Patent Application
20210213438
The present invention relates to the field of chemistry-physics, more specifically to a kit and methods to assess the adsorbent properties of the surface of a material of interest to be studied using cationic, anionic, or uncharged (no elementary charge) molecules.
The kit of the present invention comprises cationic, anionic, or uncharged molecules.
Exposing a material to a liquid phase allows the adsorption of molecules dissolved in the liquid phase into the material. This adsorption can be mediated, for example, by electrostatic interactions, by lyophobia of the dissolved molecules, among others, and depends at least in part on the composition of the liquid phase, on the composition of the molecules and on the composition of the surface of the material.
There are technologies that allow the characterization of the surface properties of a material. For example, instruments for measuring nitrogen adsorption using the BET method (Brunauer, Emmett, and Teller) allow to determine the surface area value of the surface of a material. The determination of flux potentials and light scattering under microelectrophoresis conditions allows to measure the difference in electrical potential between the sine of a liquid phase and the surface of a material.
However, none of these technologies allow to assess the chemical-physical properties of the surface of a material. For this reason, it is necessary to make available kits and methods to characterize the composition of the surface of a material with respect to its potential to adsorb cationic, anionic, or uncharged molecules. The present invention delivers a solution to this and other limitations of previous technologies.
The present invention describes a kit and methods to assess the adsorbent properties of the surface of a material of interest to be studied in terms of its ability to interact with cationic, anionic, or uncharged molecules.
The kit of the present invention comprises a solution volume containing cationic molecules, a solution volume containing anionic molecules, and a solution volume containing molecules with no elementary charge.
The material of interest of the present invention corresponds to any material to be assessed, for example, a metallic surface, a metallic nanoparticle surface, such as a volume of colloid containing a particulate material in dispersed state whose surface is modified with organic molecules of known chemical characteristics or any unknown surface to be studied. The surface of the material of interest may comprise an electrostatic component and a lyophobic component, the electrostatic component of the surface being a motif with elementary charges and the lyophobic component of the surface a nonpolar motif with no elementary charges.
The methods to assess the adsorbent properties of the surface of a material of interest of the present invention correspond to a method that defines the electrostatic component of the surface and a method that defines the lyophobic component of the surface. In order to define the electrostatic component of the surface of the material of interest, this invention provides a method that assesses the adsorption of cationic and anionic molecules, in the absence and presence of ionizable electrolytes that generate ionic strength. In order to define the lyophobic component of the surface of the material of interest, this invention provides a method that assesses the adsorption and desorption of molecules with no elementary charge.
The methods comprise obtaining test samples and control samples. The test samples comprise surface amounts of the material of interest exposed to amounts of the solution volumes contained in the kit, while control samples comprise equivalent amounts of the solution volumes contained in the kit. After sampling, the methods allow to determine the number of molecules adsorbed on the surface of the material of interest. Finally, the comparison of the results obtained with each sample allows to define the adsorbent properties of the surface of the material of interest based on its ability to interact with cationic, anionic, or uncharged molecules.
The invention will be described hereunder with reference to the accompanying drawings, in which:
The present invention describes a kit and methods to assess the adsorbent properties of the surface of a material of interest to be studied in terms of its ability to interact with cationic, anionic, or uncharged molecules.
The present invention refers to a kit, a method to assess the electrostatic component of the surface of a material of interest by assessing the adsorption ability of cationic or anionic molecules on the surface of the material of interest, and a method to assess the lyophobic component of the surface of a material of interest by characterizing the adsorption and desorption of molecules with no elementary charge from the surface of the material of interest.
As used in the present invention, the following terms will be understood as follows:
“molecule”, represents a defined chemical species.
“molecule with elementary charge”, represents an ionized molecule.
“molecule with no elementary charge”, represents a neutral molecule whose formal unit electrical charges located on non-adjacent atoms have the same sign.
All other terms used in the present invention should be interpreted in a similar way to that set forth in scientific literature. Furthermore, it is noted that in the present invention the steps of the methods may operate in sequences other than those described or illustrated in this disclosure.
The present invention describes a kit to assess the adsorbent properties of the surface of a material of interest that comprises a solution volume containing cationic molecules, a solution volume containing anionic molecules, and a solution volume containing molecules with no elementary charge.
