Liposomal Vitamin C Production with Ultrasonics

Liposomal vitamin formulations are known for their higher bioavailability and absorption rate. Vitamin C, an antioxidant, is a common supplement used in nutritional and medical drugs to support the human body’s own immune system. Ultrasonication is a reliable and safe method for the production of high-quality liposomes and nano-liposomes. The ultrasonic encapsulation process produces liposomes with a high load of active substances, such as vitamin C.

Liposomal Vitamin C Produced with Ultrasonication

The advantage of liposomal vitamins is that they are not administered in conventional tablet or powder form, but as liquid formulation with increased bioavailability. This means, the vitamins are encapsulated in the core of spherical phospholipid cells, so-called liposomes. Since liposomes have a similar phospholipid composition to the lipid membranes of human cells, they are significantly better absorbed into the bodys cells. Therefore liposomes are used in medicinal formulations and pharmaceuticals, supplements and nutraceuticals, cosmeceuticals and cosmetic products. Ultrasonication is a highly efficacious method to encapsulate molecules such as therapeutics into liposomes. Ultrasonic liposome production is characterized by a high entrapment efficiency, which means that a high percentage of active ingredients (e.g., vitamin C) are encapsulated into liposomes. Simultaneously, sonication ensures a uniformly small liposome size that can easily be absorbed by the human cells. Therefore, ultrasonically encapsulated vitamin C liposomes offer a very high bioavailability and efficacy. Overall, ultrasonic liposome formation is highly efficient and delivers superior liposome quality!

Advantages of Ultrasonic Liposome Preparation

  • Superior entrapment efficiency
  • High Encapsulation Efficiency (EE%)
  • High Stability
  • Non-thermal treatment (prevents degradation)
  • Compatible with various formulations
  • Rapid Process

 

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Ultrasonic encapsulation is used to produce large volumes of pharmaceutical-grade liposomes with a high load of active substances.

UIP1000hdT with glass flow reactor for liposome production

Step-by-Step Instructions for Ultrasonic Encapsulation of Vitamin C into Liposomes

Liposomal vitamin C is a delivering form for vitamin C that improves absorption and bioavailability. Probe-type ultrasonication is a efficient and reliable method for producing liposomes.
Following protocol shows the general steps for producing liposomal vitamin C using a probe-type ultrasonicator:
 

  1. Mix vitamin C powder with a small amount of distilled water to form a solution.
  2. Add the vitamin C solution to a solution of phospholipids (such as soy lecithin) in a ratio of 1:10 (vitamin C:phospholipids) by volume.
  3. Place the mixture in a glass beaker and insert the probe of the ultrasonicator, e.g. UP200Ht with S26d14, into the solution.
  4. Ultrasonicate the mixture using the probe-type ultrasonicator for 5-10 minutes, at an approx. power setting of 200W (frequency 26 kHz). The probe of the ultrasonicator should be moved gently through the solution during the sonication process. Avoid the contact of probe with the beaker wall (as glass beakers might break).
  5. After sonication, let the mixture sit for 10-15 minutes to allow the liposomes to form.
  6. Store in a dark glass bottle and preferably at the refrigerator for long-term stability.

It’s important to note that the exact protocol may vary depending on the specific ultrasonicator and materials used. Additionally, the stability and quality of the liposomes can be affected by factors such as pH, temperature, and the presence of other substances, so it’s important to carefully control these variables during the production process.
 

Ultrasonically Produced Liposomes – Superior Bioavailability
Ultrasonically prepared liposomes help to overcome the short half-life, low cell-membrane permeability, and poor oral bioavailability due to gastric and enzymatic degradation of these bioactive compounds. The encapsulation into a phospholipid bilayer protects the active ingredients against degradation and increases the absorption rate into the cells.
Ultrasonic probe UP200Ht is used for the liposomal encapsulation of vitamin C into phospholipid carriers. Ultrasonically prepared liposomes offer superior bioavailability.

Ultrasonicator UP200Ht for the efficient entrapment of vitamin C into liposomes.

Ultrasonic Liposome Formation

Liposomal formulation produced using high-performance ultrasonicationSince the formation of liposomes and nanoliposomes does not occur as a spontaneous process, an energy resource is required to promote the encapsulation process. Liposomes are lipid vesicles, which are formed when phospholipids, e.g. lecithin, are added to water, where they form bilayer structures when sufficient energy, e.g. by sonication, is applied. Ultrasonication assists the arrangement of the lipid molecules, so that a thermodynamically stable aqueous phase is obtained. Sonication does not only promote the formation of liposomes, it also reduces the size of liposomes resulting in nanoliposomes. The liposome size is an important factor when it comes to bioavailability and absorption rate since smaller liposomes can penetrate the cell membranes easier.

