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Nanoemulsion Preparation Process Using High-Pressure Homogenizer

Note: This nanoemulsion preparation protocol is intended for laboratory research purposes only. Process parameters may require optimization depending on materials, equipment configuration, and application conditions. Results may vary between laboratories.

Nanoemulsions are fine oil-in-water or water-in-oil dispersions with droplet sizes typically below 200 nm, widely used in pharmaceutical, cosmetic, and functional food applications due to their high stability and solubilization capability.

This guide provides a step-by-step workflow for nanoemulsion preparation using high-pressure homogenization for formulation and process development studies.

1. Why Use High-Pressure Homogenization for Nanoemulsions?

High-pressure homogenization is a widely used technique for producing nanoemulsions at laboratory scale due to its strong droplet size reduction capability and good reproducibility. Compared with conventional emulsification methods such as ultrasonication or low-shear mixing, it provides:

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Consistent nanoscale droplet size distribution

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High reproducibility for formulation development

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Improved physical stability of emulsions

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Process parameters that can be linearly scaled with appropriate system design

2. Materials & Reagents

API:

Oil-soluble active ingredient

Oil phase:

MCT (Medium Chain Triglycerides)

Emulsifiers:

Soy lecithin, glycerol, etc.

Aqueous phase:

Water for injection

Buffer & osmotic agents:

Na₂HPO₄, NaCl, etc.

pH adjusters:

NaOH, HCl

3. Step-by-Step Nanoemulsion Preparation

Step 1: Prepare Oil Phase

Dissolve the API in MCT and stir in a 60°C water bath until fully dissolved, forming a clear oil phase.

Step 2: Prepare Aqueous Phase

Add emulsifiers, phospholipids, and water for injection. Heat to 60°C and stir until no visible solids or oil droplets remain.

Step 3: Prepare Coarse Emulsion

Under 60°C and high-shear mixing (12,000 rpm), slowly add the oil phase into the aqueous phase. Continue shearing for 5–10 minutes to obtain a uniform coarse emulsion.

Step 4: High-Pressure Homogenization

Transfer the coarse emulsion into a high-pressure homogenizer, NanoGenizer-30K. Process at 15,000 psi for 2 passes at room temperature, then increase pressure to 30,000 psi until the desired droplet size distribution is achieved.

NanoGenizer High-Pressure Homogenizer 

Step 5: pH Adjustment

Add buffer solution, mix thoroughly, and adjust pH to the desired range using NaOH or HCl.

Step 6: Characterization

Analyze particle size, polydispersity index (PDI), and zeta potential using dynamic light scattering (DLS).

4. Results

After high-pressure homogenization, the nanoemulsion exhibits significantly reduced droplet size and improved physical stability at laboratory scale.

Before high-pressure homogenization: Coarse, heterogeneous emulsion

After high-pressure homogenization: Stable nanoemulsion with uniform nanoscale droplets

Nanoemulsion Before and After High-Pressure Homogenization

Particle Size Comparison Before and After High-Pressure Homogenization

5. Application Notes

This nanoemulsion preparation workflow is widely used in pharmaceutical formulation development, cosmetic research, and functional delivery system design. Process parameters such as pressure, number of passes, and temperature should be optimized according to specific formulation properties.

6. Process Development & Scale-Up Considerations

While this protocol describes a laboratory-scale workflow, high-pressure homogenization allows scalable transition of process parameters when moving toward pilot and further scale-up development, provided that system configuration and formulation conditions are appropriately optimized.

For process optimization, scale-up support, or equipment selection, please contact Genizer technical team.

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How to Prepare Liposomes: Step-by-Step Liposome Preparation Guide

Liposomes are spherical vesicles widely used in pharmaceuticals, cosmetics, and nutraceuticals for their excellent biocompatibility and capability to encapsulate both hydrophilic and lipophilic substances. This guide presents a step-by-step laboratory workflow for liposome preparation, intended for research and educational purposes.

1. Materials & Reagents

Phospholipids: Soy Phosphatidylcholine (SPC), Saturated Soy Phosphatidylcholine (SSPC), etc.

Cholesterol: Improves membrane stability and fluidity

DSPE-PEG2000: Optional, for long circulation or targeted liposomes

Lipophilic compounds: Pre-dissolved in organic solvents if required

Hydrophilic compounds / aqueous phase: PBS buffer, pure water, or compound-containing buffer

Organic solvents: Anhydrous ethanol, chloroform

2. Step-by-Step Liposome Preparation

Step 1: Dissolve Lipids

Dissolve phospholipids, cholesterol, and other lipid components in anhydrous ethanol or chloroform. Sonicate the mixture until a clear solution is obtained.

Step 2: Form Lipid Film

Evaporate the solvent using a rotary evaporator under vacuum for approximately 60 minutes to form a uniform, dry lipid film at the bottom of the flask.

Step 3: Hydration

Add the hydration medium to the flask and rotate in a water bath at a temperature above the phospholipid phase transition temperature for approximately 30 minutes to form crude liposomes (multilamellar vesicles, MLVs). 

Step 4: Particle Size Reduction

To reduce the particle size of the crude liposome suspension, a high-pressure homogenizer such as the NanoGenizer can be used. Process the suspension at 10,000–25,000 psi for 3–10 cycles until the target particle size is achieved.

Alternatively, membrane filtration extrusion with a liposome extruder is suitable for both small- and larger-scale production. Pass the crude liposome suspension through filter membranes with defined pore sizes until the desired particle size is obtained.

Tip: To learn more about choosing between a high-pressure homogenizer and a liposome extruder, read our guide here.

Step 5: Remove Unencapsulated Compounds

Free compounds can be separated from liposomes using techniques such as gel filtration, ultrafiltration, dialysis, or ultracentrifugation.

Step 6: Characterization

Particle size and zeta potential: Measure the average particle size, polydispersity index (PDI), and surface charge of liposomes using dynamic light scattering (DLS). Encapsulation efficiency: Determine drug content after disrupting the liposomes to calculate encapsulation efficiency.

