@PROFDOCDIGITALLIBRARIES
PR
CLINICAL CHEMISTRY
A REVIEWER
O
FD
O
C
BSMT STUDENT | ST. DOMINIC COLLEGE OF ASIA
,@PROFDOCDIGITALLIBRARIES
CLINICAL CHEMISTRY 1
LABORATORY AUTOMATION AND
COMPUTER SYSTEMS
OUTLINE
1. Automation
a. Continuous Flow Analyzer There are two basic approaches to automation in use
b. Discrete Analyzer today.
2. Analytical Methods
a. Spectrophotometry a. CONTINUOUS FLOW ANALYZERS
o Absorption
▪ use liquid reagents pumped through a continuous
Spectrophotometry
o Atomic Absorption system of tubing. Each sample is introduced in a
Spectrophotometry sequential manner.
b. Nephelometry ▪ Uses air bubbles to separate, samples are being
c. Turbidimetry dispense in a glass tubing and are separated by
d. Fluorometry bubbles. When they meet in the reaction chamber
PR
e. Electrochemistry with the reagents and incubation is done, then
f. Potentiometry analytic methods will be read (e.g photometric
g. Coulometry analysis, reflectance photometry, Scintillations)
h. Amperometry
3. Osmolality
a. Colligative Properties
4. Electrophoresis
O
a. Protein Electrophoresis
b. Isoenzyme Electrophoresis
5. Immunoassay
FD
6. Chromatography
a. Gel Permeation/Filtration
b. Adsorption Chromatography b. DISCRETE ANALYZERS
c. Partition Chromatography ▪ house samples and reagents in separate
d. Ion-Exchange Chromatography containers. Multiple tests can be performed on a
e. Column Chromatography single sample (random access analysis), or one
O
f. Planar Chromatography test can be selected to perform on multiple
g. Thin-Layer Chromatography samples (batch analysis). Also, parallel testing
h. High Performance Liquid and sequential testing.
C
Chromatography ▪ Random Access Analysis: Most important. Any
i. Gas Chromatography
type of samples, STAT samples can be prioritized
in any running tests.
AUTOMATION
In the clinical chemistry laboratory
context is the mechanization of chemical
analysis to minimize manual
manipulation. For example, one chemistry
analyzer uses a dry slide technology for
sample handling and measurement,
whereas another uses a closed-system
cuvette for holding and mixing sample and
reagent.
, @PROFDOCDIGITALLIBRARIES
REFLECTANCE PHOTOMETRY ➢ Colored glass filters – only selects the color
of light and filter.
Automations that produce less waste because ➢ Interference filters
polyabsorbent material is used and doesn’t ➢ Prisms – made up of quarts or glass that
use water. Also known as DRY CHEMISTRY. refracts the light. Emit rainbow color.
Wherein you used reagent pads for samples ➢ Diffraction gratings – groves that diffracts
then you are detecting the reflecting light
the light.
rather than the absorbed light.
3. The sample cell contains the solution in:
➢ Cuvettes
➢ Tubing (typical in automated equipment)
LABORATORY INFORMATION SYSTEM ➢ Plastic packs
4. The photodetector converts radiant energy to
LIS) is a system of computer software electrical energy. Three types of photodetectors
designed to handle laboratory data. The
are:
functions of an LIS include:
➢ Photocell (barrier-layer cell)
▪ Database of patient information ➢ Phototube – light bulb that has cathode and
▪ Compilation of specimen test results – anode.
PR
important for DELTA CHECKING ➢ Photomultiplier tube – multiplies radiant
(history of patient, compare past and energy detected, very sensitive.
present reports) ➢ Photodiode – detects multiple wavelengths,
▪ Production of patient reports excellent linearity. In analytes that has many
▪ Production of ancillary reports isoenzymes, absorbs same wavelength but
▪ Data storage
O
emits diff kind of light.
An LIS achieves its function via a central
computer, a number of input/output devices,
FD
and the computer software.
ANALYTICAL METHODS
a. SPECTROPHOTOMETRY
O
▪ A SPECTROPHOTOMETER is an
instrument that measures the transmitted
light of a solution and allows the operator to
C
read the absorbance of the solution on a
meter.
