Current Application of Capillary Electrophoresis in Nanomaterial Characterisation and Its Potential to Characterise the Protein and Small Molecule Corona
Abstract
:1. Introduction
2. Understanding CE as an Analytical Platform
2.1. CE Principals
2.2. CE Separation Techniques as Applied to NM Characterisation
2.3. CE Detectors for NM Detection
2.3.1. CE-UV
2.3.2. CE-MS for Elemental Analysis
3. Summary of NMs Analysed by CE to Date
3.1. Gold NPs
3.2. Silver NPs
3.3. Carbon NMs
3.4. Polystyrene NPs
3.5. Silica NPs
3.6. Iron NPs
3.7. Analysis of Mixtures of NMs
4. CE for NM Corona Characterisation
4.1. Protein Corona Characterisation
4.2. Small Molecule Corona Characterisation
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
APTES | 3-(aminopropyl)triethoxysilane |
BFS | bare fused silica |
BGE | background electrolyte |
BPEI | branched polyethyleimine |
CAPS | 3-cyclohexylamino-1-propanesulphonic acid |
CCG | chemically converted graphene |
CE | capillary electrophoresis |
CE-MS | capillary Electrophoresis–Mass Spectrometry |
CGE | capillary gel electrophoresis |
cIEF | capillary isoelectric focussing |
CNT | carbon nanotubes |
CTAB | cetrimonium bromide |
CZE | capillary zone electrophoresis |
DDABr | didodecyldimethylammonium bromide |
DLS | dynamic light scattering |
EDTA | ethylenediaminetetraacetic acid |
EOF | electroosmotic flow |
FFF | field flow fractionation |
GO | graphene oxide |
HCl | hydrochloric acid |
HPC | hydroxypropyl cellulose |
HRMS | high resolution mass spectrometry |
ICP-MS | inductively coupled plasma–mass spectrometers |
ITC | isothermal titration calorimetry |
ITP | isotachophoresis |
LC-MS | liquid chromatography–mass spectrometry |
LC-ESI-MS | liquid chromatography–electrospray ionisation–mass spectrometry |
LIF | laser induced fluorescence |
LLS | laser light scattering |
LOD | limit of detection |
LOQ | limit of quantification |
MALDI-MS | matrix assisted laser desorption ionisation–mass spectrometry |
MEKC | micellar electrokinetic chromatography |
MWNT | multi-wall nanotubes |
nESI | nanoelectrospray ionisation |
nLC-nESI-MS | nanoliquid chromatography–nanoelectrospray ionisation–mass spectrometry |
NM | nanomaterials |
NMR | nuclear magnetic resonance |
NP | nanoparticle |
PB | polybrene |
PEG | polyethylene glycol |
PVP | polyvinylpyrrolidone |
PEOS | 2-[methoxy(polyethyleneoxy)propyl]trimethoxysilane |
PEG-PHDCA | poly(methoxypolyethyleneglycol cyanoacrylate-co-hexadecylcyanoacrylate) |
PNC | particle number concentration |
REPSM | reversed electrode polarity stacking mode |
RSD | Relative Standard Deviation |
SDS | sodium dodecyl sulphate |
SDS-PAGE | sodium dodecyl sulphate-Polyacrylamide gel electrophoresis |
SERS | surface enhanced Raman spectroscopy |
spICP-MS | single particle inductively coupled plasma–mass spectrometers |
SPIONs | superparamagnetic iron nanoparticles |
SWNT | single-wall nanotubes |
TDA | Taylor Dispersion Analysis |
TEM | transmission electron microscope |
TMAOH | tetramethylammonium hydroxide |
Tris | tris(hydroxymethyl)aminomethane |