In the present invention, a cationic molecule can be methylene blue and its derivatives; tetrazolium derivatives such as 2,3,5-triphenyltetrazolium chloride (TTC), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT); phenazine derivatives such as phenazine methosulfate, safranin-O, and phenophosphranine; diarylmethane as Auramine O (basic yellow 2); triarylmethane derivatives such as malachite green, fuchsin, crystal violet. For its part, an anionic molecule can be derived from fluorescein such as erythrosine, fluorescein isothiocyanate, eosin; sulfonic acid derivatives such as 1,5-naphthalenedisulfonic acid, methyl orange (azo), acid orange (azo), tosylate, picric acid; and a molecule with no elementary charge can be neutral non-zwitterionic molecules (excluding zwitterions) such as phenazine; aromatic hydrocarbons such as anthracene, phenanthrene, naphthalene, azulene, pyrene.
The adsorption ability of these molecules will depend on the physicochemical properties of the surface of the material of interest. If the surface of the material of interest contains motifs with ionized groups, the cationic and anionic molecules in the kit will be adsorbed to the surface reversibly by electrostatic interactions. If the surface of the material of interest contains nonpolar (lyophobic) motifs, the molecules with no elementary charge in the kit will be excluded from the aqueous liquid solvent and adsorbed on the surface of the material, also reversibly. In this manner, the determination of the reversible adsorption ability of these molecules on the surface of a material of interest allows to infer the intrinsic properties of the material of interest.
The method for assessing the electrostatic component of the surface of a material of interest, of the present invention, comprises obtaining test samples and control samples, which in turn comprises and does not comprise the addition of electrolytes to generate ionic strength (
If the concentration of ionic molecules in a volume of the liquid phase free of the surface of interest is determined to be greater in the presence of the electrolytes, it is demonstrated that the adsorption of the molecules with elementary charge is reversible and mediated by an electrostatic component. This method comprises determining the amount of ionic molecules that are adsorbed to the surface of a material of interest by electrostatic interactions. In this manner, it can be inferred whether the surface of a material of interest contains motifs with non-zero elementary charges (ionized motifs). The assay was repeated three times in order to have statistical validity and each sample was assessed independently (
Schematically in
The present invention also refers to a method to assess the lyophobic component of the surface of a material of interest. In this case, the molecules with no elementary charge can be adsorbed to the surface by lyophobia. Reciprocally, it can be inferred if the surface of the material contains lyophobic motifs. If desorption is detected in the presence of an immiscible nonpolar solvent with the liquid phase of the test sample, it is demonstrated that the adsorption of the molecules with no elementary charge is mediated by lyophobia.
The method for assessing the lyophobic component comprises determining the amount of molecules with no elementary charge that are adsorbed on the surface of the material of interest. For this, the method comprises obtaining a test sample, a control sample, and a nonpolar solvent control sample (
Schematically in
Next, an example representing an embodiment of the present invention is described, where the chosen material of interest corresponds to a monolayer of 11-thioundecanoic acid (MUA) self-assembled molecules on nuclei of gold nanoparticles.
The sulfhydryl group of 11-mercaptoundecanoic acid (MUA) molecules covalently reacts with gold atoms, generating a self-assembled monolayer of MUA molecules around metallic nuclei of gold nanoparticles. This monolayer of MUA molecules contains an internal hydrophobic motif located just above the surface of the gold nanoparticles, and an external hydrophilic motif that interacts with the continuous phase of the colloid. The internal hydrophobic motif is formed by grouping of the aliphatic chains, and the external hydrophilic motif is formed by grouping of carboxylate and carboxylic groups.
To obtain the metallic nuclei, gold nanoparticles were synthesized by means of reduction by citrate ion methodology. To self-assemble the MUA molecules, a colloid volume comprising the synthesized gold nanoparticles was functionalized by reduction of citrate ions (50 ml) with a concentration of MUA molecules at 100 μM. To decrease the concentration of residual compounds in the continuous phase of the colloid, the volume of functionalized nanoparticles was subjected to ultrafiltration by centrifugation using a commercial filter with exclusion size equal to 100 kDa. The sample was centrifuged at 4,000×g for 15 min and a 1 ml retentate of colloid enriched in MUA self-assembled monolayer on gold nanoparticles was obtained. Permeate was discarded and 9 ml of 1.2 mM sodium citrate buffer was added to the retentate. The ultrafiltration and resuspension process was repeated 6 times.
