SUB

: Physical Chemistry MICELLAR CATALYSIS,

TOPIC: SOLUBILIZATION, MICRO EMULSIONS, REVERSE MICELLES, CHARACTERIZATION OF MICRO EMULSIONS.

Solubilizaation
Introduction
It has been known that the aqueous solubility of sparingly soluble or insoluble substances can be increased by adding an appropriate third component.systematic studies using surfactants led to this phenomenon being called solubilization. Solubilization plays a very important role in industrial and biological processes.Mcbain and Hutchinson defined solubilization as a particular mode of bringing into solution substances that are otherwise insoluble in a given medium, involving the previous presence of a colloidal solution whose particles take up and incorporate within or upon themselves the otherwise insoluble material . This definition is too narrow, because the increase in solubility is not always caused by direct introduction of colloid particles into the system . More often ,the enhanced solubility of the solubilizates as colloidal particles is due to the presence of a third component . Therefore, the term solubilization has come to have the following very broad definition : the preparation of a thermodynamically stable isotropic solution of a substance normally insoluble or very slightly soluble in a given solvent by the introduction of an additionally amphiphilic component or components. Numerous applications of chemical engineering for example , the dissolution of drugs into aqueous solution and their transport to the body, the preparation of agricultural chemical solutions, and the recovery of oil- depend on solubilization by suitable surfactants. In addition,studies of the physical chemistry of the bile acids and the bile salts, on one hand , and of their physiological function as solubilizers,on the other hand,make it clear that the behavior of bile salts in vitro and their functions in vivo are closely related. Solubilization will be increasingly important in the future.

PHASE RULE OF SOLUBILIZATION

Much of the published work on solubilization is on the phase-separation model of the micelle. Accordingly , solubilization has been treated as a portioning of solubilizate molecules between a micellar phase and the intermicellar bulk phase .A few papers are based on the mass-action approach and theoretical discussion from this position have also appeared . Unfortunately,papers discussing solubilization from the standpoint of the Gibbs phase rule are very few. This section examine solubilization in terms of the phase rule. If the micelles are regarded as a phase, then adding an excess solubilizate phase means there are three phases (the third in the intermicellar bulk phase).The total numbers of components is three (solvent, surfactant and solubilizate),so the presence of three phases makes the system divariant.That would mean the surfactant concentration would be constant at constant

temperature and pressure but , in fact,the maximum additive concentration (MAC) changes the total surfactant concentration.Even it were postulated that the increase in the MAC with surfactant concentration above the CMC is due to an increase in the total micellar phase, the concentration of solubilizate in the micellar phase should still remain constant , because the concentration is an intensive property of the system and is therefore homogeneous throughout the micellar phase. If , on the other hand, the micelles are regarded as a phase and the system does not contain an excess solubilizate phase, there are three degrees of freedom. The surfactant concentration is then a unique variable that determines every intensive property of the system at constant temperature and pressure. In other words , the solubilizate monomer concentration in the intermicellar bulk phase (and therefore also in the micellar phase) is set automatically by the surfactant concentration,irrespective of the total solubilizate concentration in the system. This is not only totally incorrect as theory but is contrary to the experimental evidence that the concentration of solubilizates is determined only by the amount added to the system. Clearly, the phase- separation model of micelles and the partition model of solubilization disagree with reality.This contradiction is easily solved by treating the micelles as a chemical species.

Micellar Solubilization
An important property of micelles that has particular significance in pharmacy is their ability to increase the solubility of sparingly soluble substances in water. In this context, solubilization can be defined as the spontaneous dissolving of a substance by reversible interaction with the micelles of a surfactant in water to form a thermodynamically stable isotropic solution with reduced thermodynamic activity of the solubilized material (17). If we plot the solubility of a poorly soluble compound as a function of the concentration of surfactant, as shown in Figure 4, usually what happens is that the solubility is very low until the surfactant concentration reaches the cmc. At surfactant concentrations above the cmc the solubility increases linearly with the concentration of surfactant, indicating that solubilization is related to micellization.

Figure 4: Schematic plot of the concentration of a poorly soluble compound as a function of the surfactant concentration in aqueous solution. From the thermodynamic point of view, the solubilization can be considered as a normal partitioning of the drug between two phases, micelle and aqueous, and the standard free energy of solubilization ( GS ) can be represented by the following expression (1):

(1) where R is the universal constant of the gases, T is the absolute temperature, and P is the partition coefficient between the micelle and the aqueous phase. Usually, the solubilization of a molecule by a surfactant can be evaluated based on two descriptors that are the molar solubilization capacity, , and the micelle-water partition coefficient, P (24). The value is defined as the number of moles of the solute (drug) that can be solubilized by one mol of micellar surfactant, and characterizes the ability of the surfactant to solubilize the drug. It can be calculated based on the general equation for micellar solubilization:

(2) where S tot is the total drug solubility, S W is the water drug solubility, C surf is the molar concentration of surfactant in solution, and cmc is the critical micelle concentration (25). Since above the cmc the surfactant monomer concentration is approximately equal to the cmc , the term (C surf - cmc ) is approximately equal to the surfactant concentration in the micellar form and, therefore, is equal to the ratio of drug concentration in the micelles to the surfactant concentration in the micellar form. On the other hand, the micelle-water partition coefficient is the ratio of drug concentration in the micelle to the drug concentration in water for a particular surfactant concentration, as follows:

(3) Combining Equations (2) and (3), we can relate the two solubility descriptors. Accordingly, for a given surfactant concentration:

(4) As can be seen, P is related to the water solubility of the compound, in contrary to (25). In order to eliminate the dependence of P on the surfactant concentration, a molar micelle-water partition coefficient (PM), corresponding to the partition coefficient when Csurf = 1 M, can be defined as follows:

(5) The lower is the cmc value of a given surfactant, the more stable are the micelles. This is especially important from the pharmacological point of view, since upon dilution with a large volume of the blood, considering intravenous administration, only micelles of surfactants with low cmc value still exist, while micelles from surfactants with high cmc value may dissociate into monomers and their content may precipitate in the

