Article Type : Research Article
Authors : Raoofi Asl Soofiani M and Wilczura-Wachnik H
Keywords : Micro emulsions; Bio compounds; Delivery systems; Surfactants; Micelles
In
the past and more recently, micro emulsions containing biological compounds
have increased the interest of researchers. They have shown great potential in
cosmetics, medicines, detergents. They are also used in combinations of many
natural/biocompatible compounds in the continuous or dispersed phase, which is
otherwise challenging to formulate. Natural compounds are complex mixtures that
contain compounds of different chemical nature; when combined in compounds and
have universal therapeutic activity, they act alone or in combination with
other compounds. In the present study, in this article, we intend to provide a
summary of the structure and properties of micro emulsion and micellar systems
and their applications in various industries, including pharmaceuticals and
food.
The three compound systems
containing hydrocarbons, water, and emulsifying agents, also called
surfactant's molecules depending on conditions, can form emulsion, micro
emulsions, Nano emulsions. Except for conditions under which these systems appear,
they differ with the dispersed phase's droplet size. The emulsions are systems
in which the average droplet size of the oil dispersion in water or water in
oil ranges from ~0, 1 - 100 m. In micro emulsions, the suitable droplet size is
in the range 0.0015- 0.15?m, and in Nano emulsions x10-9m. The Nano
emulsions are very stable systems, transparent, and they can be obtained by
mixing. Micro emulsions are classified as durable systems and can be obtained
by mixing also. Emulsions are cloudy and unstable systems that can be obtained
by shaking. The surfactant and co-surfactant molecules introduced into such
systems form the interfacial film stabilizing, especially micro emulsion’s
structure. Belonging to the continuous and scatter phases, three basic micro
emulsions are known: direct (oil scatter in water, o/w), reversed (water
distributed in oil w/o) bicontinuous. In 1959 Schulman et al. used it to
describe the transparent system containing oil, water, surfactant, and alcohol
[1]. Presently, such methods are called "micellar emulsion," micellar
solution, or "swollen micelles" [2,3]. A severe interest in micro
emulsion occurred at the end of the 20th century when it turned out that such
systems could be used in petro chemistry, wastewater treatment, and organic
synthesis. After several years of interest in micro emulsions and micellar
solutions, it was found that these systems can be applied in extraction,
catalysis, microreactors, and drug systems delivery.
Conventional microemulsions are
thermodynamically permanent, transparent, Newtonian, non-viscous liquids. They
have an enormous solubilization capacity for other beverages or guest molecules
like food additives, nutraceuticals, aromas, cosmetic compounds, active
compounds, and drugs [4]. Microemulsions are good vehicles for solubilization
and delivery of water and/or oil insoluble biologically active compounds [4,5].
Microemulsions 1are used in several applications, including agriculture and
food, drug solubilization, metal recovery, cosmetics, and beverages [6-12].
Microemulsion systems may be misrepresented by the title
"microemulsion." This claim is based on the description of an
emulsion, which contains the term "inherently unstable"; however, "microemulsions"
are inherently stable, by definition. In 2006 Flanagan and Singh's description
primarily differentiated their systems from the macroemulsion systems, which
were excellently studied at that course time [13]. An alternate moniker, or a
name that defines a self-assembled, isotropic structure and thermodynamically
stable, would be preferable to microemulsion. The initiation of a novel term to
describe microemulsions would be confusing in light of other words (such as
solubilized micellar solutions and swollen micelles), used to explain
self-assembled, isotropic systems, and thermodynamically stable. Generally,
although technically imprecise, it seems that the term
"microemulsion" is here to remain. In 2011 Rao and McClements
emphasize another terminology conflict: 'Some workers have challenged the
efficacy of defining systems which contain either low volume portions of water
or oil as microemulsions, suggesting that they should be referential as swollen
micelles, and reserving the name of microemulsion for systems which have
adequate dispersed phase incorporated into the surfactant micelles, such that
the micelle can be considered to have the properties of its bulk [14]. This is
compared to the opinion that there is no diversity between swollen micelles and
microemulsions. The term micellar solution should be maintained for
self-assembled surfactant micelles in solution. Given the inner nature of
'sufficient dispersed phase,' these authors agree with Malcolmson et al. that
thermodynamically stable, isotropic systems containing water, oil, and
surfactant should be named microemulsions, and surfactant solutions should be
called micelles or reverse micelles [15]. Microemulsions can dissolve large
amounts of bioactive in the internal phase and interface because their ratio is
too large. In addition to solubility, Microemulsions can act as carriers of the
active substance through human membranes (intestines), which is very important
in microemulsions systems. Of particular interest is the transport of
biologically active compounds in food to the human organism and drugs, mainly
insoluble in aqueous or poorly permeable systems. This review aims to outline
current knowledge concerning the solubilization of bio compounds in
microemulsions and to examine some new possible applications for microemulsions
as microreactors and crystallization sites. It is challenging to comprehend
general concepts to newcomers in the microemulsion field because research in
this field is seldom systematic and based on thermodynamic rules. The
researchers have used representative molecules and model systems and studied
different features of solubilization and transfer. It should be emphatic that
microemulsions have disadvantages. For example, microemulsions require
comparatively high condensation of surfactants. They are not suitable for some
uses, such as food applications. In many cases, co-solvents (alcohol and
polyols) are also needed, which are again considered inappropriate compounds.
