CCHM 321: Clinical Chemistry 1│Lecture

2022-2023 3RD YEAR, 1ST SEMESTER Prof. Kimberly Ann Pulga, RMT, MPH

THREE PHASES:
Automation and
 Pre-analytical
Terminologies
(Preliminary Term, 4th Topic) o Sample collection, transport, and
Trans Outline: processing
Topic 1: Automation Terms • focus: collection and
A. Phases Topic 3: Chemistry
B. Advantages Analyzer Operation processing
C. Disadvantages Topic 4: Types of o There’s automated delivery of sample
Topic 2: Definition of Automatic Analyzer nowadays and automated specimen
processing
AUTOMATION o Steps:
• Have the request
 Technique, method, or system of operating or • Get sample from patient
controlling a process by highly automatic • Delivery of sample from
means, as by electronic devices, reducing collection location to facility
human intervention to a minimum • Manual: use ice pack
 The modern clinical chemistry laboratory with 2-8 degrees C
uses a high degree of automation. Many steps • Note: for send out
in the analytic process that were previously tests, test is not
performed manually can now be performed available inside the
automatically, permitting the operator to lab/hospital so will
focus on manual processes and increasing have to send it out to
both efficiency and capacity other lab
 Leonard Skeggs: first one to release the first • Earliest automated transport
auto-analyzer which is based on continuous system to be introduced:
flow technology pneumatic tube
 Analytical
 Process by which instrument or machines
o Actual test is being performed
automatically perform test that would
o Processing of test involving reagents
otherwise have to be performed manually by
and machine
the MLS
 Post-analytical
 Significantly improve reproducibility,
o Release of result to requesting
accuracy, flexibility, and turn-around time of
physician
testing
o Store result to database
 Lowers cost of laboratories and eliminates
• For delta checking and
errors as tests became standard/uniform
• To check medical history of
unlike manual processes patient in the future
 1970: first commercial centrifugal analyzer o Samples are stored for repeat testing
was introduced as a spinoff technology from when needed, and not discarded
from NASA outerspace research immediately
 It was also in 1970 when automatic clinical • Stored at a pre-determined
analyzer (ACA) was introduced by DuPont time (freezer or ref temp)
(presently known as Siemens)

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CCHM321 | BSMLS 2024 CLINICAL CHEMISTRY 1 TRANSCRIBER: CRUZ, A. M. A.
2022-2023 3RD YEAR, 1ST SEMESTER Prof. Kimberly Ann Pulga, RMT, MPH

• Depending on SOP of lab, 4. Many systems are impractical for small

discarded after 1 or 2 weeks number of samples
a. Something that should not be
ADVANTAGES:
done by small laboratories or
1. Rapid results
hospitals, don’t invest in
a. Due to elimination of manual
automated machines when
steps
patients are only less than 50 a
b. Turn-around time demands can be
day
met
2. Increasing in the number of tests DEFINITION OF TERMS
performed
3. Saves time and effort  Test repertoire
4. Errors in calculations and transcription o Number of tests that can be performed
are reduced on instrument
a. There’s variation of results in o Seen on machine’s manual
manual  Selective
b. Standardized/uniform o Only performs requested tests
5. Better precision and accuracy  Dwell time
a. Due to elimination of potential o Maintain time required to obtain
errors results after the initial sampling of the
6. Eliminates the needs for personnel specimen
increase o Minimum number of time required to
a. Staff shortages can be addressed obtain the result
b. It’s fine if personnel is only  Throughput
limited as long as there’s enough o Maximum number of samples or tests
or many machines available that can be processed in an hour
7. Economical  Cost
o Labor maintenance, reagents,
DISADVANTAGES: calibration, quality control,
1. Expensive to purchase and maintain consumables and capital
a. Price ranges to millions and needs  Test
maintenance, calibration o A list of the analytes or a tests that a
2. There may be limitations in the laboratory would be able provide for
methodology that can be used patient testing
a. There are tests that are skipped in  Workload
automated o A number of test results that are
b. In semi-automated, some generated by the laboratory during a
procedures will require manual given time
procedures  Walk-away capability
3. Laboratory personnel is often o The ability of an operator to program
discouraged from making observations the instrument to perform other tasks
and using their own judgment about while the instrument processes the
potential problems tests

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CCHM321 | BSMLS 2024 CLINICAL CHEMISTRY 1 TRANSCRIBER: CRUZ, A. M. A.
2022-2023 3RD YEAR, 1ST SEMESTER Prof. Kimberly Ann Pulga, RMT, MPH

