Determination of Glucose Concentration in
Whole Blood using FTIR Spectroscopy
Y.C. Shen, A.G. Davies, E.H. Linfield
Cavendish Laboratory, University of Cambridge, Cambridge CB3 0HE, UK
P.F. Taday, D.D. Arnone
TeraView Ltd, 302/304 Cambridge Science Park, Cambridge CB4 0WG, UK
T.S. Elsey
Clinical Biochemistry, Addenbrooke’s NHS Trust, Cambridge CB2 2QR, UK
2002-9-30
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
Background
Determination of glucose in blood
diabetes
Infrared spectroscopy
no reagents
multi-analyses from a single spectrum
no damage to the samples
automation
2002-9-30
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
Whole blood:
*45% cellular components
*55% water and dissolved solids
*Clinical level of glucose: 2-30mM (5mM, 90mg/dL)
FTIR spectroscopy
? high water absorption
? complex blood matrix
2002-9-30
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
Glucose absorption
Transimmision spectra of water from John E. Bertie (Appl. Spectr. 1996)
Transmission
Water
absorptionspectrum of water
1
FTIR 700-7500cm-1
transmission
0.8
0.6
0.4
0.2
25um
50um
200um
0
0
1000
2000
3000
4000
wavenumber (cm-1)
2002-9-30
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
5000
6000
** Samples
25mm liquid cell
** FTIR measurement
700-7500cm-1, 2cm-1 resolution
** Glucose concentration prediction
Partial Least Square (PLS) model
2002-9-30
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
Results I: Glucose solution samples
2002-9-30
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
Absorption spectrum of glucose solution (60mM)
FTIR spectrum
0
0.1
spectrum
0.2
950
best-fit
1000
1050
1100
wavenumber (cm-1)
1150
2002-9-30
Determination
of glucose in whole blood
using FTIR
The spectrum
can be well
simulated
using
eight Lorentz oscillators
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
Glucose solution samples (1-8mM)
0.3
transission
0.25
0.2
0.15
0.1
0.05
0
850
1050
1250
wavelength (cm-1)
2002-9-30
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
1450
1650
Glucose determination: PLS model
Measured (mM)
6
calibration set
test set
5
4
3
2
y = 0.999x
R2 = 0.995
1
0
0
1
2
3
4
prepared (mM)
5
6
SEC=0.073mM; SEP=0.086mM
2002-9-30
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
Sensitivity:
better than 0.1mM
Specificity?
Complex blood matrix
2002-9-30
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
Results II: Glucose solution mixtures
D-glucose
2002-9-30
D-Fructose
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
Glucose in solution mixture: PLS prediction
Prepared
Measured
Fructose
Glucose
Glucose
0.0
0.0
-0.02
8.3
7.0
6.5
11.1
7.0
6.8
13.9
7.0
6.8
16.7
7.0
6.7
19.4
7.0
6.7
22.2
7.0
6.8
2002-9-30
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
Sensitivity and specificity
Results III: Whole blood samples
2002-9-30
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
Whole blood samples (2-30mM)
0.3
transmission
0.25
0.2
0.15
0.1
Glucose absorption
0.05
0
850
2002-9-30
1050
1250
1450
wavenumber (cm-1)
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
1650
Glucose in whole blood
FTIR data (mM)
30
Calibration set
Test set
20
R2 = 0.996
10
0
0
10
20
Clinical data (mM)
SEC=0.46mM; SEP=0.58mM
2002-9-30
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
30
Compare with previous results:
Sample and Spectral range
SEP
method
(cm-1)
(mM)
ATR (liquid)
750-1500
0.9, 1.1
950-1250
0.8
Dried film
on PE card
950-1185
1.3
Liquid cell
Liquid cell
2002-9-30
964-1182
950-1200
0.81
0.59
notes
adsorption of protein on
ATR crystal
KSCN was added to
blood as a reference
(doping procedure)
learning set: 7 patients
test set: 1 patient
learning set: 14 patients
test set: 14 patient
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
Summary:
we have shown that FTIR transmission
measurements, together with a PLS model, are a
convenient and precise tool for the quantitative
determination of glucose concentrations in blood.
Results IV: The second derivative spectrum
2002-9-30
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
Glucose solution samples: (2-8mM)
10
8
6
4
2
0
2nd derivative
predicted
the second derivative spectrum
0.3
0.1
-0.1
0
y = 1.000x
R2 = 0.992
2
4
6
prepared (mM)
-0.3
950
2002-9-30
8
10
1000
1050
1100
wavenumber (cm-1)
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
1150
Blood sample from a single patient (5-50mM)
4
3
2nd derivative
6
2
2R = 0.996
4
1
2
0
0
-1
5
20
35
50
-2
-3
950
2002-9-30
1000
1050
1100
1150
wavenumber (cm-1)
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
1200
Blood sample from different patients
30
R2 = 0.979
3
2
1
20
10
0
0
0
10
20
30
glucose conc (mM)
Calibration set
2002-9-30
R2 = 0.976
FTIR (mM)
p-p value
4
0
10
20
Clinical (mM)
Test set
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
30
Conclusions
(1) The IR transmission measurements are a
convenient and precise tool for the quantitative
determination of glucose concentrations in blood
(2) The second derivative spectrum is useful for
glucose determination
2002-9-30
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
Acknowledgement
EPSRC (UK)
The Royal Society
Toshiba Research Europe Ltd.
Ms. R. Woodward
Mr. P. C. Upadhya
Mr. R. Driver
2002-9-30
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy
Thank you!
2002-9-30
Determination of glucose in whole blood using FTIR
spectroscopy, YC Shen et al., THz-bridge
Conference, 2002, Capri, Italy