3.1. Headgroup pKa Measurements
NMR titration analyses were performed to measure pK
a values for tridecanoic-Glu’s primary and γ carboxylic acid functional groups.
Figure 2a,b shows NMR titration curves for the monomeric surfactant. A titration curve for the surfactant’s γ carboxylic acid in micellar form is shown in
Figure 2c.
Table 1 summarizes all pK
a values. The
Table 1 entries labeled L-Glutamic Acid were taken from the literature [
44]. Uncertainties are not reported for the literature values but are included for the experimental pK
a measurements. L-Glutamic Acid pK
as from the literature were included to allow comparison of the free solution carboxylic acid acidities to corresponding values for the tridecanoic-Glu surfactant.
The pKa analyses show that in monomeric form, the surfactant γ pKa value of 4.7 is similar to the corresponding L-Glutamic acid value of 4.3. In contrast, the pKa of the free amino acid’s primary-CO2H functional group (2.2) is considerably lower than the corresponding value for the tridecanoic-Glu monomeric surfactant (4.2). In other words, the primary carboxylic acid is more acidic in free L-Glutamic acid and less acidic when the amino acid is connected to the surfactant’s hydrocarbon chain through an amide bond.
The above observation is likely attributable to the formation of an intramolecular hydrogen bond between the primary-CO
2H hydrogen atom and the carbonyl oxygen of the surfactant’s amide bond. This H-bond decreases the acidity of the primary carboxylic acid because deprotonation of the hydrogen-bound carboxylic acid hydrogen atom places two negative oxygen atoms close to one another. Deprotonation thus destabilizes the conjugate base and, therefore, reduces the acidity of the primary-CO
2H [
45,
46]. Intramolecular hydrogen bond formation involving the γ-CO
2H functional group may affect its pK
a in a similar manner; however, the γ carboxylic acid is farther from other H-bond donor and acceptor atoms in the surfactant’s headgroup making intramolecular H-bond formation less likely. This observation may explain why the surfactant’s γ-CO
2H pK
a is more similar to the free amino acid value.
Molecular modeling and molecular dynamics simulation experiments were used to test these hypotheses and determine if the monomer’s structure allowed intramolecular hydrogen bond formation. The tridecanoic-Glu surfactant was built with the
MOE (Chemical Computing Group,
https://www.chemcomp.com/, URL accessed on 10 June 2024) software package and a conformational search was carried out. The two lowest energy structures from this search are shown in
Figure 3a,b. Note that in each structure, a hydrogen bond is observed between the primary CO
2H hydrogen atom and the carbonyl oxygen of the surfactant’s amide functional group. These structures also show an H-bond between the headgroup’s γ-CO
2H oxygen atom and the amide NH. However, no H-bond involving the γ carboxylic acid hydrogen atom was observed in these two structures. There was a higher energy structure from the conformational search (
Figure 3c), however, in which an H-bond was formed between the γ-CO
2H proton and the amide carbonyl oxygen. These results show that the structure of the tridecanoic-Glu surfactant’s amino acid headgroup allows for H-bond formation between the primary-CO
2H hydrogen atom and the amide functional group. Formation of this H-bond would be expected to reduce the primary carboxylic acid’s pK
a as observed in the NMR titrations [
45,
46]. In addition, if the surfactant also spends time in the higher energy conformation shown in
Figure 3c, the γ-CO
2H would be expected to be slightly less acidic than the free amino acid as was observed in
Table 1.
A molecular dynamics (MD) simulation was also performed to further investigate intramolecular hydrogen bond formation in the tridecanoic-Glu surfactant headgroup. In this experiment, a 100 ns MD simulation was performed with a system containing the tridecanoic-Glu monomer and approximately 2000 water molecules. A hydrogen bond analysis was then performed on the MD simulation trajectory. These experiments showed that an H-bond between the primary-CO
2H proton and the carbonyl oxygen of the amide functional group was present for 19.8% of the MD simulation. This is the same H-bond that was observed in the conformational search (
Figure 3a).
Figure S6a of the Supplemental Information plots the distance between the heavy atoms involved in the H-bond between the primary-CO
2H proton and the carbonyl oxygen of the amide functional group versus simulation time. The figure shows that the distance varies between approximately 3 and 4 Å and remains relatively constant throughout the MD simulation. Since the H-bond cutoff distance in the H-bond analysis was 3.5 Å, the percent occupancy of this H-bond is as high as 19.8%. Furthermore, an H-bond between the γ-CO
2H proton and amide oxygen was also observed in the MD simulation, but the percent occupancy was only 7.5%. This H-bond is shown in
Figure 3c.
