivery and other applications.

2) Scope

This SOP applies to all operators using high-pressure homogenizers to reduce the particle size of nanoparticles in formulations.

3) Responsibilities

• Operators: Responsible for setting up and operating the high-pressure homogenizer.
• QA: Ensures the final nanoparticles meet quality and size specifications.

4) Procedure

4.1 Preparation of Sample

4.1.1 Sample Preparation

• 4.1.1.1 Prepare the nanoparticle suspension as per formulation guidelines, ensuring that the starting particle size is suitable for homogenization.
• 4.1.1.2 Transfer the suspension into the homogenizer feed tank.

4.2 High-Pressure Homogenization

4.2.1 Equipment Setup

• 4.2.1.1 Calibrate the high-pressure homogenizer according to the manufacturer’s instructions.
• 4.2.1.2 Set the operating pressure between 500 and 1500 bar, depending on the formulation needs.

4.2.2 Particle Size Reduction

• 4.2.2.1 Pass the nanoparticle suspension through the homogenizer for multiple cycles (usually 3–5 passes) until the desired particle size is achieved.
• 4.2.2.2 Monitor the particle size during the process using dynamic light scattering (DLS) or a similar technique.

4.3 Cleaning and Maintenance

4.3.1 Cleaning of Equipment

• 4.3.1.1 Clean the homogenizer thoroughly after each batch using sterile water and a cleaning solution to remove any residual particles or contaminants.

4.4 Storage of Nanoparticles

4.4.1 Storage Conditions

• 4.4.1.1 Store the nanoparticle suspension in sealed, sterile containers at 4°C, or freeze-dry for long-term storage if necessary.

5) Abbreviations, if any

• DLS: Dynamic Light Scattering
• QA: Quality Assurance

6) Documents, if any

• High-Pressure Homogenization Process Logbook

7) References, if any

• Manufacturer’s instructions for high-pressure homogenizer

8) SOP Version

Version 1.0

Annexure

High-Pressure Homogenization Process Logbook Template

  

    
Date
    
Batch Number
    
Pressure (bar)
    
Particle Size
    
Encapsulation Efficiency
    
Operator Initials
    
QA Initials
  
  

    
DD/MM/YYYY
    
Batch Number
    
Pressure Used (e.g., 1000 bar)
    
Size in nm
    
% Encapsulation
    
Operator Name
    
QA Name
  
  

    
 
    
 
    
 
    
 
    
 
    
 
  

Standard Operating Procedure for Nanoprecipitation Method in Nanoparticle Formulation

1) Purpose

This SOP outlines the nanonprecipitation method for preparing nanoparticles, typically used to create polymer-based nanoparticles for drug delivery. The technique allows for the formation of nanoparticles by precipitation from a solution.

2) Scope

This SOP applies to laboratory personnel responsible for the formulation of nanoparticles using the nanonprecipitation method.

3) Responsibilities

• Operators: Responsible for following the nanonprecipitation protocol and ensuring accurate nanoparticle formation.
• QA: Ensures the nanoparticles meet the required quality standards.

4) Procedure

4.1 Polymer Solution Preparation

4.1.1 Polymer Selection

• 4.1.1.1 Select an appropriate polymer (e.g., PLGA) and dissolve it in a water-miscible organic solvent (e.g., acetone or ethanol).
• 4.1.1.2 Stir until the polymer is completely dissolved.

4.2 Precipitation Process

4.2.1 Addition of Polymer Solution

• 4.2.1.1 Slowly add the polymer solution into an aqueous phase containing stabilizers under gentle stirring to form nanoparticles by precipitation.
• 4.2.1.2 Maintain the stirring rate until the nanoparticles have formed, typically over a period of 10–15 minutes.

4.2.2 Particle Size Control

• 4.2.2.1 Monitor particle size using dynamic light scattering (DLS) to ensure the nanoparticles are within the desired size range.

4.3 Washing and Purification

4.3.1 Centrifugation

• 4.3.1.1 Centrifuge the nanoparticle suspension at 10,000 g to collect the nanoparticles and discard the supernatant.
• 4.3.1.2 Wash the nanoparticles with distilled water and repeat the centrifugation process.

4.4 Drying and Storage

4.4.1 Drying

• 4.4.1.1 Freeze-dry or vacuum-dry the nanoparticles, depending on the formulation requirements.