 Bar code o All samples are loaded at the same

o A means of providing positive sample time, and a single test is conducted on
identification each sample
 Selectivity  Parallel Testing
o The lowest value that can be reliably o More than one test is analyzed
be detected by a method without concurrently on a given clinical
providing a false positive result specimen
 Specificity  Random Access Testing
o The ability to measure only the o Any test can be performed on any
analyte requested sample in any sequence
 Shelf life  Sequential Testing
o The term used to define reagent o Multiple tests analyzed one after
stability before use another on a given specimen
o Reagent stability (until when it is
stable) CHEMISTRY ANALYZER OPERATION
 Carry over/ Specimen carry over 1. Identification and Preparation
o This occurs when previous samples - Collection and preparation of sample
have a higher or lower results. This 2. Chemical Reaction
occurs in system that reuse cuvettes - Sample and reagent measurements
that are insufficiently washed after and mixing
each testing cycle. - Incubation
o Cuvettes are used in 3. Data Collection and Analysis
spectrophotometer, some are - Monitoring or Sensing the reaction
reusable and when not washed result
properly could affect the result of the - Quantitating the reaction result
next test - Visualizing the results
o The transport quantity of the analyte
or reagent from one specimen reagent IDENTIFICATION AND PREPARATION
to another can contaminate the 1. Sample This information can be
identifica- entered manually.
subsequent one
tion
o In CC, absorbance is measured most
2. Determine The LIS communicates to
of the time test(s) to the analyzer which test(s)
 Maintenance time (downtime) perform have been ordered.
o The time the analyzer is not in use CHEMICAL REACTION
 Open Reagent System 3. Reagent One or more reagents can be
o A system other than manufacturer’s systems dispensed into the reaction
reagents can be utilized for and cuvet.
measurement delivery
 Closed Reagent System 4. Specimen A small aliquot of the
o A system where the operator can only measure- sample is introduced into the
use the manufacturer’s reagent ment and reaction cuvet.
 Batch Testing deliver

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CCHM321 | BSMLS 2024 CLINICAL CHEMISTRY 1 TRANSCRIBER: CRUZ, A. M. A.
2022-2023 3RD YEAR, 1ST SEMESTER Prof. Kimberly Ann Pulga, RMT, MPH

5. Chemical The sample and reagents are necessarily carried to equilibrium

reaction mixed and incubated. since samples and standards are
phase treated exactly alike.
DATA COLLECTION AND ANALYSIS o Uses only 1 cuvette
6. Measure- Optical readings may be o Liquids are pumped through a system
ment initiated before or after all of continuous tubing
phase reagents have been added. o Samples flow through a common
7. Signal The analyte concentration is reaction vessel or pathway
processing estimated from a calibration
o Air bubbles at regular intervals
and data curve that is stored in the
handling analyzer. o A heating bath maintains the required
8. Send The analyzer communicates temperature of the reaction to allow
result(s) to results for the ordered tests complete color development
LIS to the LIS. o Reaction rate is controlled by
Note: LIS – Laboratory Information System temperature
o Mixing samples and reagents: by
TYPES OF AUTOMATIC ANALYZER using a glass coil inserted into the
flow path
 Continuous Flow Analyzer
• Example: Simultaneous
o Developed by Leonard Skeggs in
Multiple analyzer (SMA) or
1957
o This system has won wide acceptance Technicon
in both routine and research • Disadvantage: All test are
performed in parallel
laboratories
o Features
• Technicon Autoanalyzer II
• Use of Plastic Tubings
– capable of running 3
• of different diameters
different tests at 60-80
• has a peristaltic pump
samples/hour
per continuous
• SMA 6/60 – capable of
pumping of samples
running 6 tests at 60 samples
and reagents; this
per hour
maneuver reduces the
• SMA 12/60 – capable of
pipetting steps of
running 12 tests at 60 samples
manual procedures
per hour
• Introduction of Air Bubbles
• SMAC – capable of running
• separates the sample
40 tests at 120 samples per
hour and the reagent
streams into segments
Note: SMA – Simultaneous Multiple Analyzer • separates also one
o Principle: All samples are carried sample from the next
through the same analysis pathway. • for continuous
All samples automatically pass from scrubbing of tubing
one-step to another without waiting to • to prevent cross-
bring the samples to the same stage of contamination or carry
completion. The reactions are not