Factors influencing solubilization

It is very enlightening and useful to consider the factors influencing solubilization. The total solubilizate concentration [ ] is a function of , [R], [M] or [ ]. These values determine the general behavior of solubilization as follows : 1. The MAC increases with increasing total surfactant concentration. As long as the value of and [R] remain constant because solubilization is small, the MAC increases linearly above the CMC. 2. The MAC of a hydrophobic solubilizate per mole of surfactant increases with alkyl chain length for a series of homologous surfactants. This effect can be attributed to the increase in the number of surfactant molecules available for micellization caused by (1) the decrease in CMC ; (2) an increase in due to the increasing volume of the hydrophobic part of the micelles; and (3) the fortified hydrophobicity of the micelle palisade layer brought about by the closer packing of the surfactant molecules at the micellar surface.The MAC also depends on the hydrophobicity of the solubilizate :the MAC decreases with the increase in the hydrophobicity or alkyl chain length of the solubilizates as a result of a decreases in [R].However increases. 3. The MAC of some solubilizates increases with temperature owing to an increase in the monomeric solubilizate concentration [R]. However, the value usually decrease with temperature. In this case, the MAC depends largely on the temperature dependence of [R] and . 4. The MAC is increased by addition of excess salt. This effect is due to (1) a decrease in CMC, (2) an increase in micelle size and/or a change in micelle shape, and (3) fortified hydrophobicity of the palisade layer owing to reduced repulsion between hydrophilic groups. These factors lead to the higher value.However, the salt effect is relatively small for micelles with small aggregation numbers, for example bile salts.

The MAC is determined in exactly the same way as solubility, by a measurement requiring complete separation of micellar solution from the solubilizate phase. Other useful information on solubilization is available from liquid chromatography,gel filtration,NMR,dialysis,potentiometry, spectroscopy,mixed micelle formation, kinetics and volumetric studies.

LOCATION OF SOLUBILIZATES IN MICELLES

The position of solubilizates in micelles,as well as in living membranes,provides very important information concerning the physiochemical properties and physiological functions of both solubilizate and micelle or membrane. This property can be investigated using probe molecules, the molecular spectrum of which indicates the surrounding conditions. The absorption spectrum of a molecule depends on the dielectric constant of the medium surrounding the molecule.The dielectric constant of a micelle ranges from 2 for the liquid hydrocarbon in the inner core to 80 for the water of the outer micellar surface. The following generally accepted rules for solubilizate position are derived from many works : (1) Nonpolar aliphatic hydrocarbons locate in an inner hydrophobic micellar core. (2) Semipolar and polar compounds such as alcohols, acids, and amines locate at the so-called palisade layer of the micelle with the polar group of the micellar surface and the nonpolar hydrocarbon groups in the micellar core.(3) Aromatic hydrocarbons such as benzene, toluene, and naphthalene sit in the micellar core and at the micellar surface (the two- state model. The fraction of solubilizates occupying each of these two sites depend on the solubilizate concentration in the intermicellar bulk phase.the fraction in the inner core increases with increasing concentration in the bulk phase because the increase in the chemical potential of the solubilizate enables the solubilizate bto move toward the micelle core. Recently the postulated Laplace pressure increase in the micellar interior has often been employed to explain the diminished free energy change of transfer per methylene group from the aqueous bulk into the micellar interior, compared with the free energy change from the aqueous bulk into the bulk liquid hydrocarbon. In addition, the decreased free energy change of micelle formation per methylene group has been attributed to partial crystallinity of the alkyl chain in the micellar interior caused by the increased pressure.An interfacial tension does exist just at the boundary between two bulk phases. If the postulated pressure increase exists,it can take place only in the case where micelles can be a separate phase. However, according to the phase rule, micellesare not a separate phase but a chemical species.This model implies partial crystallinity of the alkyl chains in the solubilized state in the micellar core, with the hydrophilic group of solubilizate molecules anchored to the micellar surface, resulting in greatly reduced alkyl chain mobility and thus reduced free energy change of transfer per methylene group.In other words the Laplace pressure is unnecessary to elucidate the above phenomenon, which were investigated by the solubilization of p-n-alkylbenzoic acids with different alkyl chain lengths into dodecyl sulfonic acid micelles.

Micellar Catalysis
The special properties of surfactants are important in a wide variety of applications in chemistry, biology, engineering, materials science, and other areas. These molecules are said to be amphipathic; that is, they have distinct hydrophilic (polar) and hydrophobic (nonpolar) regions. The polar region, called the headgroup, may be either neutral, cationic, anionic, or zwitterionic. The hydrophobic tail has one or more chains of varying length, composed usually of a hydrocarbon. Common examples are: Polyoxyethylene(6) octanol, CH3(CH2)7(OCH2CH2)6OH (neutral) Cetyltrimethylammonium bromide, CH3(CH2)15(CH3)3N+Br- (C16TAB, cationic) Sodium dodecyl sulfate, CH3(CH2)11OSO3-Na+ (SDS, anionic) N-dodecyl-N,N-dimethylglycine, CH3(CH2)11(CH3)2N+CH2COO- (zwitterionic).

Surfactants dissolve completely in water at very low concentrations, but above a certain level, the critical micelle concentration (CMC), the molecules form globular aggregates, called micelles. As shown in Figure 1, the hydrophobic tails group together to create a nonpolar interior with the headgroups located at the surface of the glob in contact with the aqueous environment. Micelles vary in size and shape, but are commonly rough-surfaced spheres with aggregation numbers on the order of 50-100.

Figure 1.. This is a cross-section of an overall roughly spherical structure. The Br- counterions are distributed in the surface layer of the micelle and in the surrounding solution. The hydrophobic tails are actually tangled randomly, and some of them protrude on the surface to help shield the positive charges from each other. Surfactants are best known to us as detergents. Grease on skin, dishes, clothes, etc. is attracted to the hydrophobic micelle interior and is subsequently rinsed away. This ability to solubilize nonpolar materials has made aqueous surfactant systems increasingly popular alternatives to organic solvents in various applications. An example is emulsion polymerization, in which water-insoluble molecules are polymerized in an aqueous micellar environment. This procedure alleviates problems due to high viscosities and removal of solvent and monomer.

