microsenzorii stocastici ca noi instrumente de masurare a ...uefiscdi.gov.ro/userfiles/file/pn...
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Microsenzorii stocastici ca noi
instrumente de masurare a
substantelor de importanta biologica
PN-II-ID-PCE-2011-3-0570
Raluca-Ioana van Staden
Laboratorul de Electrochimie si PATLAB Bucuresti
Institutul National de Cercetare pentru Electrochimie si Materie Condensata
PATLAB
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Obiective PATLAB
O1 Identificarea celor mai bune materiale nanostructurate pentru designul microsenzorilor stocastici; O2 Evaluarea diferitilor parametrii care pot afecta caracteristicilor de raspuns ale microsenzorilor stocastici cum ar fi, natura matricei, pH-ul si taria ionica, electrolitii; O3 Aplicarea noilor microsenzori stocastici la determinarea moleculelor de importanta biologica, cum ar fi hormoni tiroidieni, biomarkeri pentru obezitate, etc.; O4 Dezvoltarea de instrumente pentru scanarea substantelor de importanta biologica; O5 Dezvoltarea unei baze de date pentru analiza calitativa utilizand microsenzori stocastici; O6 Integrarea tinerilor cercetatori in echipa laboratorului.
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Test de scanare pentru obezitate
PATLAB
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Electroactive
material
Matrix Calibration
equation
Linear
conc.
range
toff
(s)
Sensitivity (mg/s . mL)
P DP 1/ton=0.02 + 1.96 x 102 x c
R = 0.9965 0.125pg/mL-0.125µg/mL 6.4 1.96x105
MD DP 1/ton=0.06 + 1.69 x 102 x c
R = 0.9966 1.25pg/mL-0.125µg/mL 3.9 1.69x105
MD Graphite
1/ton=0.04 + 1.30 x 102 x c
R = 0.9791 1.25pg/mL-0.125µg/mL 5.2 1.30x105
MD Graphene
1/ton=0.03 + 2.26 x 103 x c
R = 0.9993 0.125pg/mL-12.5pg/mL 6.1 2.26x106
MD CNT 1/ton=0.05 + 0.07 x 102 x c
R = 0.9091 12.5pg/mL-1.25µg/mL 3.9 7.00x103
MD C60
1/ton=0.02 + 1.21 x 102 x c
R = 0.9998 1.25pg/mL-0.125µg/mL 7.4 1.21x105
Leptin PATLAB
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PAI-1
Electroactive
material
Matrix Calibration
equation
Linear
conc.
range
toff
(s)
Sensitivity (mg/s . mL)
P DP 1/ton=0.04 + 3.9 x c
R = 0.9626 5ng/mL – 5µg/mL 7.2 3.90x103
MD DP 1/ton=0.06 + 3.8 x c
R = 0.9966 50ng/mL – 5µg/mL 3.6 3.37x103
MD Graphite
1/ton=0.01 + 9.43 x 103 x c
R = 0.9998 50pg/mL – 5ng/mL 9.9 9.43x106
MD Graphene
1/ton=0.07 + 4.50 x 102 x c
R = 0.9745 50pg/mL – 50ng/mL 2.6 4.50x105
MD CNT 1/ton=0.05 + 3.31 x 102 x c
R = 0.9695
500pg/mL –
50ng/mL 2.8 3.31x105
MD C60
1/ton=0.02 + 1.2 x 101 x c
R = 0.9996 50ng/mL – 5µg/mL 5.6 1.20x104
PATLAB
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Scanarea probelor de sange pentru leptin.
Sample Leptin, pg/mL
P/DP MD/DP -CD/DP MD/C60 MD/
Graphite
MD/
Graphene
MD/CNT
1 10.1 15.0 11.7 15.6 9.87 10.0 10.4
2 14.0 19.0 12.0 10.0 10.0 14.0 16.0
3 1.00 1.00 1.46 1.73 1.35 1.60 1.36
4 10.0 10.0 14.8 19.0 11.7 10.0 14.0
5 3.00 3.00 3.49 2.80 3.30 3.73 3.30
PATLAB
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Scanarea probelor de sange pentru PAI-1.
Sample PAI-1, pg/mL
P/DP MD/DP -CD/DP MD/C60 MD/
Graphite
MD/
Graphene
MD/CNT
1 108 119 99.0 106 107 105 109
2 105 109 97.0 103 136 105 100
3 146 180 130 120 135 100 136
4 466 441 410 440 488 420 490
5 508 500 538 573 563 510 560
PATLAB
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Este necesara o metoda de scanare a salivei
copiilor pentru BPA.
