CC1: Clinical Chemistry 1

L5: Analytic Techniques (Instrumentation)│ Mr. Marlex Lee Sanchez, RMT, AMT, MSMLS
College of Our Lady of Mt. Carmel (P) – College of Allied Health Professions
Medical Technology Department │ 1st Semester – A.Y. 2023-2024

Analytic Techniques o h = constant (6.62 x 10-27 erg sec) - Planck's

• Provide the foundation for all measurements made in Constant
clinical chemistry o v = frequency
• Because frequency of a wave is inversely
Basic Disciplines (Analytical Chemistry) proportional to the wavelength, thus the energy of
1. Spectrometry electromagnetic radiation is inversely proportional to
i. Spectrophotometry (Colorimetry) wavelength.
ii. Atomic Absorption
iii. Mass Spectrometry
2. Luminescence
i. Fluorescence
ii. Chemiluminescence
3. Electroanalytic Methods
i. Electrophoresis
ii. Potentiometry
ELECTROMAGNETIC RADIATION
iii. Amperometry
4. Chromatography
i. Gas
ii. Liquid
iii. Thin layer

ELECTROMAGNETIC RADIATION
Light Energy and Radiant Energy Spectrum

ENERGY
• transmitted via electromagnetic waves that are
characterized by their frequency and wavelength
• or measure light intensity without consideration of
wavelength COLORIMETRY
• Radiant energy that passes through an object will be
partially reflected, absorbed, and transmitted. I. COLORIMETRY
• method of chemical analysis based on the
comparisons of colors produced by an unknown and
a standard solution when reacted with similar chemical
reagents.
• Two primary considerations in colorimetric analysis:
1. Quantity of color
2. Intensity of color

WAVELENGTH FORMS OF COLORIMETRY:

• the distance between two successive peaks and it
is expressed in terms of nanometer (nm
1. Visual colorimetry
• involves the comparison of an unknown solution
with a series of colored standard solutions using the
naked eye. The unknown is diluted until its color
matches the standard.
2. Photoelectric colorimetry
• measurement is done by an instrument which
measures light intensity by converting light energy to
electric energy.

PHOTOELECTRIC COLORIMETRY
• Isolation of discreet portions of the spectrum for
ELECTROMAGNETIC ENERGY purposes of measurement
• Described as photons of energy traveling in waves Principle: Selected light passing through a solution to a greater
or lesser extent strikes a photocell or phototube which generates
The relationship between wavelength and energy (E) İs current registered by a galvanometer.
described by Planck's Formula:
• E = hv
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Medical Technology Department │ 1st Semester – A.Y. 2023-2024

TYPES OF PHOTOELECTRIC COLORIMETER: 5. Sample cell /Cuvet

a. Filler Photometer
• So called because the selection of wavelength of color
is accomplished by placing a colored glass filter in
the path of the light beam. This is also called an LIGHT / RADIANT SOURCE
abridged spectrophotometer.
b. Spectrophotometer
• Involves measurement of the light transmitted by a
solution to determine the concentration of the light-
absorbing substances in the solution.
A. SPECTROPHOTOMETRY
6. Photodetector
7. Meter or Read-out device
• Provides polychromatic light and must generate
sufficient radiant energy or power to measure the
analyte of interest.
• Provides radiant energy in the form of visible or non-
visible light that may pass through the
monochromator. The light of proper wavelength is
made incident on the analytical cell.