The presence of micelles can have marked effects on chemical reactions. The thermodynamic favorability of, for example, an acid dissociation can be shifted significantly. Of particular interest in this experiment is the alteration in chemical kinetics. Reaction rates can be either accelerated or decelerated, depending on the chemical system, the type and concentration of the surfactant, and other factors, such as pH, ionic strength, etc. The effect of surfactants on reaction kinetics is often called micellar catalysis.

Contributing Factors in Micellar Catalysis

A side from inherent interest in micelles as catalytic entities, a good deal of consideration of micellar reactions as models for certain aspects of enzyme catalyzed reactions has been developed. The same statement is also true for catalysis of organic reactions by polysoaps. In many respects, micelles fail as models for enzyme catalyzed reactions. Functional micelles have been developed which match certain enzymatic reaction velocities but nonfunctional micelles are much less effective catalysts. Neither functional nor non-functional micelles exhibit the degree of specificity associated with enzymatic reactions and neither class of micellar reaction is subjected to the kind of control to which the enzymes are. Nonetheless. In one important respect, non-functional micelles are suitable models for enzymatic catalysis. Enzymes and micelles derive significant portion of their catalytic ability from the same sources. This matter has been discussed, in revealing detail by Jencks (Jencks 1975). The fact that micelles are catalysts for a number of reactions is equivalent to that there is a decrease in the standard free energy of the transition state relative to reactants in the aqueous phase. The question is: to what factor or factors may one attribute this diminution in standard free energy difference between reactants and transition state? In dealing with reactions in homogeneous systems, it is customary to discuss this question in terms of the BrnstedBjerrum equation: G* = RT ln f*/fA fB (1.10) In terms of this equation, one analyzes rate changes in terms of effects on activity coefficients for substrate and transition state. While this is a straightforward procedure, for the most part, for homogeneous systems that

there are two significant difficulties in carrying out such analysis for reactions occurring on the surface of a micelle or, for that matter, on the surface of an enzyme. First, an important contributor to catalysis in micellar or enzymatic systems for second order and higher order reactions derives from a decrease in entropy of the reactants by virtue of their binding to the catalyst surface. That is to say, if the substrates are confined to the micellar surface the volume available to them is much decreased from that available in the bulk aqueous phase. This is equivalent to recognie that the two or more reactants will be much more concentrated with respect to each other as a consequence of the binding reactions. This entropic contribution to the reaction rate is not easily understood in terms of the BronstedBjerrum equation. Second, the activity coefficient of a molecule in the micellar phase may not be revealing in attempting to account for an increase in reaction rates. This derives from the fact that there may be different microenvironments for different parts of the absorbed molecule. The fact that an organic substrate, for example, associates with micelles with an equilibrium constant, greater than unity, requires that its activity coefficient decreases on going from the aqueous phase to the micellar phase. However, the overall decrease in the activity coefficient of the reactant may be accompanied by an increase in the activity coefficient at the site of chemical reaction. This consideration is true for both enzymatic and micellar reactions. It, too, is not evident on the basis of the BrnstedBjerrum equation. Understanding of catalysis for organic reactions in the presence of micelles requires that one separate the two factors indicated above: the entropic contribution reflecting concentration effects and the effects on the relative activity coefficients at the site of reaction for substrates and transition states, a consequence of the nature of the microenvironment in which the reaction occurs. Perhaps the simplest means of accomplishing this end is to begin by considering unimolecular reactions for which the entropic contribution cannot be important. Rate changes, whether it be catalysis or inhibition, must necessarily reflects the changes in the nature of the medium in which the reaction occurs. If it is assumed that all of the substrate is in the micelle and those activity coefficients for both substrate and transition state in the aqueous phase is unity, the extent of catalysis is given by the simple equation: K/K0 = fA/f# (1.11) in which the activity coefficients refer to the micellar system. Clearly, catalysis may result from destabilization of the reactant, an increase in fA, or stabilization of the transition state, reflected in a decrease in f. As a pertinent example of micellar catalysis for a unimolecular reaction, let us consider the decarboxylation of 5nitrobenzisoxazole3--carboxylate, a reaction probed in considerable detail by Bunton and his coworkers (Bunton 1970, 1971, 1973):

5nitrobenzisoxazole3carboxylate This reaction is catalyzed by cationic, nonionic, and zwitterionic surfactants.The decarboxylation of 5nitrobenzisoxazole3carboxylate is a reaction in which a negatively charged localized in the substrate, is delocalized in the transition state. Consequently, it might be anticipated that the reaction would be accelerated in less polar environments. The work of Kemp and Paul, prior to the initiation of studies in micellar systems, established that this is the case (Kemp 1970). Consequently, the most logical explanation for the fact that micelles are effective catalysts for this reaction is substrate destabilization in the less polar environment, provided by the micellar surface. This destabilization is most probably electrostatic in nature, since, a consideration indicated above suggests that the carboxylate function is probably not significantly desolvated at the micellar surface. This reaction shows one additional notable feature. The rates are modestly increased upon the addition of certain salts. This is contrary to the observations made for many bimolecular reactions in micellar systems for which salts are almost uniformly inhibitory. In this particular case, the catalytic effect of added salts must reflect some alteration in the shape and properties of the micelles. (Kunitake et al 1977). have investigated the polysoapcatalyzeddecarboxylation of the same substrate (Kunitake 1977). Partially, laurylatedpoly(4vinylpyridine) and poly(2 ethyllvinylimidazoles) are moreeffective as catalysts for this reaction than simple cationic surfactants. Theaddition of hydrophilic salts elicits complex kinetic behavior. Such salts first diminishes then, at higher concentrations, increase the rate of the polysoap dependent decarboxylation. Like the micellar reaction, catalysis, observed in the presence of polysoaps probably reflects destabilization of the carboxylate moiety of the substrate in the nonpolar environment. Bovine serum albumin was observed by the same workers to be noncatalytic for this reaction (Kunitake 1977). Klotz and coworkers have observed that partially laurylated polyethyleneimines are even more potent catalysts for 5 nitrobenzisoxazole 3 fold was observed. The reaction obeys the Michaelis Menten kinetics pattern typical of enzymatic reactions. A second example of micellar catalysis which must derive from medium effects, as opposed to entropic ones, was provided carboxylate decarboxylation (Suk 1976). A maximum catalytic effect of 1300 by the unimolecular hydrolysis of phosphate esters.

