Liposomal Ingredients: Are You Getting What You Paid For?
By By Mike Montemarano, Associate Editor | 02.21.24
Liposomal encapsulation is a nascent technology in the supplements space, and proving liposomes were created requires a battery of technical, expensive tests.
Liposomes are hollow, microscopic spheres made of lipid bilayers which can be used as nano carriers for active ingredients within an aqueous interior, which gives them both hydrophobic and hydrophilic characteristics. These structures, formed from phospholipids, can enhance the bioavailability and absorption rates of both water-soluble and fat-soluble ingredients by severalfold, and protect ingredients as they pass through the digestive tract until they reach the target absorption site.
Liposomes have been investigated as potential delivery systems for pharmaceutical drugs since the 1970s, but the first successful commercial products only hit the market decades later, in the 1990s.
Until very recently, liposomal technology only was used for highly-stabilized active pharmaceutical ingredients, which are by requisite much more pure than food or cosmetic ingredients. But the past decade or so saw major strides in the development and production of processing equipment which could reliably create phospholipids potentially suitable for dietary ingredient applications. They’re now used in a wide range of ingredients, like vitamins, antioxidants, omega-3s, and several herbal ingredients.
According to Sebastian Balcombe, founder and CEO of liposomal ingredient supplier Specnova, natural ingredients are much more difficult to encapsulate into liposomes than any pharmaceutical product, and if a supplier claims that an ingredient is, in fact, liposomal, they have a major burden of proof to deal with. Likewise, finished product companies claiming their ingredients are liposomal should be careful about which ingredient or supplier they select.
While Specnova hasn’t tested all competing liposomal ingredients, an overwhelming majority of the commercially-available products it has tested, which claim to be liposomal, were not. “Most products we have tested have no liposomes at all.” Balcombe said. “Some have micelles [a thin, spherical monolayer of lipids], but typically we find a slurry of sunflower lecithin and the active ingredients with no liposomes or micelles. It is no surprise that these products perform no differently than non-liposomal products.”
There aren’t any third-party programs available to the dietary supplements industry to prove that nutraceutical liposomes have been successfully created, and there isn’t a universally-validated set of tests to run in order to prove that they exist, contain active ingredients within, and meet all other criteria.
What Testing Methods Should Be Done?
“Liposomal validation is highly technical and expensive, and you have to know where and how to have the testing done,” Balcombe said. “Currently, there are no set standards in the industry, which is why we created TruLiposome Validation. These standards ensure that every batch of liposomal ingredients is truly liposomal. Our liposomal ingredients are tested using 4-6 different methods. We provide third-party documentation for all of our ingredients, and so should all legitimate liposome manufacturers. If a manufacturer can’t provide this proof, then it’s doubtful that the ingredient is truly liposomal.”
Specnova looks for four criteria in its liposomal ingredients in order to prove that they are effective.
First, in order to be absorbed by the small intestines effectively, a liposome should be between 100 and 200 nanometers. Specnova uses dynamic light scattering analysis (DLS) to determine the average size of liposomes in each batch. “Each active ingredient, when wrapped in a liposome, will have a different size. For example, liposomal NMN should be around 175 nm.”
Second, liposomes should be tested for the strength of their encapsulation by calculating the zeta potential, a measurement used on suspensions and emulsions to determine electrochemical equilibrium at the particle-liquid surface. As is the case with particle size, zeta potential will vary depending on what ingredient is being encapsulated, Balcombe said. “LipoVantage NMN was measured at -41.4 meV, confirming a strong encapsulation of the active ingredient and stable colloidal suspension. Each ingredient has a different zeta potential.”
In order to confirm the spherical shape of liposomes, images of them must be created using a Scanning Electron Microscope (SEM).
Last, liposomes must be well-dispersed and free of leakage. “A cryo transmission electron microscope (TEM) is used to ensure liposomes are spherical in shape, well dispersed, and there is minimal liposomal debris which might indicate vesical rupture. The core of the liposomal structure is well-defined by the dark portion of the structure,” Balcombe said.
In order to maintain brand credibility, and establish the credibility for the entire liposomal ingredients segment, “every liposomal ingredient should come with a portfolio/dossier of all the critical testing that was done,” Balcombe said. “If the testing for that particular batch cannot be produced, don’t trust it. It is also important to know that having only SEM testing or only SEM and TEM testing is not enough.”
“There are four criteria that must be met to validate a true liposome, and these four discreet tests must be completed,” Balcombe continued.
Once liposomes are created and validated, Specnova utilizes a proprietary DualHydrogel technology, in which it encapsulates each unit (active ingredient and liposome) in another protective layer made up of a specific blend of natural ingredients. “This technology is customized for the active,” Balcombe said. “This additional layer protects the liposome and increases its stability, resulting in better bio-absorption and bioavailability.”
Performance Testing
Once validation testing is complete, Balcombe noted that there’s plenty of value in testing the performance of liposomal ingredients to show improved bioavailability and performance compared to before encapsulation.