▪ The components of a spectrophotometer SINGLE BEAM SPECTROPHOTOMETER
include the following:
1. The light source provides radiant energy. ➢ One sample cell and reference cell. Read for the
➢ Tungsten lamps are the typical source reference or standard.
in most spectrophotometers. Visible in DOUBLE BEAM SPECTROPHOTOMETER
near eye, near infrared.
➢ Deuterium (and hydrogen) lamps are ➢ Two photodetector or chopper (reflecting mirror)
used in spectrophotometers that that splits of the incident light that is being read
examine the ultraviolet (UV) spectrum. by the monochromator. Calibrate before
Isotope of Hydrogen proceeding on reading the sample.
➢ Mercury lamps are used in high-
performance liquid chromatography ▪ Photons – Particles of light or Electromagnetic
spectrophotometers. radiant energy is described in terms of wavelike
2. The monochromator isolates the properties, specifically as photons, which are
wavelength of interest. Examples include: discrete packets of energy traveling in waves.
, @PROFDOCDIGITALLIBRARIES
▪ Wavelengths - A wavelength (λ) of ABSORPTION SPECTROPHOTOMETRY
electromagnetic energy is the linear distance ▪ is defined as the measurement of radiant
between successive wave peaks and is energy absorbed by a solution. This
usually measured in units of nanometers measurement can be related to the
(10−9 m). concentration of a substance in the solution.
▪ Frequency is the number of wave peaks per ➢ Every solution has an ability to absorb and
given unit of time. The higher the frequency, transmit light, and only transmitted light can
the shorter the wavelength. be measured.
▪ Amplitude is the height of the peak. ➢ Transmittance is defined as the proportion
of incident light that is transmitted and is
usually expressed as a percentage:
%T = I/I0 × 100
where I is the transmitted radiant energy, and
I0 is the original incident radiation.
Transmittance varies inversely and
logarithmically with the concentration of the
solution.
PR
➢ Absorbance is calculated as follows:
A = 2 − log% T
The absorbance is the critical measure
used in the calculation of concentration
▪ The electromagnetic spectrum has a large (Beer’s law).
O
range of wavelengths. Gamma rays and x-
rays have very long wavelengths, whereas ATOMIC ABSORPTION
UV rays inhabit the portion of the SPECTROPHOTOMETRY
electromagnetic spectrum from 10 to 400 ▪ Measures concentration through the detection
FD
nm. The visible spectrum lies between 400 of absorbance of electromagnetic radiation by
and 800 nm. Violet light has the longest atoms instead of molecules. It is used to
wavelength of the visible spectrum, followed measure concentration of metals that are not
by blue, green, yellow, orange, and red easily excited. If it is molecules then it is easily
(VIBGYOR; ROY G. BIV). The infrared excited so it is easily destroyed.
O
spectrum lies above 800 nm, and the ▪ Principle. An element of interest is
shortest wavelengths are microwaves. dissociated (e.g calcium can be detected
➢ Excitation - Interactions of light with easily by atomic absorption because it is not
C
matter occur when a photon intercepts bound, while ionized calcium will need to be
an atom, ion, or molecule. The photon is disintegrated first, also called as atomizers or
absorbed, and the energy of the photon spray/aerosols) from its chemical bonds in the
changes the matter (excitation). flame; then it is in an unexcited state. At this
low energy, the atom can absorb radiation at a
TYPES OF SPECTROPHOTOMETRY narrow specific bandwidth. A wavelength of
light (emitted by a light source) specific for the
BEER’S LAW states that the concentration atom is absorbed by the low-energy atoms in
of a substance is directly proportional tothe the flame, resulting in a decrease in the
amount of radiant energy absorbed: intensity of the light measured by the detector.