After the described process, an enriched colloid in MUA monolayer on gold nanoparticles (AuNP@MUA colloid) was obtained. The colloid presented a maximum absorbance value of the plasmon band of 60/cm (
Subsequently, in order to estimate the surface area value of the MUA monolayer, the number of nanoparticles contained in a volume of AuNP@MUA colloid was determined and was related to the surface area value of a sphere representative of AuNP@MUA nanoparticles.
To determine the number of particles contained in a volume of AuNP@MUA colloid, the diameter of the metallic nuclei without MUA monolayer was used and was related to the value of the maximum absorbance of the plasmon band as described by Haiss et al (2007). Dynamic light scattering measurements indicated that the value of the diameter of the metallic nuclei according to the number of particles was 10.53 nm. The maximum absorbance value of the plasmon band was 60/cm. This indicates that the AuNP@MUA colloid contained 4.51×1011 nanoparticles per microliter of dispersion.
To calculate the surface area of a sphere representative of AuNP@MUA nanoparticles, the diameter value obtained from the AuNP@MUA colloid by means of dynamic light scattering measurements was used. The diameter value was 13.55 nm, and the replacement of this value in the simple relation to determine the surface area of a sphere indicates that the surface area of a representative sphere of AuNP@MUA nanoparticles is 576.8 nm2.
Finally, the linear relationship between the number of particles and the surface area of the representative particle defines the AuNP@MUA colloid as a substance that contains 2.60 cm2 of MUA monolayer per microliter of colloidal dispersion.
Method to Assess the Electrostatic Component of the Surface of the MUA Monolayer by Determining the Adsorption of Molecules with Elementary Charge Mediated by Electrostatic Interactions.
The adsorption of methylene blue molecules on the surface of the MUA monolayer was determined, in the presence and absence of the NaCl electrolyte. In order to obtain the test sample, 364 cm2 of MUA monolayer surface were exposed to 300 pmol of methylene blue, in a final reaction volume of 150 μl. For this, 140 μl of AuNP@MUA colloid (maximum absorbance OD of the plasmon band=30/cm) were mixed with 5 μl of Milli-Q water and 5 μl of a standard solution of 60.00 μM methylene blue dissolved in Milli-Q water. To obtain the control sample, 140 μl of particle-free solution obtained from a volume of the AuNP@MUA colloid (maximum absorbance OD of the plasmon band=0/cm) were mixed with 5 μl of Milli-Q water and with 5 μl of a standard 60.00 μM methylene blue solution dissolved in Milli-Q water. The electrolyte test sample and electrolyte control sample were obtained in an identical manner to the previous ones, but using 5 μl of 300 mM NaCl instead of 5 μl of Milli-Q water. To separate the MUA monolayer from the aqueous solvent, all samples were centrifuged at 24,000×g for 15 min. A volume of 130 μl free of AuNP@MUA nanoparticles was recovered from all samples. The amount of methylene blue was measured in all recovered volumes by reversed-phase high-performance liquid chromatography (RP-HPLC) and UV visible detection using a diode array detector. The methylene blue signal was detected at a wavelength of 665 nm (mAU). The amount of methylene blue was determined as value of integration area of the signal corresponding to the elution peak (mAU·min). Test samples, control samples, electrolyte test samples, and electrolyte control samples were independently obtained and processed. Each sample was measured independently. The assay was repeated three times in order to have statistical validity.
The results of concentration determination in the recovered volumes indicated that 124.5 pmol of methylene blue is adsorbed on the surface of the MUA monolayer in the absence of NaCl, while 31.5 pmol of methylene blue was adsorbed in the presence of 10 mM NaCl. The difference between these values indicates that, under these experimental conditions, at least 93.0 pmol of methylene blue are adsorbed to the surface of the MUA monolayer by means of electrostatic interactions (
In parallel, the adsorption of methyl orange molecules on the surface of the MUA monolayer was determined, in the presence and absence of the NaCl electrolyte. All the samples were obtained in an identical manner to the samples containing methylene blue, but a 60 μM methyl orange standard solution was used instead of a 60 μM methylene blue standard solution. The samples were processed in an identical manner to the samples containing methylene blue. The amount of methyl orange was measured in all recovered volumes by RP-HPLC and UV visible detection using a diode array detector. The methyl orange signal was detected at a wavelength of 432 nm (mAU). The amount of methyl orange was determined as value of integration area of the signal corresponding to the elution peak (mAU·min). Test samples, control samples, electrolyte test samples, and electrolyte control samples were independently obtained and processed. Each sample was measured independently. The assay was repeated three times in order to have statistical validity.