*http://wakeup-world.com/2014/05/11/10-things-to-know-about-bpa-bisphenol-a/ *http://www.stats.org/stories/2009/science_suppressed_BPA_intro_jun12_09.html
*
BPA poate contribui la obezitatea copiilor
PATLAB
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Caracteristicile de raspuns ale microsenzorilor stocastici
Matricea Materialul
Electroactiv
Ecuatia de calibrare si
coeficientul de corelatie*
Domeniul
linear de
concentratie
(mol/L)
toff (s) Sensibilitatea
(s mol L-1)
Graphene
MD
1/ton= 0.04 + 3.62 x 1011 x C
r=0.9999 10-15 – 10-13 6.6 3.62 x 1011
CNT 1/ton= 0.10 + 6.37 x 1011 x C
r=0.9999 10-15 – 10-13 8.2 6.37 x 1011
C60 1/ton= 0.04 + 1.33 x 1010 x C
r=0.9994 10-14 – 10-12 9.2 1.33 x 1010
PATLAB
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Microsenzor
bazat pe
1 2 3 4 5 6 7 8 9 10 11 12
MD/C60 0.11 0.52 0.86 0.38 0.73 0.40 0.01 0.27 0.22 0.17 0.02 0.01
MD/CNT 0.14 0.53 0.87 0.39 0.74 0.46 0.01 0.26 0.23 0.15 0.01 0.03
MD/Graphene 0.15 0.56 0.80 0.36 0.85 0.56 0.01 0.20 0.18 0.20 0.03 0.02
ELISAa 0.12 0.56 0.87 0.45 0.74 0.56 -b 0.25 0.34 0.43 -b -b
Determinarea BPA (µg/mL) in saliva
copiilor
PATLAB
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time (sec)
0 200 400 600 800 1000 1200 1400
I (A
)
-400x10-12
-300x10-12
-200x10-12
-100x10-12
0
100x10-12
200x10-12
300x10-12
400x10-12
toff
ton
Time (sec)
0 200 400 600 800 1000 1200 1400
I (A
)
-300x10-12
-200x10-12
-100x10-12
0
100x10-12
200x10-12
300x10-12
400x10-12 toff
ton
time (sec)
0 200 400 600 800 1000 1200 1400
I (A
)
-150x10-12
-100x10-12
-50x10-12
0
50x10-12
100x10-12
150x10-12
ton
toff
MD/C60 MD/GRAPHENE
MD/CNT
Diagrame obtinute pentru determinarea BPA in saliva copiilor
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PATLAB
Test de scanare pentru DHT, E2, T2
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Caracteristicile de raspuns ale microsenzorilor stocastici
Microsenzori bazati pe DP
si
Ecuatiile de calibrare si
coeficientii de corelatie (r)
Domeniul linear de
concentratie
(mol/L)
toff
(s)
Sensibilitatea
(s/mol L-1)
DHT
P 1/ton=0.05 + 1.86 x 1010 x c
r=0.9853 10-14 – 10-12 4.0 1.86 x 1010
MD 1/ton=0.08 + 2.65 x 1011 x c
r=0.9996 10-15 – 10-13 1.8 2.65 x 1011
-CD 1/ton=0.05 + 6.92 x 108 x c
r=0.9825 10-13 – 10-11 5.1 6.92 x 108
T2
P 1/ton=0.065 + 1.27 x 106 x c
r=0.9942 10-10 – 10-8 5.3 1.27 x 106
MD 1/ton=0.061 + 1.95 x 108 x c
r=0.9989 10-12 – 10-10 3.5 1.95 x 108
-CD 1/ton=0.069 + 1.27 x 107 x c
r=0.9999 10-11 – 10-9 3.1 1.27 x 107
E2
P 1/ton=0.03 + 2.51 x 109 x c
r=0.9987 6.59 x 10-14 – 6.59 x 10-12 7.4 2.51 x 109
MD 1/ton=0.02 + 3.50 x 105 x c
r=0.9966 6.59 x 10-11 – 6.59 x 10-8 8.5 3.50 x 105
-CD 1/ton=0.06 + 1.83 x 109 x c
r=0.9976 6.59 x 10-14 – 6.59 x 10-12 5.2 1.83 x 109
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Determinarea hormonilor (pg/mL) in saliva
copiilor
aELISA (metoda standard). bELISA nu a putut determina analitul.