TYPES:
A. Continuum Source: emits radiation that changes in
intensity; widely used in the laboratory.
Example:
I. Tungsten lamp (commonly used light source in the
visible and near infrared region)
II. Deuterium Lamp (provide UV radiation in analytic
spectrometers)
III. Xenon Lamps (covers both UV and visible region)

B. Line Source: emits limited radiation and wavelength

TWO TYPES OF SPECTROPHOTOMETERS:
A. Single-beam spectrophotometer Example:
• Simplest type of absorption spectrometer I. Mercury and Sodium Vapor Lamps (UV and visible
• It is designed to make one measurement at a time at region)
one specified wavelength II. Hollow Cathode Lamp (AAS)
B. Double-beam spectrophotometer III. LASER
• An instrument that splits the monochromatic light into ENTRANCE SLIT
two components - one beam passes through the
sample, and the other through a reference solution or
• It minimizes unwanted or stray light and prevents
the entrance of scattered light into the monochromator
blank.
system
• The additional beam corrects for variation in light
source intensity Stray Light
1. Double-beam-in-space: Uses 2
photodetectors; for the sample beam and
• Refers to any wavelengths outside the band
transmitted by the monochromator, it does not
reference beam
originate from the polychromatic light source; it causes
2. Double-beam-in-time: Uses 1 photodetector
absorbance error
and alternately passes the monochromatic light
through the sample cuvet and then reference
MONOCHROMATOR
cuvet using a chopper or rotating sector mirror
• A system for isolating radiant energy of a desired
BASIC COMPONENTS OF SPECTROPHOTOMETER wavelength and excluding that of other wavelengths.

TYPES OF MONOCHROMATORS
1. Prisms
• A wedge-shaped piece of glass or quartz
which separates white light into a continuous
spectrum by refraction, that is, shorter
wavelengths are bent or refracted, more than
longer wavelengths.
2. Diffraction grating
• Most commonly used; better than prism
Parts:
made by cutting grooves (parallel grooves)
1. Light/Radiance Source or slits into an aluminized surface of a flat
2. Entrance Slit piece of crown glass - Wavelengths are bent
3. Monochromator as they pass a sharp Corner
4. Exit Slit

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3. Filters d. Photodiode
• Simple; least expensive, not precise but • It is not as sensitive as PMT but with
useful. excellent linearity: most useful as a
• Made by placing a semi-transparent silver simultaneous multichannel detector
films on both sides of a dielectric such as
magnesium fluoride METER OR READ-OUT DEVICE
4. Holographic gratings • It displays output of the detection system

EXIT SLIT TYPES OF METERS:

• It controls the width of light beam (bandpass)
• Allows only a narrow fraction of the spectrum to
reach the sample cuvette
Bandpass
• the total range of wavelengths transmitted
CUVET
• Also called absorption cell / analytic cell / sample
cell
• It holds the solution whose concentration is to be
measured
TYPES OF CUVET:
a. Borosilicate glass - suitable for measurement in the
visible region.
b. Quartz or silica - suitable for measurements below
340 nm.
c. Plastic - designed for disposable, single use
application.
d. Soft glass
PHOTODETECTOR
• Electron tubes capable of amplifying current by
converting transmitted energy into an equivalent
amount of photoelectric energy.
• Detects the amount of light that passes through the
sample in the cuvet.
TYPES OF DETECTORS:
a. Barrier-layer cell or Photocell or Photovoltaic cell
• Composed of a film of light sensitive
material like selenium on iron plate with
transparent layer of silver.
• It requires no external voltage source.
b. Photoemissive tube or Phototube
• Possess a photosensitive material that
gives off electrons when light energy strikes
it.
• It is consisting of two electrodes (cathode
and anode) sealed in an evacuated glass to
prevent scattering of photoelectrons by
collision with gas molecules.
• It requires an outside voltage for operation.
a. Direct reading systems - the output of the photocell
is used to drive a sensitive meter directly with no
further amplification.
b. Null point system - the output of the detector is
balanced against the output of a reference circuit.
c. Digital readouts - provide a visual numerical display
of absorbances or converted values of concentrations.
Examples:
• galvanometer
• ammeter
• light-emitting diode (LED) display
BEER'S LAW
• This law forms the mathematical basis of
colorimetry.
• It states that the concentration of a substance is
directly proportional to the amount of light absorbed
or inversely proportional to the logarithm of the
transmitted light.
• It may also be stated that the absorbance (A) or the
optical density (O.D.) of any colored solution is equal
to the products of the molar absorptivity E (fraction of
a specific wavelength of light absorbed by a given type
of molecule); b is the length of light path through the
solutions; and c the concentration of absorbing
molecules.
• The mathematical expression of Beer's Law is as
follows:
o A = E x b x c or 2-log%T
% Transmittance
• Ratio of the radiant energy transmitted (T), divided by
the radiant energy incident on the sample (I).

c. Photomultiplier tube (PMT) BLANKING TECHNIQUE

• Has a series of electrodes to internally • A blank is a sample that contains everything except
amplify the photosignal before leaving the for the analyte of interest.
tube.