3. Typical Liposome Preparation Results

After particle size reduction, the liposome suspension becomes more uniform and transparent, with an average particle size of 50–150 nm and a narrower distribution.

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Lipid Nanoparticles(LNPs): Advantages, Applications, and Preparation

What Are Lipid Nanoparticles?

Lipid nanoparticles (LNPs) are nanoscale delivery systems composed of lipid-based materials, typically 50–150 nm in diameter. They are widely used in pharmaceutical, biotechnology, and industrial formulation applications to encapsulate and transport therapeutic molecules. By forming a core stabilized by a lipid shell, LNPs provide effective protection for sensitive compounds and enable controlled release and efficient cellular delivery.

Advantages and Applications of LNPs

Advantages of LNPs:

High stability: Protects sensitive drugs from degradation.

Improved cellular uptake: Lipids help drugs enter cells.

Optimized biodistribution: Ensures targeted delivery and longer circulation.

Versatility: Can carry nucleic acids, small molecules, or hydrophobic drugs.

Scalable production: Suitable for lab research and industrial manufacturing.

Applications of LNPs:

mRNA vaccines: Protect RNA and enhance immune response.

Gene therapy: Deliver RNA or DNA safely to target cells.

Small molecule drugs: Improve solubility, stability, and targeted delivery.

Cosmetics and nutraceuticals: Controlled release of active ingredients.

Preparation of Lipid Nanoparticles

To achieve consistent performance, lipid nanoparticles must be produced using preparation methods that provide reproducible particle size and stability while supporting scale-up. Common LNP preparation techniques include thin-film hydration, solvent injection, double emulsion methods, microfluidic mixing, and mechanical size-reduction processes. Among these approaches, high-pressure homogenization is widely adopted for scalable production. By using the diamond interaction chamber to generate mechanical forces such as high shear, cavitation, impact, and high-frequency oscillation, the high-pressure homogenization method avoids complex microstructures and disposable components, making it ideal for both laboratory process development and large-scale manufacturing.

High-Pressure Homogenization Process

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Polymer Nanoparticles: From Formulation to Scalable Manufacturing

What Are Polymer Nanoparticles?

Polymer nanoparticles are widely used in modern drug delivery and advanced material systems. Owing to their small size and tunable surface properties, they enable controlled drug release, targeted delivery, and improved bioavailability. Beyond pharmaceuticals, polymer nanoparticles are also applied in diagnostics, functional coatings, nanocomposites, and other functional materials where stability and consistency are critical.

As interest in nanomedicine and functional materials continues to grow, the challenge is no longer just how to design polymer nanoparticles—but how to manufacture them reliably and at scale.

Figure 1. Schematic illustration of polymer nanoparticles

As polymer-based nanotechnology continues to evolve, increasing attention is being paid not only to formulation design, but also to scalable and reproducible manufacturing strategies.

Why Are Polymer Nanoparticles Difficult to Manufacture?

Although polymer nanoparticles offer clear functional advantages, producing them with consistent quality remains challenging.

Traditional preparation methods—such as emulsion–solvent evaporation and nanoprecipitation—often present several limitations:

Broad particle size distributions

Residual organic solvents

Limited batch-to-batch reproducibility

Difficult and time-consuming scale-up

In practice, formulations that perform well at the laboratory scale frequently require substantial re-optimization when transferred to pilot or manufacturing volumes. Taken together, these challenges make it difficult to translate promising lab-scale formulations into stable, repeatable production processes.

As a result, there is growing demand for processing technologies that can provide both formulation flexibility and manufacturing reliability.

What Is High-Pressure Microfluidic Homogenization?

High-pressure microfluidic homogenization is an advanced particle processing technology that combines precisely engineered microfluidic channels with high-pressure energy input. Unlike conventional microfluidic mixing, which mainly relies on controlled flow and diffusion, this process is energy-driven.

Within the interaction chamber, intense shear, impact, and cavitation forces are generated, actively breaking down polymer particles to the nanoscale. All critical processing occurs inside this chamber, enabling highly controlled and repeatable particle size reduction.

Figure 2. Internal microchannel structure of a high-pressure microfluidic interaction chamber

This approach offers several key advantages:

Narrow and uniform particle size distribution

Straightforward scalability from laboratory to pilot and production volumes

Excellent batch-to-batch reproducibility

Gentle processing conditions, which may reduce the need for excessive stabilizers and help preserve sensitive polymer structures

Because the same physical processing principles can be maintained across different throughput levels, high-pressure microfluidic homogenization is well suited for applications that require reliable scale-up without compromising particle quality.

Bridging Research and Real-World Production

One of the central challenges in polymer nanoparticle development is ensuring that successful laboratory formulations can be translated into robust manufacturing processes.

Figure 3. High-pressure microfluidic homogenizer used for polymer nanoparticle production

As polymer nanoparticle applications continue to expand across pharmaceuticals and advanced materials, scalable and reproducible manufacturing has become just as critical as formulation design. Technologies that enable seamless scale-up play an essential role in bringing polymer nanoparticle innovations from the laboratory to reliable industrial production.

High-pressure microfluidic homogenization systems, such as those developed by Genizer, are designed to maintain consistent processing conditions across laboratory, pilot, and manufacturing scales. By preserving key parameters such as particle size distribution and formulation performance, these systems help bridge the gap between early-stage research and real-world production requirements.

Explore Genizer Homogenizers

NanoGenizer High-Pressure Microfluidic Homogenizer – Lab Scale for Nanoparticle and Liposome Applications

PilotGenizer High-Pressure Homogenizer – Pilot & Production Volumes

WILL Series High-Pressure Homogenizer – Production Scale

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E. coli Cell Disruption by High-Pressure Homogenization

What is E. coli cell disruption and how to disrupt E. coli cells?

Escherichia coli (E.coli) cell disruption is essential in microbial research, protein purification, and biotechnological applications. disruption is essential in microbial research, protein purification, and biotechnological applications. Among the available methods, high-pressure homogenization provides efficient, reproducible, and scalable E. coli cell disruption, making it ideal for laboratory and industrial purposes.