UV | ultraviolet |
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Technique | Property Measured | Advantages of Determining via CE | Refs |
---|---|---|---|
Dynamic Light Scattering (DLS) | Hydrodynamic diameter of a particle Zeta potential Electrophoretic mobility | Less expensive More versatile Able to analyse polydisperse samples or complex samples Accurate for the sizing of non-spherical particles | [4,5] |
Field Flow Fractionation (FFF) | Separation technique that can separate materials over a wide colloid range | Less expensive Typically less than 20 min run times Very little loss of sample to capillary whereas significant losses to the FFF membrane can occur if improper sample preparation and method development is performed | [6,7,8] |
Transmission Electron Microscopy (TEM) | Microscopy technique allowing for size and shape determination of electron dense materials | Less expensive Non-destructive Rapid (typically <20 min per sample) Minimal sample preparation required Relatively larger sample volume/amount analysed making analysis more representative Clear cut results with no user interpretation required thus reducing bias | [9,10] |
Ultra-Violet Visible Light Spectroscopy (UV/VIS) | Spectroscopy technique able to quantitatively determine different analytes and biological macromolecules | Can be used as a separation technique | [5] |
NP Characteristic Driving Separation | NP Properties that Can Be Calculated Using CE-UV/LIF/LLS | NP Properties that Can Be Calculated Uniquely by CE-(SP) ICP-MS |
---|---|---|
Size | Relative size (using calibration curve) | Elemental composition |
Shape | Zeta potential | Size |
Cross sectional area | Surface charge density | Size distribution |
Surface charge/functionalisation | Concentration | Trace level concentrations |
Capping material | Particle number concentration (spICPMS) | |
Composition |
NM Composition | NP Diameters (nm) | Capillary Material/Dimensions | Injection Pressure and Duration | Separation Voltage and Temperature | Separation Time (min) | Background Electrolyte | pH | Detection | Result | Ref |
---|---|---|---|---|---|---|---|---|---|---|
Gold | Not defined | BFS 50 µm × 30 cm | 100 mBar 3 s | Not provided 25 °C | 35 | 30 mM sodium phosphate in 20% EtOH | 10.0 | LIF 485/550 nm UV-Vis 214 nm | Baseline separation of a range of NPs | [41] |
Gold | 5.3, 12.1, 40.1, 59.9 | BFS 75 µm i.d. × 33.5 cm | 50 mBar 5 s | 20 kV 25 °C | 4 | 70 mM SDS, 10 mM CAPS | 10.0 | UV-Vis 520 nm | Size vs. Mt R2 0.985. Good correlation between TEM and CE for size measurements | [38] |
Gold | 3.5, 6.5, 10.5 | BFS 50 µm i.d. ×100 cm | 4.9 kPa 100 s | 10 kV 20 °C | 25 | 20 mM NH4AC, 20 mM TRIS, 10 mM CAPS | 8.5 | ELSD | Non-baseline separation of NPs. Ability to distinguish between 3 NP sizes covering just a 7 nm size difference. Good correlation between CE and TEM | [42] |
Gold | 5, 10, 20, 40 | Polyamide coated BFS µm i.d. × 36.5 cm to UV and 45 cm to C4D | 50 mBar 12 s | 20 kV | 15 | 20 mM PIPES | 7.4 | UV-Vis 210/220/235 nm ICP-MS C4D | CE-ICP-MS LOD of 2 × 10−15 M. Conductivity not suitable as a detector for AuNPs | [14] |