4.4.2 Storage

• 4.4.2.1 Store the dried nanoparticles in sterile containers at 4°C or room temperature.

5) Abbreviations, if any

• PLGA: Poly(lactic-co-glycolic acid)
• DLS: Dynamic Light Scattering

6) Documents, if any

• Nanoprecipitation Process Logbook

7) References, if any

• Guidelines for nanoprecipitation methods in nanoparticle formulation

8) SOP Version

Version 1.0

Annexure

Nanoprecipitation Process Logbook Template

  

    
Date
    
Batch Number
    
Polymer Type
    
Particle Size
    
Encapsulation Efficiency
    
Operator Initials
    
QA Initials
  
  

    
DD/MM/YYYY
    
Batch Number
    
Polymer Name
    
Size in nm
    
% Encapsulation
    
Operator Name
    
QA Name
  
  

    
 
    
 
    
 
    
 
    
 
    
 
  

Standard Operating Procedure for Supercritical Fluid Techniques in Nanoparticle Preparation

1) Purpose

This SOP outlines the use of supercritical fluid techniques for the preparation of nanoparticles, which allow for controlled particle size and purity. The supercritical fluid method is commonly used to prepare drug nanoparticles with enhanced solubility.

2) Scope

This SOP applies to all laboratory personnel involved in the use of supercritical fluids for the preparation of nanoparticles.

3) Responsibilities

• Operators: Responsible for following the supercritical fluid technique protocol accurately.
• QA: Ensures that nanoparticles prepared using supercritical fluids meet the required quality standards.

4) Procedure

4.1 Setup of Supercritical Fluid Apparatus

4.1.1 Equipment Preparation

• 4.1.1.1 Ensure that the supercritical fluid apparatus is cleaned, calibrated, and properly set up according to the manufacturer’s guidelines.
• 4.1.1.2 Check for leaks and ensure that all safety protocols are in place for handling supercritical CO₂ or other fluids.

4.1.2 Solvent and Drug Preparation

• 4.1.2.1 Dissolve the drug or active ingredient in a solvent appropriate for the supercritical fluid process (e.g., ethanol, acetone).
• 4.1.2.2 Set up the drug solution and supercritical fluid for introduction into the apparatus.

4.2 Supercritical Fluid Process

4.2.1 Introduction of Supercritical Fluid

• 4.2.1.1 Slowly introduce the supercritical fluid (e.g., CO₂) into the system at the specified temperature and pressure conditions.
• 4.2.1.2 Ensure that the flow rates of both the drug solution and supercritical fluid are consistent and as per the process requirement.

4.2.2 Particle Formation

• 4.2.2.1 As the supercritical fluid expands, nanoparticles are formed as the solvent evaporates and the drug precipitates.
• 4.2.2.2 Collect the formed nanoparticles in the collection chamber.

4.3 Washing and Drying

4.3.1 Washing with Solvent

• 4.3.1.1 Wash the nanoparticles using a suitable solvent to remove any residual supercritical fluid or impurities.

4.3.2 Drying of Nanoparticles

• 4.3.2.1 Dry the nanoparticles under vacuum or in a desiccator to ensure complete removal of solvent residues.

4.4 Characterization

4.4.1 Particle Size Analysis

• 4.4.1.1 Use dynamic light scattering (DLS) or similar techniques to measure the particle size of the nanoparticles.
• 4.4.1.2 Record the particle size and ensure it meets the required specifications.

4.5 Storage of Nanoparticles

4.5.1 Storage Conditions

• 4.5.1.1 Store the nanoparticles in sterile, sealed containers and maintain them at room temperature or 4°C depending on the stability of the formulation.

5) Abbreviations, if any

• DLS: Dynamic Light Scattering
• CO₂: Carbon Dioxide
• QA: Quality Assurance

6) Documents, if any

• Supercritical Fluid Process Logbook

7) References, if any

• Relevant literature on supercritical fluid techniques in nanoparticle preparation

8) SOP Version

Version 1.0

Annexure

Supercritical Fluid Process Logbook Template

  

    
Date
    
Batch Number
    
Fluid Used
    
Particle Size
    
Encapsulation Efficiency
    
Operator Initials
    
QA Initials
  
  

    
DD/MM/YYYY
    
Batch Number
    
Fluid (e.g., CO2)
    
Size in nm
    
% Encapsulation
    
Operator Name
    
QA Name