Figure S6b of the Supplemental Information plots the distance between the heavy atoms involved in the H-bond between the γ-CO
2H proton and amide oxygen versus simulation time. Here, the change in distance varied over a relatively broad range of 3 to 7 Å and the distance is much more variable than for the H-bond with the 19.8% occupancy. This plot, therefore, is consistent with a lower H-bond percent occupancy. Finally, the MD simulation also yielded an H-bond between the γ-CO
2H proton and primary-CO
2H oxygen with a percent occupancy of 5.0%.
Therefore, like the conformational analysis, the MD simulation showed that the surfactant headgroup structure allowed multiple opportunities for intramolecular hydrogen bond formation. The H-bond formed most often though was between the primary-CO
2H proton and the amide carbonyl oxygen. NMR titration analyses showed that this proton’s pK
a was significantly different than the corresponding proton in free L-Glutamic acid where H-bond formation of this type is not possible. H-bonds involving the γ-CO
2H proton were also observed in the MD simulation, although these H-bonds formed less often than the H-bond shown in
Figure 3a.
Table 1 also presents NMR titration pK
a values for the tridecanoic-Glu surfactant in micellar form. In these experiments, the surfactant concentration was 50.0 mM, which is well above the CMC values given in
Table 2. These CMC data will be discussed in more detail below. In NMR titrations with the surfactant micelles, only the γ pK
a value could be measured. pK
a experiments were performed by lowering the solution pH and observing the change in chemical shift of the headgroup Hα and Hγ protons. However, in micellar form, the surfactant precipitated from the solution before a significant change in the Hα chemical shift was observed. Therefore, only the γ pK
a value is reported. Precipitation likely occurred in the micellar solutions because the Glutamic acid headgroup’s primary carboxylate began to protonate below pH 6. The monomer’s charge then changed from predominantly −1 to neutral, making the surfactant both less hydrophilic and less water-soluble. In the micellar solutions, the surfactant concentration was also relatively high (50.0 mM), causing the less hydrophilic, protonated surfactant to precipitate from the solution. At the lower sub-micellar concentrations used in the monomer NMR titrations, the surfactant likely remained water-soluble even after protonation of the primary carboxylate began to occur.
Table 1 shows that the surfactant’s γ pK
a value was larger when the surfactant was in micellar form (pK
a = 6.9) than when the surfactant was monomeric (pK
a = 4.2). In other words, the γ-CO
2H proton is less acidic when the surfactant is in micellar form and more acidic in monomeric form. This increase in pK
a and decrease in acidity is likely caused by intermolecular H-bond formation between different surfactant unimers when they are close to one another at the surface of the micelle.
The
Table 1 pK
a values can also be used to predict how the charges of the micellar surfactant molecules change with solution pH. Since the pK
a of the micellar γ-CO
2H is 6.9, if the pH is in the 7.0 range, the solution will contain populations of both protonated and deprotonated side chains. At pH 7, however, the primary carboxylic acid will be ionized since its pK
a would be expected to be lower than the γ-CO
2H. Therefore, at neutral pH, the solution will contain a mixture of −1 and −2 headgroup charges. At pH 6.0, however, the γ carboxylic acid is predominately protonated and the surfactant charge is −1. At pH values well above 7.0, the γ carboxylic acid is predominantly deprotonated and the surfactant charge is −2. The effect of headgroup charge on the tridecanoic-Glu surfactants’ CMC will now be presented. The binding of cationic counterions to the anionic micelle surface as a function of pH will also be examined.
3.2. Critical Micelle Concentration Measurements
CMC measurements were performed in solutions with the diamine counterions 1,4-diaminobutane, 1,6-diaminohexane, trans-1,4-cylcohexanediamine, and trans-1,2-cyclohexanediamine. As described above, CMC values were measured by progressively diluting tridecanoic-Glu-diamine mixtures with deionized water and recording the solution’s conductivity after each dilution. Initial surfactant concentrations in the conductivity experiments were typically 20.0 mM and final concentrations were 3.0 mM. Solution pH measurements were performed at these concentrations to confirm that the pH did not change significantly during the conductivity experiments. In a tridecanoic-Glu-1,4-diaminobutane mixture, the pH was 7.07 at a surfactant concentration of 20.0 mM. The pH was 6.82 after the 20.0 mM solution was diluted to 3.0 mM with deionized water. In three trials of this experiment, pH decreases of 1.4% to 3.6% were observed. The pH of the solutions likely remained relatively constant during the conductivity experiments because of buffering provided by the mixture of the acidic surfactant and basic diamine.