Gold | 5.3, 40.1 | BFS 75 µm i.d. × 25 cm | 50 mBar 2 s | 20 kV 25 °C | 3 | 70 mM SDS, 10 mM CAPS | 10.0 | UV-Vis 520 nm | REPSM increases sensitivity. Addition of NaH2PO4 for reduced migration time | [32] |
Gold | 5, 20 | BFS 75 µm i.d. × 55 cm | 5 s | 28 kV | 6 | 50 mM TRIS | 9.2 | UV-Vis 520 nm | Separation of the two NP sizes. Ability to separate AuNPs from polystyrene NPs | [45] |
Gold | 5.2, 5.9, 7.2, 8.6, 14.6 | BFS 75 µm i.d. × 27 cm | 50 mBar 5 s | 20 kV 25 °C | 10 | 6 mM NH4Ac/acetic acid | 5 | UV-Vis 520 nm | Good correlation between size and mobility R2 0.9745 | [39] |
Gold | 5.3, 9.8, 19.0, 29.3, 41.2 | BFS 75 µm i.d. × 43.1 cm | 10 s | 20 kV 20 °C | Not defined | 70 mM SDS, 10 mM CAPS | 10.0 | UV-vis 546 nm | Good correlation between size and mobility R2 0.99. Mobility RSD below 1% for 5.3 and 19 nm AuNPs | [31] |
Gold | 5, 10, 21.5, 30.2, 41.2 | BFS 75 µm i.d. | 50 mBar 3 s | 18 kV 15 °C | <5 | 70 mM SDS, 10 mM CAPS | 11.0 | UV-vis | Strong linear relationship between NP size and mobility R2 0.992. Electrophoretic mobility RSD <0.8%. Good correlation between CE and SEM methods for NP size determination | [60] |
Gold | 5, 40, 60 | BFS 75 µm i.d. × 33.5 cm | 50 mBar 50s | 20 kV 25 °C | 4 | 70 mM SDS, 10 mM CAPS | 10 | UV-vis 520 nm | Baseline separation of NPs with a R2 0.99 for linearity of mobility and NP size. REPSM method utilized to improve sensitivity | [61] |
Gold | 10, 30, 60 | Polyimide coasted fused silica capillary | 50 mBar 5 s | 30 kV | <11 | 70 mM SDS 10 mM CAPS | 10 | spICP-MS | Determination of Mt, size, PNC in a single analysis. Non-baseline CE separation due to broad particle size distribution. Strong linear relationship between particles injected and particles detected R2 ≥ 0.998 | [52] |
Gold Gold/Silver | 17.2 60.1 | BFS 75 µm i.d. × 25 cm | 50 mBar 50 s | 20 kV 25 °C | 5 | 40 mM SDS, 10 mM CAPS | 10.0 | UV-vis 520 nm | Baseline separation between the AuNPs. REPSM method utilized to improve sensitivity | [30] |
Gold and silver | Au: 5, 10, 20, 50 Ag: 7, 30 | Polyamide coated fused silica capillary 75 µm i.d. × 70 cm | 50 mbar 3 s 20 kV 8 s | 29 kV | <10 | 60 mM SDS, 10 mM CAPS | 10 | ICP-MS | Distinguished between AuNPs and AgNPs. Strong correlation between mass spectrometer response and NP size R2 = 0.999 | [36] |
Silver | 17, 49.7 | BFS 75 µm i.d. × 40 cm | 50 mBar 1 s | 20 kV 15 °C | <20 | 20 mM SDS, 10 mM TRIS | 8.5 | UV-Vis 350, 395 440 nm | Baseline separation of the 2 NPs. Non-baseline separation of NP (sphere) and NM (rod) | [63] |
Silver and gold | 10, 20, 40 10 20 | BFS 75 µm i.d. × 60 cm | 50 mBar 15 s | 25 kV 25 °C | 10 | 10 mM Tris, 10 mM H3BO3, 10 mM Na2B4O7 | 9.0 | ICP-MS | Non-baseline separation of the 3 NPs however, good linear relationship between size and mobility R2 0.9982 Size determination compared favourably to DLS and TEM | [46] |