How does High-Pressure Homogenization work for E. coli cell disruption?

During high-pressure homogenization, E.coli cells pass through an interaction chamber containing micro-scale channels under high pressure. Intense shear, impact, turbulence, cavitation, and sudden pressure drops lead to cell disruption, rupturing cell walls and membranes and releasing intracellular contents into the solution.

This method, also known as cell lysis, enables efficient and reproducible E.coli cell disruption while preserving protein activity.

Compared with chemical lysis or sonication, high-pressure homogenization provides uniform and reproducible E.coli cell disruption and is particularly suitable for high-density cultures. Typical operating pressures range from 12,000–20,000 psi. More than three passes achieve >99.99% disruption, while five passes generally result in complete lysis and consistent protein extraction.

To evaluate the effectiveness of high-pressure homogenization, the following experiments were conducted.

Experimental Setup for E. coli Cell Disruption

1. Experimental Method

E. coli samples were homogenized 3–5 times using the NanoGenizer-30K high-pressure homogenizer equipped with RT-cooling diamond interaction chambers (H30Z-RT & H10Z-RT) and a heat exchanger. The temperature was maintained at 4 °C using a chilled water bath to preserve protein activity. 

2. Disruption Rate Test Method

2.1 Viable Bacteria Counting

After 24 hours of cultivation, the E. coli fermentation broth was centrifuged at 4,000 rpm for 15 minutes, the supernatant was discarded, and the cells were resuspended in buffer. Then, 100 µL of an appropriate dilution was applied to a plate containing ampicillin and chloramphenicol for viable bacteria counting. The remaining bacterial solution was divided into two portions and homogenized at 15,000 psi for 3 passes and 5 passes, respectively. After homogenization, 100 µL of each bacterial solution was taken, appropriately diluted, plated, and the viable bacteria were counted.

2.2 Protein Extraction

Bacterial samples were homogenized 3 or 5 times at 15,000 psi using the NanoGenizer-30K. The homogenate was centrifuged at 11,000–13,000 rpm for 15 minutes, and the supernatant was collected for protein extraction and quantification. This process ensures consistent recovery of intracellular proteins.

3. Experimental Results 3.1 Appearance after Homogenization

3 passes: solution contained numerous flocculent particles and layering; bacterial fragments relatively large. 5 passes: solution more uniform, indicating thorough homogenization.

E. coli suspension after high-pressure homogenization

3.2 Pre-Homogenization Count

Diluted 10³ times: ~500 colonies → estimated concentration 5×10⁸ CFU/mL.

E. coli suspension before high-pressure homogenization

3.3 Post-Homogenization (3 passes)

Diluted 10³ times: ~20 colonies → ~2×10⁴ CFU/mL.

E. coli suspension after 3 passes of high-pressure homogenization

3.4 Post-Homogenization (5 passes)

Diluted 10, 10², and 10³: no colonies observed → 100% cell lysis.

E. coli suspension after 5 passes of high-pressure homogenization

3.5 Protein Extraction

Bacterial proteins were effectively extracted with similar efficiency using Z-type interaction chambers (3 or 5 passes) or Y-type interaction chambers (5 passes).

Protein extraction from E. coli after high-pressure homogenization

Conclusion

3 passes of high-pressure homogenization achieved >99.99%  E. coli cell disruption (lysis).

5 passes achieved complete cell disruption. 

Protein extraction yields remained consistent across different passes and chamber types.

NanoGenizerHigh Pressure Homogenizer for E. coli cell Disruption – NanoGenizer

Laboratory-Scale High-Pressure Homogenizer – NanoGenizer (up to 45,000 psi)

The NanoGenizer supports precise pressure control, efficient cooling, and reproducible results, making it ideal for microbial cell disruption (including bacteria, yeast, algae, mammalian, and insect cells) and protein extraction. It is also suitable for related applications such as nanoemulsions, fat emulsions, liposomes, nano dispersions, deagglomeration, and lipid nanoparticles, etc.

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Graphene Exfoliation and Dispersion Using High Pressure Homogenizers

What Is Graphene and Its Applications

Graphene is a two-dimensional carbon material valued for its excellent electrical conductivity, mechanical strength, thermal stability, and chemical resistance. These properties make graphene ideal for energy storage, electronics, functional coatings, conductive inks, and advanced composite materials.

Challenges in Graphene Production 

Graphene Exfoliation using NanoGenizer High Pressure Homogenizer  

Liquid-Phase Exfoliation with High Pressure Homogenizers

High pressure homogenizers offer a reliable way to exfoliate and disperse graphene. Graphite suspensions are forced through engineered microchannels under ultra-high pressure, generating intense shear, cavitation, and particle collisions. This efficiently exfoliates graphite into few-layer or single-layer graphene while preserving its sheet structure and high aspect ratio. The result is uniform exfoliation and stable dispersions suitable for further processing. Below is an example of graphene before and after homogenization using the NanoGenizer High Pressure Homogenizer equipped with Diamond Interaction Chamber:

Graphene after 2 passes through the Diamond Interaction Chamber This visual demonstrates how the high pressure homogenizer transforms aggregated graphite into a well-dispersed, uniform graphene suspension.  

Industrial and Lab Applications

High pressure homogenizers can produce graphene dispersions with particle sizes typically in the 100–500 nm range, depending on pressure and number of passes. They are suitable for lab-scale R&D, pilot trials, and industrial-scale production. Many clients use these dispersions in conductive inks, coatings, batteries, and polymer composites, with reproducible performance and long-term stability.

Advantages of High Pressure Homogenizers

Process Precision:  Layer number and particle size can be finely tuned, ensuring reproducible graphene dispersions.

High-Quality Product:  Maintains intact crystal structure with low defect density, resulting in stable dispersions.

Scalability:  Supports both laboratory and industrial scales, enabling continuous, high-throughput production.

Environmentally Friendly:  Water-based processing avoids strong acids, bases, or harmful chemical dispersants.