Microsenzor
bazat pe DP
si
1 2 3 4 5 6 7 8 9 10 11 12
F F F F F F M M M M M M
T2
-CD/DP 3.1 0.3 1.2 1.3 3.3 1.7 2.3 2.1 1.0 1.1 1.7 1.1
P/DP 3.3 0.3 2.2 1.7 3.9 1.9 2.7 1.9 1.3 1.4 1.7 1.3
MD/DP 3.2 0.3 1.9 1.1 3.3 1.1 2.2 1.8 1.1 1.2 1.6 1.5
ELISAa -b 0.7 1.1 0.4 0.6 0.9 1.0 1.4 1.1 0.5 0.6 0.3
DHT
-CD/DP 0.3 0.3 0.3 0.2 0.1 0.3 0.3 0.3 0.3 0.1 0.2 0.2
P/DP 0.2 0.2 0.4 0.2 0.1 0.4 0.2 0.2 0.2 0.2 0.2 0.1
MD/DP 0.2 0.3 0.3 0.1 0.1 0.4 0.3 0.3 0.2 0.1 0.1 0.3
ELISAa -b -b 0.2 0.4 0.2 0.5 0.4 0.7 0.8 0.4 1.1 0.3
E2
-CD/DP 1.9 2.9 1.1 1.1 2.3 2.2 1.8 1.7 0.2 1.0 0.8 0.9
P/DP 1.7 2.3 1.0 1.0 2.2 2.0 1.4 1.5 0.6 1.3 1.0 1.0
MD/DP 1.1 2.7 1.2 1.5 2.6 1.5 1.6 1.4 0.7 1.0 0.8 1.6
ELISAa 1.8 1.7 1.3 1.1 1.5 2.0 1.3 1.7 0.5 1.1 1.0 0.9
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Analiza DHT, E2, T2 in saliva utilizand microsenzorul bazat pe P/DP
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Analiza DHT, E2, T2 in saliva utilizand microsenzorul bazat pe MD/DP
DHT T2 E2
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Analiza DHT, E2, T2 in saliva utilizand microsenzorul bazat pe -CD/DP
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HEPATITA B
Scanarea sangelui pentru
hepatita B si cancer !!!
PATLAB
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Biomarkeri
• Cancer de san:
- CA15-3
• Hepatita B:
- HBvAg
- Acid D-pipecolic (D-PA)
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Caracteristicile de raspuns ale senzorului
multimode bazat pe MD/DP, determinate in
modul stocastic.
Biomarker toff
(s)
Domeniul linear de
concentratie
Sensibilitate Ecuatie de calibrared
CA15-3 3.3 5 – 500 a 44mU s/mL 1/ton= 20.11 + 0.044xC;a
R= 0.9608
HBVAg 1.9 10-10-10-8 b 3.9 x 105 g s/mL 1/ton= 0.05 + 3.9x105xC;b
R= 0.9997
D-PA 4.9 10-12-10-10 c 3.0 x 108 mol s/L 1/ton= 0.02 + 3.0x108xC;c
R= 0.9488
aU/mL; bg/mL; cmol/L; d<ton> = s.
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Caracteristicile de raspuns ale senzorului
multimode bazat pe MD/DP, determinate in
modul DPV.
Biomarker E
(mV)
Linear
concentration
range
Sensitivity Equation of calibration
CA15-3 780 5 – 500 a 5pA U/mL H= 0.3 + 5.0xC;a,d
R= 0.9382
HBVAg 526 10-12- 5x10-9 b 2.9nA g/mL H= 3.1 + 2.9xC;b,e
R= 0.9976
D-PA 350 10-11-10-8 c 155.4pA mol/L H= 0.2 + 0.16xC;c,e
R= 0.9915
aU/mL; bg/mL; cmol/L; d<H>=pA; e<H>=nA
E(mV)
0.68 0.70 0.72 0.74 0.76 0.78 0.80 0.82 0.84 0.86 0.88
I(A
)
-5.0e-11
0.0
5.0e-11
1.0e-10
1.5e-10
2.0e-10
2.5e-10
3.0e-10
3.5e-10
E(mV)
0.25 0.30 0.35 0.40 0.45 0.50 0.55 0.60 0.65
I(A
)
-2.0e-10
0.0
2.0e-10
4.0e-10
6.0e-10
8.0e-10
1.0e-9
1.2e-9
E(mV)
0.1 0.2 0.3 0.4 0.5 0.6
I(A
)
0.0
2.0e-10
4.0e-10
6.0e-10
8.0e-10
1.0e-9
1.2e-9
1.4e-9
1.6e-9
1.8e-9
D-PA 1x10-12mol/L
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Analiza calitativa a biomarkerilor in probele de
sange.