Although phosphate monoanions are readily incorporated into micelles,formed from cationic surfactants. This does not result in an appreciablealteration in the rate of hydrolysis (Bunton 1968). Those phosphate ester dianions in which the leaving group contains strong electron attracting groupsalso hydrolyze via unimolecular elimination of metaphosphate. In this case,however, cationic micelles are good catalysts for hydrolysis (Bunton 1968,70 ,Buist 1970,). For example, the rate of hydrolysis of 2,5dinitrophenyl phosphate dianion is approximately 25 times more rapid in the presence of an optimal concentration of hexadecyltrimethylanunonium bromide than in water.The loss of the metaphosphate anion from a phosphate ester dianion involves dispersal of two negative charges. Consequently one may argue that the catalytic driving force involves destabilization of the substrate relative to the transition state by the relatively nonpolar micellar surface, an explaination essentially the same as that invoked in the case of decarboxylation reactions. This conclusion is consistent with the fact that the rate of hydrolysis of phosphate ester dianions is significantly increased with a decrease in solvent polarity in the absence of micelles. These examples of catalysis by micelles for unimolecular reactions indicate that the utilization of binding forces between substrate and micelle can bring reactive functionalities of the substrate into an environment in which reactivity is augmented. We now turn attention to the case of bimolecular reactions in which not only medium effects but entropic effects resulting from concentration of reactants may be important. In one of the earliest thorough studies of micellar catalysis, Duynstee and Grunwald established that rate and equilibrium constants for addition of hydroxide ion to stable triphenyl methyl cationic dyes, such as Crystal Violet, is subjected to catalysis by cationic surfactants (Duynstee and Grunwald 1959). This conclusion has been confirmed and amplified in several subsequent investigations (Bunton 1972-1973, 1976, Albrizio 1972). Facilitation of this reaction may reflect (i) concentration of hydroxide ion in the presence of the cationic dye by the cationic miceilar surface and (ii) destabilization of the cationic dye by the cationic micellar surface that the latter effect is important & suggested by two considerations. First, equilibrium constants for incorporation of the cationic dye and the corresponding alcohol into the micellar phase indicate a strong electrostatic effect in this process.

The magnitude of the effect is about the same as that for the equilibrium constant for addition of hydroxide ion to the cationic dyes in the presence of micelles (Duynstee 1959).

A Kinetic Model For Micellar Catalysis

It has been tempting to provide an explicit quantitative explanation to account for the principal features of micelle catalyzed reactions. These include the shape of rate concentration profiles, dependence of catalytic parameters on the nature of the surfactant, particularly on the length of the hydrocarbon chain which determines the cmc, dependence of catalytic parameters on the hydrophobicity of the substrate, and inhibition of the reaction by salts. The first kinetic model for micelle catalyzed reactions was proposed by Menger and Portnoy (1967):

employing certain simplifying assumptions, this kinetic scheme provides the following rate law: in which K is the equilibrium constant for association of substrate, 5, with micelles, Dn and Km is the concentration of micelles: Cm(CDcmc)/N, CD being the total surfactant concentration and is the micelle aggregation number. This equation predicts a s increase in rate constant with increasing surfactant concentration. Such behavior is seen for unimolecular reactions in the presence of micelles and this simple equation gives a good account of the data. On the other hand, reactions which are secondorder, or higher order, usually exhibit an optimal rate at some surfactant concentration above which the rates decrease with increasing concentration. This fact has led to a search for a more satisfactory kinetic treatment for these more complex cases. An important advance was made by Berezin and coworkers who treated the case of reaction of two uncharged organic molecules. The equation which they derived is:

in which V is the molar volume of the surfactant, P is the partition coefficient of the substrate between the two phases, and the other quantities have been identified earlier. This equation accounts well for data which was intended to

explain. On the other hand, it is not readily applicable to understand some of the features of reactions between ions and organic molecules. Perhaps the most generally satisfactory theory was that developed by Romsted in the author's laboratory (50). Romsted's theory depends on the following assumptions. First, one can write an equilibrium constant for the interaction of the substrate with the micelles. This assumption is common to all kinetic treatments of micellar catalysis but may fail in cases in which micelle structure and properties change as a function of some parameter in the reacting system. Second, and crucial, is the assumption that the Stern layer is always saturated with respect to counterions. In this respect, the Romsted treatment differs from all previous ones. This assumption is equivalent to the statement that the ground state for ions is the ion bound to the micellar surface, and not the ion free in the bulk phase. This assumption was introduced into the kinetic treatment in the form of an equilibrium constant describing counterion exchange on the micellar surface:

in which I is taken to be a reactive and X an unreactive counterion. Mathematical analysis yields the following equation to describe the rate constants for secondorder reactions in the micellar phase:

in which the degree of binding of counterions to the Stern layer and S is the molar density of micellar phase. In the case of first order reactions, this simply reduces to the equation of Menger and Portnoy (eq. 1.14). The utility of (1.17), lies largely in the fact that it accounts quantitatively for the basic features of reaction kinetics in micellar systems. Let us briefly consider few examples, more detailed analyses are available (Romsted 1975,1977 ). First, one of the repeated observations for secondorder reactions in the presence of micelles is that plots of observed rate constants against surfactant concentration pass through maxima. Computergenerated plots, based on eq. 10 mimic this behavior (Romsted 1975,1977). This fact can be understood in terms of two competing effects, both of which are integrated into eq. 1.17. On the one hand, with increasing surfactant concentration, the relative concentrations of organic substrate and ionic reactant in the Stern layer increase rapidly; this tends to accelerate the reaction, accounting for the ascending limb of the curve. On the other hand, increasing surfactant