Attempts were made to use GRAS (generally recognized as safe) surfactants as
components (mainly phospholipids), but these systems also have problems.
Surfactants show unique properties when present at low concentration in a system containing unmixed liquids (i.e., water and oil). This exceptional property involves the accumulation (adsorption) of the particles of the surfactant on the surfaces (liquid/gas, usually air) or the interfaces (among two immiscible liquids) of the system. Thus, an altering to a marked degree of the level of interfacial free forces is observed. Measuring a liquid's surface tension, we estimate the interfacial free force per unit of the liquid's boundary and the air above it. Surfactants usually reduce interfacial free force, and in this way, stabilize the system liquid/air. The system consisting of two immiscible liquids like water and oil can be merged by introducing additional external energy (stirring, shaking, mixing). Finally, an unstable emulsion can be formed. Such unstable emulsion easily collapses to the two initial simple liquid phases. The surfactants were introduced into such an unstable system extending interface and significantly decreasing surface tension between the liquid phase. As a result, the system becomes to be stable and macroscopically transparent [16]. This unique property of surfactant molecules is because of a characteristic molecular structure called amphipathic. Amphipathic molecules consist of two parts: lyophilic/hydrophilic (polar) and lyophobic/hydrophobic/lipophilic (non-polar) (Figure 1).
Figure 1: Scheme of the surfactant molecule
The surfactants are classified into
a few groups depending on the type of polar head. There are anionic, cationic,
amphoteric, zwitterion surfactants. Rosen has given the general structural
features and physicochemical characteristic of surfactants in 2004 [16] (Figure
2).
Except for surfactant, also so-called co-surfactant molecules introduced into such systems form the interfacial film stabilizing, especially microemulsion's structure. The interfacial film's curvature depends on the surfactant type and structure, as well as on temperature, oil to water phase ratio, and such factors as salinity, electrolyte presence in the systems, and others. The ionic surfactants containing single hydrocarbon toil (e.g., SDS sodium dodecyl sulfate) can aggregate to the micellar structures in the co-surfactant presence. Contrary to them, nonionic surfactants or ionic surfactants with double hydrocarbon toils (i.e., AOT) aggregate without the presence of co-surfactants. This ability of surfactant molecules to form the interfacial film between the dispersed droplets and dispersion phases relying on reducing the level tension between the water phase and the oil phase is the microemulsions micellar's fundamental property systems. This lowering level tension is sometimes enough to increase the level area so much that it is unnecessary to introduce external energy to the system, and the microemulsion forms spontaneously.
Figure
2: Examples
of surfactants belonging to different types.
The theories of "microemulsions" formation and stabilization are explained in brief here. For more details and information, the reader is directed to the research [17]. The mixed film theory considers the interfacial film as a duplex film, having different properties on the water and oil side of the interface, which is zero interfacial tension. The solubilization theory states microemulsions as solutions with solubilized water or hydrocarbon: Which are considered to be one-phase systems. Microemulsions' thermodynamic theory suggests that the free force of formation, ?Gm, includes various terms, like interfacial free force. When ?Gm is of a very slight or little negative value, microemulsion formation can be facilitated [17]. Regardless of the mechanism of microemulsion stabilization, the reduction of the interfacial free energy to a meager amount is critical in simplifying "microemulsions" formation [13].
Isotropic regions in a phase diagram are constructed based on the three major components in a system compound of the oil phase, surfactants phase (and possibly co-surfactant), and aqueous phase. Thermodynamically stable isotropic regions will form along with some phase diagrams. Mostly, at low oil concentrations (<30%), oil-in-water (o/w) microemulsions will form. Conversely, at low aqueous concentrations, water-in-oil (w/o) systems are developed. However, many different determining systems, such as micellar, reverse micellar, lamellar, and bicontinuous phases may exist at different oil-water-surfactant concentrations inside the specific o/w and w/o microemulsion areas. When there is no oil or water, micelles or reverse micelles may form the surfactant molecules through spontaneous self-assembly. All of them can exist, either alone or in composition with other systems, as transparent one-phase microemulsions. The boundary may be quite indistinct between microemulsion formation areas and areas outside the scope of one phase transparency. It may also coexist in balance with an excess of oil or water to single-phase microemulsions. These phases usually appear when there is an insufficient surfactant, and these multiphase systems are known as Winsor systems, and are illustrated in Figure 3. The Winsor type systems contain an o/w microemulsion, which balances with an excess oil phase. Contrariwise, Winsor type II comprises a w/o microemulsion in balance with a lower different water phase. Winsor type III includes a microemulsion phase in balance with an excess aqueous phase and an additional oil phase. In 1988 Kunieda et al. proposed extending Winsor's classification with the fifth category of the degree in microemulsions [18,19]. Finally, the single-phase systems already described (o/w, w/o, and bicontinuous) are Winsor type IV microemulsions [13] (Figure 3).