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CCHM321 | BSMLS 2024 CLINICAL CHEMISTRY 1 TRANSCRIBER: CRUZ, A. M. A.
2022-2023 3RD YEAR, 1ST SEMESTER Prof. Kimberly Ann Pulga, RMT, MPH

over by the previous

specimen
• Removal of Proteins by
dialysis
• The flow-through of cuvette
in interference filter
• Record Read Out
o Parts:
• Sampler
• Pumps and Manifolds  Discrete Sampling analyzer
• Dialyzer o Principle: Each sample reaction is
• Heating bath handled in separate compartment and
• Detector-Recorder does not come into contact with
o Different tests that can be performed another sample. The samples and
in the machine (Brand SEAL standards are handled on a batch basis
AA500): and must be brought before
• Ammonia proceeding to the next procedure. All
• Nitrate reactions must be carried out until
• Nitrite equilibrium is reached
• Phosphate • Dupont ACA
• Silicate • ABBOTT ABA-100
• Total Nitrogen biochromatic analyzer, ABA-
• Total Phosphorus 200 and VP analyzer
• Chloride • BECKMAN ASTRA 8 and
• Cyanide ASTRA 4
• Phenol • Beckman DSA
• Sugars • American Monitor KDA
• Nicotine o All samples must reach equilibrium
before proceeding to next procedure
(difference with Continuous)
o Most popular and versatile analyzer-
measures only the tests requested on a
sample
o Requires only 2-6 microliter of the
sample – minimum volume
o Capable of running multiple-tests-
one-sample-at-a-time (sequential
testing)
o Each sample-reagent mixture handled
separately in its own reaction vessel
o Software controlled
o For dry slide technology (reflectance
photometry), the spreading layer

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CCHM321 | BSMLS 2024 CLINICAL CHEMISTRY 1 TRANSCRIBER: CRUZ, A. M. A.
2022-2023 3RD YEAR, 1ST SEMESTER Prof. Kimberly Ann Pulga, RMT, MPH

permits a rapid uniform spreading o Brand: Thermo Scientific Gallery

layer over the reagent
o Mixing: magnetic driven teflon
stirring bar (Beckman), forceful
dispensing, magnetic stirring bars
(Astra), rotating paddle and ultrasonic
energy (Paramax)
o Examples: Vitros, Beckman ASTRA,
Roche Cobas Integra 800
o Major Advantage: Random Access
Capability- Allows STAT samples to
be easily tested
o Brand: BluVision

 Centrifugal Analyzers
o 1 cuvette per test or 1 cuvette per
several test
o All samples are loaded at the same
time
o Only 1 sample is being test; slow
procedure
o It uses the force generated by
centrifugation to transfer specimen
and reagents
o Liquids are placed in separate cuvet
for measurement at the perimeter of
spinning rotor (1000 rpm)
o It uses acceleration and decelaration
of the rotor to transfer the reagents
and sample from one chamber to
another
o Mixing: Cobas-Bio (Roche), IL-
Monarch
o Major Advantage: Batch Analysis
(Discrete batch type system)

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CCHM321 | BSMLS 2024 CLINICAL CHEMISTRY 1 TRANSCRIBER: CRUZ, A. M. A.
2022-2023 3RD YEAR, 1ST SEMESTER Prof. Kimberly Ann Pulga, RMT, MPH

o Principle: As the rotor is accelerated,

centrifugal force moves the reagents
and sample to a mixing chamber and
then through a small channel into the
cuvette. As the filled cuvette rotates
past a fixed light beam, the
absorbance of the reaction is
measured spectrophotometrically
• Centifichem
• RotoChem  Thin Film Analyzers (Dry Slide Technology)
o Easy to maintain (Brand: EasyRA) in o Can be
less than 5 minutes • Discretionary type
• Simple automated cleaning • Non-discretionary type
• Visual component check o Principle: A 16 mm2 chip that
contains several very thin layers
accepts a meter drop of serum,
spreads it evenly into a reagent layer,
and then confines the colored product
to the fixed area for reflectance
spectrophotometry
• Spreading layer – top layer
• Reagent layer
• Indicator layer
• Support layer – bottom layer
o Kodak EktaChem
o Vitros
o Beckman ASTRA
o Roche Cobas Integra 80
o Brand: Filmetrics F40
o Dry Slide Technology – Reflectance
photometry

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CCHM321 | BSMLS 2024 CLINICAL CHEMISTRY 1 TRANSCRIBER: CRUZ, A. M. A.
2022-2023 3RD YEAR, 1ST SEMESTER Prof. Kimberly Ann Pulga, RMT, MPH

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CCHM321 | BSMLS 2024 CLINICAL CHEMISTRY 1 TRANSCRIBER: CRUZ, A. M. A.