Equipment for Graphene Exfoliation and Dispersion

Whether you are developing graphene formulations in the lab or scaling up to industrial production, high pressure homogenizers provide a reliable path from R&D to manufacturing.  

Explore Our Products:

Lab-scale and Industrial High Pressure Homogenizers for graphene exfoliation and dispersion → [View Products]

Get in Touch:

Contact us to discuss your graphene processing requirements → [Get a Quote]

Email: [email protected]  

Related Article:

What is High Pressure Homogenizer?

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Liposome & Lipid Nanoparticle Preparation and Production Equipment

Liposomes are artificially synthesized vesicles composed of phospholipids and cholesterol that mimic biological cell membranes. Their particle sizes typically range from 50–200 nm and can be adjusted according to preparation methods. Liposomes can encapsulate both hydrophilic and hydrophobic compounds and are widely studied for drug delivery, gene delivery, and vaccine formulation research.

As liposome- and LNP-based formulations continue to expand in clinical and commercial applications, they are often categorized as complex injectable dosage forms. Their development requires dedicated processing equipment and carefully controlled production parameters. Among these parameters, particle size control remains one of the most critical factors for ensuring batch-to-batch consistency and predictable delivery behavior.

Common techniques for reducing liposome and lipid nanoparticle size include ultrasonication, shearing, high-pressure homogenization (HPH), and membrane extrusion. Genizer focuses on shearing, extrusion, and HPH-based technologies, supporting scalable preparation from laboratory research through pilot and industrial production.

Particle Size Reduction Technologies

1. Shearing

Shearing applies mechanical forces as samples pass through narrow gaps at high rotational speeds, resulting in particle size refinement. Excessive energy may disrupt lipid bilayer structures, so shearing is typically used as a pretreatment step or for specific formulations such as multilamellar vesicles or passively loaded liposomes.

2. High-Pressure Homogenization (HPH)

High-pressure homogenization is a widely adopted method for producing liposomes and lipid nanoparticles with controlled particle size. As formulations pass through high-pressure interaction chambers, they are subjected to shear, impact, and turbulence, resulting in efficient size reduction while maintaining formulation consistency across different processing scales.

3. Extrusion

Liposome Extrusion Mechanism

Extrusion involves forcing lipid vesicles through membranes at temperatures above the phospholipid phase transition point. Particle size is governed by membrane pore size, applied pressure, and processing temperature. Proper optimization enables narrow particle size distributions and high reproducibility, making extrusion particularly suitable for applications requiring uniform liposome populations.

Liposome Extrusion Size Distribution

Genizer Liposome and Lipid Nanoparticle Equipment

Hand Driven Liposome Extruder: Designed for small-scale laboratory preparation with processing volumes from 0.1–2.5 mL

Pneumatic Liposome Extrusion System: Suitable for medium-scale and continuous operation, supporting volumes from 10 mL to 10 L

High-Pressure Sanitary Pump with Online Liposome Extrusion System: Scalable from laboratory to industrial production, enabling controlled particle sizes in the range of 80–250 nm and supporting higher throughput via modular configurations

High-Pressure Homogenizer (HPH): Available for laboratory-scale applications (NanoGenizer) and large-scale manufacturing (WILL Series)

Additional Advantages

Scalable configurations supporting continuous processing from research to production

Temperature control options, including pre-heating and rapid post-process cooling, to support particle size stability

These systems can be configured individually or in combination according to specific formulation and production requirements, enabling efficient and reproducible liposome and lipid nanoparticle preparation.

Applications of Liposomes and Lipid Nanoparticles

Liposome- and LNP-based formulations are commonly applied in:

Targeted drug delivery to improve circulation time and formulation stability

Vaccine delivery systems and adjuvant research

Anticancer, anti-infective, and immunotherapy development

By adjusting particle size, polydispersity, and processing parameters, uniform and stable lipid-based nanoparticles can be obtained with improved encapsulation performance and formulation robustness.

Explore Genizer Liposome Equipment

Select the appropriate system for your laboratory or production requirements:

Hand Liposome Extruder

Pneumatic Liposome Extruder

High Pressure Sanitary Pump + Online Liposome Extruder

High-Pressure Homogenizer (HPH)

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Genizer NanoMixer: Achieve Precision Mixing for Advanced Nanotechnology Applications

Creating stable and effective liquid formulations in biotechnology and pharmaceuticals hinges on achieving a uniform dispersion of active ingredients. The Genizer NanoMixer Microfluidic Mixing System is designed to meet this critical need, offering a compact, programmable, and highly precise solution for laboratory-scale preparation of sensitive nano-formulations.

Unlike traditional methods that rely on compressed air or hydraulic oil, this electrically driven benchtop instrument gives researchers unparalleled control over the mixing process, making it an indispensable tool for developing next-generation treatments and products .

Key Features of the Genizer NanoMixer

Precision Programmable Control: Manage flow rates from 0 to 10 mL/min per pump with an inlet accuracy of ±0.1 mL. This ensures consistent and reproducible results, batch after batch.

Flexible Mixing Configurations: Choose from three models to match your project's needs: the NanoMixer for dual-component mixing (A+B=AB), the NanoMixer-Triplex for three components (A+B+C=ABC), and the NanoMixer-Quad for complex four-component formulations (A+B+C+D=ABCD).

Sanitary & Compliant Design: Built with 316L stainless steel and other FDA-compliant, sanitary-grade materials, the system adheres to GMP standards. It supports sterile processes and is easy to clean with a flush-to-clean system.

Compact and User-Friendly: Its integrated design (70×36×30 cm) saves valuable lab space. The system features a digital interface for monitoring pressure and flow rate, simplifying operation.

Who Can Benefit from the NanoMixer?

This system is versatile and supports a wide range of advanced applications, including:

mRNA, DNA, and siRNA Delivery Systems: Precisely formulate lipid nanoparticles (LNPs) for genetic medicine and vaccine development.

