Cobas-C-311-Operator Manual-En PDF
Cobas-C-311-Operator Manual-En PDF
Cobas-C-311-Operator Manual-En PDF
COBI CD
Compendium of Background Information
cobas c 311 analyzer
Document information
Intended use This document is intended to provide background information for a better
understanding of the hardware, test principles and calibration methods of the
cobas c 311 analyzer.
Instrument approvals The cobas c 311 analyzer meets the protection requirements laid down in IVD
Directive 98/79/EC. Furthermore, our instruments are manufactured and tested
according to the following international standards:
o IEC 61010-1: 2001
o IEC 61010-2-010: 2003
o IEC 61010-2-081: 2001
o IEC 61010-2-101: 2002
o UL 61010-1: 2001
o CAN/CSA C22.2 No. 61010-1-04
o EN 61326-2-6:2006
Compliance is demonstrated by the following marks:
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cobas c 311 analyzer
Contact addresses
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cobas c 311 analyzer
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cobas c 311 analyzer
Table of contents
1 Photometric technology
Calculating data alarms Part D
General photometer characteristics A-5
7 Calculating data alarms
Introduction D-5
Test principles Part B Prozone effect D-5
Linearity verification (>Lin) D-7
2 ISE unit - Ion selective electrode principles Sensitivity limit check (Sens.E) D-9
Introduction B-5 Duplicate limit check (Dup.E) D-9
Calculation of unknown sample concentrations B-5 Technical limit check (>Test) D-11
Repeat limit check (>Rept) D-12
3 Photometric principles Reaction limit check (>React) D-12
Types of photometric assays B-9
Comprehensive assay descriptions B-12
Reaction cell and calibration data B-21
Quality control Part E
Endpoint assays B-24
Rate assays B-30 8 Applying QC rules
Prozone check B-39 Introduction E-5
Summary of assay techniques B-44 Rule 1: 1-2SD E-6
Rule 2: 1-2.5SD (Q2.5SD alarm) E-6
4 Serum index principles Rule 3: 1-3SD (Q3SD alarm) E-7
Introduction B-49 Rule 4: 2-2SA (S2-2Sa alarm) E-8
Definition of serum indices B-49 Rule 5: R-4SD (R4SD alarm) E-9
Measurement of serum indices B-49 Rule 6: 2-2SW (S2-2Sw alarm) E-10
Evaluating serum indices B-51 Rule 7: 4-1SA (S4-1Sa alarm) E-11
Serum index data alarms B-51 Rule 8: 4-1SW (S4-1Sw alarm) E-12
Rule 9: 10XA (S10Xa alarm) E-13
Rule 10: 10XW (S10Xw alarm) E-14
Calibration Part C
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Roche Diagnostics
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cobas c 311 analyzer
We have included the files necessary to install Adobe Acrobat Reader in this CD. If this
software is not installed on your computer, proceed as follows:
1 Close all running applications.
2 Change to the folder \reader on the CD-ROM.
3 Double-click on AdbeRdr707_en_US.exe to start the installation routine for
Adobe Acrobat Reader.
4 Follow the instructions on screen.
5 It is recommended that you restart your computer after the installation process
has finished.
Operator’s Manual Contains information about safety, hardware components and operating the analyzer
as well as maintenance and troubleshooting. A table of contents at the beginning of
the manual as well as at the beginning of each chapter, and an index at the end of this
manual help you to find information quickly.
Online Help Contains a detailed description of the software of the cobas c 311 analyzer. In
addition to the software description, the whole Operator’s Manual is included in the
Online Help. This makes it possible to retrieve information from both Online Help
and Operator’s Manual using the search functions available for electronically stored
documents.
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cobas c 311 analyzer
The software of the cobas c 311 analyzer has a context sensitive Online Help feature to
aid you in operating the instrument. "Context sensitive" means that wherever you are
located within the cobas c 311 software, choosing the Help feature displays Help text
or a screenshot relating to that area of the software. The Online Help offers a quick
and convenient way to find information, such as explanations of screens and dialog
boxes and how to perform particular processes.
F1 Help There are two ways to enter the Online Help: via the Help icon in the bottom left of
the screen or by pressing F1 on the keyboard. The context sensitive entry displays
information relating to your current location in the software.
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Measurement technology A
Photometric technology
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1 Photometric technology cobas c 311 analyzer
Table of contents
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cobas c 311 analyzer 1 Photometric technology
General photometer characteristics
A B C D E F G H I J K L M
When the light beam enters the photometer, it strikes a diffraction grating, which
separates the light into its constituent wavelengths and reflects them onto a fixed
array of 12 photodiodes. Each photodiode is permanently positioned to detect light at
a different wavelength.
Absorbance readings are taken each time a reaction cell rotates past the photometer.
When the reaction cell passes through the photometer lightpath, absorbance at the 12
wavelengths for each individual assay is measured.
Most Roche Diagnostics photometric tests use two wavelength readings to calculate
results. The end product of a chemical reaction absorbs the most light at one
particular wavelength. However, using the difference between readings at two
wavelengths (bichromatic system) eliminates the effect of interferences sometimes
found when using a single wavelength (monochromatic system) and compensates for
most of the photometric noise which improves the photometric resolutions.