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• Reagent blank corrects absorbance caused by the
color of the reagents - the absorbance of reagents is
automatically subtracted from each unknown reading.
B. FLAME EMISSION PHOTOMETRY (FEP)
• It measures the light emitted by a single atom
burned in a flame
Principle: excitation of electrons from lower to higher energy
state
• Light source: Flame (also serves as the cuvet)
• Method: Indirect Internal Standard Method
• Internal standard: Lithium/Cesium - corrects
variations in flame and atomizer
• Flickering light: Indicates changes in the fuel reading
of the instrument
• Flame photometry: It is used in clinical chemistry for
the determination of electrolytes.
The following electrolytes produces the following colors:
1. Sodium - yellow
2. Potassium - violet
3. Lithium - red
4. Magnesium - blue
5. Calcium - red
6. Rubidium - red
COMPONENTS OF EFP:
1. Aspirator - draws sample into flame.
2. Atomizer - breaks up the solution into finer
droplets so that the atom will absorb heat energy from
the flame and get excited.
• Settling agent - minimizes changes in
atomizer flow rate due to differences in
viscosity of the sample.
• Viscosity effect is reduced by a dilution of
1:100 or 1:200.
3. Flame - provides energy for excitation.
C. ATOMIC ABSORPTION SPECTROMETRY (AAS)
COMPONENTS OF AAS:
1. Hollow cathode lamp as light source
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• Contains argon or neon gas at a pressure of a
few millimeters of mercury. An argon-filled lamp
produces a blue to purple glow during operation
and the neon produces a reddish-orange glow
inside the lamp.
• Quartz or special glass that allows transmission
of the proper wavelength is used as a window.
2. Burner - for absorption of energy.
3. Mechanical rotating chopper or nebulizer -
modulates the light beam coming from Hollow
Cathode Lamp and sprays the Sample into the
flame.
4. Prism or Diffraction gratings as monochromator
5. Photomultiplier
6. Meter
INTERFERENCE IN AAS
1. Chemical interference
• Refers to the situation when the flame cannot
dissociate the sample into free atoms so that
absorption can occur (e.g., calcium-phosphate
complexes).
Remedy: Extraction techniques and the introduction of
competing cations to release the element to be measured from
complexing or chelating anions.
2. Ionization interference
• Results when atoms in the flame become excited,
instead of only being dissociated and then emits
energy of the same wavelength that is being
measured.
Remedy:
a. Addition of easily ionized substance that will absorb
most of the flame energy so that the substance of
interest will not become excited.
b. Reduction of flame temperature.
3. Matrix interference may be due to:
a. Enhancement of light absorption by organic solvents.
b. Light absorption caused by formation of solids from
sample droplets as the solvent is evaporated in the
flame.
c. Refractory oxides of metals formed in the flame.
II. VOLUMETRIC (Titrimetric)
• This involves the determination of a substance by
reacting it with a measured volume of known
standardized solution. This process is commonly
called titration.
o V1 x C1 = V2 x C2
Where:
• V1 - volume of titrant (ml) required to reach the end
point
• C1 -known concentration of titrant expressed in
equivalents/L or mEq/L
• V2 - volume of unknown (ml) to be measured
• C2 - calculated concentration of unknown in
equivalents/L or mEq/L
Examples:
I. Schales and Schales Method (Chloride Test)
Clinical Chemistry 1
Medical Technology Department │ 1st Semester – A.Y. 2023-2024