The results of concentration determination in the recovered volumes indicated that methyl orange does not adsorb on the surface of the MUA monolayer, neither in the absence nor in the presence of 10 mM NaCl (
The comparison of the results obtained with methylene blue and methyl orange shows that the surface of the MUA monolayer adsorbs cationic molecules by electrostatic interactions, and does not adsorb anionic molecules. This indicates that the composition of the surface of the MUA monolayer contains motifs ionized with electron elementary charges and does not contain motifs ionized with proton elementary charges.
Method to Assess the Lyophobic Component of the Surface of the MUA Monolayer by Determining the Adsorption of Hydrophobic Molecules with No Elementary Charge.
The adsorption of phenanthrene molecules on the surface of the MUA monolayer and the desorption of the same molecules were determined after exposure of the surface to the organic solvent ethyl acetate. To obtain the test sample, 728 cm2 of MUA monolayer surface were exposed to 600 pmol of phenanthrene, in a final reaction volume of 150 μl. For this, 140 μl of AuNP@MUA colloid (maximum absorbance OD of the plasmon band=60/cm) were mixed with 5 μl of Milli-Q water and 5 μl of a standard solution of 120.00 μM phenanthrene dissolved in ultra-pure ethanol. To obtain the control sample, 140 μl of particle-free solution obtained from a volume of the same AuNP@MUA colloid (maximum absorbance OD of the plasmon band=0/cm) were mixed with 5 μl of Milli-Q water and 5 μl of a standard 120.00 μM phenanthrene solution dissolved in ultra-pure ethanol. The nonpolar solvent control sample was obtained after mixing 145 μl of ethyl acetate with 5 μl of a standard 120.00 μM phenanthrene solution dissolved in ultra-pure ethanol. To concentrate the surface of MUA monolayer, all samples were centrifuged at 24,000×g for 15 min. A volume of 130 μl free of AuNP@MUA nanoparticles was recovered from the control sample and the test sample. The remaining volumes of control sample (20 μl) and enriched MUA monolayer surface test sample (20 μl) were independently mixed with 150 μl ethyl acetate. Subsequently, a volume of 125 μl of ethyl acetate was recovered from each mixture. The amount of phenanthrene was measured in all recovered volumes and in the nonpolar solvent control sample by RP-HPLC and UV visible detection using a diode array detector. The phenanthrene signal was detected at a wavelength of 251 nm (mAU). The amount of phenanthrene was determined as value of integration area of the signal corresponding to the elution peak (mAU·min). Test samples, control samples, and nonpolar solvent control samples were independently obtained and processed. Each sample was measured independently. The assay was repeated three times in order to have statistical validity.
The results of phenanthrene concentration determination indicated that, after exposure to 4.00 μM phenanthrene, the concentration of phenanthrene dissolved in the aqueous liquid phase of the AuNP@MUA colloid is 0.31 μM. On the other hand, the concentration of phenanthrene determined in the volumes of ethyl acetate recovered from the control samples and test samples was 0.24 and 1.15 μM.
Material balance calculations indicate that under these experimental conditions the surface of the MUA monolayer adsorbs 553.5 pmol of phenanthrene, while in the presence of ethyl acetate at least 136.5 pmol of phenanthrene are desorbed (
Analysis of the data indicates that the surface of the MUA monolayer adsorbs molecules with no elementary charge, and does so reversibly. This indicates that the composition of the MUA monolayer surface contains a lyophobic motif that allows adsorption of phenanthrene molecules.
| Number | Date | Country | Kind |
|---|---|---|---|
| 1473-2018 | Jun 2018 | CL | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CL2019/050044 | 5/28/2019 | WO | 00 |