For each reaction cell, a waterblank is measured and then absorbance readings are
taken 57 times (57 measure points) in 10 minutes.
Choice of wavelengths Bichromatic analysis uses two wavelengths: One that is at or near the peak absorbance
of the chromogen produced by the reaction, and a second wavelength at which little
or no absorbance of the desired chromogen occurs.
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1 Photometric technology cobas c 311 analyzer
General photometer characteristics
Any absorbance ( A 2 ) that occurs, due to interference from other substances in the
sample, is measured at the secondary wavelength. This amount is then subtracted
from the total absorbance ( A 1 ) occurring at the primary wavelength to yield the net
absorbance ( A C ).
A1 Observed
Chromophore
C
A
Absorbance
A2
Interferent
λ1 λ2
Wavelength
The optimum measure points for each test are part of the application parameters,
which are available via download.
The assay code and calibration type programmed from the application parameters
determine how final results are calculated for each test.
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Test principles B
This chapter provides you with an overview of the ion selective electrode test
principles and result calculation used by the cobas c 311 analyzer.
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2 ISE unit - Ion selective electrode principles cobas c 311 analyzer
Table of contents
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cobas c 311 analyzer 2 ISE unit - Ion selective electrode principles
Introduction
Introduction
Es Electromotive force (voltage) of the unknown sample for the specific ion
E IS Electromotive force (voltage) of the internal standard for the specific ion
S Slope
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2 ISE unit - Ion selective electrode principles cobas c 311 analyzer
Calculation of unknown sample concentrations
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3 Photometric principles cobas c 311 analyzer
Table of contents
Photometric principles
This chapter provides you with an overview of the photometric test principles and
assay techniques used by the cobas c 311 analyzer.
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cobas c 311 analyzer 3 Photometric principles
Types of photometric assays
There are four different assay types. The assay types are divided in endpoint assays
and rate assays:
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3 Photometric principles cobas c 311 analyzer
Types of photometric assays
Measure points Independent of the programmed application parameters, the photometer measures
the absorbance of a reaction mixture in fixed intervals of 3 to 24 seconds. Not all of
these measurements are used for the calculation of the result. Therefore, the
numbering of the photometer measure points differs form the numbering of the
measure points used in calculations.
The figure below represents an endpoint assay programmed for two measure points
( mp1 and mp2 ).
In this example, the application parameters define the 6th photometer measure point
( mp6 ) to be mp 1 and the 24th photometer measure point ( mp24 ) to be mp2 . In
other words, mp 6 of the instrument is set to be mp1 of the test calculation, and mp24
of the instrument is set to be mp2 of the test calculation.
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cobas c 311 analyzer 3 Photometric principles
Types of photometric assays
The Analyze tab on the Utility > Application screen displays the assay type and
measure points among other application parameters for a selected test.
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3 Photometric principles cobas c 311 analyzer
Comprehensive assay descriptions
In the following section one example of an endpoint assay and one example of a rate
assay is given, along with detailed explanations of the application parameters and
result calculations.
e For extended programming and calculation examples, see:
Example of a 2 Point End assay on page B-12
Example of a Rate A assay on page B-17
A 2 Point End assay is an endpoint assay with sample blank measurement and can be
programmed for two or more reagents. 2 Point means there are readings at two
measure points, mp1 and mp2 :
o mp 1 is the sample blank which is measured before or shortly after the final
reagent is added.
o mp 2 measures the absorbance of the final reaction product; it is set after addition
of final reagent and after the reaction is completed.
2 Point End assay graph A graphic representation of a 2 Point End assay using reagents dispensed at R1 and R2
timing is shown below.
R2
Absorbance
R1
Amp2
S
Amp1
C1 C2 C3
mp 1 mp 2 Time
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Comprehensive assay descriptions
Example data The following data from the Utility > Application screen are used for this example:
Test GLUC2
Assay type 2 Point End
Time 10 min
Points 6, 24
2nd wavelength 700 nm
Primary wavelength 340 nm
Conc. value for Std (1) 0.0
In the later sections, the entries for the Assay/Time/Point text boxes on Utility >
Application > Analyze are shown as follows:
Assay/Time/Point: [ 2 Point End ] [ 10 ] [ 6 ] [ 24 ] [ 0 ] [ 0 ]
This means:
o The assay type is 2 Point End.
o The reaction time is 10 minutes.
o The sample blank absorbance (sample plus first reagent) is determined by the 6th
photometer measurement of the respective reaction cell.
o The absorbance of the sample plus first and second reagents is determined by the
24th photometer measurement of the respective reaction cell.
Dilution factor After the mixture of sample and R1 reagent is measured as sample blank, it is diluted
by the addition of R2 reagent. Therefore, the readings cannot be subtracted, unless a
correction for the dilution is taken into account. A dilution factor ( d ) is calculated as
follows and applied to the sample + R1 absorbance:
V samp + V R1
Equation B-2 d = ---------------------------------------------
-
V samp + V R1 + V R2
d Dilution factor
V R1 R1 volume
V R2 R2 volume
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3 Photometric principles cobas c 311 analyzer
Comprehensive assay descriptions
Reaction monitor The two measure points for this assay’s calculation are set at the 6th and 24th
photometer measurements; the first is the sample blank reading, the second is the
final absorbance reading (endpoint), as indicated in the Reaction Monitor below.
