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Brno, 23. 11. 2017
Mgr. Jana Krenkova, Ph.D.
Nanoparticle modified monolithic materials
Control of surface chemistry
• Preparation from functional monomers
• polymerization
• grafting
F. Svec, J. Chromatogr. A 2010, 1217, 902–924
Control of surface chemistry
• Modification of reactive monoliths
F. Svec, J. Chromatogr. A 2010, 1217, 902–924
• Attachment of nanomaterials
Control of surface chemistry
Why nanomaterials?
selectivity – chromatography, sample preparation
surface area
ligands for immobilization of bioactive molecules
Nanomaterials
materials of which a single unit is sized (in at least one dimension)
between 1 and 100 nanometers
Modification of monoliths with nanoparticles
simple and straightforward approach
distribution of NPs throughout the monolithic matrix
limited accessibility of NPs for desired interactions
• embedding (encapsulation) of NPs in the monolithic matrix
• attachment of nanoparticles to the surface of preformed monoliths
multistep approach
independent optimization of individual steps
improved accessibility of NPs for desired interactions
Latex nanoparticles
monolith:
poly(BMA-co-EDMA-co-AMPS)
Hilder et al. J. Chrom. A 2004 1053, 101
Ion exchange chromatography - carbohydrates
BMA – butyl methacrylate, EDMA – ethylene dimethacrylate,
AMPS - 2-acrylamido-2-methyl-1-propanesulfonic acid
latex particles (60 nm diameter) with
tertiary amine functionality
Peaks: d(+)galactose (1), d(+)glucose (2),
d(+)xylose (3), d(+)mannose (4), maltose (5),
d(−)fructose (6), sucrose (7).
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Gold nanoparticles
Colors of various sized
monodispersed gold
nanoparticles
HAuCl4 + trisodium citrate dihydrate
Au+3 ions are reduced to neutral gold atoms, where citrate
ions act as both a reducing agent and a capping agent.
Citrate synthesis
Gold nanoparticles
cystamine
TCEP
gold nanoparticles
attachment
Y. Lv et al. J. Chromatogr. A 2015, 1261, 121
1 mm
Gold nanoparticles
Cys-GMA monolith
functionalized with
15 nm gold nanoparticles
Gold nanoparticles
Cao et al. Anal. Chem. 2010, 82, 7416-7421
Thiol-containing peptide enrichment
Ligand exchange - various chromatographic modes
ion exchange
reversed-phase
Peaks:
myoglobin (1)
ribonuclease A (2)
cytochrome c (3)
Gold nanoparticles
Lectin immobilization
Erythrina cristagalli lectin - galactose-selective lectin (ECL)
H. Alwael et al. Analyst 2011, 136, 2619
Gold nanoparticles
Lectin immobilization
Erythrina cristagalli lectin - galactose-selective lectin (ECL)
Peaks:
(1) ribonuclease B
(2) insulin chain B
(3) insulin
(4) cytochrome C
(5) desialylated transferrin
(6) BSA
(7) carbonic anhydrase
(8) enolase
(9) desialylated thyroglobulin
before extraction
after extraction
after a galactose wash step
RP-LC/UV separation
of protein mixture
H. Alwael et al. Analyst 2011, 136, 2619
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Fullerenes
[6,6]-phenyl-C61-butyric acid 2-hydroxyethylmethacrylate ester
molecule of carbon in the form of a hollow sphere, ellipsoid, tube, and many other
shapes
C60 in solution
spherical fullerenes - buckyballs
cylindrical fullerenes - carbon nanotubes or buckytubes
Fullerenes
Conditions: column 53 mm x 100 μm
i.d., flow rate 0.15 μL/min, UV
detection at 254 nm; (A) mobile phase
50:50 vol % acetonitrile-water; (B)
mobile phase 50:50 vol % acetonitrilewater;
(C) mobile phase 47.5:2.5:50
vol % acetonitrile-tetrahydrofuranwater;
peaks in order of elution:
uracil, benzene, toluene,
ethylbenzene, propylbenzene,
butylbenzene, and amylbenzene
Reversed-phase chromatography
Separation of alkylbenzenes
110 000 plates/m
for the retained
benzene
S.D. Chambers et al. Anal. Chem. 2011, 83, 9478
poly(GMA-co-EDMA) poly(GMA-co-EDMA)
containing 1 wt% PCB-HEM
Carbon nanotubes
Reversed-phase chromatography
Conditions: column, 180mm × 100 mm ID, mobile phase
45% acetonitrile–5% THF–50% water, flow rate 1mL/min,
UV detection at 254 nm; peaks: uracil (1), benzene (2),
toluene (3), ethylbenzene (4), propylbenzene (5),
butylbenzene (6), and amylbenzene (7).