The critical micelle concentrations of tridecanoic-Glu at pH 6.0 and 7.0 are shown in
Table 2. The CMC values show that at pH 6.0, the 1,4-diaminobutane, 1,6-diaminohexane, and trans-1,4-cylcohexanediamine CMC values were similar to one another, ranging from 3.1 to 3.4 mM. As discussed above, at this pH, the surfactant headgroup charge is predominantly −1. When the pH increased from 6.0 to 7.0 and the tridecanoic-Glu headgroups had populations of both −1 and −2 charges, the surfactant’s CMC with 1,4-diaminobutane, 1,6-diaminohexane, and trans-1,4-cylcohexanediamine counterions all roughly double from 3.1 to 3.4 mM to 6.2 to 6.8 mM. These CMC trends can be rationalized based on changes in surfactant headgroup charge with pH. At pH 6.0, when the headgroup charge is −1, there is less repulsion between the surfactant headgroups and micelles form at a lower concentration. As pH is increased to 7.0, the population of −2 surfactant unimers increases. The larger headgroup charge increases the repulsion between surfactant monomers and thus raises the CMC [
47].
At each pH investigated, the tridecanoic-Glu CMC was lowest in solutions containing trans-1,2-cyclohexanediamine counterions. For example, CMC values in solutions containing trans-1,2-cyclohexanediamine were 1.3 mM and 2.1 mM at pH 6 and 7, respectively. These values were lower than the CMCs for all other counterions in the study at both of the pH values. One difference between trans-1,2-cyclohexanediamine and the other three counterions is that in the former, the amine functional groups are on adjacent carbon atoms. This arrangement may allow the trans-1,2-cyclohexanediamine counterion to bind simultaneously to both carboxylic acid functional groups in the tridecanoic-Glu headgroup. An energy-minimized structure from
MOE showing this mode of binding is shown in
Figure 4c. With the other counterions in
Table 1, the amine functional groups are farther apart, making simultaneous binding to both headgroup carboxylic acids less likely. If the trans-1,2-cyclohexanediamine counterion binds to the surfactant headgroup in this manner, the counterion may more effectively reduce headgroup repulsion at the micelle surface, thus leading to lower surfactant CMC values [
47]. This mode of trans-1,2-cyclohexanediamine binding to the micelles is revisited below when micelle radii are discussed.
Finally, it is well known that a surfactant’s CMC is affected by the length of its hydrocarbon chain, with longer chains generally leading to lower CMC values [
14,
47,
48,
49]. In order to investigate whether tridecanoic-Glu behaved in this manner, CMC measurements were made with the surfactant undecanoic-Glutamic Acid (undecanoic-Glu) in solutions containing the four counterions listed above. This surfactant also contains a Glutamic acid headgroup, but its hydrocarbon chain has eleven carbon atoms, compared to the thirteen-atom hydrocarbon chain in tridecanoic-Glu. CMC values for undecanoic-Glu with each diamine counterion at pH 6.0 and 7.0 are shown in
Table 2. As expected, the CMC values are higher for the undecanoic-Glu surfactant. For example, while tridecanoic-Glu at pH 6.0 had CMC values in the 3.1 to 3.4 mM range, CMC values for und-Glu ranged from 17.5 to 22.1 mM at this pH. As observed for tridecanoic-Glu, the undecanoic-Glu CMC values were also generally smaller at pH 6.0 and larger at pH 7.0. This effect can be attributed to changes in the surfactant headgroup charge with pH as discussed above.
Differences between the tridecanoic-Glu and undecanoic-Glu CMC values can be rationalized as follows. The molecules have the same headgroup, therefore favorable hydrogen-bonding interactions and unfavorable headgroup repulsions should be similar in both surfactants. The longer alkyl chain in tridecanoic-Glu, however, increases the overall hydrophobicity of the surfactant. Tridecanoic-Glu molecules therefore experience enhanced hydrophobic effects in solution compared to undecanoic-Glu, causing micelles to form at lower concentrations. An analogous effect was observed by Brycki, et al. in an investigation of divalent, dimeric alkylammonium surfactants. In this study, the CMC was 158.5 mM when the surfactant hydrocarbon chain contained four carbon atoms. The CMC decreased steadily to 0.033 mM when the length of the hydrocarbon chain was increased to eighteen carbon atoms [
48]. A similar change in CMC with increasing alkyl chain length was also observed by Bustelo, et al. in a study of histidine-containing amino acid-based surfactants [
14].