Silver | 20, 40, 60 | Polyamide coated fused silica capillary, 75 µm i.d. × 70 cm | 50 mBar 3 s REPSM up to 150 s | 20 kV | <30 | 60 mM SDS 10 mM TRIS | 10 | spICP-MS | REPSM method used to improve sensitivity. Good correlation between NPs detected and injection time R2 > 0.99. Good linearity for mobility and separation voltage R2 > 0.99. LOD < 1 µg/L | [53] |
Silver | Citrate capped: 20, 40 60 PVP capped: 40, 60 PEG coated: 40 BPEI coated: 40 | Polyimide coated fused silica 75 µm i.d. × 70 cm | 50 mBar 10 s then −20 kV REPSM | 20 kV | <30 | 60 mM SDS 10 mM CAPS | 10 | spICP-MS | Separation of NP coating by CE prior to spICP-MS detection. REPSM method used to improve sensitivity | [54] |
Fullerenes | C3 and DF1 | BFS MEKC BFS dynamic coating for CZE 50 µm × 30 or 50 cm | 0.5 PSI 20 s | +22 kV BFS −22 kV dynamic coating | 15 | MEKC: 150 mM SDS, 10 mM Sodium tetraborate CZE: 10 Mm Sodium tetraborate | 9.2 | DAD 250 nm | LODs of between 0.6 and 6 µg/mL | [64] |
Carbon | nd | BFS 50 µm × 50 cm | 0.5 PSI 5 s | 25 kV 22 °C | 60 | 80 mM glycine | 9.9 | DAD 230 nm | Separation of different carbon NMs achieved | [65] |
PVP stabilized SWNT | nd | BFS 75 µm × 37.5 cm | 500 mBar 2 s | 15 kV | <35 | 50 mM Trizma base 0.5% SDS | nd | Raman | Separated SWNT based upon length, diameter and cross-sectional area | [66] |
SWNT | Length 0.2–1.2 0.5–2.5 2–4 1.8–10 | BFS 75 µm × 75 cm (UV) 25 cm Ramen | 100 mBar 30 s | 5 kV | 20 | 50 mM Trizma base 0.5% SDS | UV 360 nm Ramen | Separated SWNT based upon length. Improved size selectivity than FFF and size exclusion chromatography | [67] | |
Graphene oxide (GO) and reduced graphene oxide (CCG) | nd | Polyimide coated BFS 75 µm × 41.5 cm | 50 mBar 5 s | 15 kV 22 °C | 15 | 250 µM tetrapropylammonium hydroxide | 10.4 | UV GO 230 nm CCG 270nm | Ability to differentiate GO and CCG demonstrated | [68] |
SWNT MWNT | SWNT 1.2–1.5 nm and 0.7–1.2 nm diameter 2–5 and 2–20 µm MWNT 20–50 nm diameter, −20 µm | BFS 75 µm × 47 cm | 0.5 PSI 10 s | 15 kV | 10 | 5 mM NH4AC with 0.025% HPMC | 8.03 | DAD 240 nm | Distinguished SWNTs and MWNTs based upon size and volume. Mt reproducibility RSD 2.7–5.4%. Peak area reproducibility RSD 3.7–7.8% | [69] |
Fullerenes | C60 C70 | BFS 75 µm × 28 cm | 20 mBar and gravity fed | 10 kV 20 °C | 26 | 10 mM borate phosphate with 100 mM SDS | 9.5 | UV 254 nm | Separation of C60 and C70 | [70] |
Graphene oxide (GO) | nd | BFS 75 µm × 50 cm | 200 mbar 40 s | 10 kV | 45 | 50 mM borate | 11 | UV 280 nm | GO sheets separated based upon size and stacking | [71] |
Silica | 20, 50, 100 | BFS 50 µm × 50 cm | 50 mBar 10 s | 27 kV 20 °C | <20 | 3 mM NH4AC 1% MeOH | 6.9 | ELSD | Strong linear relationship between peak area and NP concentration R2 0.999 LOD 1.08 ng/nL Peak area RSD <6%. Near baseline separation of the 3 NP sizes | [34] |
Silica | 7, 12, 22 | BFS 75 µm × 29.2 cm | 0.1 PSI 0.2 6 s | 7 kV 15, 20, 25 °C | 40 | 20, 30, 40, 50, 60 mM Borate | nd | UV/TDA | Zeta potential, surface charge density and hydrodynamic sized determined | [47] |
Polystyrene | 20, 50, 155, 300 | BFS 75 µm i.d. × 55 cm | 5 s | 28 kV | 6 | 50 mM TRIS | 9.2 | UV 520 and 254 nm | Baseline separation of the 4 NP sizes. Ability to separate polystyrene NPs from AuNPs | [45] |