II. EDTA Titration Method (Calcium Test) V. ELECTROPHORESIS

III. TURBIDIMETRY

• Turbidimetric measurement determines the amount of

light blocked (reduction of light) by a particulate
matter as light passes through a cuvet.
• A light blocked by a suspensions of particles ina
cuvet depends on:
a. Number of particles present
b. Cross sectional area of each particle
c. Depth of tube
• Solutions requiring quantitation by turbidimetry are
measured using visible photometers or visible
spectrophotometer
• Use:
o protein measurements (CSF, Urine)
o bacterial growth in broth cultures
o antimicrobial test (broth method)
o and to detect clot formation
• It refers to the migration of charged solutes or
IV. Nephelometry particles in liquid medium under the influence of an
electrical field.
• It separates proteins on the basis of their electric
charge densities
• The acidic and basic amino acids determine the net
charge on a protein (electrophoretic mobility)
• During electrophoresis, proteins are negatively
charged (anions) and they move towards the anode

Terms Associated with Electrophoresis:

1. Iontophoresis - migration of small ions.
2. Isotacthophoresis - a technique in which sample
components ultimately separate into adjacent zones
that all migrate at the same rate.
3. Isoelectric point (pl) - pH where net charge on a
molecule is zero.
• Nephelometric measurement determines the amount 4. Ampholytes or zwitterion - a molecule which can
of light scattered by the small particles or colloids either be positively or negatively charged.
in the sample cuvet. 5. Anode - a positive electrode where negative charge
particles (usually acidic) migrate.
Variables to control this method are: 6. Cathode - negative electrode where positive charge
a. Number of the particles particles (usually basic) migrate.
b. Size of the particles
c. Wavelength of the incident light Components of Electrophoresis System:
1. Support medium - site of electrophoretic separation;
• Light scattered by particles is measured at an angle, hold the sample and provide a path for migration of proteins;
typically 15-90 degrees to the beam incident on the act either as sieve (separation on basis of size) or charge
cuvet in system (separation on basis of net electric charge).
• Nephelometry is used in the: • e.g., agarose (electrical charge), cellulose acetate
o determination of lipoproteins and (molecular size), polyacrylamide gel (charge and
triglycerides; molecular size)
o Antigen- antibody complexes 2. Electrophoresis chamber
3. Power supply - converts alternating ine current for the
operation of the system.

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4. Detecting system FORMS OF CHROMATOGRAPHY

Use: serum proteins (applicable to CSF) 1. PLANAR - mixtures are separated on a planar surface of the
stationary phase.
Stains for Visualization of Fractions (Bands): a. Paper Chromatography
1. Amido black • The stationary phase is a layer of water or a
2. Ponceau S polar solvent coated onto the paper fibers.
3. Oil Red O
• The type of paper used is selected based on
4. Sudan Black
its homogeneity, wet strength, thickness and
5. Fat Red 7B
level of impurities and on the migration rate of
6. Coomassie Blue
mobile phase through it.
7. Gold/Silver stain – very sensitive even to nanogram
quantities of proteins • Used for tractionation of sugar and amino
acid Sorbent: Whatman paper (stationary
Methods of Quantitating Electrophoretic Samples: phase)
1. Densitometry - measurement of amount of light passing b. Thin-layer chromatography (TLC)
through the fractions. • A thin layer of the support particles is spread
2. Elution method - involves cutting the support into zones on a flat plate of glass or plastic.
containing the individual fractions and eluting the adsorbed • Using a capillary tube, syringe or mechanical
dye by means of suitable solvents such as basic buffers, applicator, aliquots of samples are applied on
weak alkali (0.1 mol/L NaOH) or alcohol solutions. the plate along a line that is parallel to the
bottom edge of the plate and a few
Types Of Electrophoresis: centimeters above it.
I. Agarose Gel Electrophoresis (AGE) • Used: Semiquantitative drug screening
II. Cellulose Acetate Electrophoresis (CAE) test; TDM
III. Polyacrylamide Gel Electrophoresis (PAGE) • Sorbent: thin plastic plates impregnated with
IV. Starch Gel Electrophoresis a layer of silica gel or alumina
c. High-performance thin layer chromatography
New Approaches to Electrophoresis: (HPTLC)
A. Isoelectric focusing (IEF) – separates molecules by • Uses smaller stationary phase particles,
migration through pH gradient thinner layer, and controlled flow rates.
B. Two-dimensional (2-D) electrophoresis
C. Capillary electrophoresis

