CHEMICAL BIOSENSORS

Concentration of Electrical
biochemical analysts Biosensors signal

Chemical input Chemical factory Chemical output

Living body
Metabolite Waste
made-up of
food product
active cells
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Important Critical-Care Analytes
• Blood gas and related parameters
– Partial pressures (PO2 , PCO2 , pH)
– SO2 (oxygen saturation)
– Hematocrit, total hemoglobin (Hb)
• Electrolytes, Na+ , K+ , Ca++ , Cl-
• Metabolytes
– Glucose, urea
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 Blood Gas and Acid-Base
Physiology
• Requirement: Fast and accurate measurement
of PO2, PCO2, pH and SO2
• O2 is carried out as:
– 98% by hemoglobin - O2 saturation (SO2)
– 2% dissolved in plasma [ HbO2 ]
So2 (%) = x100
[total Hb]
• This determines the total amount of O2 in a
given volume of blood
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Oxyhemoglobin dissociation curve
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 Oxyhemoglobin Dissociation Curve
(ODC)
• Arterial PO2 → efficiency of alveolar ventilation.
– In young adults 90-100 mmHg
• SO2 at 38°C and PO2 60 mmHg is about 85%
• ↓ in PO2 indicates insufficient alveolar ventilation
• PCO2 indicates sufficiency of ventilation.
– In arterial blood 35-40 mmHg
– ↑ due to insufficient alveolar ventilation
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 Acid-Base Status
• Indicated by [H+] concentration as:
– pH= - log10[H+] Normal range: 7.38 - 7.4
• pH ↑ (alkalosis) due to
– metabolic (vomiting etc)
– Respiratory (↑ CO2 excretion)
• pH ↓ (acidosis) due to
– metabolic
• abnormal loss of bicarbonate,
• ↑ production of fixed acid as in diabetes
– Respiratory (↓ CO2 excretion) 6
 Types of Chemical Biosensors
• Electrochemical sensors
• Chemical fibrosensors
• Noninvasive blood gas monitoring

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 Electrochemical Sensors
• Electrodes are used for measurements
– Extremely high source impedance; 10 - 100 MΩ
– Extremely high dependence to temperature
– Low signal amplitude
– Generally, need for calibration solutions
• Measurement of pH (concentration of H+)
• Measurement of PCO2
• Measurement of PO2
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 Measurement of PH
• Use Glass electrodes that generate an electric potential
when solutions of different PH are placed on the two sides
of the membrane
• Hydrogen ion travels from high concentration to low
concentration
• A charge is generated by the electrode for every ion
travelling
• The voltmeter should be able to detect changes in the 0.1
mV range

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 Measurement of pH
60 mV/pH unit - Physiological range is 0.06 pH units → 0.1 mV
Calibration solutions with pH near 6.8 and near pH 7.9

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 Measurement of PCO2
H2O + CO2 ⇔H2CO3 ⇔ H+ + HCO3- ; [CO2] = a(PCO2) ;
a =0.0301 mmol/l per mmHg PCO2
k' =[H+][HCO3-]/[H2CO3] and [H2CO3] α [CO2]
k = [H+][HCO3-]/ a(PCO2); taking log of both sides
pH = log [HCO3-] - log k - log a - log PCO2

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 The PO2 Electrode
At cathode:
O2 + 2H2O + 4e- → 2H2O2 + 4e- → 4OH -
4OH - + 4KCl → 4KOH + 4Cl -
Hydroxyl ions are buffered by electrode
At anode: 4Ag + 4Cl - → 4AgCl + 4e-
Electrons required for the reaction produced

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 Current against polarized voltage
and oxygen concentration

1.5 1.5
Air
17%
1.0 1.0

Current, µA
Current, µA

12%

0.5 7% 0.5

1.5%

0 0.2 0.4 0.6 0.8 1.0 0 5 10 15 20

Voltage, V Oxygen concentration, %

(a) (b) 13
 Properties of the PO2 Electrode
• Polarizing voltage of 0.6 to 0.8 volt required
– Flow of external current proportional to PO2
– Sensitivity around 10 nA/mm Hg
• Calibration by two gases of known PO2
– CO2-N2 mix with no O2 (background current) to set 0
– O2-CO2-N2 mixture with known O2 concentration.