Figure
3:
Winsor classification system, showing oil- and water-rich phases possible in
microemulsion systems. Redrawn with permission.20 Copyright (1997) C.R.C.
Press.13
The microemulsion formation
procedure depends on what type we want to obtain: water in oil (w/o) or oil in
water (o/w). Usually, the process of microemulsion formation consists of two
steps: (1) water, surfactant (and optionally co-surfactant) are placed together
in a reactor and stirred until surfactant dissolves. (2) After, the mixture is
titrated with oil until visually turbidity appeared. The opposite situation is
also possible: (1) the mixture of water and oil is titrated with a surfactant
solution until the whole mixture's turbidity disappears. The quantitative
presentation of all mixtures (phase diagram) is usually shown on the Gibbs
triangle. In such a ternary phase diagram, the surfactant and co-surfactant are
classified as a single component. The literature in which authors present the
typical and characteristic structures possible in a whole range of
three-component phase diagram consisting of water/oil/surfactant is wide. Among
many are single papers and books [20-23].
·
Phase titration
manner
·
Phase inversion
manner
There are the following steps:
·
Dilution of the
combination of an oil-surfactant with water > {W/O}
·
Dilution of the
water-surfactant mix with oil. [O/W]
· Mixing of all
elements at once, in some systems, the instruction of components addition may
specify whether a microemulsion forms or not
Nonionic surfactants like polyoxyethylene are
susceptible to temperature. With temperature increasing, polyoxyethylene groups
become dehydrated and consequently alter critical packing parameters, which
results in phase inversion. The equipment used for the microemulsion
preparation is a colloidal mill, rotor-stator, homogenizer.
Mother Nature has been a source of
medicinal agents for many centuries, and today 65% of the world's population
relies on nature and its plants for their primary health care [24]. A bioactive
compound is a substance with biological activity that can negatively or
positively affect a living organism, depending on the kind of importance, the
dose, and their bioavailability [25]. In general, it claimed that bioactive
natural compounds in sufficient quantities could prevent or treat various
diseases worldwide [26]. Plants have many bioactive compounds; the significance
of these biologically active constituents are alkaloids, flavonoids, tannins,
and phenolic compounds [27]. These secondary metabolites have protective
functions such as antibacterial, antiviral, antifungal, insecticide, and
vegetarians with reduced their appetite for such plants [24]. Many researchers
claim that health benefits may be derived from incorporating bioactive
combinations in food products, also known as functional foods. These bioactive
compounds boost the brain, heart, and immune system's health and decrease the risk
of chronic diseases. For example, antioxidants are protecting human cells,
reducing risk, and are known as anti-cancer agents [28]. The unsaturated fatty
acids like omega-3, 6, and 9 are another example of bioactive combinations.
They are also known for heart disease debarment, immune system function, and
cancer prevention. Also, ?-carotene, curcumin, tocopherols, essential oils are
the most lipophilic biological active compounds incorporated to fortify food.
Microemulsions are exciting fields of utilization as they can act as carriers
or transfer systems for bioactive compounds as flavors, antioxidants,
anti-cancer, and antimicrobial agents [29-31]. In general, they lead to
improved consumer health.
