Vapor-phase synthesis of molecularly imprinted polymers on nanostructured materials at room-temperature

VAPOR-PHASE SYNTHESIS OF MOLECULARLY IMPRINTED POLYMERS ON NANOSTRUCTURED MATERIALS AT ROOM-TEMPERATURE TIZIANO DI GIULIO1, ELISABETTA MAZZOTTA1 ,COSIMINO MALITESTA1, MARTINA CORSI2, GIUSEPPE BARILLARO2 1Dipartimento di Scienze e Tecnologie Biologiche e Ambientali (Di.S.Te.B.A.), Università del Salento, Lecce, Italy 2Dipartimento di Ingegneria dell’Informazione, Università di Pisa, Pisa, Italy OBJECT OF THE RESEARCH Novel, effective route enabling the preparation of artificial receptors on nanostructured materials, here used as optical transducer, and demonstrate its versatility room temperature vapor-phase synthesis of MIPs MIPs Imprinted polypyrrole Nanostructured material Nanoporous silica Target molecules Hemoglobin Doxorubicin Quercetin NANOPOROUS SILICA (nPSiO2) as optical transducer (interferometer) Nanostructured material Nanoporous silicon Thickness: 4m Porous diameter: 50 nm Porosity: 75% reflecte d light FFT Signal modification is produced by: A) insertion/grafting; B) removal or C) adsorption of compounds on porous silica layer A B C A B C m=2nd/λm EOT (effective optical thickness): 2nL light n: refractive index L: thickness MIPs Imprinted polypyrrole Target (=template) removal imprinted cavities MIP MOLECULAR MEMORY Target selective rebinding MIPs Imprinted polypyrrole Target (=template) removal imprinted cavities MIP MOLECULAR MEMORY Target selective rebinding Integration of MIPs on the transducer surface is a key issue for sensing applications, being still challenging on nanostructured materials MIPs Imprinted polypyrrole Target (=template) removal imprinted cavities MIP MOLECULAR MEMORY Target selective rebinding Integration of MIPs on the transducer surface is a key issue for sensing applications, being still challenging on nanostructured materials Diffusion-limited transport of molecules within the nanosized structures could lead to an uneven growth rate of the MIP, resulting in a nonuniform coating of the inner surface over depth PPy VAPOR-PHASE DEPOSITION ON PSiO2 ✸Higher diffusion coefficient of species in vapor phase (about 10-5 vs10-2 cm2 s-1) can facilitate the diffusion of monomer vapors within the nanostructure ✸This allows overcoming diffusion-limited transport enabling the controlled and reliable polymerization also in nanostructured materials with high aspect ratio PSiO2 nanopore PSiO2PSiO2 PPy FeCl3 Pyrrole (v) Target molecules Human hemoglobin Clinical relevance of human haemoglobin detection for several diseases, including anemia, cardiovascular risk, and coronary artery disease Doxorucin Quercetin Antibiotic and anticancer drug derived from the Streptomyces peucetius bacterium. It has widespread use as a chemotherapeutic agent One of the most important bioflavonoid in human dietary supplements will antioxidant properties (acting in reducing blood pressure, risk of heart disease, preventing neurological diseases, boosting immunity, and controlled the blood sugar level) PPY-BASED MIP FOR HUMAN HEMOGLOBIN (HHb) PPY-BASED MIP FOR QUERCETIN (QU) Hydroxyl compund Imidazole Carbamate Active Intermediate Imidazole N,N'-Carbonyl Diimidazole (CDI) Imidazole Carbamate linkage Amine Containing Compound Imidazole Carbamate Active Intermediate QU APTES on PSiO2 PPY-BASED MIP FOR QUERCETIN (QU) Hydroxyl compund Imidazole Carbamate Active Intermediate Imidazole N,N'-Carbonyl Diimidazole (CDI) Imidazole Carbamate linkage Amine Containing Compound Imidazole Carbamate Active Intermediate QU APTES on PSiO2 QU-CDI intermediate PPY-BASED MIP FOR QUERCETIN (QU) Hydroxyl compund Imidazole Carbamate Active Intermediate Imidazole N,N'-Carbonyl Diimidazole (CDI) Imidazole Carbamate linkage Amine Containing Compound Imidazole Carbamate Active Intermediate QU APTES on PSiO2 QU-CDI intermediate Polymer deposition Target removal MIP 0 5 10 15 20 25 30 35 40 400 450 500 550 600 650 700 750 800 Reflectance (%) Wavelength (nm) bare nPSiO2 APTES functionalization CDI/Quercetin anchoring 0 5 10 15 20 25 30 35 40 400 450 500 550 600 650 700 750 800 Reflectance (%) Wavelength (nm) CDI/Quercetin anchoring PPy synhtesis Template removal PPY-BASED MIP FOR QUERCETIN (QU) 0 50 100 150 200 250 300 350 400 450 500 APTES functionalization Target anchoring PPy synthesis Template removal EOT-EOTPSiO2 (nm) PSiO2functionalization steps 0 5 10 15 20 2.5 5 10 20 EOT-EOT0(nm) [Quercetin] (M) 2h 1h 30min PPY-BASED MIP FOR QUERCETIN (QU) 0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 2.50 5.00 10.00 20.00 EOT-EOT0(nm) Quercetin concentration (M) Quercetin Vanillic acid Ferulic acid Gallic acid Caffeic acid MIP vs NIP 0 10 20 30 40 50 60 70 80 050 100 150 200 250 EOT-EOT0(nm) [Quercetin] (M) y = 0.5746x + 6.3025 R² = 0.992 y = 0.1586x + 1.3794 R² = 0.9663 0 10 20 30 40 50 60 020 40 60 80 100 EOT-EOT0(nm) [Quercetin] (M) 15 20 25 30 35 40 0 1 5 10 20 30 60 EOT-EOT0(nm) Time (days) 9820 9825 9830 9835 9840 9845 9850 Blank QU solution Blank QU solution Blank QU solution EOT-EOT0(nm) MIP - repeatability and stability 0 5 10 15 20 25 30 red wine white wine Concetration foound (M) Real matrix SENSORS RESULTS HPLC RESULTS PPY-BASED MIP FOR QUERCETIN (QU) HPLC analysis quercetin signal quercetin signal white wine red wine Quercetin concentrations detected by the sensor in real matrices (red and white wine) are in agreement with those recorded by HPLC analysis CONCLUSIONS ▪A NOVEL ROUTE FOR THE SYNTHESIS OF MIPs IS PROPOSED THAT LEVERAGES VAPOR-PHASE POLYMERIZATION ON NANOSTRUCTURED MATERIALS ▪THE PROPOSED STRATEGY PAVES THE WAY TOWARDS THE USE OF MIPs IN NANOMATERIALS. ITS VERSATILITY IS DEMONSTRATED BEING APPLIED TO THE IMPRINTING OF THREE DIFFERENT MOLECULES ▪IT IS EXTENDABLE TO OTHER NANOMATERIALS/TRANSDUCERS TAKING ADVANTAGE FROM THE WIDE USE OF PPy IN THE IMPRINTING OF (MACRO)MOLECULES AND THE SUBSTRATEINDEPENDENT DEPOSITION MECHANISM THANKS FOR YOUR KIND ATTENTION! European Union Horizon Europe programme under grant agreement No 101046946 (RESORB) 0 10 20 30 40 50 60 70 80 050 100 150 200 250 EOT-EOT0(nm) [Quercetin] (M) y = 0.5699x + 6.3347 R² = 0.9901 0 10 20 30 40 50 60 020 40 60 80 100 EOT-EOT0(nm) [Quercetin] (M) 0 5 10 15 20 25 30 15 30 45 60 120 EOT-EOT0(nm) Incubation time (min) y = 0.1474x - 0.2286 R² = 0.9991 0 5 10 15 20 25 30 35 050 100 150 200 250 Area (mAU.min) Quercetin (uM) HPLC calibration curve 20 M 50 M The mobile phases consisted of A (water–acetonitrile–acetic acid, 67:32:1 v/v/v) and B (water–acetic acid, 99:1 v/v). The gradient elution conditions were as follows: 0 min (20% A + 80% B); 4 min (30% A + 70% B); 8 min (40% A + 60% B); 12 min (65% A + 35% B); 16 min (80% A + 20% B); 20 min (95% A + 5% B); 21.8 min (97% A + 3% B); 24 min (100% A) and 30 min (100% A). The flow rate was 0.8 mL min1 and the injection volume 20 uL A reversed-phase C18 BIOBASIC (10 x 2.5 cm) column was used. 100 M 200 M