3.3. Micelle Radii and Counterion Binding
NMR diffusion experiments were used to investigate how solution pH affected the hydrodynamic radii of the tridecanoic-Glu micelles and the mole fraction of cationic counterions bound to the anionic micelle surface. The counterions in the diffusion experiments were the same as those used for the CMC studies (
Table 2). Before discussing these results, it should be noted that the hydrodynamic radius, R
h, measured with NMR diffusion experiments is the radius of the particle diffusing in solution. R
h, therefore, includes both the micelle and micelle-bound counterions. NMR diffusion experiments do not directly probe the radius of only the micelle. MD simulation experiments with the entire micelle would directly measure this radius and provide the micelle’s aggregation number. These MD simulations are underway. Finally, since R
h includes both the micelle and counterions, comparing R
h values at pHs where the mole fraction of bound counterions is high to pHs where these values are low provides insight into the structure of the micelle–counterion complex [
15,
16,
17,
26]. This method will be employed below to interpret the results shown in
Figure 5.
Figure 5a plots the hydrodynamic radii of the tridecanoic-Glu micelles and the mole fraction of 1,4-diaminobutane counterions bound to the micelle surface versus solution pH.
Figure 5a shows that at pH 6, the micelle radius is ≈20 Å and the f
b,counterion value is 0.72. As pH is increased, the micelle radius decreases from 20 Å at pH 6 to 12 Å at pH 7. In the same pH range, the f
b,counterion value increases from 0.72 to 0.85. As pH is further increased from 7.0 to 13.0, the micelle radius decreases further from 12 Å to 8 Å. In the same pH range, the f
b,counterion values remain relatively constant until pH 9.7 and then f
b,counterion decreases sharply from 0.80 at pH 9.7 to only 0.10 at pH 13.0. These changes can be rationalized by changes in the surfactant headgroup and counterion charges with solution pH.
Below pH 7, the tridecanoic-Glu surfactants predominantly have a −1 charge because the primary carboxylic acid is deprotonated but the γ-CO
2H is not. Since repulsion between the headgroups at the micelle surface is less when the monomers are −1 (compared to −2 at higher pH), more monomers pack into the micelles below pH 7.0, making the micelle radii larger. pK
a values for all the counterions investigated are given in
Supplemental Information Table S1. For 1,4-diaminobutane, these are pK
a1 = 9.63 and pK
a2 = 10.8 [
44]. Therefore, below pH 7.0, the 1,4-diaminobutane counterion charge is +2. The +2 counterions are strongly attracted to the micelle surface below pH 7.0, leading to the relatively high f
b,counterion value observed in
Figure 5a.
From pH 7.0 to 9.7, the γ carboxylic acid in the tridecanoic-Glu headgroup deprotonates and the monomer charge changes to predominantly −2. Repulsion between these −2 headgroups is now greater than when the monomer charge was −1. Increased headgroup repulsion at the micelle surface likely leads to fewer monomers aggregating into micelles. Therefore, as observed in
Figure 5a, the micelle radius decreases when the headgroup charge changes from −1 to −2. In addition, in the pH range 7.0 to 9.7 when the headgroup charge is −2, the +2 1,4-diaminobutane counterions are more strongly attracted to the micelle surface than at lower pH when the headgroup charge was −1. Therefore, the mole fraction of micelle-bound counterions increases to 0.9 and reaches its maximum value in this pH range. pK
a1 of the 1,4-diaminobutane counterion is 9.63, so above pH 9.7, the amine deprotonates and the counterion has a +1 charge. The +1 diamine is now less attracted to the −2 headgroups at the micelle surface, causing the mole fraction of micelle-bound counterions to decrease sharply as the pH is raised above pH 9.7. Finally, pK
a2 for 1,4-diaminobutane is 10.8, so above pH 11, the counterion is predominately neutral and the mole fraction of micelle-bound counterions is relatively low. A model of changes in the 1,4-diaminobutane binding to the tridecanoic micelles with solution pH is shown in
Figure 6a.