Microemulsions as a delivery system of
natural compounds/bio-compounds
Microemulsions have found wide
applications in different human life areas, such as oil recovery, cosmetics,
pharmaceutics, the food industry, and others. Besides, to use in everyday life,
microemulsions have been applied in the science as simple models mimicking
merely some structural aspects of biomembranes, in organic synthesis as mini
reactors, and new areas appear [32-35]. In the past, oil recovery was the main
area of microemulsions applications [36]. Then, due to studies on
microemulsions' physicochemical properties, cosmetics, and the food industry,
applications have been developed [37]. In the previous century, pharmaceutics
discovered new potential applications for microemulsions. In 1984 Siegel's
discovery of inverted micellar structures had led to increased interest in
micellar systems as model structures in the survey of certain aspects of small
molecules' interaction with micelles [38]. These specific micellar systems'
specific properties are focused on scientists' investigations looking for
developing excellent vehicles for drugs, foods, and pharmaceutics. The
literature focused on microemulsions and micellar systems are pretty wide. It
Is describes the outcomes of UV-vis, calorimetric, NMR investigations on
biologically active compounds and systems mimicking structural aspects of parts
of natural membranes like normal and reversed micelles [39,40]. Generally,
chemists' main attention focuses on finding the privileged location of natural
compounds in the structure of micellar systems and estimating interactions
between them [41,42]. The complete investigations evidenced that the strong
limitations should be respected in the practical use of microemulsions or
micellar systems in delivering drugs, bio-compounds, or foods [43-45]. This is
due to the standards for acceptable doses of surfactants in products intended
for consumption. So, microemulsions and micellar systems still are useful for
studies on different aspects of transportation, diffusion, releasing of drugs,
food compounds. However, there was necessary to develop or find new compounds
for building a new generation of vehicles [43,46]. The similarity between
surfactants and phospholipids in the capability to aggregate micelles and microemulsion
formation has shifted drug transfer systems and bio-compounds to a new area. In
1965 Bangham et al. published the first description of liposomes, the first
structure of closed bilayer phospholipid systems [47]. This was the beginning
of the progress of new systems models for membranes, particularly bio-membranes
[48,49].
In 1971 Gregoriadis and Ryman
brought active investigations on physicochemical properties of membranes
obtained from phospholipids and the discovery by Gregory Gregoriadis possibility
for using them as drug transfer systems only synthetic pharmaceutics but the
bio-compounds too [50-53]. Reviewed application of microemulsions, and many
focused on microemulsions in food and years after Allen and Cullis in 2013
described the history of liposomes synthetically from concept to the clinical
applications [54]. It is worth underlining that in recent years, scientists
have focused attention on microemulsions containing natural oils as potential
carriers of hydrophobic bioactive compounds. There are wide literature: books,
review papers as by Xavier-Junior et al., in 2017 [55].
Nanoemulsions are systems with a typical particle size
between conventional emulsions and microemulsions, ranging from 50–200 nm [56].
The small droplet size of nanoemulsions can affect properties such as particle
stability, appearance, rheology, texture, and shelf life [57]. These systems
are described as approaching thermodynamic equilibrium [58]. Unlike
microemulsions, nanoemulsions are not thermodynamically stable. They are
kinetically stable and require much energy to produce. The droplets' net
attractive forces are reduced by reducing the droplets' size, so the nano
molecules show a lower tendency to droplet aggregation than conventional
emulsions [59].
The term
bioavailability means the natural digestion, absorption, and metabolism of a
particular nutrient. In general, the body absorbs a small number of compounds
such as active peptides, carotenes, lycopene, curcumin, essential oils, omega-3
fatty acids, vitamins, and drugs. Because microemulsions are so small in size
and therefore have a high surface-to-volume ratio, microemulsions and their
components are absorbed faster from the gut compared to emulsions. The advanced
dissolution of lycopene in O / W microemulsions increases bioavailability and
adsorption in human tissues [60]. Studies show that microemulsions can be used
to develop the oral bioavailability of drugs, including peptides [61-63]. The bioavailability
of ?-carotene increased compared to the usual dispersion when administered in
the shape of a microemulsion using a combination of surfactant monostearate and
its polyoxyethylene form [64]. Medium-chain triglycerides act as absorption
enhancers and increase the bioavailability of oils in human tissues [65]. The
increase in adsorption in different compounds depends on the kind of
emulsifier, pH, the particle size of the dispersed phase, the kind of lipid
phase, and the grade of solvability of the compound [66]. The possibility of
using microemulsions to enhance the attraction of different compounds,
including oils, vitamins, peptides, and protein drugs, is often examined in
pharmacological studies [67,68].
Today,
research studies on the utilization of microemulsions as carriers of natural
compounds are expanding. Indeed, microemulsions containing natural compounds
present many advantages, as previously mentioned in this review. Of course,
there are also problems with the utilization of microemulsions as a food
delivery system. Difficulties in applying microemulsions are two-fold:
restriction in the choice of appropriate surfactant and low solubilization of
high molecular weight triglycerides. For example, the utilization of food-grade
microemulsions in food belongs to the permissible limits of both the surfactant
kind and surfactant concentration. Therefore researchers need to use
surfactants that have been identified as GRAS for fundamental and applied
research. Dilute microemulsion systems that increase the solvability of
lipophilic compounds in aqueous phases are being developed, which increases the
application of microemulsions as a transfer system for bioactive compounds in
various industries. It will also be interesting to create scientific research
in vivo animals and humans after consuming functional foods fortified with
bioactive compounds through microemulsion. Such studies would reveal the
synergy among the bioactive compounds provided by the natural compound's
resource and their carrier, such as microemulsion systems. That could also
create new opportunities for collaboration between pharmaceuticals sciences and
the food sciences for the next few years.