Polystyrene | 55 and 70 | BFS 75 µm i.d. × 66.5 cm | 17 mBar 6 s | 7 kV 25 °C | 35 | 12.7 mM Borate | 9.2 | UV-Vis/TDA | CE-TDA correlated with TDA and DLS readings | [51] |
Polystyrene | 39, 72, 132, 308, 488, 683 | 0.5 mM CTAB treated BFS 50 µm i.d. × 47.6 cm | 30 kV 1 s | 30 kV | 5 | 1 mM ACES | 5.8 | UV-Vis 225 nm | Separation of the 6 NP sizes. Linear relationship between NP size and mobility R2 0.903 calculated manually from data presented. Mt RSD of 1.4% | [33] |
Polystyrene | 100, 180 800 | BFS 30 µm i.d. × 10 cm | HPLC injector used | 10 kV and pressure 1.1–3 kgf/cm2 | 2 | 10 mM Borate | 8.2 | UV-Vis 210 nm | Electrophoretic mobility was augmented by applying pressure to capillary | [35] |
Polystyrene | 50, 102, 204, 404, 600 | BFS 75 µm i.d. × 50 cm | 1.38 kPa 10 s | −30 kV 30 °C | <15 | 5 nM phosphate buffer | 9 | UV-Vis 200 nm | Separation of the 50, 102, 204 and 404 nm NPs | [72] |
Iron | HNO3 stabilized: 6.8, 8.9, 10.6 Citrate stabilized: 7.0, 8.9, 10.3 TMAOH stabilized: 6.4, 7.9 | 50 µm i.d. × 26.5 cm HPC coated BFS PB coated BFS DDABr coated BFS BFS | 30 mBar 3 s | BFS and HPC coated 10 kV PB and DDABr coated −10 kV 25 °C for all | <15 PB <5 PB and DDABr BFS not defined | HPC coated BFS for HNO3 stabilized FeNP 10.5 mM alanine and 10 mM HCl PB and DDABr coated 10 mM HCl Citrate stabilized FeNP on BFS: 5.7 mM TMAOH and 2.4 mM citrate TMAOH stabilized FeNP: 5 mM TMAOH | 2.9 2.0 6.1 nd | UV-Vis 200 and 254 nm | Characterized mobility of FeNP in BFS capillary with different coatings. Size based separation for <11 nm FeNPs | [73] |
Iron | All the same undefined size with different surface charge densities | 50 µm i.d. × 26.5 cm DDABr coated BFS | 20 mBar 2 s | −10 kV 25 °C | Not defined | 106.6 mM Tris 100 mM HCl | 8 | UV-Vis 200 and 254 nm | Separation driven by surface charge density. Surface charge density determined in a more reproducible manner than the ninhydrine colorimetric assay | [74] |
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Chetwynd, A.J.; Guggenheim, E.J.; Briffa, S.M.; Thorn, J.A.; Lynch, I.; Valsami-Jones, E. Current Application of Capillary Electrophoresis in Nanomaterial Characterisation and Its Potential to Characterise the Protein and Small Molecule Corona. Nanomaterials 2018, 8, 99. https://doi.org/10.3390/nano8020099
Chetwynd AJ, Guggenheim EJ, Briffa SM, Thorn JA, Lynch I, Valsami-Jones E. Current Application of Capillary Electrophoresis in Nanomaterial Characterisation and Its Potential to Characterise the Protein and Small Molecule Corona. Nanomaterials. 2018; 8(2):99. https://doi.org/10.3390/nano8020099
Chicago/Turabian StyleChetwynd, Andrew J., Emily J. Guggenheim, Sophie M. Briffa, James A. Thorn, Iseult Lynch, and Eugenia Valsami-Jones. 2018. "Current Application of Capillary Electrophoresis in Nanomaterial Characterisation and Its Potential to Characterise the Protein and Small Molecule Corona" Nanomaterials 8, no. 2: 99. https://doi.org/10.3390/nano8020099