Automation in Clinical Chemistry
Outline
• Terminology and Definitions
• Components of Automated Clinical Chemistry
Analyzer
• Advantages and disadvantages of manual
spectrophotometric analysis and automated
analyzers
• Steps for installing new analyzer
Terminology and definitions
• Automation: mechanization of the steps in a test procedure
– the process of using a machine to perform steps in laboratory
testing with only minimal involvement by the analyst
• Spectrophotometer: measuring the light of specific
wavelength transmitted through the solution in a cuvette to
determine the concentration of an chemical reaction
product or analyte.
• Calibration-to transform the instrument response
measurement to a predicted output.
• Turnaround time: time for a test result to be reported once
the laboratory receives the specimen.
Terminology and Definitions
• Random access: Different tests offered in variation of
order;
• Discrete analysis: Each test reaction in a separate
cup, cuvette or container
• Batch analyzer: Many samples analyzed quickly in a
group but perform the same tests on all samples
Definitions
• Standard: a pure solution with known concentration
– Primary
• A pure solution verified by the reference method
– Secondary
• A pure solution verified by a highly accurate method
• Calibrator
– A substance or mixture dissolved in a matrix used
to calibrate an instrument
• QC samples
– Similar in makeup to patient samples used for
verifying validity
Principle of Current Automated Systems
• Basic spectrophotometric
– AMS Autolab
– Human Humastar80
• Electrochemistry: Ion Selective Electrodes
– AMS Autolab
– Humastar
• Immunoassay
– Fluorometric or Fluorescent polarization
– Chemiluminescent
– Nephelometric
Basic Instrument Components
(Spectrophotometric analyzer)
Basic spectrophotometer detection system
components include:
1. Light sources (UV and visible)
2. Wavelength selector (monochromator)
3. Sample containers (cuvettes)
4. Detector
5. Signal processor and readout
Automated Chemistry Analyzer Components
• Sample cup holder
• Sample pump and pipettor
• Reagent pump and pipettor
• Test cuvettes: built in, flowthrough
• Mixer
• Incubator
• Detection system
– Photomultiplier tube
• Read –out device
• Waste container
Sample and reagent Pipettor and Pump
• Sample Pipettor
and Pump
– Probe or pipettor
– Wash solution
• Reagent Pump and
Pipettor
– Deliver reagent
volume accurately
Sample Cuvettes in Automation
• Cuvettes: contains test reaction where
photometer readings can be taken, it is optically
clear
– May be placed in instrument by tech
– May be built into the instrument and self-washing
– May be flow-through cuvettes
• Optical cell for test solution
• Solution in cuvette absorbs some light and transmits
remaining light based on concentration of analyte/
product.
Detector in Automation
• The photomultiplier tube
– Commonly used detector in UV-Vis spectroscopy
– Photomultiplier tubes are electron tubes that
amplify current
• Photodiode arrays
– Example of a multichannel photon detector. These
detectors are capable of measuring all elements of
a beam of dispersed radiation simultaneously
– Diodes discharge energy when they are struck by
light
Steps automated
• Automation can handle all or some of the steps of
analytical techniques:
– Specimen identification, specimen volume
measuring, sample pretreatment
– Reagent volume measuring, sample and reagent
mixing, incubation
– Reaction timing, reaction analysis, calculation, and
result presentation on a visual screen and/or in
print.
Advantages of Automation versus Manual Testing
• Sample and reagent volumes are less with
automation.
• Time (less time-consuming with automation)
– Pipetting, incubation, etc. is automated
– Turn-around time
– Frees up technician to do other work
• Human error is less; however, not error free
– Accuracy and precision of results improves
– Built in error detection with trouble-shooting guide
• Transcription errors are less with automation.
Automation
• Disadvantages compared with manual testing:
– Cost of instrumentation (lease and maintenance
contracts)
• More daily and weekly maintenance required
– Need for back-up instrumentation or retain manual
methods (get used to quick turn-around)
– Reagent suppliers (contracts)
• Open systems allow many vendors for provision of
supplies and reagents
• Closed systems allow only manufacturer supplies and
reagents which may be more costly
Semi-Automation vs Automation Chemistry Analyzers
• In Semi-automation there are steps performed by
the technologist:
– prepares sample for testing
– loads sample cup onto analyzer
– manually inputs Patient ID and Test(s) Requested
into instrument via keyboard
• In full automation:
– specimens are placed directly on the analyzer
– Barcode reader inputs Patient ID and test request
Steps for using a New Analyzer
• A new automated instrument
has just been received in the
laboratory.
• The technologist must now
set up, calibrate and validate
the instrument, comparing it
to an existing reliable method
and running QC.
Initial Instrument Setup
• Out-of-the box steps: check for:
– Visual damage
– Loose parts or connections
– All parts and accessories
– Computer boards are properly seated
• Pre-installation Steps: Check for the following items:
Proper electrical wiring and voltage, and
uninterrupted power, Temperature controlled,
Adequate distilled water supply
• Establish a working procedure
Initial Instrument Calibration
• Follow manufacturers recommendation for
reagent and control preparation
– Make a habit of reading package inserts
• Place reagents on instrument as required
• Place calibrator(s) on instrument in proper
order sequence
• Run prepared calibrator(s) on both automated
and manual methods
Instrument Calibration
• 3-5 standards are analyzed for analyte
• Absorbance is measured
• Absorbance versus concentration graph is
plotted
• Slope and intercept is determined
• Linearity is determined
• Concentration curve is stored in analyzer
– QC samples verify concentration curve
Calibration Curve
1.00
0.80
absorbance
0.60
0.40
0.20
0.00
0 20 40 60 80 100 120
concentration
Automation Analyzer Calibration
Once an analyte is proved to follow Beer's Law at a
specific wavelength (i.e. linear plot of ABS vs.
concentration with a zero intercept is obtained),
an unknown (test) concentration relates to a single
standard and is calculated in the analyzer by:
ABS STD conc STD
=
ABS test conc test
ABS test
conc test = X conc STD
ABS STD
Calibration:
• Deviations from Beer's Law: Variation from
linearity of the ABS vs. concentration curve
will occur when:
Concentration exceeds linearity
Incident radiant energy is not monochromatic
Sample absorption is significant compared to the
solute absorbance
Sample blank may be necessary
Eg. lipemia or hemolysis
Instrument calibration
• If calibration is unsuccessful, then
troubleshoot
– 1st determine if it is instrument or reagent
problem
– Verify proper preparation of calibrator
– If all trouble shootings fail, contact manufacturer
for support
Ongoing Automation Analyzer Calibration
• Determine QC results by standard curve or factor
• If controls within stated value for analyzer and
methodology, then move to instrument validation
• Test calibration will be repeated
– When there is a new lot number of reagent or
– In accordance with manufacturer’s
recommendations or
– On more frequent basis as indicated by quality
control results
Implementation Plan of new Analyzers
• Implementation only after Method Evaluation**
– Establish a working procedure [Write a Standard
Operating Procedure (SOP)]
– Establish the quality control system
– Establish reference ranges
• Healthy range, specific patient population studies
– Instruct laboratory personnel
– Introduce the method into routine use
– Monitor routine performance
• Instrument Maintenance
Instrument Maintenance
• This should be done by the technologists who
have received prior training.
• It includes for example, systematic and routine
cleaning, simple adjustment or replacement of
instrument and equipment parts.
– Daily
– Weekly
– Monthly
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