Biomarker Sample 1 Sample 2 Sample 3
Stochastic
mode
Standard
method
Stochastic
mode
Standard
method
Stochastic
mode
Standard
method
CA15-3 + + + + + +
HBVAg + + + + + +
D-PA + + + + + +
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E(mV)
0.0 0.2 0.4 0.6 0.8 1.0
I(A
)
-2e-10
0
2e-10
4e-10
6e-10
8e-10
1e-9
Determinarea cantitativa a biomarkerilor direct
din sange.
Biomarker Sample 1 Sample 2 Sample 3
M1 M2 Std M1 M2 Std M1 M2 Std
CA15-3a 10.2 10.8 9.6 43.7 42.9 42.0 87.2 87.9 86.0
HBVAgb 420 419 417 327 327 324 529 528 524
D-PAc 325 326 320 219 216 211 625 627 620
CA15-3 HBVAg
D-PA
Sample 2
t(s)
0 100 200 300 400
I(A
)
-1e-10
-8e-11
-6e-11
-4e-11
-2e-11
0
2e-11
4e-11
6e-11
8e-11
1e-10 D-PA HBVAg CA15-3
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Diseminarea rezultatelor
Capitol in carte:
Mechanism of potential development for potentiometric sensors, based on modeling of interaction between electrochemically active compounds from the membrane and analyte
R.I. Stefan-van Staden
Chapter 4, pp.131-154 in
Chemical Sensors Simulation and Modeling
Volume 5: Electrochemical Sensors (Ghenadii Korotcenkov, Ed)
Published in Sensors Technology Series (Joe Watson, Ed.)
Momentum Press, LLC, New York, 2013.
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Diseminarea rezultatelor 1. Stochastic sensors based on nanostructured materials used in the screening of
whole blood for hepatitis B. R.I. Stefan-van Staden, I. Moldoveanu J Electrochem Soc, 161(2), B3001-B3005, 2014
2. New tool for screening of whole blood for early detection of breast cancer antigen (CA153). R.I. Stefan-van Staden, J.F. van Staden J. Mod.Med.Chem., 1(2), 86-91, 2013
3. Enantioselective Surface Plasmon Resonance Sensor Based on C60 Fullerene-Glutathione Self-Assembled Monolayer (SAM). R.I. Stefan-van Staden. Chirality, 26(3), 129-131, 2014
4. New Multimode Sensors based on Nanostructured Materials for Simultaneous Screening of Biological Fluids for Specific Breast Cancer and Hepatitis B Biomarkers. R.I. Stefan-van Staden, I. Moldoveanu J Electrochem Soc, 161(4), B45-B48, 2014
5. A new genetic test for obesity, based on stochastic sensors. Raluca-Ioana Stefan-
van Staden, Livia Alexandra Gugoasa. J Electrochem Soc, 161(9), B167-B170, 2014
6. Screening of children saliva samples for bisphenol A using stochastic, amperometric and multimode microsensors. R.I. Stefan-van Staden, L.A. Gugoasa, B. Calenic, J.F. van Staden, J Legler Analytical Chemistry Research, 1, 000, 2014
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Diseminarea rezultatelor
1. Analysis of L-thyroxine and 3,3’,5-triiodo-L-thyronine using potentiometric
microsensors. Iuliana Moldoveanu, Raluca-Ioana Stefan-van Staden, Jacobus
Frederick van Staden, Gabriel Lucian Radu U.P.B. Sci. Bull., In Press.
2. Amperometric microsensors based on inulins for the assay of L-T3 and L-T4.
Livia Alexandra Gugoasa, Raluca-Ioana van Staden, Grzegorz Bazylak,
Jacobus Frederick van Staden, Gabriel-Lucian Radu U.P.B. Sci. Bull., In Press.
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Implicarea tinerilor
• Iuliana Moldoveanu – doctorand
• Livia Alexandra Gugoasa – doctorand
• Ionela Raluca Comnea – doctorand
• Ramona Georgescu - doctorand
• Dr Bogdan Calenic – postdoc
• Oana Elena Stoica – asistent tehnician
• Anita Girbea – masterand
• Radu Constantin Iacomin - masterand