▪ Components
A = abc or ebc
1. The light source (hollow cathode lamp) –
where a (or e) is molar absorptivity (a eg. lead, iron
constant for a given molecule); b is the 2. Flame (produced by a burner head) –
length of the path traveled by the light; and c disassociates aerosols from atomizers
is the concentration of absorbing molecules. 3. Monochromator
4. Photodetector (photomultiplier tube)
PR
CLINICAL CHEMISTRY
A REVIEWER
O
FD
O
C
BSMT STUDENT | ST. DOMINIC COLLEGE OF ASIA
,@PROFDOCDIGITALLIBRARIES
CLINICAL CHEMISTRY 1
LABORATORY AUTOMATION AND
COMPUTER SYSTEMS
OUTLINE
1. Automation
a. Continuous Flow Analyzer There are two basic approaches to automation in use
b. Discrete Analyzer today.
2. Analytical Methods
a. Spectrophotometry a. CONTINUOUS FLOW ANALYZERS
o Absorption
▪ use liquid reagents pumped through a continuous
Spectrophotometry
o Atomic Absorption system of tubing. Each sample is introduced in a
Spectrophotometry sequential manner.
b. Nephelometry ▪ Uses air bubbles to separate, samples are being
c. Turbidimetry dispense in a glass tubing and are separated by
d. Fluorometry bubbles. When they meet in the reaction chamber
PR
e. Electrochemistry with the reagents and incubation is done, then
f. Potentiometry analytic methods will be read (e.g photometric
g. Coulometry analysis, reflectance photometry, Scintillations)
h. Amperometry
3. Osmolality
a. Colligative Properties
4. Electrophoresis
O
a. Protein Electrophoresis
b. Isoenzyme Electrophoresis
5. Immunoassay
FD
6. Chromatography
a. Gel Permeation/Filtration
b. Adsorption Chromatography b. DISCRETE ANALYZERS
c. Partition Chromatography ▪ house samples and reagents in separate
d. Ion-Exchange Chromatography containers. Multiple tests can be performed on a
e. Column Chromatography single sample (random access analysis), or one
O
f. Planar Chromatography test can be selected to perform on multiple
g. Thin-Layer Chromatography samples (batch analysis). Also, parallel testing
h. High Performance Liquid and sequential testing.
C
Chromatography ▪ Random Access Analysis: Most important. Any
i. Gas Chromatography
type of samples, STAT samples can be prioritized
in any running tests.
AUTOMATION
In the clinical chemistry laboratory
context is the mechanization of chemical
analysis to minimize manual
manipulation. For example, one chemistry
analyzer uses a dry slide technology for
sample handling and measurement,
whereas another uses a closed-system
cuvette for holding and mixing sample and
reagent.
, @PROFDOCDIGITALLIBRARIES
REFLECTANCE PHOTOMETRY ➢ Colored glass filters – only selects the color
of light and filter.
Automations that produce less waste because ➢ Interference filters
polyabsorbent material is used and doesn’t ➢ Prisms – made up of quarts or glass that
use water. Also known as DRY CHEMISTRY. refracts the light. Emit rainbow color.
Wherein you used reagent pads for samples ➢ Diffraction gratings – groves that diffracts
then you are detecting the reflecting light
the light.
rather than the absorbed light.
3. The sample cell contains the solution in:
➢ Cuvettes
➢ Tubing (typical in automated equipment)
LABORATORY INFORMATION SYSTEM ➢ Plastic packs
4. The photodetector converts radiant energy to
LIS) is a system of computer software electrical energy. Three types of photodetectors
designed to handle laboratory data. The
are:
functions of an LIS include:
➢ Photocell (barrier-layer cell)
▪ Database of patient information ➢ Phototube – light bulb that has cathode and
▪ Compilation of specimen test results – anode.
PR
important for DELTA CHECKING ➢ Photomultiplier tube – multiplies radiant
(history of patient, compare past and energy detected, very sensitive.
present reports) ➢ Photodiode – detects multiple wavelengths,
▪ Production of patient reports excellent linearity. In analytes that has many
▪ Production of ancillary reports isoenzymes, absorbs same wavelength but
▪ Data storage
O
emits diff kind of light.
An LIS achieves its function via a central
computer, a number of input/output devices,
FD
and the computer software.
ANALYTICAL METHODS
a. SPECTROPHOTOMETRY
O
▪ A SPECTROPHOTOMETER is an
instrument that measures the transmitted
light of a solution and allows the operator to
C
read the absorbance of the solution on a
meter.
