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DQCM Beating the Standard Coagulometer in
the Domain of Sensitivity Range and
Information for Hemostasis of Human Plasma
Dr. Munawar Hussain
Biosensor Research Group
19 May 2014 1
19 May 2014 2
Instrumentation: qCell T
Photograph with permission from 3T Analytik
19 May 2014 3
Instrumentation: qCell T
• 10 MHz QCM (mass and visco-elastic sensitive transducer)
• Equally suitable for real time liquid and gas phase sensing
• Equipped with efficient thermostat with fast temperature control
• Powerful pumping system that eliminates any chance of bubble/
fluctuation in liquid phase analysis
• Fast and excellent baseline stability in few seconds
• The only DQCM instrument suitable for Hemostasis because of
excellent baseline stability
• User friendly software with latest features for data handling
Literature
19 May 2014 4
* Y. Chunyan, Q. Ling, F. Weiling, Sensors. 2013, 13(6), 6946.
* R. S. Lakshmanan, V. Efremov, S. M. Cullen, A. J. Killard, Sens. Act. B. 2014, 192, 23.
• Some reports on fibrinogen concentration calculation
using QCM*
• No method for cumulative study of aPTT with fibrinogen
concentration in a single set of measurement.
• No study on comparison of aPTT with standard
coagulometer.
Experimental
19 May 2014
5
VP-DVB
polymerization
under UV
λmax 350 nm
UV hardened
QCMs
aPTTqCell T
measurementsMeasurements
Spin coated QCMs coated
with thin film
UV λmax 350 nm
5000 rpm
Standard
Coagulometer
19 May 2014 6
DQCM Exemplary aPTT Curve
-750
-600
-450
-300
-150
0
150
300
450
600
0 20 40 60 80
Δ f
/H
zΔ
Γ/
Hz
t/ s
aPTT Coagulation
aPTT CoagulationDamping
Negative Control withPlasma
Negative Control withPlasma Damping
Negative Control withoutPlasma
Negative Control withoutPlasma Damping
aPTT Starter
CaCl2 Addition
aPTT Point
Total Coagulation
19 May 2014 7
aPPT for Heparin Induced Plasma
-1200
-800
-400
0
400
800
1200
1600
0 100 200 300 400 500 600
Δ f
/H
zΔ
Γ/
Hz
t/ s
Plasma
Plasma Damping
Plasma + 0.25 IU/mlHeparin
Plasma + 0.25 IU/mlHeparin Damping
Plasma + 0.5 IU/mlHeparin
Plasma + 0.5 IU/mlHeparin Damping
Plasma + 1 IU/ml Heparin
Plasma + 1 IU/ml HeparinDamping
aPTT Starter
Addition
19 May 2014 8
DQCM Vs Standard Coagulometer
25
50
75
100
125
150
25 50 75 100 125 150
tQC
M/
s
tCoag / s
Plasma
+ 0.25 IU ml-1 Heparin
+ 0.5 IU ml-1 Heparin
19 May 2014 9
aPTT for Heparin 1 IU/mL Plasma
0
50
100
150
200
250
300
1 2 3 4 5 6 7 8 9 10 11
tQC
M/
s
Samples Containing Heparin 1 IU/ml
tCoag / s Not Applicable
19 May 2014 10
Bland Altman Plot
-10.00
-5.00
0.00
5.00
10.00
25 50 75 100 125 150
tQC
M -
tCoa
g /
s
(tQCM + tCoag)/2 / s
Plasma
+ 0.25 IU ml-1 Heparin
+ 0.5 IU ml-1 Heparin
Mean-2SD
Mean
Mean +2SD
19 May 2014 11
Fibrinogen Gradient Curves for PPP
19 May 2014 12
Fibrinogen Calibration Curves
y = 20.755x2 - 30.875x - 59.939
R² = 0.9908
y = 6.4611x2 + 30.514x - 35.362
R² = 0.9983
y = 23.023x2 - 47.686x - 26.988
R² = 0.9767
y = 17.278x2 - 46.529x + 34.267