You can move the focus from one measure point to the next using the scroll bar below
the graph. The absorbance at the measure point that has the focus is displayed in the
Abs. field above the graph. Alternatively, the absorbance values of all measure points
are listed on the Reaction Monitor report also:
Reaction Monitor 06/02/07 17:54
The values on the Reaction Monitor report (as well as those in the Abs. field on the
Reaction Monitor window) are absorbance × 104. Moreover, these values are already
corrected for the water blank value, determined during the cell blank measurement.
e See Cell Blank Measurement report on page B-21.
The real time water blank values displayed in the CB1-3 column of the Reaction
Monitor report serve to verify the integrity of the reaction cell immediately before
sampling.
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Comprehensive assay descriptions
Reaction absorbance To determine the reaction absorbance A x , the sample blank value is corrected for
dilution and then subtracted from the endpoint absorbance:
Equation B-3 A x = Amp 24 – d ⋅ Amp 6
Calculation of concentration The calculation of the unknown concentration of the analyte in the sample uses the
following endpoint reaction formula:
Equation B-4 C x = [ K ( A x – A b ) + C b ] ⋅ IFA + IF B
K and A b are displayed on the Working Information window. Select Calibration >
Status > Calibration Result > Working Information to display this window.
When the test's concentration value for Std (1) is programmed with a decimal, the
displayed K factor includes an extra digit for each number to the right of the decimal
point.
A b is the absorbance of the first standard solution, Std (1), which is a blank
calibrator. This value is also displayed on the Working Information window in the
S1 Abs. field.
e See Working Information window on page B-22.
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Comprehensive assay descriptions
C b , the concentration of the analyte in the first standard solution Std (1), is displayed
on the Others tab of the Utility > Application screen. This C b value controls the
number of digits of the displayed K and the rounding of the final results. When the
test's C b value is programmed with a decimal, K includes an extra digit for each
number to the right of the decimal point.
e See Others tab on page B-23.
Example values The following values are used for this example:
Cb 0.0
The result is rounded to 5.3 on the report because C b , the concentration value for
Std (1), the blank calibrator, contains one zero to the right of the decimal point as
displayed on Utility > Application > Others.
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Comprehensive assay descriptions
For rate assays, the time course of the reaction is followed by measuring the
absorbance as a function of time. That is, measurements are taken as the reaction
proceeds. Rate assays use these measurements because their concentration
calculations are based on the determination of the rate of change in absorbance, v :
dA
Equation B-5 v x = --------x-
dt
A Rate A assay is programmed for multiple measure points. This means, there is a
measuring window and every photometric measurement within this window is taken
into account for the rate calculation—beginning with the reading at the first
programmed measure point ( mpinitial ) through the reading at the second
programmed measure point ( mpfinal ).
The absorbance values are converted into the rate of change in absorbance ( v ) by
least squares analysis. There is no need for a dilution factor because all readings are
taken after the addition of the last reagent.
Rate A assay graph A graphic representation of a Rate A assay using a reagent dispensed at R1 and R2 or
R3 timing is shown below.
Absorbance limit
vx
Absorbance
Blank
S, R1 R2/R3
C1 C2 C3
mp 1 mp 2 Time
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COBI CD · Version 1.0 B-17
3 Photometric principles cobas c 311 analyzer
Comprehensive assay descriptions
Example data The following data from Utility > Application screen are used for this example:
Test ALTL
Assay Rate A
Time 10 min
Points 12, 31
2nd wavelength 700 nm
Primary wavelength 340 nm
Conc. value for Std (1) 0.00
In the later sections of this chapter, the entries for the Assay/Time/Point text boxes on
Utility > Application > Analyze are shown as follows:
Assay/Time/Point: [ Rate A ] [ 10 ] [ 12 ] [ 31 ] [ 0 ] [ 0 ]
This means:
o The assay type is Rate A.
o The reaction time is 10 minutes.
o The initial absorbance reading is the 12th photometer measurement of the
respective reaction cell.
o The final absorbance reading is the 31st photometer measurement of the
respective reaction cell.
Reaction monitor The rate of change in absorbance is calculated by least squares analysis of the
absorbance values measured within the measuring window, as indicated in the
reaction monitor below:
The values on the reaction monitor report are reaction absorbance × 104. Moreover,
these values are already corrected for the water blank value determined during the cell
blank measurement.
e See Cell Blank Measurement report on page B-21.
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Comprehensive assay descriptions
The absorbance values measured between the initial and the final absorbance reading
( mp12 through mp 31 ) represent a change over 4.05 minutes. The mathematical
analysis results in a rate of change in absorbance of -0.0503 per minute.
Reaction Monitor 16/02/07 16:22
Result calculation The calculation of the unknown concentration of the analyte in the sample uses the
following rate reaction formula:
Equation B-6 C x = [ K ( v x – v b ) + C b ] ⋅ IF A + IF B
K Calibration factor
vx Rate of change in absorbance (expressed in 104/min)
K and v b are displayed on the Working Information window. Select Calibration >
Status > Calibration Result > Working Information to display this window.