concentration (for ionic surfactants) requires that the unreactive counterion concentration also increase while the reactive ion concentration remains constant. Since there are a limited number of ionic binding sites in the Stern layer, this requires that the concentration of the reactive ion in the vicinity of bound organic substrate decrease. This accounts for the descending limb observed at high surfactant concentrations. Second, it has been observed in a number of cases that increasing substrate hydrophobicity results in larger maximal rate increases which are attained at progressively lower surfactant concentrations (Table 1.4 for example). Computergenerated plots based on eq. 10 reproduce this behavior very nicely (Romsted 1975,1977 ). The increase in substrate hydrophobicity is reflected in eq. 10 in terms of an increase in K, the equilibrium constant for incorporation of the organic substrate into the Stern layer. The greater the binding constant, the less surfactant required to incorporate the substrate into the micellar pseudophase. This leads to faster rate increases as a function of surfactant concentration. In turn, this means that less uncreative counterion will be present, accounting for the fact that greater maximal rate increases are observed. Third, it is frequently observed that increasing surfactant hydrophobicity also leads to greater maximal rate increases which are attained at lower surfactant concentrations (Table 1.4 for example). This is accounted for in just the same way as that employed above: increasing hydrophobicity (in substrate or surfactant) leads to increases in Ka and hence, to a greater concentration of reactive ion in the Stern layer. Hence, eq. 10 accounts well for this observation, too. Fourth, a particularly nice success of eq. 10 is that it accords with the important observation of (Bunton and Wolfe) that secondorder rate constants for specific acid catalyzed hydrolysis of nitrobenzaldehyde diethyl acetal in the presence of sodium dodecyl sulphate decrease with increasing acid concentration. Note that eq. 10 predicts that the observed secondorder rate constants are inversely related to the reactive ion concentration, accounting for the observation. This realization was first stated by Berezin and his coworkers (Berezin 1973). Finally, there are many examples of inhibition of reactions in micellar systems through increasing concentrations of unreactive counterions; the extent of inhibition increases with increasing affinity of the unreactive counterions for the Stern layer. This phenomenon finds a ready explanation in terms of eq. 10 since a competition between reactive and unreactive ions for sites in the Stern layer, and hence in the vicinity of bound organic substrate, has been built into the model explicitly (eq. 9). These examples should suffice to indicate that the theory of Romsted, and to a significant extent that of Berezin, is adequate to account for the salient features of micellar catalysis in a qualitative way at least. No one would argue that the theoretical treatments available are the last word; quite the contrary, it seems certain that improvements will be forthcoming regularly. However, in addition to provide chemically rational explanations for the dependence of the kinetics of these reactions on a number of variables, the equations derived are of predictive value. Efforts to examine these predictions will certainly lead to additional insight into reaction kinetics in micellar

systems.

MICROEMULSIONS
The design and development of new drug delivery systems with the intention of enhancing the efficacy of existing drugs is an ongoing process in pharmaceutical research. It is necessary for a pharmaceutical solution to contain a therapeutic dose of the drug in a volume convenient for administration. Dispersions of oil and water are commonly employed in the pharmaceutical industry. These dispersions can be classified in three major categories: 1. Microemulsions 2. Micellar Solutions 3. Conventional Emulsions (or) Macroemulsions

(Figure 1. Structures of Micellar solution, Microemulsion and Emulsion) Out of these three, Microemulsions have attracted much interest in recent years in terms of their drug delivery potential. Microemulsions are liquid dispersions of water and oil that are made homogenous, transparent (or translucent) and thermodynamically stable by the addition of relatively large amounts of a surfactant and a co-surfactant and having diameter of the droplets in the range of 100 1000 A (10 100 nm).

(Figure 2:- Microemulsion Structure) They appear to represent a state intermediate between thermodynamically stable solubilized solutions i.e. micelles containing solubilized oils and ordinary emulsions which are relatively unstable. Three types: 1. O/W Microemulsion 2. W/O Microemulsion 3. Bicontinuous Microemulsion

(Figure 3: O/W, W/O and Bicontinuous Microemulsions) History Of Microemulsion Microemulsions were not really recognized until the work of Hoar and Schulman in 1940, who generated a clear single phase solution by titrating a milky emulsion with hexanol. The term microemulsion was first used even later by Schulman et al. in 1959 to describe a multiphase system consisting of water, oil, surfactant and alcohol, which forms a transparent solution. There has been much debate about the word microemulsion to describe such systems. Although not systematically used today, some prefer the names micellar emulsion or swollen micelles. Microemulsions were probably discovered well before the studies of Schulman: Australian housewives have used since the beginning of last century water/eucalyptus oil/soap flake/white spirit mixtures to wash wool, and the first commercial microemulsions were probably the liquid waxes discovered by Rodawald in 1928. Interest in microemulsions really stepped up in the late 1970s and early 1980s when it was recognized that such systems could improve oil recovery and when oil prices reached levels where tertiary recovery methods became profit earning. Nowadays this is no longer the case, the other microemulsion applications are discovered, e.g., catalysts, preparation of submicron particles, solar energy conversion, liquid-liquid extraction (mineral, proteins, etc.). Together with classical applications in detergency and lubrication, the field remains sufficiently important to continue to attract a number of scientists. From the

fundamental research point of view, a great deal of progress has been made in the last 20 years in understanding microemulsion properties.