Ultrasonic Liposome Size Reduction

Ultrasonic dispersion is a simple and efficient method for reducing the size of liposomes and manufacturing nanoliposomes. In order to prepare smaller liposomes, hydrated vesicles are sonicated for a few minutes with a probe-type ultrasonicator in a temperature-controlled reactor. As a non-thermal, purely mechanical method, ultrasonic size reduction does neither degrade the phospholipids nor the bioactive compounds.

The ultrasonic method ensures the formation of liposomes with specific features by promoting the encapsulation of active ingredients and by adjusting their size and lamellarity through controlled processing steps. Hielscher sonicators are renowned for best results in liposome formation.

After the formation of a lipidic film subsequent rehydration, sonication is used to promote the entrapment of active ingredients in the liposome. Additionally, sonication achieves the desired liposome size.

Ultrasonic Processors for Liposome Production

Hielscher Ultrasonics systems are widely used in the pharmaceutical and supplement production to formulate high-quality liposomes loaded with vitamins, antioxidants, peptides, polyphenols and other bioactive compounds. To meet its customers demands, Hielscher supplies ultrasonicators from the compact hand-held lab homogeniser and bench-top ultarsonicators to fully industrial ultrasonic systems for the production of high-volumes of liposomal formulations. A broad range of ultrasonic sonotrodes and reactors are available to ensure an optimal setup for your liposome production. The robustness of Hielscher’s ultrasonic equipment allows for 24/7 operation at heavy duty and in demanding environments.
The table below gives you an indication of the approximate processing capacity of our ultrasonicators:

Batch Volume Flow Rate Recommended Devices
1 to 500mL 10 to 200mL/min UP100H
10 to 2000mL 20 to 400mL/min UP200Ht, UP400St
0.1 to 20L 0.2 to 4L/min UIP2000hdT
10 to 100L 2 to 10L/min UIP4000hdT
n.a. 10 to 100L/min UIP16000
n.a. larger cluster of UIP16000

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Literature/References

Facts Worth Knowing

Vitamin C

Vitamin C, also known as L-ascorbic acid or ascorbate, is a water-soluble antioxidant with the chemical formula of C6H8O6. As an anitioxidant, vitamin C acts as an electron-donor to various enzymatic and non-enzymatic reactions. Furthermore, vitamin C is a cofactor in many enzymatic reactions which regulate essential biological functions such as wound healing and collagen synthesis. Scurvy is the most well-known form of severe vitamin C deficiency that occur due to impaired collagen synthesis.
As a vitamin, ascorbic acid is essential to the human body, which means the body cannot synthesise vitamin C, but it must be consumed with food. Foods rich in vitamin C include citrus fruits, camu camu, acerola, kale, roseship, black currant, guava as well as other fruits and vegetables.
Whilst a well-balanced diets easily delivers adequate amounts of vitamin C. However, dietary supplements such as vitamin drops, capsules, tablets, powders and gummies are frequently used to ensure a sufficient vitamin supply. The recommended daily vitamin C intake by the European Food Safety Authority is 110 mg/day for male and 95 mg/day for female adults.
In medicine, vitamin C is administered in high doses under medical supervision in order to support cancer treatments and to strengthen the immune system. For instance, studies found that a regular intake of vitamin C supplements reduce the length and severity of cold and flu.

What is a Liposome?

Liposomes are spherical vesicles of microscopical size, which can range from 30nm to several micrometers. A reliable way to synthesise liposomes under controlled conditions is the ultrasonic encapsulation. Mostly, liposomes are composed of phospholipids, especially phosphatidylcholine, but they can also include other lipid compounds, such as egg phosphatidylethanolamine.
Being vesicles made from phospholipids, liposomes act as microcontainers that encapsulate bioactive compounds such as vitamins, antioxidants, polyphenols, peptides, drug substances (e.g., vaccines, medicinal compounds). The phospholipid bilayer encapsulates the bioactive substance and transports it into cells. Since the liposome bilayers are made from similar phospholipids as the cell membranes, liposomes can cross the cell membranes easily and deliver the bioactive substances into the cells. This makes liposomes a highly potent drug carrier with a high bioavailability and absorption rate. The amphiphilic properties of the phospholipid bilayers make the liposomes soluble in both aqueous and polar liquids.

High performance ultrasonics! Hielscher's product range covers the full spectrum from the compact lab ultrasonicator over bench-top units to full-industrial ultrasonic systems.

Hielscher Ultrasonics manufactures high-performance ultrasonic homogenizers from lab to industrial size.

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