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Comprehensive assay descriptions
When the test's concentration value for Std (1) is programmed with a decimal, the
displayed K factor includes extra digits for each number to the right of the decimal
point. v b is displayed in the S1 Abs. field of the Working Information window.
e See Working Information window on page B-22.
C b , the concentration of the analyte in the first standard solution, Std (1), is displayed
on the Others tab of the Utility > Application screen.
e See Others tab on page B-23.
Example values The following values are used for this example:
Cb 0.00
Applying these values to the rate reaction formula (Equation B-6) yields:
C x = { – 42,04 ⋅ [ – 0,0503 – ( – 0,0001 ) ] + 0,0 } ⋅ 1 + 0
C x = – 42,04 ⋅ ( – 0,0502 )
C x = 2,110
The result is displayed as 2.11 on the report because C b , the concentration value for
Std (1), the blank calibrator, contains two zeroes to the right of the decimal point as
displayed on Utility > Application > Others.
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Reaction cell and calibration data
The following three sections explain the Cell Blank Measurement report, the Working
Information window, and information given on the Others tab. These three sections
are frequently referred to in other parts of this document which describe result
calculations of the various types of assays:
Both the Working Information window and the Others tab of the Utility >
Application screen display calibration information for individual tests and
calibrators, respectively. The Cell Blank Measurement report contains data necessary
for the calculation of absorbance values, which are the basis for all other calculations.
e For more information, see:
Cell Blank Measurement report on page B-21
Working Information window on page B-22
Others tab on page B-23
Reaction absorbance in a cell is measured against the cell's water blank value (current
cell blank). This cell blank report is requested as part of weekly maintenance. The
values on this report are stored and compared to the real time water blank values that
display on the Reaction Monitor report.
e See Reaction monitor on page B-14.
If the difference between the current real time water blank values and the previous cell
blank measured by the Cell Blank maintenance function is greater than 0.1 Abs, an
alarm is issued.
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3 Photometric principles cobas c 311 analyzer
Reaction cell and calibration data
S1 Abs. The Working Information window displays the current calibration curve and values
for the application selected under Calibration > Status > Calibration Result. For
endpoint assays based on an RCM or Linear calibration, the value under S1 Abs.
equals the blank calibrator’s absorbance value × 104. For rate assays it is the rate of
change in absorbance of the reaction with the blank calibrator.
S1 Abs. is subtracted from the reaction absorbance of all other samples including
calibrators Std(2) through Std(6), controls, STAT and routine samples.
K factor The K factor—as well as S1 Abs.—is used in the result calculation of every measured
test. Given a linear calibration curve, the two main types of assays use the following
formulas for result calculation:
Equation B-7 C x = K ⋅ ( A x – A b ) + C b for endpoint assays
K Calibration factor
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cobas c 311 analyzer 3 Photometric principles
Reaction cell and calibration data
Others tab
Use this tab to display test parameters such as calibrator codes, calibrator set points,
calibrator positions, and pipetting volumes.
When the test’s Std (1) concentration (blank calibrator concentration) is
programmed with a decimal, the displayed K factor on the Working Information
window gets the same number of decimal places. This also determines the decimal
placement in displayed results, as shown in the table below:
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COBI CD · Version 1.0 B-23
3 Photometric principles cobas c 311 analyzer
Endpoint assays
Endpoint assays
In the following sections the various types of endpoint assays are explained in detail.
After a brief listing of assay characteristics, a graphical representation of the
absorbance in the course of the reaction is given, as well as an example of result
calculation.
e For details on the various types of endpoint assays, see:
1 Point assay on page B-24
2 Point End assay on page B-27
1 Point assay
Assay characteristics:
o Called 1 Point because only one measure point is designated in the Application
screen.
o Addition of one or more reagents is possible.
o No sample blanking required.
o The absorbance reading for this type of assay can be taken during any disk
rotation after addition of the final reagent.
c 311 1 ≤ mp1 ≤ 57
1 ≤ time ≤ 10
Cell blank = (C1 + C2 + C3) / 3
Reaction volume = 100-250 µL
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cobas c 311 analyzer 3 Photometric principles
Endpoint assays
1 Point assay with R1 timing A graphic representation of a 1 Point assay using a reagent dispensed at R1 timing is
shown below. The figure below shows an increase in absorbance as the reaction
occurs. A decrease in absorbance as the reaction occurs is also possible.
Absorbance
S, R1
Amp 1
C1 C2 C3
Time
mp 1
1 Point assay with R1 and A graphic representation of a 1 Point assay using reagents dispensed at R1 and R2 or
R2 or R3 timing R3 timing is shown below.
Absorbance
R2, R3
S, R1
Amp 1
C1 C2 C3
Time
mp 1
S Pipetting of sample
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COBI CD · Version 1.0 B-25
3 Photometric principles cobas c 311 analyzer
Endpoint assays
Entries on Utility > The following data from Utility > Application are used for this calculation example:
Application > Analyze
Test CHO2I
Assay/Time/Point [ 1 Point ] [ 10 ] [ 57 ] [ 0 ] [ 0 ] [ 0 ]
Calculation of concentration The calculation of the concentration of the analyte in the sample uses the following
equation:
Equation B-9 C x = [ K ( A x – A b ) + C b ] ⋅ IFA + IF B
Table B-3 Definitions and values for quantities used in the calculation
(a) See Reaction Monitor report above.