Important Characteristics Of Microemulsions

Particle size < 200 nm Thermodynamically stable Optically clear Surface area increased High solubilizing capabilities

Preparation of microemulsion

On the basis of the solubility studies, oleic acid was selected as the oil phase. Tween 80 and PEG 400 were selected as surfactant and cosurfactant,respectively. Distilled water was used as an aqueous phase. Surfactant and cosurfactant (Smix) were mixed at different mass ratios (1:1,2:1,3:1,4:1). These ratios were chosen in increasing concentration of surfactant with respect to cosurfactant and increasing concentration of cosurfactant with respect to surfactant for a detailed study of the phase diagrams. Predetermined amounts of the drug were dissolved in the required quantity of oil. Surfactant and co-surfactant were added to the above mixture as a fixed ratio. Distilled water was added gradually with continuous stirring, which resulted in the formulation of a transparent and homogenous microemulsion. Parameters optimized for the preparation of microemulsion were the type and concentration of the oil phase, surfactant and cosurfactant.

Advantages Of Microemulsion Based Systems

Microemulsions exhibits several advantages as a drug delivery system :

1.Microemulsions are thermodynamically stable system and the stability allows selfemulsification of the system whose properties are not dependent on the process followed. 2.Microemulsions act as supersolvents of drug. They can solubilize hydrophilic and lipophilic drugs including drugs that are relatively insoluble in both aqueous and hydrophobic solvents. This is due to existence of microdomains of different polarity within the same single-phase solution. 3.The dispersed phase, lipophilic or hydrophilic (oil-in-water, O/W, or water-in-oil, W/O microemulsions) can behave as a potential reservoir of lipophilic or hydrophilic drugs, respectively. The drug partitions between dispersed and continuous phase, and when the system comes into contact with a semi-permeable membrane, the drug can be transported through the barrier. Drug release with pseudo-zero-order kinetics can be obtained, depending on the volume of the dispersed phase, the partition of the drug and the transport rate of the drug. 4.The mean diameter of droplets in microemulsions is below 0.22 mm; they can be sterilized by filtration. The small size of droplet in microemulsions e.g. below 100 nm, yields very large interfacial area, from which the drug can quickly be released into external phase when absorption (in vitro or in vivo) takes place, maintaining the concentration in the external phase close to initial levels. 5.Same microemulsions can carry both lipophilic and hydrophilic drugs. 6.Because of thermodynamic stability, microemulsions are easy to prepare and require no significant energy contribution during preparation. Microemulsions have low viscosity compared to other emulsions. 7.The use of microemulsion as delivery systems can improve the efficacy of a drug, allowing the total dose to be reduced and thus minimizing side effects. 8.The formation of microemulsion is reversible. They may become unstable at low or high temperature but when the temperature returns to the stability range, the microemulsion reforms.

Disadvantages Of Microemulsion Based Systems

1.Use of a large concentration of surfactant and co-surfactant necessary for stabilizing the nanodroplets. 2.Limited solubilizing capacity for high-melting substances 3.The surfactant must be nontoxic for using pharmaceutical applications

4.Microemulsion stability is influenced by environmental parameters such as temperature and pH. These parameters change upon microemulsion delivery to patients. Comparison With Emulsions (Macroemulsions) Emulsions (Macroemulsions)

Microemulsions

Figure 4: - Emulsions 1.

1. 1.

1.

1.

1.

Figure 5: - Microemulsions 1. They constantly evolve between various Emulsions consist of roughly spherical structures ranging from droplet like droplets of one phase dispersed into the swollen micelles to bicontinuous other. structure. Droplet diameter: 1 20 mm. 1. 10 100 nm. Most emulsions are opaque (white) 1. Microemulsions are transparent or because bulk of their droplets is greater translucent as their droplet diameter are than wavelength of light and most oils less than of the wavelength of light, have higher refractive indices than water. they scatter little light. 1. Microemulsion droplet may disappear Ordinary emulsion droplets, however within a fraction of a second whilst small exist as individual entities until another droplet forms spontaneously coalesance or ostwald ripening occurs. elsewhere in the system. They may remain stable for long periods 1. More thermodynamically stable than of time, will ultimately undergo phase macroemulsions and can have essentially separation on standing to attain a infinite lifetime assuming no change in minimum in free energy. They are composition, temperature and pressure, kinetically stable thermodynamically and do not tend to separate. unstable. They are lyophobic. 1. They are on the borderline between

1. Require intense agitation for their formation.

lyophobic and lyophilic colloids. 1. Generally obtained by gentle mixing of ingredients.

Factors To Be Considered During Preparation Of Microemulsion

Three important conditions: Surfactants must be carefully chosen so that an ultra low interfacial tension (< 10-3 mN/m) can be attained at the oil / water interface which is a prime requirement to produce microemulsions. 1.Concentration of surfactant must be high enough to provide the number of surfactant molecules needed to stabilize the microdroplets to be produced by an ultra low interfacial tension. 2.The interface must be flexible or fluid enough to promote the formation of microemulsions.

Phase Behaviour
The phase behaviour of simple microemulsion systems composing oil, water and surfactant can be studied with the aid of ternary phase diagram (at fixed pressure and temperature) in which each corner of the diagram represents 100% concentration of the particular component. Generally, pharmaceutical microemulsions contain additional components such as a cosurfactant and/or drug. The co-surfactant is also amphiphilic with an affinity for both the oil and aqueous phases and partitions to an appreciable extent into the surfactant interfacial monolayer present at the oil-water interface. A large number of drug molecules are by themselves surface active and they are expected to influence phase behaviour. For four or more components, pseudo ternary phase diagrams are used to study the phase behaviour. In this diagram a corner will typically represent a binary mixture of two components such as surfactant/co-surfactant, water/drug or oil/drug. The number of different phases present for a particular mixture can be visually assessed.