Changes in the micelle radii and mole fraction of micelle-bound 1,6-diaminohexane counterions are plotted versus solution pH in
Figure 5b. Many of the trends discussed above for 1,4-diaminobutane are also seen with 1,6-diaminohexane. At pH 6.0, the micelle radius is 17 Å and the mole fraction of micelle-bound counterions is 0.5. In the pH range of 6.0 to 7.0, the radii decrease to 9.0 Å and the f
b,counterion values increase to 0.6. From pH 7.0 to 11.0 the micelle radii and f
b,counterion values remain relatively constant. Finally, above pH 11.0, f
b,counterion decreases to 0.4.
The change in micelle radius in
Figure 5b from pH 6.0 to 7.0 likely occurs, as in the 1,4-diaminobutane solutions, because the surfactant headgroup charge changes from −1 to −2 in this pH range. The f
b,counterion value also increases from pH 6.0 to 7.0 because the +2 counterions are strongly attracted to the −2 surfactant headgroups. However, unlike in the 1,4-diaminobutane solutions, the f
b,counterion values do not decrease at pH 9.7 but rather remain constant until pH 11.0. This behavior can be attributed to the two counterions having different ionization constants. The pK
as for 1,6-diaminohexane are pK
a1 = 10.76 and pK
a2 = 11.86 [
44]. Therefore, the counterion’s charge remains predominately +2 up to pH 11.0. In
Figure 5b, the 1,6-diaminohexane f
b,counterion values remain constant up to pH 11.0 as well. When the counterion deprotonates above pH 11, the counterion charge is reduced and the f
b,counterion values are reduced as well. In other words, the reduction in the counterion’s f
b,counterion values occur at a higher pH for 1,6-diaminohexane and a lower pH for 1,4-diaminobutane because of the former counterion’s charge remains +2 over a larger pH range. A model of 1,6-diaminohexane binding to the tridecanoic-Glu micelles is shown in
Figure 6b.
Another notable difference between the binding of 1,4-diaminobutane and 1,6-diaminohexane counterions to the tridecanoic-Glu micelles is that the maximum f
b,counterion value for 1,4-diaminobutane was 0.9, while the maximum f
b,counterion for 1,6-diaminohexane was 0.6. A similar result was reported by Maynard-Benson, et al. in a study of linear diamine counterions binding to undecanoic L-norleucine micelles [
17]. Previous studies have shown that these linear diamine counterions bind parallel to the surface of amino acid-based micelles, allowing the two amine function groups to interact with multiple surfactant unimers. Maynard-Benson, et al. suggested that f
b,counterion values were larger for 1,4-diaminobutane and smaller for 1,6-diaminohexane because the spacing between the amine functional groups in the former counterion was optimal for the counterion to bridge between two surfactant monomers [
17]. A similar effect likely explains why the maximum f
b,counterion values were also larger for 1,4-diaminobutane counterions than for 1,6-diaminohexane when these two diamines bound to the tridecanoic-Glu micelles.
Finally, in
Figure 6, 1,4-diaminobutane and 1,6-diaminohexane are shown to bind parallel to the micelle surface with both amine functional groups interacting with different surfactant monomers. These counterions have been shown to bind to micelles formed by other amino acid-based surfactants in an analogous manner [
17,
26]. The results plotted in
Figure 5a suggest that 1,4-diaminobutane also binds to tridecanoic-Glu micelles in a parallel fashion. As previously discussed, above pH 9.7, the 1,4-diaminobutane counterion deprotonates, its charge decreases, and the counterion is less attracted to the anionic micelle surface. However, above pH 9.7, as the 1,4-diaminobutane f
b,counterion values decrease, the micelle hydrodynamic radii remain relatively constant. This result suggests that 1,4-diaminobutane binds parallel to the tridecanoic-Glu micelle surface because the micelle R
h values are similar when f
b,counterion is both high and low. Comparing the R
h values in
Figure 5a,b when, 1,4-diaminobutane and 1,6-diaminohexane are micelle-bound, respectively, shows that above pH 7, the R
h values for micelles with both counterions are very similar. In other words, the R
h values are not appreciably larger when 1,6-diaminohexane counterions with a longer alkyl chain are micelle-bound compared to when 1,4-diaminobutane counterions with a shorter alkyl chain are bound to the tridecanoic-Glu micelles. This result suggests that 1,6-diaminohexane, like 1,4-diaminobutane, binds parallel to the micelle surface as shown in
Figure 6.