(b) Displayed on Working Information window. For explanations, see Working Information window on
page B-22.
(c) Displayed on Utility > Application > Others. For explanations, see Others tab on page B-23.
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Endpoint assays
Assay Characteristics:
o Called 2 Point because there are readings at two measure points, mp1 and mp2 ,
which are designated on Utility > Application > Analyze.
o Allows for two or more reagent additions.
o Performs sample blank measurement.
o The first absorbance reading for this type of assay can be taken during any disk
rotation. Usually it is taken before or shortly after the final reagent is added.
o The second absorbance reading can be taken during any disk rotation after the
final reagent is added.
R2/R3
Absorbance
R1 Amp2
S
Amp1
C1 C2 C3
Time
mp 1 mp 2
S Pipetting of sample
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COBI CD · Version 1.0 B-27
3 Photometric principles cobas c 311 analyzer
Endpoint assays
The following data from the Utility > Application screen are used for this example:
Test GLUC2
Assay/Time/Point [ 2 Point End ] [ 10 ] [ 6 ] [ 24 ] [ 0 ] [ 0 ]
The result calculation is based on a calculated value for the absorbance of the final
reaction product A x . To determine this value the sample blank reading is corrected
for dilution and subtracted:
Equation B-10 A x = Amp 2 – d ⋅ Amp 1 with
V samp + V R1
d = ---------------------------------------------
-
V samp + V R1 + V R2
Assuming absorbance values on the reaction monitor report are the following:
d Dilution factor
V R2 Volume of reagent R2 50 µL
Table B-4 Definitions and values for quantities used in the calculation
(a) See Reaction Monitor report above.
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cobas c 311 analyzer 3 Photometric principles
Endpoint assays
Calculation of concentration The calculation of the concentration of the analyte in the sample uses the following
equation:
Equation B-11 C x = [ K ( A x – A b ) + C b ] ⋅ IFA + IF B
Table B-5 Definitions and values for quantities used in the calculation
(a) Displayed on Working Information window. For explanations, see Working Information window on
page B-22.
(b) Displayed on Utility > Application > Others. For explanations, see Others tab on page B-23.
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3 Photometric principles cobas c 311 analyzer
Rate assays
Rate assays
The following sections explain in detail the various types of rate assays. After a brief
listing of assay characteristics, a graphical representation of the absorbance in the
course of the reaction is given, as well as an example of result calculation.
e For details on the various types of rate assays, see:
Rate A assay on page B-30
Rate A assay with sample blank correction on page B-33
2 Point Rate assay on page B-36
Rate A assay
Assay Characteristics:
o One or more reagent additions are possible.
o Rate of change in absorbance is calculated by least squares method.
o Substrate depletion is monitored for linearity.
vx
Absorbance
S, R1
C1 C2 C3
mp 1 mp 2 Time
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cobas c 311 analyzer 3 Photometric principles
Rate assays
Rate A assay with R1 and A graphic representation of a Rate A assay using reagents dispensed at R1 and R2 or
R2 or R3 timing R3 timing is shown below.
vx
Absorbance
S, R1 R2/R3
C1 C2 C3
mp 1 mp 2 Time
S Pipetting of sample
The following data from the Utility > Application screen are used for this example:
Test ALTL
Assay/Time/Point [ Rate A ] [ 10 ] [ 12 ] [ 31 ] [ 0 ] [ 0 ]
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COBI CD · Version 1.0 B-31
3 Photometric principles cobas c 311 analyzer
Rate assays
Calculation of concentration The calculation of the unknown concentration of the analyte in the sample uses the
following equation:
Equation B-12 C x = [ K ( v x – v b ) + C b ] ⋅ IF A + IF B with
v x = v (mp 2,mp 1)
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Rate assays
Assay Characteristics:
o Assay with sample blank measurement.
o One or more reagent additions are possible.
o Rate of change in absorbance is calculated by least squares method.
o Substrate depletion is monitored for linearity.
S, R1 v (mp 3,mp 4) R3
v (mp 1,mp 2)
Absorbance
C1 C2 C3
mp 3 mp 4 mp 1 mp 2 Time
S Pipetting of sample
mp 1 , mp 2 Measure point 1 and 2 (initial and final measure points for rate reaction)
mp 3 , mp 4 Measure point 3 and 4 (initial and final measure points for sample blank)
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Rate assays
Test CREJ2
Assay/Time/Points [ Rate A ] [ 10 ] [ 27 ] [ 37 ] [ 15 ] [ 23 ]
V samp + V R1
d = ---------------------------------------------
-
V samp + V R1 + V R2
v (mp 3,mp 4) Rate of change in absorbance between mp 3 and mp 4 – 1.507 × 10-2 min-1
v (mp 1,mp 2) Rate of change in absorbance between mp 1 and mp 2 3.093 × 10-2 min-1
d Dilution factor
V R1 Reagent 1 volume 90 µL
V R2 Reagent 2 volume 47 µL
Table B-7 Definitions and values for quantities used in the calculation
The rate of change in absorbance between 15th and 23rd measurement of the reaction
cell (sample blank) is multiplied by the following to correct for dilution:
10µL + 90µL - = 100
d = --------------------------------------------------- --------- = 0,6803
10µL + 90µL + 47µL 147
Therefore:
v x = 0,0309 – 0,6803 ⋅ ( – 0,0015 ) = 0,0319
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cobas c 311 analyzer 3 Photometric principles
Rate assays
Calculation of concentration The calculation of the unknown concentration of the analyte in the sample uses the
following equation:
Equation B-14 C x = [ K ( v x – v b ) + C b ] ⋅ IF A + IF B
Roche Diagnostics
COBI CD · Version 1.0 B-35
3 Photometric principles cobas c 311 analyzer
Rate assays
Assay Characteristics:
o Rate assay measures rate of change in absorbance.