VI. CHROMATOGRAPHY
• It is used to separate or purify small amounts of
closely related compounds from one another in a
mixture.
• This is based on one or more of the four
physicochemical principles of absorption, partition,
ion-exchange and exclusion (molecular sieving).

A chromatographic system consist of a

MIXTURE OF PHASES:
1. Mobile Phase - a liquid or gas moves through the
system.
2. Stationary Phase - a liquid or solid is fixed or
motionless during the process of chromatography.

BASIS OF SEPARATION:
1. Rate of diffusion
2. Solubility of the solute
3. Nature of the solvent
4. Sample volatility/solubility
5. Distribution between 2 liquid phases 2. COLUMN - a tube or column is fitted with particles of the
6. Molecular size (molecular sieving stationary phase and the mobile phase is passed through the
7. Hydrophobicity of the molecule resultant chromatographic bed.
8. Ionic attraction a. Gas Chromatography
9. Differential distribution between two immiscible liquids • a form of chromatography in which volatile
10. Selective separation of substance solutes are separated by passing them
11. Differences in adsorption and desorption of solutes through a column of stationary phase using

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an inert gas (e.g., nitrogen, helium, • Requires interface methods to convert nonvolatile to
hydrogen, or argon) as the mobile phase. volatile compound’s
• Because the gaseous mobile phase "carries" Ex: electrospray (ES) and Atmospheric Pressure Chemical
the solute molecules through the column, it is Ionization (APCI)
often referred to as the carrier gas. Use: TDM, toxicology, studies of drug metabolites
Types:
1. Gas-solid chromatography - the stationary phase is
particles of sorbent that have large surface area.
2. Gas-liquid chromatography - a non-volatile liquid is
coated onto particles of column packing or directly on
the wall of a chromatographic column.

MASS SPECTROMETRY (MS)

• Measuring the mass-to-charge ratio (m/z) of one or
more molecules present in a sample
• Used to identify unknown compounds via
molecular weight determination, to quantify known
compounds, and to determine structure and
chemical properties of molecules.
• Consequently, it is an extremely useful instrument for
determining the elemental composition and structure of
both inorganic and organic compounds.

GAS CHROMATOGRAPHY/MASS SPECTROMETRY

(GC/MS)
• GC/MS is a powerful analytical technique that
combines the resolving power of the gas
chromatograph with the exquisite specificity and
sensitivity of the mass spectrometer.
1. Gel/Gel Permeation / Gel Filtration / Size Exclusion /
• GC/MS is used primarily for the analysis of drugs
(gold standard for drug testing).
Separation Mechanisms Used in Liquid Chromatography
Molecular Sieve Chromatography
• Separates molecules based on difference in their size
and shape
A. Hydrophilic Gel (Gel Filtration) - separates
enzymes, antibodies and proteins (dextran,
agarose)
B. Hydrophobic Gel (Gel Permeation) - separation
of TAG and FA (sephadex)

2. Ion exchange Chromatography

• The differences in the sign and magnitude of ionic
b. Liquid Chromatography charges are the basis for separation.
• Based on the distribution of solutes • This technique is most useful for separation of
between a liquid mobile phase and a inorganic ions, amino acids, nucleotides and
stationary phase. proteins.