Hypothetical Phase Regions of Microemulsion Systems

Hypothetical Phase Diagram From above figure, we can see that, With high oil concentration surfactant forms reverse micelles capable of solubilizing water molecules in their hydrophilic interior. Continued addition of water in this system may result in the formation of W/O microemulsion in which water exists as droplets surrounded and stabilized by interfacial layer of the surfactant / co-surfactant mixture. At a limiting water content, the isotropic clear region changes to a turbid, birefringent one. Upon further dilution with water, a liquid crystalline region may be formed in which the water is sandwiched between surfactant double layers. Finally, as amount of water increases, this lamellar structure will break down and water will form a continuous phase containing droplets of oil stabilized by a surfactant / co-surfactant (O/W microemulsions)

Characterization Of Microemulsions
Microemulsions have been characterized using a wide variety of techniques. The characterization of microemulsions is a difficult task due to their complexity, variety of

structures and components involved in these systems, as well as the limitations associated with each technique but such knowledge is essential for their successful commercial exploitation. Therefore, complementary studies using a combination of techniques are usually required to obtain a comprehensive view of the physicochemical properties and structure of microemulsions. At the macroscopic level viscosity, conductivity and dielectric methods provides useful information. (A) Phase Behavior Studies Phase behavior studies are essential for the study of surfactant system determined by using phase diagram that provide information on the boundaries of the different phases as a function of composition variables and temperatures, and, more important, structural organization can be also inferred. Phase behaviour studies also allow comparison of the efficiency of different surfactants for a given application. In the phase behaviour studies, simple measurement and equipments are required. The boundaries of one-phase region can be assessed easily by visual observation of samples of known composition. The main drawback is long equilibrium time required for multiphase region, especially if liquid crystalline phase is involved. Other useful means and ways of representing the phase behaviour are to keep the concentration of one component or the ratio of two components constant. As the number of components increases, the number of experiments needed to define the complete phase behaviour becomes extraordinary large and the representation of phase behaviour becomes extremely complex. One approach to characterize these multicomponent systems is by means of pseudoternary diagrams that combine more than one component in the vertices of the ternary diagram. (B) Scattering Techniques for Microemulsions Characterization Small-angle X-ray scattering (SAXS), small-angle neutron scattering (SANS), and static as well as dynamic light scattering are widely applied techniques in the study of microemulsions. These methods are very valuable for obtaining quantitative informations on the size, shape and dynamics of the components. The major drawback of this technique is the dilution of the sample required for the reduction of interparticular interaction. This dilution can modify the structure and the composition of the pseudophases. Nevertheless, successful determinations have been carried out using a dilution technique that maintains the identity of droplets. Small-angle X-ray scattering techniques have been used to obtain information on droplet size and shape. 23-27 Static light scattering techniques have also been widely used to determine microemulsion droplet size and shape. In these experiments the intensity of scattered light is generally measured at various angles and for different concentrations of microemulsion droplets. Dynamic light scattering, which is also referred as photon correlation spectroscopy (PCS), is used to analyse the fluctuations in the intensity of scattering by the droplets due to Brownian motion. The selfcorrelation is measured that gives information on dynamics of the system. 28-32 (C) Nuclear Magnetic Resonance Studies

The structure and dynamics of microemulsions can be studied by using nuclear magnetic resonance techniques. Self-diffusion measurements using different tracer techniques, generally radio labeling, supply information on the mobility of the components. The Fourier transform pulsed-gradient spin-echo (FT-PGSE) technique uses the magnetic gradient on the samples and it allows simultaneous and rapid determination of the self-diffusion coefficients (in the range of 109 to 10-12 m2s-1), of many components. (D) Interfacial Tension The formation and the properties of microemulsion can be studied by measuring the interfacial tension. Ultra low values of interfacial tension are correlated with phase behaviour, particularly the existence of surfactant phase or middle-phase microemulsions in equilibrium with aqueous and oil phases. Spinning-drop apparatus can be used to measure the ultra low interfacial tension. Interfacial tensions are derived form the measurement of the shape of a drop of the lowdensity phase, rotating it in cylindrical capillary filled with high-density phase. (E) Viscosity Measurements Viscosity measurements can indicate the presence of rod-like or worm-like reverse micelle. Viscosity measurements as a function of volume fraction have been used to determine the hydrodynamic radius of droplets, as well as interaction between droplets and deviations from spherical shape by fitting the results to appropriate models (e.g. for microemulsions showing Newtonian behaviour, Einsteins equation for the relative viscosity can be used to calculate the hydrodynamic volume of the particles). (F) Predicting Microemulsion Type A well-known classification of microemulsions is that of Winsor who identified four general types of phase equilibria: Type I The surfactant is preferentially soluble in water and oil-in-water (O/W) microemulsions form (Winsor I). The surfactant-rich water phase coexists with the oil phase where surfactant is only present as monomers at small concentration. Type II The surfactant is mainly in the oil phase and water-in-oil (W/O) microemulsions form. The surfactant-rich oil phase coexists with the surfactant-poor aqueous phase (Winsor II) Type III A three-phase system where a surfactant-rich middle-phase coexists with both excess water and oil surfactant-poor phases (Winsor III or middle-phase microemulsion). Type IV A single-phase (isotropic) micellar solution, that forms upon addition of a sufficient quantity of amphiphile (surfactant plus alcohol). (G) Simple tests used are Dye Solubilization

A water soluble dye is solubilized within the aqueous phase of the W/O globule but is dispersible in the O/W globule. A oil soluble dye is solubilized within the oil phase of the O/W globule but is dispersible in the W/O globule. Dilutability Test O/W microemulsions are dilutable with water whereas W/O are not and undergo phase inversion into O/W microemulsion. Conductance Measurement O/W microemulsion where the external phase is water are highly conducting whereas W/O are not, since water is the internal or dispersal phase. To determine the nature of the continuous phase and to detect phase inversion phenomena, the electrical conductivity measurements are highly useful. A sharp increase in conductivity in certain W/O microemulsion systems was observed at low volume fractions and such behaviour was interpreted as an indication of a percolative behaviour or exchange of ions between droplets before the formation of bicontinuous structures. 37 Dielectric measurements are a powerful means of probing both structural and dynamic features of microemulsion systems. (H) Electron Microscope Characterization Transmission Electron Microscopy (TEM) is the most important technique for the study of microstructures of microemulsions because it directly produces images at high resolution and it can capture any co-existent structure and micro-structural transitions. There are two variations of the TEM technique for fluid samples. 1. The cryo-TEM analyses in which samples are directly visualized after fast freeze and freeze fructose in the cold microscope. 2. The Freeze Fracture TEM technique in which a replica of the specimen is images under RT conditions. Applications Of Microemulsions