“We test our liposomal ingredients using either a Caco-2 permeability model or a dissolution model,” he said. “Results obtained from these methods can positively predict in vivo absorption across the intestinal tract into the bloodstream. Once we see the improved bioavailability in the Caco-2 or dissolution model, we then move on to a randomized, double-blind, placebo-controlled clinical study, the gold standard of the industry.”
Liposomes have been investigated as potential delivery systems for pharmaceutical drugs since the 1970s, but the first successful commercial products only hit the market decades later, in the 1990s.
Until very recently, liposomal technology only was used for highly-stabilized active pharmaceutical ingredients, which are by requisite much more pure than food or cosmetic ingredients. But the past decade or so saw major strides in the development and production of processing equipment which could reliably create phospholipids potentially suitable for dietary ingredient applications. They’re now used in a wide range of ingredients, like vitamins, antioxidants, omega-3s, and several herbal ingredients.
According to Sebastian Balcombe, founder and CEO of liposomal ingredient supplier Specnova, natural ingredients are much more difficult to encapsulate into liposomes than any pharmaceutical product, and if a supplier claims that an ingredient is, in fact, liposomal, they have a major burden of proof to deal with. Likewise, finished product companies claiming their ingredients are liposomal should be careful about which ingredient or supplier they select.
While Specnova hasn’t tested all competing liposomal ingredients, an overwhelming majority of the commercially-available products it has tested, which claim to be liposomal, were not. “Most products we have tested have no liposomes at all.” Balcombe said. “Some have micelles [a thin, spherical monolayer of lipids], but typically we find a slurry of sunflower lecithin and the active ingredients with no liposomes or micelles. It is no surprise that these products perform no differently than non-liposomal products.”
There aren’t any third-party programs available to the dietary supplements industry to prove that nutraceutical liposomes have been successfully created, and there isn’t a universally-validated set of tests to run in order to prove that they exist, contain active ingredients within, and meet all other criteria.
What Testing Methods Should Be Done?
“Liposomal validation is highly technical and expensive, and you have to know where and how to have the testing done,” Balcombe said. “Currently, there are no set standards in the industry, which is why we created TruLiposome Validation. These standards ensure that every batch of liposomal ingredients is truly liposomal. Our liposomal ingredients are tested using 4-6 different methods. We provide third-party documentation for all of our ingredients, and so should all legitimate liposome manufacturers. If a manufacturer can’t provide this proof, then it’s doubtful that the ingredient is truly liposomal.”
Specnova looks for four criteria in its liposomal ingredients in order to prove that they are effective.
First, in order to be absorbed by the small intestines effectively, a liposome should be between 100 and 200 nanometers. Specnova uses dynamic light scattering analysis (DLS) to determine the average size of liposomes in each batch. “Each active ingredient, when wrapped in a liposome, will have a different size. For example, liposomal NMN should be around 175 nm.”
Second, liposomes should be tested for the strength of their encapsulation by calculating the zeta potential, a measurement used on suspensions and emulsions to determine electrochemical equilibrium at the particle-liquid surface. As is the case with particle size, zeta potential will vary depending on what ingredient is being encapsulated, Balcombe said. “LipoVantage NMN was measured at -41.4 meV, confirming a strong encapsulation of the active ingredient and stable colloidal suspension. Each ingredient has a different zeta potential.”
In order to confirm the spherical shape of liposomes, images of them must be created using a Scanning Electron Microscope (SEM).
Last, liposomes must be well-dispersed and free of leakage. “A cryo transmission electron microscope (TEM) is used to ensure liposomes are spherical in shape, well dispersed, and there is minimal liposomal debris which might indicate vesical rupture. The core of the liposomal structure is well-defined by the dark portion of the structure,” Balcombe said.
In order to maintain brand credibility, and establish the credibility for the entire liposomal ingredients segment, “every liposomal ingredient should come with a portfolio/dossier of all the critical testing that was done,” Balcombe said. “If the testing for that particular batch cannot be produced, don’t trust it. It is also important to know that having only SEM testing or only SEM and TEM testing is not enough.”
“There are four criteria that must be met to validate a true liposome, and these four discreet tests must be completed,” Balcombe continued.
Once liposomes are created and validated, Specnova utilizes a proprietary DualHydrogel technology, in which it encapsulates each unit (active ingredient and liposome) in another protective layer made up of a specific blend of natural ingredients. “This technology is customized for the active,” Balcombe said. “This additional layer protects the liposome and increases its stability, resulting in better bio-absorption and bioavailability.”
Performance Testing
Once validation testing is complete, Balcombe noted that there’s plenty of value in testing the performance of liposomal ingredients to show improved bioavailability and performance compared to before encapsulation.
“We test our liposomal ingredients using either a Caco-2 permeability model or a dissolution model,” he said. “Results obtained from these methods can positively predict in vivo absorption across the intestinal tract into the bloodstream. Once we see the improved bioavailability in the Caco-2 or dissolution model, we then move on to a randomized, double-blind, placebo-controlled clinical study, the gold standard of the industry.”