Nanoemulsions and Liposomes: Create uniform and stable vesicles for drug delivery and cosmetic applications.

Cell Disruption and Nanoparticle Suspensions: Achieve consistent particle size for various research and development needs.

Why Choose Genizer?

The Genizer NanoMixer transforms complex nano-formulation processes into a reliable, streamlined workflow. By providing a tool that combines precision, flexibility, and compliance, Genizer empowers scientists and researchers to accelerate innovation from the lab bench onward.

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E. coli cell disruption Using NanoGenizer High Pressure Homogenizer

Efficient E. coli cell disruption is essential for microbial research, laboratory experiments, and protein extraction. Designed for lab-scale use, the NanoGenizer-30K high-pressure homogenizer delivers reliable and reproducible cell lysis, achieving over 99.99% disruption and consistent protein recovery.

1.  Experimental Method

E. coli samples were homogenized 3–5 times using the NanoGenizer high-pressure homogenizer equipped with RT-cooling diamond interaction chambers (H30Z-RT & H10Z-RT) and a heat exchanger. The temperature was maintained at 4°C using a chilled water bath. 

This setup ensures consistent cell lysis, which is essential for effective E. coli disruption and high-yield protein recovery.  

2.  Disruption Rate Test Method

2.1 Viable Bacteria Counting 

1)  After 24 hours of cultivation, centrifuge the E. coli fermentation broth at 4,000 rpm for 15 minutes. Discard the supernatant and resuspend the cells in buffer. 2)  Take 100 µL of an appropriate dilution and apply it to a plate containing ampicillin and chloramphenicol for viable bacteria counting.  3)  Divide the remaining bacterial solution into two portions and homogenize each at 15,000 psi for 3 passes and 5 passes, respectively. 4)  After homogenization, take 100 µL of each bacterial solution, appropriately dilute it, coat plates containing ampicillin and chloramphenicol, and count the viable bacteria. Viable count reduction is a key metric for selecting a high-pressure homogenizer for microbial disruption and protein recovery.

2.2 Protein Extraction 

1)  Homogenize bacterial samples 3 or 5 times at 15,000 psi using NanoGenizer-30K.

2)  Centrifuge the homogenate at 11,000–13,000 rpm for 15 minutes.

3)  Collect the supernatant and quantify protein content.

High-pressure homogenization efficiently extracts proteins while preserving their activity.

3.   Experimental results

3.1 Appearance after Homogenization

3 passes: solution contained numerous flocculent particles and layering; bacterial fragments relatively large.

5 passes: solution more uniform, indicating thorough homogenization.

3.2 Pre-Homogenization Count

Diluted 10³ times: ~500 colonies → estimated concentration 5×10⁸ CFU/mL.

3.3 Post-Homogenization (3 passes)

Diluted 10³ times: ~20 colonies → ~2×10⁴ CFU/mL.

3.4  Post-Homogenization (5 passes), no viable bacteria were detected 

3.5 Protein Extraction 

The bacterial proteins were effectively extracted with similar efficiency using Z-type interaction chambers (3 or 5 passes) or Y-type interaction chambers (5 passes).

4. Conclusion

1) 3 passes resulted in >99.99% E. coli lysis.

2) 5 passes achieved 100% lysis.

3) Protein extraction yields were consistent results across 3 and 5 passes.

The NanoGenizer-30K provides a fast, efficient, and reproducible solution for lab-scale E. coli lysis and protein recovery.

FAQ -  E. coli Cell Disruption Using High-Pressure Homogenizer

1. Why is a high-pressure homogenizer effective?

Forces generated in the diamond interaction chamber (shear, cavitation, impact) efficiently disrupt cells, releasing proteins consistently.

2. Recommended pressure for E. coli lysis?

12,000–20,000 psi typically suffices; the NanoGenizer-30K supports up to 30,000 psi.

3. How many passes are required to achieve complete lysis?

3 passes → >99.99% lysis; 5 passes → 100% lysis. The optimal number of passes depends on strain type, culture density, and protein sensitivity.

4. Does high pressure homogenization affect protein activity?

No, NG-30K features RT cooling interaction chambers and a heat exchanger connected to a cooling bath system set to maintain 4°C, protecting temperature-sensitive proteins during high-pressure homogenization.

5. Advantages of NanoGenizer:

Minimal sample volume (5 mL)

High-pressure capability (up to 45,000 psi)

Diamond interaction chamber for precise control

User-friendly touchscreen

Adjustable sample intake (5–16 mL)

Automatic stop function

Real-time cooling

Compact and portable

Versatile reservoir options

6. Can NanoGenizer process other microbes? 

Yes. It can process other bacteria, yeast, algae, and mammalian cells, suitable for biotechnology, biopharma, and protein purification.

Explore Related Products

 PilotGenizer Series – Pilot-scale homogenizers for scale-up studies

UltraGenizer Series – High-pressure homogenizers for nanoemulsion and liposome applications

Contact us at [email protected] to find the right model, request a quote, or schedule a demonstration.

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Ease of Use and the Liposome HandExtruder

One of Genizer’s most compact pieces of equipment is the HandExtruder. The HandExtruder is popular in academic and small labs because of its small size and low dead volume. Researchers utilize it to get the desired size of liposome or lipid nanoparticle. Particle size can impact circulation time, uptake and stability. All of these are important aspects in different uses for liposomes. The affordability of the HandExtruder makes it an accessible entry point to liposome extrusion for academics. The HandExtruder has been cited in academic work in pharmaceutical research. One example is a study where our Handextruder, with 200nm membranes, was used to create liposomes to study cancer treatment (link). In another study, the HandExtruder was used in the preparation of vaccination nanoparticles (link). Beyond pharmaceuticals, reasearchers use the HandExtruder in diverse fields like agriculture (link), and protocell research (link). As time has gone on, we continue to learn from client feedback.

After talking with our clients at trade shows, we learned of an issue some clients experienced with the HandExtruder. The small syringe end was becoming uncomfortable after multiple passes. We decided to add a more ergonomic option. Our team developed syringe pads to increase the surface area on your palm. These pads increase the ease to which researchers can get liposomes down to size. The syringe pads can be added or removed.