▪ The components of a spectrophotometer SINGLE BEAM SPECTROPHOTOMETER
include the following:
1. The light source provides radiant energy. ➢ One sample cell and reference cell. Read for the
➢ Tungsten lamps are the typical source reference or standard.
in most spectrophotometers. Visible in DOUBLE BEAM SPECTROPHOTOMETER
near eye, near infrared.
➢ Deuterium (and hydrogen) lamps are ➢ Two photodetector or chopper (reflecting mirror)
used in spectrophotometers that that splits of the incident light that is being read
examine the ultraviolet (UV) spectrum. by the monochromator. Calibrate before
Isotope of Hydrogen proceeding on reading the sample.
➢ Mercury lamps are used in high-
performance liquid chromatography ▪ Photons – Particles of light or Electromagnetic
spectrophotometers. radiant energy is described in terms of wavelike
2. The monochromator isolates the properties, specifically as photons, which are
wavelength of interest. Examples include: discrete packets of energy traveling in waves.
, @PROFDOCDIGITALLIBRARIES
▪ Wavelengths - A wavelength (λ) of ABSORPTION SPECTROPHOTOMETRY
electromagnetic energy is the linear distance ▪ is defined as the measurement of radiant
between successive wave peaks and is energy absorbed by a solution. This
usually measured in units of nanometers measurement can be related to the
(10−9 m). concentration of a substance in the solution.
▪ Frequency is the number of wave peaks per ➢ Every solution has an ability to absorb and
given unit of time. The higher the frequency, transmit light, and only transmitted light can
the shorter the wavelength. be measured.
▪ Amplitude is the height of the peak. ➢ Transmittance is defined as the proportion
of incident light that is transmitted and is
usually expressed as a percentage:
%T = I/I0 × 100
where I is the transmitted radiant energy, and
I0 is the original incident radiation.
Transmittance varies inversely and
logarithmically with the concentration of the
solution.
PR
➢ Absorbance is calculated as follows:
A = 2 − log% T
The absorbance is the critical measure
used in the calculation of concentration
▪ The electromagnetic spectrum has a large (Beer’s law).
O
range of wavelengths. Gamma rays and x-
rays have very long wavelengths, whereas ATOMIC ABSORPTION
UV rays inhabit the portion of the SPECTROPHOTOMETRY
electromagnetic spectrum from 10 to 400 ▪ Measures concentration through the detection
FD
nm. The visible spectrum lies between 400 of absorbance of electromagnetic radiation by
and 800 nm. Violet light has the longest atoms instead of molecules. It is used to
wavelength of the visible spectrum, followed measure concentration of metals that are not
by blue, green, yellow, orange, and red easily excited. If it is molecules then it is easily
(VIBGYOR; ROY G. BIV). The infrared excited so it is easily destroyed.
O
spectrum lies above 800 nm, and the ▪ Principle. An element of interest is
shortest wavelengths are microwaves. dissociated (e.g calcium can be detected
➢ Excitation - Interactions of light with easily by atomic absorption because it is not
C
matter occur when a photon intercepts bound, while ionized calcium will need to be
an atom, ion, or molecule. The photon is disintegrated first, also called as atomizers or
absorbed, and the energy of the photon spray/aerosols) from its chemical bonds in the
changes the matter (excitation). flame; then it is in an unexcited state. At this
low energy, the atom can absorb radiation at a
TYPES OF SPECTROPHOTOMETRY narrow specific bandwidth. A wavelength of
light (emitted by a light source) specific for the
BEER’S LAW states that the concentration atom is absorbed by the low-energy atoms in
of a substance is directly proportional tothe the flame, resulting in a decrease in the
amount of radiant energy absorbed: intensity of the light measured by the detector.
▪ Components
A = abc or ebc
1. The light source (hollow cathode lamp) –
where a (or e) is molar absorptivity (a eg. lead, iron
constant for a given molecule); b is the 2. Flame (produced by a burner head) –
length of the path traveled by the light; and c disassociates aerosols from atomizers
is the concentration of absorbing molecules. 3. Monochromator
4. Photodetector (photomultiplier tube)