Pharmaceutical Applications Parenteral delivery Oral drug delivery

Microemulsions in biotechnology Parenteral Delivery Parenteral administration (especially via the intravenous route) of drugs with limited solubility is a major problem in industry because of the extremely low amount of drug actually delivered to a targeted site. Microemulsion formulations have distinct advantages over macroemulsion systems when delivered parenterally because of the fine particle microemulsion is cleared more slowly than the coarse particle emulsion and, therefore, have a longer residence time in the body. Both O/W and W/O microemulsion can be used for parenteral delivery. The literature contains the details of the many microemulsion systems, few of these can be used for the parenteral delivery because the toxicity of the surfactant and parenteral use. An alternative approach was taken by Von Corsewant and Thoren43 in which C3-C4 alcohols were replaced with parenterally acceptable co-surfactants, polyethylene glycol (400) / polyethylene glycol (660) 12hydroxystearate / ethanol, while maintaining a flexible surfactant film and spontaneous curvature near zero to obtain and almost balanced middle phase microemulsion. The middle phase structure was preferred in this application, because it has been able to incorporate large volumes of oil and water with a minimal concentration of surfactant. Oral Delivery Microemulsion formulations offer the several benefits over conventional oral formulation for oral administration including increased absorption, improved clinical potency,and decreased drug toxicity.44 Therefore, microemulsion have been reported to be ideal delivery of drugs such as steroids, hormones, diuretic and antibiotics. Pharmaceutical drugs of peptides and proteins are highly potent and specific in their physiological functions. However, most are difficult to administer orally. With on oral bioavailability in conventional (i.e. non-microemulsion based) formulation of less than 10%, they are usually not therapeutically active by oral administration. Because of their low oral bioavailability, most protein drugs are only available as parenteral formulations. However, peptide drugs have an extremely short biological half life when administered parenterally, so require multiple dosing. A microemulsion formulation of cyclosporine, named Neoral has been introduced to replace Sandimmune, a crude oil-in-water emulsion of cyclosporine formulation. Neoral is formulated with a finer dispersion, giving it a more rapid and predictable absorption and less inter and intra patient variability.

Microemulsions in Biotechnology Many enzymatic and biocatalytic reactions are conducted in pure organic or aqua-organic media. Biphasic media are also used for these types of reactions. The use of pure apolar media

causes the denaturation of biocatalysts. The use of water-proof media is relatively advantageous. Enzymes in low water content display and have 1. Increased solubility in non-polar reactants 2. Possibility of shifting thermodynamic equilibria in favour of condensations 3. Improvement of thermal stability of the enzymes, enabling reactions to be carried out at higher temperatures. Many enzymes,including lipases, esterases, dehydrogenases and oxidases often function in the cells in microenvironments that are hydrophobic in nature. In biological systems many enzymes operate at the interface between hydrophobic and hydrophilic domains and these usually interfaces are stabilized by polar lipids and other natural amphiphiles. Enzymatic catalysis in microemulsions has been used for a variety of reactions, such as synthesis of esters, peptides and sugar acetals transesterification; various hydrolysis reactions and steroid transformation. The most widely used class of enzymes in microemulsion-based reactions is of lipases.

Other Applications Microemulsion in enhanced oil recovery Microemulsions as fuels Microemulsions as lubricants, cutting oils and corrosion inhibitors Microemulsions as coatings and textile finishing Microemulsions in detergency Microemulsions in cosmetics Microemulsions in agrochemicals Microemulsions in food Microemulsions in environmental remediation and detoxification Microporous media synthesis (microemulsion gel technique) Microemulsions in analytical applications Microemulsions as liquid membranes Novel crystalline colloidal arrays as chemical sensor materials

REVERSE MICELLES

In a non-polar solvent, it is the exposure of the hydrophilic head groups to the surrounding solvent that is energetically unfavourable, giving rise to a water-in-oil system. In this case the hydrophilic groups are sequestered in the micelle core and the hydrophobic groups extend away from the centre. These inverse micelles are proportionally less likely to form on increasing headgroup charge, since hydrophilic sequestration would create highly unfavorable electrostatic interactions. Reverse micelles are nanometer-sized (1-10 nm) water droplets dispersed in organic media obtained by the action of surfactants. Surfactant molecules organize with the polar part to the inner side able to solubilize water and the apolar part in contact with the organic solvent. Proteins can be solubilized in the water pool of reverse micelles. Studies on the structurefunction relationships of proteins in reverse micelles are very important since the microenvironment in which the protein is solubilized has physico-chemical properties distinct from a bulk aqueous solution. Some of the unique characteristics of reverse micelles make them very useful for biotechnological applications. Charge and hydrophilic/hydrophobic characteristics of the protein and the selection of surfactant can be used to achieve selective solubilization of proteins. This has been used to extend the classical liquid-liquid extraction with solvents to protein bioseparation. For biocatalysis the presence of a bulk organic solvent allow synthetic reactions to be performed via the control of water content and the solubilization of hydrophobic substrates. This is accomplished with a higher interfacial area (about 100 m2/mL) than the conventional biphasic systems, minimizing mass transfer problems

REFERENCES
Y.Moroi,Micelles : Theoretical and Applied Aspects,(1992) Plenum Press,New York. www.scribd.com/doc/.../Rosen-M-J-Surfactants-AndInterfacial-Phenomena en.wikipedia.org/wiki/Solubilization www.chem.ufl.edu/~itl/4411L_f98/micelle/micelle.html en.wikipedia.org/wiki/Microemulsion www.chm.bris.ac.uk/eastoe/Surf_Chem/3%20Microemulsion s.pdf arxiv.org/pdf/1108.2794