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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
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.
Test de scanare pentru obezitate
PATLAB
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
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
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
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
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
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
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
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
PATLAB
Test de scanare pentru DHT, E2, T2
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
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
Analiza DHT, E2, T2 in saliva utilizand microsenzorul bazat pe P/DP
Analiza DHT, E2, T2 in saliva utilizand microsenzorul bazat pe MD/DP
DHT T2 E2
Analiza DHT, E2, T2 in saliva utilizand microsenzorul bazat pe -CD/DP
HEPATITA B
Scanarea sangelui pentru
hepatita B si cancer !!!
PATLAB
Biomarkeri
• Cancer de san:
- CA15-3
• Hepatita B:
- HBvAg
- Acid D-pipecolic (D-PA)
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.
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
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 + + + + + +
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
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.
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
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.
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