If you would like to add syringe pads, please let our team know when you order. We’ll get you re-sizing liposomes or lipid nanoparticles with ease!

Liposome HandExtruder

Description: Hand driven extruder is for formulation of 0.25mL-2.5mL liposome and lipopolyplex; Temperature control is available.

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Precision & Power: Genize Sanitary High Pressure Pumps for Critical Processes

In the world of pharmaceutical manufacturing, bioprocessing, and liposome development, the transfer and processing of sensitive fluids demand unparalleled precision, hygiene, and reliability. Genizer's range of Sanitary High Pressure Pumps is engineered to meet these rigorous demands, providing the controlled force essential for applications from early-stage R&D to full-scale GMP production. Whether driving liposomes through extrusion membranes or enabling sanitary transfer, Genizer pumps deliver consistent performance where it matters most.

The Genizer Pump Advantage: Core Strengths

Sanitary & Compliant Design: Constructed with FDA-approved materials and designed to meet GMP sanitary standards, all pumps ensure product integrity and are suitable for regulated environments.

Precision Control & Monitoring: Advanced models feature digital displays and programmable controls for accurate management of pressure (up to 2,000 bar), flow rate, temperature, and volume, guaranteeing process reproducibility.

Scalable Solutions: Our portfolio covers every stage of development, from manual benchtop handling to fully automated, high-capacity production, ensuring seamless scale-up.

Robust & Efficient Performance: Designed to be strong, silent, and energy-efficient, these pumps deliver high performance (e.g., >30 L/hr at 300 bar) with minimal downtime.

The Genizer Pump Portfolio: Find Your Perfect Match

HandPump (Sanitary Stainless)

Description: A manual, portable, hand-driven pump for the sanitary transfer of small fluid samples under pressure. It is the ideal companion for Genizer online liposome extruders during small-scale testing.

Ideal For: Quick, portable preparation of small-volume liposomes and lipopolyplex for drug and gene delivery research.

NanoPump (Sanitary Stainless)

Description: An electrically-powered, integrated sanitary piston pump capable of reaching 2,000 bar with controllable flow. It combines a small footprint with intelligent programming and noiseless operation.

Ideal For: High-pressure liposome extrusion and sanitary liquid delivery in lab and pilot settings. Its CE and RoHS compliance makes it suitable for global markets.

PilotPump & ProdPump (Sanitary Stainless)

Description: These sanitary high-pressure piston dual pumps are designed for transferring large fluid samples with continuous, controllable pressure and flow. They are mobile and built for robust performance.

Ideal For: Continuous pilot-scale and full-scale manufacturing production of liposomes and lipopolyplex when integrated with Genizer online extruders. They are built to comply with FDA and GMP standards for pharmaceutical production.

Sanitary High Pressure Pump

Description: A high-performance pump reaching up to 2,000 bar, featuring a comprehensive digital display for monitoring pressure, temperature, flow rate, volume, and time.

Ideal For: Applications requiring the highest level of process control and monitoring for sanitary liquid delivery and liposome extrusion.

Your Partner in Process Excellence

Genizer pumps are more than just components; they are the heart of reliable and compliant processes. By providing the precise pressure and sanitary conditions required, they enable scientists and engineers to achieve reproducible results and accelerate innovation in drug delivery, vaccine development, and beyond.

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Achieve Precision & Scale in Liposome Development with Genizer Extruders

In the fields of drug delivery, vaccine development, and genetic medicine, the precision and reproducibility of lipid nanoparticles (LNPs) and liposomes are paramount. Genizer's comprehensive portfolio of Liposome Extruders is engineered to meet this critical need, providing researchers and manufacturers with reliable, scalable, and compliant solutions from initial discovery to large-scale GMP production.

Why Choose Genizer Liposome Extruders?

Our extrusion technology is designed to transform heterogeneous lipid suspensions into homogeneous, unilamellar vesicles with narrow size distribution in a simple, one-step process. The core advantages that define our systems include:

Unmatched Size Control: Produce uniform liposomes and LNPs with precise diameters, typically achieved in just 3-10 passes through our defined pore-size, track-etched membranes.

Minimal to Zero Dead Volume: Maximize the yield of your valuable samples, from precious 0.25 mL research formulations to 200 L commercial batches, minimizing waste and cost.

Precise Temperature Control: Critical for handling heat-sensitive compounds, our systems (like the HandExtruder-RT and Jacketed Extruder) maintain optimal temperatures throughout the extrusion process to ensure lipid stability and integrity.

Scalability from Bench to Plant: Our range is designed for seamless scale-up. Start with the manual HandExtruder for small-scale R&D, move to the Online Extruder for pilot studies, and integrate the Multi Online System with sanitary pumps for full-scale GMP manufacturing.

Regulatory Compliance: Constructed from pharmaceutical-grade 316L stainless steel, our systems are easy to clean, sterilize, and are designed to comply with FDA and GMP sanitary standards, ensuring your process is audit-ready.

Explore the Genizer Extruder Portfolio:

HandExtruder: The perfect entry point for basic research. Manually operated for samples from 0.25mL to 2.5mL, it offers minimal dead volume and optional temperature control for fast, easy formulation.

Jacketed Liposome Extruder: Compressed air-driven for consistent pressure up to 500 psi. Its jacketed design provides excellent temperature control for processing sensitive drug and gene delivery formulations.

Online Liposome Extruder: Engineered to be connected inline with a sanitary pump or homogenizer. It is ideal for continuous processing from pilot to production scale, offering no dead volume and full temperature control.

Multi Online Liposome Extruders System: The solution for high-volume manufacturing. This system can control temperature and produce up to 200L per batch of therapeutic liposomes, such as those for anticancer drugs.

Sanitary Pump Systems (NanoPump & PilotPump): The intelligent force behind the extrusion. These PLC-controlled pumps deliver highly accurate pressure and feature full monitoring capabilities, ensuring a reproducible and successful process.