R² = 0.9757
y = -10.879x2 + 1.7767x - 158.05
R² = 0.9671
y = 1.1489x2 - 82.923x + 51.613
R² = 0.9892
y = -188.9ln(x) - 6.9266
R² = 0.9978
y = -13.209x2 + 41.728x - 146.64
R² = 0.9982
-600
-400
-200
0
200
400
600
0 1.5 3 4.5 6
Δ f
/ H
z
Δ Γ
/ H
z
Plasma Fibrinogen / (g/L)
Δ f /Δ Γ [From Start Point till aPTT Point]
Pl Poor
Pl 0.3 million
Pl 0.15 million
Pl 0.05 million
(A)
-750
-600
-450
-300
-150
0
150
300
450
600
0 20 40 60 80
Δ f
/H
zΔ
Γ/
Hz
t/ s
19 May 2014 13
Fibrinogen Calibration Curves
y = 102.37x2 - 376.61x + 514.95
R² = 0.9772
y = 51.687x2 - 63.232x + 20.824
R² = 0.9995
y = 26.626x2 + 99.158x - 59.395
R² = 0.9982 y = 70.278x2 - 378x + 778.15
R² = 0.9929
y = -33.352x2 + 94.794x - 316.41
R² = 0.9871
y = -9.4979x2 - 53.289x + 55.646
R² = 0.9923
y = -5.2748x2 - 56.587x - 15.148
R² = 0.9753
y = -20.803x2 + 152.62x - 494.77
R² = 0.9972
-1000
-500
0
500
1000
1500
2000
0 1.5 3 4.5 6
Δ f
/ H
z
Δ
Γ/
Hz
Plasma Fibrinogen / (g/L)
Δ f /Δ Γ [From aPTT Point till Total Coagulation]
Pl Poor
Pl 0.3 million
Pl 0.15 million
Pl 0.05 million
(B)
-750
-600
-450
-300
-150
0
150
300
450
600
0 20 40 60 80
Δ f
/H
zΔ
Γ/
Hz
t/ s
19 May 2014 14
Fibrinogen Calibration Curves
y = 123.12x2 - 407.49x + 455.01
R² = 0.9811
y = 58.149x2 - 32.718x - 14.538
R² = 0.9994
y = 49.649x2 + 51.471x - 86.383
R² = 0.9953
y = 87.556x2 - 424.53x + 812.42
R² = 0.9933
y = -44.231x2 + 96.57x - 474.46
R² = 0.9831
y = -8.349x2 - 136.21x + 107.26
R² = 0.9995
y = 6.8344x2 - 209.23x + 123.44
R² = 0.9802
y = -34.012x2 + 194.35x - 641.41
R² = 0.9994
-2000
-1500
-1000
-500
0
500
1000
1500
2000
2500
0 1.5 3 4.5 6
Δ f
/H
z
Δ Γ
/ H
z
Plasma Fibrinogen / (g/L)
Δ f /Δ Γ [From Start Point till Total Coagulation]
Pl Poor
Pl 0.3 million
Pl 0.15 million
Pl 0.05 million
(C)
-750
-600
-450
-300
-150
0
150
300
450
600
0 20 40 60 80
Δ f
/H
zΔ
Γ/
Hz
t/ s
19 May 2014 15
Summary:
Excellent correlation at lower and mid range aPTT with
respect to standard mechanical coagulometer
Plasma-1 IU/mL heparin mechanical coagulometer N/A.
aPTT+Fibrinogen data from a single set of measurements.
Sensing layers are stable can be reused (10 times tested)
qCell T a suitable instrument for coagulation
DQCM has bright future of relevant clinical applications
Thanks for your kind attention
19 May 2014 16
19 May 2014 17
19 May 2014 18
Fibrinogen Gradient Curves
20
30
40
50
60
70
0 1.5 3 4.5 6
aP
TT
/ s
Plasma Fibrinogen / (g/L)
[Platelets Poor Plasma]
Coagulometer
QCM
WHO Fibrinogen Standard
in Healthy Human Plasma
(A)
20
30
40
50
60
70
0 1.5 3 4.5 6
aP
TT
/ s
Plasma Fibrinogen / (g/L)
[Platelets 0.3 million]
Coagulometer
QCM
WHO Fibrinogen Standard
in Healthy Human Plasma
(B)
20
30
40
50
0 1.5 3 4.5 6
aP
TT
/ s
Plasma Fibrinogen / (g/L)
[Platelets 0.15 million]
Coagulometer
QCM
WHO Fibrinogen Standard
in Healthy Human Plasma
(C)
20
30
40
50
60
0 1.5 3 4.5 6
aP
TT
/ s
Plasma Fibrinogen / (g/L)
[Platelets 0.05 million]
Coagulometer
QCM
WHO Fibrinogen Standard
in Healthy Human Plasma
(D)