Separation of alkylbenzenes
poly(GMA-co-EDMA)
poly(GMA-co-EDMA)
containing 0.25 wt%
entrapped MWNT
Multi-wall nanotubes
S.D. Chambers et al. J. Chromatogr. A 2011, 1218, 2546
Metal-organic frameworks (MOF)
compounds consisting of metal ions or clusters coordinated to organic molecules to
form one-, two-, or three-dimensional structures
MOFs in monolithic materials
• preparation in situ
• admixing preformed MOFs
properties
superlative porosity
wide chemical tunability
high stability
applications
gas storage
gas separation
catalysis
Metal-organic frameworks (MOF)
monolith containing carboxylic acid
functionalities
Preparation in situ
synthesis of MIL-100
inorganic metal ions - FeCl3
organic ligand - 1,3,5-benzenetricarboxylic acid (BTC)
A. Saeed et al. Adv. Funct. Mater. 2014, 24, 5790
high specific micropore surface area of 389 m2/g
(original polymer monolith – surface area of 106 m2/g)
Metal-organic frameworks (MOF)
A. Saeed et al. Adv. Funct. Mater. 2014, 24, 5790
layer-by-layer growth (30 cycles)
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after enrichment
before enrichment
Phosphopeptide enrichment (IMAC) - MALDI/MS
Metal-organic frameworks (MOF)
A. Saeed et al. Adv. Funct. Mater. 2014, 24, 5790
analysis of nonfat milk
Titanium and zirconium oxide nanoparticles
Metal oxide affinity
chromatography (MOAC)
MALDI mass spectrum obtained from in vitro
phosphorylated ERK1 digest before (A) and
after enrichment with poly(DVB)-TiO2/ZrO2
microcolumns (B)
M. Rainer et al. Proteomics 2008, 8, 4593
Iron oxide nanoparticles
before attachment of NPs
after attachment of NPs
• 200 mL polypropylene pipette tips
• modification of the inner wall by methyl methacrylate/ethylene dimethacrylate
• poly(2-hydroxyethyl methacrylate-co-ethylene dimethacrylate)
5 mL monolithic bed prepared by UV-initiated polymerization
• UV-photografting of [3-(methacroylamino)propyl]trimethylammonium
chloride on the pore surface of the monolith
• attachment of 20 nm citrate stabilized iron oxide NPs
on the quaternary amine functionalized monolith
Fe3+ + Fe2+ + OH- Fe3O4 + H2O
g-Fe2O3
oxidation
n(Fe2+)/n(Fe3+)=1/2
pH>10
Krenkova J., Foret F. Anal. Bioanal. Chem. 2013, 405, 2175
Hydroxyapatite nanoparticles
• 200 mL polypropylene pipette tips
• modification of the inner wall by methyl methacrylate/ethylene dimethacrylate
• poly(2-hydroxyethyl methacrylate-co-ethylene dimethacrylate) with embedded
hydroxyapatite NPs (nanorods – 50 x 150 nm)
5 mL monolithic bed prepared by UV-initiated polymerization
Krenkova J., Foret F. Anal. Bioanal. Chem. 2013, 405, 2175
hydroxyapatite nanoparticles
morphology: rods - 50 x 150 nm
surface area: 115 m2/g
Phosphopeptide enrichment – MALDI/MS analysis
b-casein: 224 amino acid residues
5 phosphorylation sites
after enrichment using the
hydroxyapatite NP modified tipbefore
enrichment
after enrichment using the
titanium dioxide tip
after enrichment using the
iron oxide NP modified tip
m/z m/z
m/z m/z
*-phosphopeptides
Krenkova J., Foret F. Anal. Bioanal. Chem. 2013, 405, 2175
Institute of Analytical Chemistry of the CAS
Veveří 967/97
602 00 Brno
www.iach.cz
Dana Moravcova – moravcova@iach.cz
Jana Krenkova – krenkova@iach.cz