o Called 2 Point because there are 2 measure points (or duplicate readings at mp1
and mp2 ).
o The first absorbance reading for this type of assay can be taken during any disk
rotation after the final reagent is added.
o This reaction is monitored for substrate depletion, but not for linearity.
Amp 2
R2/R3
S, R1 Amp1
C1 C2 C3
mp 1 mp 2 Time
S Pipetting of sample
Roche Diagnostics
B-36 COBI CD · Version 1.0
cobas c 311 analyzer 3 Photometric principles
Rate assays
Test CO2-L
Assay/Time/Point [ 2 Point Rate ] [ 10 ] [ 2 ] [ 18 ] [ 0 ] [ 0 ]
Figure B-27
The result calculation is based on a calculated value for the rate of change in
absorbance of the reaction mixture v x . To determine this value, readings are
subtracted and divided by the time between measure points 1 and 2:
Equation B-15 v x = ( Amp 2 – Amp 1 ) ⁄ t
Table B-9 Definitions and values for quantities used in the calculation
(a) See Reaction monitor above.
Roche Diagnostics
COBI CD · Version 1.0 B-37
3 Photometric principles cobas c 311 analyzer
Rate assays
Calculation of concentration The calculation of the unknown concentration of the analyte in the sample uses the
following equation:
Equation B-16 C x = [ K ( v x – v b ) + C b ] ⋅ IF A + IF B
Table B-10 Definitions and values for quantities used in the calculation
(a) Displayed on Working Information window. For explanations, see Working Information window on
page B-22.
(b) Displayed on Utility > Application > Others. For explanations, see Others tab on page B-23.
Therefore:
C x = – 730,0 ⋅ [ – 0,0351 – ( – 0,0019 ) ] + 0,0
C x = – 730,0 ⋅ – 0,0332
C x = 24,24 (24.2 on reports and Data Review screen)
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B-38 COBI CD · Version 1.0
cobas c 311 analyzer 3 Photometric principles
Prozone check
Prozone check
Prozone checks applying the antigen readdition method compare the absorbance
before and after a final reagent addition at R2 or R3 timing, as indicated below:
R3
Absorbance
Apmp 2
R2
S, R1
Apmp 1
C1 C2 C3
S Pipetting of sample
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COBI CD · Version 1.0 B-39
3 Photometric principles cobas c 311 analyzer
Prozone check
To the right of the Prozone Limit field there are nine boxes:
[ lower limit ] [ upper limit ] [ pmp1 ] [ pmp2 ] [ 0 ] [ 0 ] [ comp. ] [ 0 ] [ 0 ]
o The first two boxes indicate the lower and upper prozone limits (in Abs × 104).
o The next four boxes are for the prozone measure points ( pmp ):
O 3rd entry: First prozone measure points ( pmp 1 )
O 4th entry: Second prozone measure points ( pmp2 )
O 5th entry: Set to zero for this method
O 6th entry: Set to zero for this method
Appropriate values are: 2 ≤ pmp1 < pmp2 ≤ 57. If all entries are set to zero,
prozone check is not performed.
o The seventh box (Inside/Outside) indicates in which case a data alarm (>Proz) is
issued: If the entry is set to Inside, an alarm is issued in case the obtained check
value lies inside the defined range between the lower and upper prozone limits
(first two boxes).
Vice versa, if the entry is set to Outside, an alarm is issued in case the obtained
check value lies outside the defined range.
o The eighth and ninth boxes are not used (set to zero) for this method.
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B-40 COBI CD · Version 1.0
cobas c 311 analyzer 3 Photometric principles
Prozone check
Prozone check value calculation The calculation of the prozone check value uses the following equation:
Equation B-17 PC = Apmp 2 – d ⋅ Apmp 1 with
V samp + V R1
d = ---------------------------------------------
-
V samp + V R1 + V R2
d Dilution factor
V R1 R1 volume
V R2 R2 volume
Calculation example
This section provides an example of a 2 Point End assay with prozone check (antigen
readdition method) with the calculation of the prozone check value.