Your Partner in Advanced Formulation

Whether you are developing the next breakthrough mRNA vaccine, a targeted cancer therapeutic, or a novel cosmetic delivery system, Genizer provides the extrusion technology you can trust. Our systems are built to deliver the precision, reliability, and compliance required to accelerate your product from the lab to the clinic.

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Precision Nano Solutions: Genizer Homogenizers Simplify Advanced Manufacturing

Genizer High Pressure Homogenizers transform complex nanotechnology into reliable industrial processes. Our patented Dual-Y Diamond Interaction Chamber enables effortless creation of nanoemulsions and nanoparticles in the 20-200nm range – the critical size for enhanced drug delivery, deeper skincare penetration, and efficient battery materials.

Why Customers Choose Genizer:

Nanoscale Made Simple

Achieve 50nm particles in 1-3 passes (vs. 5-10+ passes with conventional machines)

Built-in cooling system prevents heat damage to sensitive compounds like mRNA or vitamins

Unmatched Consistency

Maintain ±5nm particle size deviation across 100+ production batches

Automatically adjust pressure (1,500-45,000 PSI) for different viscosities

Scale Up Seamlessly

Identical results from lab (MicroLab®) to factory (Industrial Titan series)

Quick-change parts reduce downtime by 70%

Real-World Applications:

Pharma: Produce stable lipid nanoparticles for vaccines

Cosmetics: Create clear vitamin C serums with 3x faster skin absorption

Batteries: Disperse silicon anode materials evenly for longer battery life

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E. coli Model for Protein Amplification

Escherichia coli is a bacterium that most of us know first from recalls of foods like meat and lettuce. Although E. coli is a commensal (non-harmful) gut bacterium, the wrong circumstance and strain can make infection uncomfortable and dangerous. E. coli is gram negative, meaning it has a double-layered cell membrane that makes it more difficult to treat with antibiotics.

Our homogenization equipment is regularly used to disrupt E. coli. In “Identification and Biosynthesis of pro-inflammatory sulfonolipids” authors Hou et al. use the NanoGenizer to break apart E. coli after using it for protein expression. Many other researchers have done the same (https://www.genizer.com/u_file/2206/file/6bffa6a456.pdf, https://www.genizer.com/u_file/2312/file/298f8a65e6.pdf , https://www.genizer.com/u_file/2312/file/3ca3c35d8a.pdf)

What makes this common cause of diarrheal illnesses a welcome tool in so many research labs?

First, we need to understand the research problem E. coli is being used to address in many labs.

Researchers often need large amounts of proteins for a variety of purposes. One common example is for pharmaceutical production at a large scale. However, having to obtain that protein from its natural source can be time-consuming, resource intensive and complicated. Instead, researchers will often take the gene that encodes that protein and transform it into a bacterium. There, the bacteria will make that protein more quickly. However, not just any bacteria are suited for this purpose, and that’s what makes E. coli a laboratory star.

Speed: E. coli can double as fast as every 20 minutes. Optimal conditions for quickly producing large quantities of protein.

Density: E. coli can grow densely, reducing the amount of space required for protein production.

Materials: E. coli can grow well on affordable and easily obtainable materials, making it accessible to researchers.

Universality: The wide breadth of research done using E. coli as a model makes it easier to work with.

Tractability: Tractability refers to the ease of editing a genome. E. coli is naturally genetically tractable and can pick up new genes and add them to itself. This eases the process of adjusting its genome for researchers. As a pathogen, this feature also helps E. coli evade antibiotics.

So researchers, go forth and enjoy this model organism in the laboratory, if not always in your kitchen!

Sources: https://www.ncbi.nlm.nih.gov/books/NBK564298/ https://pmc.ncbi.nlm.nih.gov/articles/PMC4029002/ https://www.ncbi.nlm.nih.gov/books/NBK562895/

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Scaling with Genizer: Homogenizers

At Genizer, we want our equipment to be with you every step of the way! That’s why we make equipment that squeezes into an academic lab all the way up to manufacturing. Here’s the High-Pressure Homogenization equipment that can take you all the way to capacity.

HandGenizer

For a more portable solution to high-pressure homogenization, we have the HandGenizer. Well suited to long-term storage, the HandGenizer is great in an academic R&D context. Its’ distinctive manual option and compact size set the HandGenizer apart, though it is best suited to small, even solutions like nanoemulsions.

NanoGenizer

Next is the NanoGenizer. Our most popular model, the NanoGenizer fits easily in a benchtop with a length under 28 inches (70 cm). The Nanogenizer has a max flow of 100mL/min, and a max pressure of 45,000 psi. Ideal for R&D, laboratory, and small-scale production. The NanoGenizer is quiet and easy to use. It also comes in Dual and Quad options for additional capacity, and the NanoGenizer and all larger units have a MixGenizer option, for in-chamber mixing.

UltraGenizer

Our half-step up from the NanoGenizer is the UltraGenizer. The UltraGenizer comes with the unique ability to reach 60,000 psi! At a pressure of 30kpsi, it can reach 25L/Hr. It also has a higher runtime of 4-6 hours, giving a much higher processing capacity.

PilotGenizer & ProdGenizer

WillNano

Finally, our WillNano ManuGenizer series is ideal for your manufacturing needs! The Willnano offers up to 4 pumps to reach your desired flow rate. At 30kpsi, these machines can reach up to 480L/hr. And if that still doesn’t meet your needs, there’s always more than one.

We also offer many sanitary certifications and IQ/OQ documentation. With our scalable options, we know you can find the right size with Genizer!