Types: 3. Partition Chromatography (Liquid-Liquid

1. High Performance liquid Chromatography (HPLC) -
it uses pressure for fast separations, controlled
temperature, in-line detectors and gradient elution
Use: fractionation of drugs, hormones, Iipids, carbohydrates and
proteins separation and quantitation of various hemoglobin
associated with specific diseases
2. Liquid Chromatography - Mass Spectrometry - for
detecting nonvolatile substances in body fluids. Utilized
to confirm positive results from screening of illicit
drugs.
Chromatography)
• Separation based on the relative solubility of the
solute molecule in the stationary and mobile phase.
• For separation of TDM and their metabolites
4. Affinity Chromatography
• Separation based on interaction that occurs
between biochemical species: specific enzyme-
substrate, hormone-receptor for antigen-antibody
complexes.
• Uses the lock-and-key binding that is widely present in
biologic system

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• For separation of lipoproteins, CHO and glycated
hemoglobins; antibodies
5. Adsorption Chromatography (Liquid-Solid
Chromatography)
• Separation based on the electrostatic, hydrogen-
bonding or dispersive interactions between a
molecule and solid support or adsorbent.
• The compounds are adsorbed to a solid support such
as silica or alumina
VII. FLUOROMETRY / MOLECULAR LUMINESCENCE
SPECTROPHOTOMETRY
• the determination of the characteristics and amount
of luminescence produced by substances when
examined under controlled conditions.
• Fluorescence is the property of some chemical
compounds to absorb light energy and then re-emit
some of this energy in light of a longer wavelength than
the light originally absorbed.
Principle: It determines the amount of light emitted by a
molecule after excitation by electromagnetic radiation
• Light source: mercury arc or xenon lamp
• Light detector: PMT or phototube
• Uses 2 monochromators (either filters, prisms or
gratings) - the wavelength that is best absorbed by the
solution to be measured is selected by the primary
filter: the incident light is prevented from striking the
photodetector by the secondary filter.
Use: porphyrins, Mg, Ca, and catecholamines
Fluorometry
amounts of energy lost in the form of photons
when electronically excited product molecules
relax to their stable ground state.
Principle: The chemical reaction yields an electronically
excited compound that emits light as it returns to its ground
state, or that ransters its energy to another compound,
which then produces emission
• Light is created from a chemical or electrochemical
reaction, and not from absorption of electromagnetic
energy
• Photodetector: PMT (luminometer)
Use: immunoassays
• Involves the oxidation of an organic compound
(dioxetane, luminol, acridinium ester) by an oxidant
(hydrogen peroxide, hypochlorite or oxygen) which
occur in the presence of catalyst such as enzymes
(alkaline phosphatase, horseradish peroxidase), metal
ions (CU2+ or Fe3+) and hemin
IX. OSMOMETRY
• a technique for measuring the concentration of a solute
particle which, in turn, is related to the osmotic
pressure of a solution.
• Measurement of an aqueous solution such as serum,
plasma, or urine
Principles: it based on measuring changes in the colligative
properties of solutions that occur owing to variations in particle
concentration

Colligative properties:
1. Osmotic pressure
2. Vapor pressure
3. Boiling point
4. Freezing point

TYPES OF OSMOMETER:
1. Vapor pressure osmometer
• Osmolality measurement is not related directly to a
change in vapor pressure but to the decrease in the
dew point temperature of the pure solvent
• Caused by the decrease in vapor pressure of the
solvent by solutes
2. Colloid osmotic pressure osmometer (COP)
• Directly measures the contribution of
macromolecules to the osmolality.
VIII. CHEMILUMINESCENCE 3. Freezing point depression osmometer or Cryoscope
• The emission of light that occurs as a result of
certain chemical reactions that produce high
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• Used to determine the concentration of osmotically 2. COULOMETRY

active particles in physiological fluids because its • Is the measurement of the amount of electricity (in
measurement is simple and convenient. coulombs) at a fixed potential
• Is an electrochemical titration in which the titrant is
X. ELECTROCHEMISTRY TECHNIQUES electrochemically generated and the endpoint is
• The measurement of current or voltage generated detected by amperometry
by the activity of a specific ion. • It follows Farady's law
• Electrochemistry is concerned with chemical • Use: Chloride Test (CSF, serum and sweat)
transformations that involve proton or electron transfer • Interference: bromide, cyanide, cysteine
and result in the flow of electricity.
• Analytical electrochemistry for the clinical laboratory
includes
A. potentiometry,
B. amperometry,
C. coulometry,
D. voltammetry,
E. conductometry and
F. polarography.