The following data from the Utility > Application screen are used for this example:
Test ALBU2
Assay/Time/Point [ 2 Point End ] [ 10 ] [ 6 ] [ 15 ] [ 0 ] [ 0 ]
Prozone Limit [ -32000 ] [ 1000 ] [ 24 ] [ 30 ] [ 0 ] [ 0 ] [ Inside ] [ 0 ] [ 0 ]
Prozone check value calculation The calculation of the prozone check value uses the following equation:
Equation B-18 PC = Apmp 2 – d ⋅ Apmp 1 with
V samp + V R1 + V R2
d R3 = ------------------------------------------------------------
-
V samp + V R1 + V R2 + V R3
V R1 R1 volume 100 µL
V R2 R2 volume 20 µL
V R3 R3 volume 26 µL
Table B-11 Definitions and values for quantities used in the calculation
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COBI CD · Version 1.0 B-41
3 Photometric principles cobas c 311 analyzer
Prozone check
Therefore:
PC = Apmp 2 – d ⋅ Apmp 1
Prozone checks applying the reaction rate method compare the rate of change in
absorbance at two different times after final reagent addition, as indicated below:
v (pmp 3,pmp 4)
∆A (pmp 4,pmp 3)
v (pmp 1,pmp 2)
Absorbance
S, R1
C1 C2 C3
S Pipetting of sample
Roche Diagnostics
B-42 COBI CD · Version 1.0
cobas c 311 analyzer 3 Photometric principles
Prozone check
Prozone check value calculation The calculation of the prozone check value uses the following equation:
Equation B-19 PC = [ v (pmp 3,pmp 4) ⁄ v (pmp 1,pmp 2) ] × 100 with
Roche Diagnostics
COBI CD · Version 1.0 B-43
3 Photometric principles cobas c 311 analyzer
Summary of assay techniques
Rate A with sample blank 1 ≤ mp 3 < mp 4 < mp 1 < mp 2 ≤ 57, C x = [ K ( v (mp 3,mp 4) – v b ) + C b ] ⋅ IF A + IF B
mp 3 + 2 < mp 4 ,
mp 1 + 2 < mp 2
Roche Diagnostics
B-44 COBI CD · Version 1.0
cobas c 311 analyzer 3 Photometric principles
Summary of assay techniques
vx
Absorbance
Absorbance
R2, R3
S, R1
Amp1 S, R1 R2/R3
C1 C2 C3 C1 C2 C3
mp 1 Time mp 1 mp 2 Time
1 Point assay Rate A assay
S, R1 v (mp ,mp )
3 4 R2/R3
R2/R3
v (mp 1,mp 2)
Absorbance
R1
Absorbance
Amp 2
S
Amp 1
C1 C2 C3
C1 C2 C3
mp 1 mp 2 Time
mp 3 mp 4 mp 1 mp 2 Time
Amp 2
R2/R3
S, R1 Amp 1
C1 C2 C3
mp 1 mp 2 Time
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COBI CD · Version 1.0 B-45
3 Photometric principles cobas c 311 analyzer
Summary of assay techniques
Roche Diagnostics
B-46 COBI CD · Version 1.0
cobas c 311 analyzer 4 Serum index principles
Table of contents
This chapter provides you with an overview of the serum index test principles used by
the cobas c 311 analyzer.
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COBI CD · Version 1.0 B-47
4 Serum index principles cobas c 311 analyzer
Table of contents
Roche Diagnostics
B-48 COBI CD · Version 1.0
cobas c 311 analyzer 4 Serum index principles
Introduction
Introduction
The icterus index, I, is reported in icterus units that are linear, up to 60 mg/dL, and
semi-quantitative. For example, an icterus index of 20 is equivalent to a known
bilirubin concentration of approximately 20 mg/dL.
The hemolysis index, H, is reported in hemolysis units that are linear, up to
1000 mg/dL, and semi-quantitative. For example, a hemolysis index of 500 is
equivalent to a known hemoglobin concentration of approximately 500 mg/dL.
The lipemia index, L, is reported in lipemia units corresponding to mg/dL of
Intralipid® (Kabi-Pharmacia, Inc.), an artificial lipid material. These units are linear,
up to 2000 mg/dL, and semi-quantitative. For example, an L index of 1000 is
equivalent to a 1000 mg/dL Intralipid solution.—Hence, the L index provides an
estimate of sample’s turbidity, not its concentration of triglycerides.
The analyzer takes an aliquot of the patient sample, dilutes it with 0.9% NaCl, and
then measures the absorbances at three pairs of wavelengths:
o For measurement of lipemia (L), wavelengths 700/660 nm are used because this
range is free from influence by hemolysis and icterus (see Figure B-31 below).
o Hemolysis (H) is measured at 600/570 nm and correction is made for absorption
due to lipemia.
o Icterus (I) is measured at 505/480 nm and correction is made for absorption due
to lipemia and hemolysis.
Shown below are example absorption spectra of turbid serum, hemolytic solution,
and bilirubin solution.