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NanoGenizer Lab Homogenizer for Cell Disruption NanoGenizer High Pressure Homogenizer for Nanomaterials NanoGenizer, a lab-scale microfluidic high pressure homogenizer, offers effective performance on particle size reduction and cell disruption. It utilizes interaction high-pressure microfluidic jet technology, and equipped with highly modular power and processing units. NanoGenizer is suitable for rare and valuable samples because of its minimum dead volume design and its continuous flow rate, which enables it to process up to 120 mL/min. With successful applications in various nanotechnology settings, the NanoGenizer has gained increasing recognition due to its notable shear rate, excellent repeatability, and guaranteed scalability.   NanoGenizer provides efficient solutions for various nanomaterials, including: liposomes, nano-emulsions, nano-crystals, micelles, lipid nanoparticles, cosmetic nano-encapsulation materials, nano-particle dispersion, graphene, carbon nanotube, and more. As more organizations recognize this, NanoGenizer is becoming an essential high pressure homogenizer for laboratories working on high-end nanomaterial preparation.

Core Technology: Interaction Chamber with Microfluidic Jet NanoGenizer can be divided into a power unit (including the power system, control system, and high-pressure pump system) and a processing unit (including the microfluidic diamond interaction chamber, material inlet and outlet, and heat exchanger). The diamond interaction chamber is a reaction component with a specially-designed fix geometry, where high pressure, high speed micro-jets occur and samples are processed. After being pressurized and accelerated by the intensifier, the sample’s micro-jet can reach speeds of 1,000 m/s, faster than the speed of sound (340 m/s). When the high-speed bullet-type micro-jet passes through the diamond interaction chamber’s micro-channels, it undergoes complex physical effects, such as high-frequency shearing, high-energy impact, cavitation and pressure drop, and more. After that, the material is homogenized and nano-sized.   The diamond interaction chamber is the core area where the high-speed micro-jet, high-shear, high-energy collision, and other effects occur. It has fixed Y- or Z-shaped diamond micro-channels inside, which guarantees a consistent effect: accurate and repeatable particle size distribution.

Features of the NanoGenizer High Pressure Homogenizer High Shear Rate After being pressurized by the power unit, the liquid or solid-liquid mixture sample’s micro-jet can reach speeds up to 1,000 m/s inside the diamond interaction chamber (DIXC). The minimum dimension of the inside of the DIXC can be as small as 50 μm. As shown in Figure 4, the NanoGenizer’s shear rate is the highest among comparable technologies.

The NanoGenizer high pressure homogenizer’s shearing force on the material is unmatched by conventional homogenizers or other homogenizing equipment.   Fixed Internal Structure and Consistent Reaction Pressure

The diamond interaction chamber has a fixed internal shape (Figures 5 and 6) and does not move with pressure changes. When the sample passes through the DIXC, the applied pressure is at a constant peak during each cycle’s process (see the blue curve in Figure 7). However, the traditional homogenizing valve has a dynamically adjustable structure. The pressure of the material changes dynamically when it passes through a conventional homogenizer’s valve, which operates at peak pressures for mere moments of each cycle (the black curve in Figure 7). The particle size of the material processed by the microfluidic diamond interaction chamber is reduced efficiently, and the distribution is more uniform than can be achieved with a traditional homogenizing valve.

Interaction Effect The design of the Y-type DIXC takes full advantage of the interaction collision of the materials themselves. The instantaneous relative velocity of the two jets is doubled, resulting in an explosion effect. The collision between the materials greatly reduces the materials’ wear and shear to the interaction chamber, prolonging the chamber’s service life and avoiding the risk of stainless steel particles falling off.  The patented DIXC-RT with cooling function (see Figure 3) protects the bioactivity of the samples.   Guaranteed Scalability 

The single-slotted DIXC (shown in Figure 2 and Figure 3) is suitable for lab-scale high pressure homogenization equipment. The multi-slotted DIXC (shown in Figure 8 and Figure 9) is composed of multiple diamond micro-channels copied in parallel, ensuring the treatment effect’s consistency. The increased number of channels also allows for larger volumetric flow rate.   Genizer LLC is one of two companies in the world that supplies high pressure homogenizers equipped with original microfluidic diamond interaction chambers. In recent years, Genizer’s microfluidic diamond interaction chambers have been equipped by more and more homogenizer manufacturer.   Easy to Operate and Intelligent The highly modular and integrated control design of NanoGenizer makes it extremely simple to install and operate. Operators can complete routine operations by clicking the three buttons—"pressure rate,” “start,” and “stop”—on the touch screen. Customers find the equipment simple to operate: They only need to set a pressure on the screen and then click to start or stop. There is no need to screw the valve forcefully or waste any sample to adjust the pressure. To process a particularly small volume, you can set the sample volume processed by one stroke, the stroke cycle times, and the automatic stop time on the setup interface. Compared with other high pressure equipment, the NanoGenizer’s operation is elegant and efficient.

Small Minimum Sample NanoGenizer’s highly modular design enables simple installation and maintenance. The dead volume is no more than 1 mL, far below the dead volume of other high pressure homogenizers, which is typically more than 10 mL. Therefore, when a 10 mL sample is processed in traditional homogenizers, it is almost impossible get product from the outlet. NanoGenizer's modular pipeline design can achieve a minimum single processing volume of 5 mL by setting the volume autonomously, making it the best equipment option for preparing and developing rare, costly samples. NanoGenizer’s continuous flow rate is 7 L/h, suitable for conventional small batches.   Software and Hardware Options Temperature Control: Jacketed interaction Chamber; jacketed inlet cylinder; 3D heat exchanger Suspension Preparation: Hollow magnetic stirrer Corrosion Resistance: Titanium cylinder option Precision Sampling: Precision sampling as accurate as 0.1 mL (e.g., 10.0mL, 10.1mL, and 10.2mL sampling are all possible) Ports: Data log, USB port, and multi-sensor ports for temperature and pressure     Scientific researchers who have used NanoGenizer are impressed by its high processing efficiency, easy operation, and intelligent control. NanoGenizer is currently the one of the most selected homogenizers for nanomaterials in the world.

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Genizer, located in Technology Link in Greater Los Angeles, is dedicated to advancing homogenizer nanotechnology. We provide high pressure homogenizers, liposome extruders, sanitary heat exchangers, diamond interaction chambers, and high pressure gauges compatible with other brands of high pressure homogenizers and high pressure pumps.