Figure 5c plots micelle radii and f
b,counterion values for trans-1,4-cyclohexanediamine counterions binding to tridecanoic-Glu micelles. These data closely resemble the corresponding plot in
Figure 5a for 1,4-diaminobutane. For example, with both counterions, the micelle radii decrease and f
b,counterion values increase when the surfactant monomer charge changes from −1 to −2. The mole fraction of micelle-bound counterions is also constant in the pH range of 7.0 to 9.4 when the counterion charge is +2. When the counterion deprotonates, f
b,counterion values then decrease. This decrease in f
b,counterion occurs at a slightly lower pH with trans-1,4-cyclohexanediamine because the counterion’s pK
a1 of 9.4 is lower than the corresponding 1,4-diaminobutane value of 9.63 [
44].
One notable difference, however, between the behavior of the trans-1,4-cyclohexanediamine and 1,4-diaminobutane-containing solutions is that in the former the micelle radii are larger throughout the pH range investigated. This difference is illustrated in
Figure 4a where the micelle hydrodynamic radii for solutions containing both counterions are plotted on the same graph. As discussed above, the results in
Figure 5a suggest that 1,4-diaminobutane counterions bind parallel to the tridecanoic-Glu micelle surface. Fletcher, et al. investigated the binding of trans-1,4-cyclohexanediamine to amino acid-based undecyl-LL-Leucinevalanate micelles. This study showed that trans-1,4-cyclohexanediamine bound to the micelles in a perpendicular fashion with one amine functional group interacting with the anionic micelle surface and the rest of the molecule extending out into free solution [
26]. This binding model is shown in
Figure 4b. It is likely that the trans-1,4-cyclohexanediamine counterion interacts with the tridecanoic-Glu micelles in a similar manner given the larger micelle hydrodynamic radii measured for this counterion.
Figure 5d plots f
b,counterion values and micelle radii versus pH for solutions containing trans-1,2-cyclohexanediamine. The decrease in micelle radii with increasing pH observed for solutions containing trans-1,2-cyclohexanediamine is comparable to that observed with the other counterions. This decrease is likely caused by changes in headgroup charge that occur when the headgroup’s γ-carboxylic acid functional groups deprotonate. One notable difference, though, between trans-1,2-cylcohexanediamine and trans-1,4-cyclohexanediamine binding to the micelles is that the maximum f
b,counterion values are 0.52 and 0.80, respectively. In other words, the trans-1,2-cyclohexanediamine counterion’s maximum f
b,counterion value is smaller than the corresponding 1,4 isomer. This difference is likely attributable to the amine functional groups in the two isomers having different pK
a values. The ionization constants for trans-1,2-cyclohexanediamine are pK
a1 = 6.47 and pK
a2 = 9.94 [
50]. Corresponding values for trans-1,4-cylcohexanediamine are 9.94 and 10.8, respectively. The pK
a1 value for the 1,2 isomer is smaller than the 1,4 isomer because in the former, the amine functional groups are on adjacent carbon atoms and thus deprotonation allows an intramolecular hydrogen bond to form. Therefore, in the pH range of 7.0 to 10.0, the charge of the trans-1,2-cyclohexanediamine counterions are predominately +1 compared to +2 for trans-1,4-cyclohexanediamine. The +1 trans-1,2-cyclohexanediamine counterions are thus less attracted to the anionic micelle surface than the +2 counterions of the 1,4 isomer and as a result, the f
b,counterion values for the 1,2 isomer are smaller than the 1,4 isomer in the pH range shown in
Figure 5.
Figure 4a compares the radii of the tridecanoic-Glu micelles in solutions containing trans-1,2-cyclohexanediamine and trans-1,4-cyclohexanediamine counterions. Throughout the pH range investigated, the micelles were larger in solutions containing the 1,2 isomer. Recall from above, tridecanoic-Glu CMC values were also lower in solutions containing trans-1,2-cyclohexanediamine and larger in solutions containing the 1,4-isomer. The CMC difference was attributed to the 1,2-isomer forming simultaneous hydrogen bonds with the two carboxylate functional groups in the surfactant’s headgroup. This interaction is shown in
Figure 4c. At the micelle surface, the simultaneous binding of the trans-1,2-cyclohexanediamine counterion to both carboxylate functional groups may neutralize the headgroup negative charge more effectively compared to counterions that only interact with one of the carboxylates. The corresponding reduced repulsion between the monomer headgroups may then allow more monomers to aggregate, thus increasing the radii of the tridecanoic-Glu micelles.