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 Chemical Fibrosensors
• Advantages:
– Small size
• multiple sensors in one catheter
– No electric hazard
– No electrical interference
– No reference electrode
• Disadvantages:
– Sensitive to ambient light
– Limited dynamic response
– Long-term stability of reagent 15
 Properties Measured
• Transmission of an incident light at a specific
wavelength through the sample and
measurement of absorption of it by the sample.
– Normally measurement is done at two different
wavelengths
• Addition of reagents to the sample and
measurement of the fluorescence (emission of
light at a different wavelength) of the reagent
that is modulated by the concentration of the
analyte in the sample.
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Fluorescence Optical pH and
PCO2 Sensors (Irreversible)

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 Basic

Emission
Acidic spectrum

pH-sensitive dye is excited at 410

and 460 nm and fluoresces at 520 nm 18
The emission spectrum of
oxygen-sensitive dye

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 Fluorescence Optical PO2 Sensor

As the concentration of oxygen in the oxygen sensitive dye increases,

more of incident light is absorbed by the oxygen leading to a decrease
in the intensity of the fluorescent light from the dye known as oxygen
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quenching.
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Noninvasive Blood Gas Monitoring
• Advantages:
– Safe
– Fast and real-time measurement is possible
– Not-painful
• Limitations:
– Measurement through the skin
– Normal gas diffusion through skin is low but
increases if skin is heated above 40°C
– Tissue properties change in time and among
individuals
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 Techniques Available
• Pulse oximetry for transcutenous So2
measurement
• Transcutenous arterial oxygen tension -
tcPo2
• Transcutenous arterial carbon dioxide
tension - tcPco2

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 Principle of pulse oximeter
(Transcutaneous Arterial Oxygen
Saturation Monitoring)

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 Two wavelength approach
• Reflective and transmission mode
• Transmission mode:
– Two wavelengths (660 and 940 nm)
– DC component is used to normalize
– AC comp. is used to represent the light absorption
• Used for continuous determination of So2 in:
– administration of anesthesia
– pulmonary function tests
– intensive care units
– Neonatal monitoring, etc..
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Non-invasive patient monitor with
ECG, BP, pulse oximetry & temp.
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A transcutaneous oxygen sensor

Heating the skin

promotes
arterializations27
 Transcutanous Arterial Oxygen
Tension (tcPo2) Monitoring
• Hyperemia (transmission of gases through)
of the skin by:drugs, cream, abrasion
(removal of dead layer by sand paper),
heating to over 40°C
• O2 that passes through the skin is detected
by amperometric techniques (conventional
Clark type Po2 electrode)
• Continuous monitoring requires frequent
change of location of electrodes (especially
in infants) to avoid skin burning 28
A transcutaneous carbon dioxide
sensor

Heating the skin

promotes arterializations
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 Transcutanous Arterial CO2
Tension (tcPco2) Monitoring
• Similar to tcPo2 monitoring but slower in
response
• CO2 that passes through the skin and pH
electrode is used to sense the CO2 concentration
• Used in neonates and adults who are not in
hypothermia to determine:
– Effectiveness of mechanical ventilation
– Effectiveness of circulation to a limb and response to
therapy 30
 Blood-Glucose Sensors
• Important in long-term management of
diabetes that approach 5% of population in
developed countries
• Normal range 80 to 90 mg/100 ml during
the first hour after meal. Increases to 300 to
700 mg/100 ml in case of insulin deficiency
→ hyperglycemia.

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 Glucose Measured By
• Electroenzymetric approach
Glucose oxidase
Glucose + O2 ———→ Gluconic acid + H2O2
– As O2 used by glucose, it's concentration in
the O2 electrode decreases
– Differential approach improves the quality of
measurement
• Optical approach
• Infrared absorption spectroscopy
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In the enzyme electrode,
when glucose is present it
combines with O2, so less
O2 arrives at the cathode

In the dual-cathode enzyme

electrode, one electrode
senses only O2, and the
difference signal measures
glucose independent of O2
fluctuations

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 Optical Approach in Measurement
of Glucose Concentration

The affinity sensor measures glucose concentration by detecting

changes in fluorescent light intensity caused by competitive
binding of a fluorescein-labeled indicator 34
The optical system for a
glucose affinity sensor uses
an argon laser and a fiber-
optic catheter

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 Attenuated Total Reflection
(ATR) and Infrared (IR)
Absorption Spectroscopy
• Due to vibrational and rotational oscillations of
molecules, every substance has a specific
resonance absorption peaks (that may overlap) -
fingerprints.
• No heating of tissue
• No effect of thickness of sample - layer
thickness more than 3 times the test wavelength
is sufficient 36
The infrared absorption spectrum of anhydrous D-glucose has
a strong absorption peak at 9.7 µm

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