Generally, amperometry and potentiometry are the most

commonly used techniques and these centers on blood gas
measurements.
1. POTENTIOMETRY
• Potentiometry is the measurement of the electrical
potential difference between two electrodes in an
electrochemical cell.
• An electrochemical or galvanic cell always consists of
two electrodes (electron or metallic conductors) that
are connected by an electrolyte solution (ion
conductor).
• Follows the Nernst Equation
• Reference electrode: calomel and silver-silver
chloride
• Use: pH and pC02 tests
THE ION SELECTIVE ELECTRODE (ISE)
• The ion selective electrode is carried out using an
electrode designed specifically for the detection of
the component of interest.
• Selectivity is created in part by the use of porous
organic polymer (membrane/barrier) which allows only
one ion of a specific size to pass through.
• A wide variety of polymeric materials are employed in
these electrodes, either as simple membranes to
separate ions or as part of an ion complexing. liquid
membrane system.
Types of ISE
1. Direct ISE (without sample dilution)
2. Indirect ISE (with sample dilution)
ISE Membrane:
A. Glass aluminum silicate (sodium)
B. Valinomycin gel (potassium)
C. Organic liquid membrane ion exchangers (calcium
and lithium)
D. Gas and enzyme electrodes
3. AMPEROMETRY
• the measurement of the current flow produced by
an oxidation-reaction
• Use: pO2, glucose, chloride, and peroxidase
determinations
4. POLAROGRAPHY
• The measurement of differences in current at a
constant voltage
• It follows the llkovic equation
5. VOLTAMMETRY
• Measurement of current after which a potential is
applied to an electrochemical cell
• It allows sample to be pre concentrated, thus utilizing
minimal analyte
• Anodic stripping voltammetry - for lead and iron
testing (in ug/dl)
AUTOMATION
A. Continuous-Flow Analysis
• Type of analysis which uses air bubbles in the
sample and reagent streams.
• Air is injected into each stream as a series of small
bubbles which travel along the reaction system.
• The air bubbles minimize diffusion of reagents and
mixing between samples, preserving the integrity of
each individual reaction.
Disadvantage: Systems can only analyze for one constituent;
to measure another components of the instrument had to be
reconfigured for the new purpose
B. Centrifugal Analysis
• Sample and reagent are placed in a rotor. When
centrifugal force is applied, the rotor spins and causes
the two components to flow into a reaction chamber
where they combine to form a product.

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C. Dry-Slide Technology
• Slides contain all the materials necessary for a
single analysis. No reagents were needed to prepare
the slide for use.
• Reflectance Spectroscopy: Light of a selected
wavelength shines through the bottom of the. The light
beam reflects off the underside of the spreading layer
and passes through the reagent and indicator layers to
a photodetector.
o Any absorbance which takes place must
be due to the colored material formed by
reactions in the slide

D. Random Access Analysis

• Instrument analyses patient samples only for those
constituents specifically ordered and stat samples
can be carried out by momentarily interrupting the
normal sequence of patient analyses. The instrument
is also capable of incorporating new tests into the
analytical scheme.

1. Batch Testing - all samples are loaded at the same time

and a single test is conducted on each sample
2. Parallel Testing - more than one test is analyzed
concurrently on a given clinical specimen
3. Sequential Testing - multiple tests analyzed one after
the another on a given specimen
4. Open reagent system - reagents from other manufacturers
may be used
5. Closed reagent system - operator can only use
manufacturer's reagents

10 | B . L . , D . P . , L . L . , M . K . , M . I .
MT – II