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COBI CD · Version 1.0 B-49
4 Serum index principles cobas c 311 analyzer
Measurement of serum indices
Lipemic serum
Bilirubin solution
Absorbance
Hemolytic solution
Figure B-31 Example absorption spectra of a turbid (lipemic) serum, a hemolytic solution, and
a bilirubin solution
Calculation of serum indices To obtain the serum indices L, H, and I from the sample’s absorbance values, the
analyzer uses the following formulas:
1
Equation B-20 L = --- ⋅ ( Abs 1 )
C
1
Equation B-21 H = --- ⋅ ( Abs 2 – B ⋅ Abs 1 )
A
1
Equation B-22 I = ---- ⋅ ( Abs 3 – E ⋅ Abs 2 – F ⋅ Abs 1 )
D
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cobas c 311 analyzer 4 Serum index principles
Evaluating serum indices
Once the serum indices are determined, refer to the Limitations section of the
application’s package insert to assess the results. This indicates the index up to which
potential interference is within the Roche Diagnostics specification or when the
hemolyzed, icteric, or lipemic sample may not be used with the respective application.
Results which fall outside the permitted range are also flagged.
Upper limits for serum indices can be defined individually for each test. Limit values
are loaded with the application and displayed in the serum index boxes (L, H, and I)
on the Range tab of the Utility > Application screen. If a measured serum index value
is greater than the corresponding value in the L, H, or I box, an alarm is issued. When
serum index limits are set to 0, the serum index check will be neglected.
The following example shows how a data alarm is issued when a test-specific limit
value for a serum index is exceeded in a patient sample.
Example For this example the application GLUC2 is programmed with a L index limit of 10, H
index limit of 10, and I index limit of 60.
Note that these are just example values! Real values are downloaded or can be retrieved
from package inserts.
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COBI CD · Version 1.0 B-51
4 Serum index principles cobas c 311 analyzer
Serum index data alarms
Figure B-32 Utility > Application > Range with limits set for all three serum indices
If the obtained L or H index is greater than 10 and/or the I index is greater than 60, a
data alarm (>Index) is issued.
The measurement of the sample yielded an L index of 31, H index of 0, and I index of
7. The results are displayed on the Data Review screen.
Figure B-33
For GLUC2 the limit of 10 for the L index is exceeded. Therefore a >Index data alarm
is attached to the result.
Roche Diagnostics
B-52 COBI CD · Version 1.0
Calibration C
Roche Diagnostics
COBI CD · Version 1.0 C-3
5 ISE unit - Ion selective electrode calibration cobas c 311 analyzer
Table of contents
Roche Diagnostics
C-4 COBI CD · Version 1.0
cobas c 311 analyzer 5 ISE unit - Ion selective electrode calibration
ISE calibration
ISE calibration
The following ISE calibrators are used depending on the calibration method:
o Std (1) or S1: ISE Low, a water-based solution
o Std (2) or S2: ISE High, a water-based solution
o Std (3) or S3:
o For global use: ISE Comp., a serum-based solution, is used for full
calibrations.
o For use in US only: ISE High (compensated) with compensated set points is used
for full calibrations.
The following table displays all calibration methods and the corresponding
calibrators.
Additionally, an internal standard (IS) will be measured. The calibration interval for
all ISE applications is 24 hours.
For each calibration, the standard solutions are aspirated into the electrode cartridges,
and—after equilibration occurs at the electrode membrane—the electromotive force
(EMF, voltage) is measured.
The slope of the calibration will be calculated based upon these readings and the
assigned value of the standards.
Roche Diagnostics
COBI CD · Version 1.0 C-5
5 ISE unit - Ion selective electrode calibration cobas c 311 analyzer
Slope calculation
Slope calculation
The slope is calculated in millivolts (mV) from the aqueous high and low standards.
The slope is calculated according to the following formula:
EH – EL
Equation C-1 S = --------------------
-
CH
log ⎛⎝ ------⎞⎠
CL
S Slope
Due to factors such as the condition of the electrodes, the measured slope may deviate
from this ideal slope. Therefore, the slope obtained should fall within the following
ranges:
Na+ 50 to 68 mV
K+ 50 to 68 mV
Cl- -40 to -68 mV
S Slope
The calculated value of the internal standard, as well as the voltage, is shown on the
Calibration report.
Roche Diagnostics
C-6 COBI CD · Version 1.0
cobas c 311 analyzer 5 ISE unit - Ion selective electrode calibration
One-point calibration
One-point calibration
Compensation overview
Since the standard low and high are aqueous, a protein-based ISE Standard 3 is used
for compensating the differences in the electrode response between aqueous solutions
and a human serum matrix.
In the US, the ISE Standard High (compensated) with compensated set points is used
for ISE Standard 3 to correct differences between aqueous standards and human
serum matrix.
These differences are very small for regularly maintained ISE units. However, under
certain conditions these differences may become more prominent, thus requiring
compensation.
The ISE unit is in an optimal condition when the compensation values (C. Value) are stable
and negligible low.
S Slope
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COBI CD · Version 1.0 C-7
5 ISE unit - Ion selective electrode calibration cobas c 311 analyzer
Reference electrode
Reference electrode
A 1M KCl solution is measured concurrently with each sample analysis. The reference
electrode is used for this purpose. The voltage of the reference electrode serves as a
reference point for all measurements. That is, all reported voltages are readings from
which the voltage of the reference electrode has been subtracted.
Roche Diagnostics
C-8 COBI CD · Version 1.0
cobas c 311 analyzer 6 Photometric calibration
Table of contents
Photometric calibration
This chapter provides you with an overview of the calibration types used by the
cobas c 311 analyzer for photometric assays. The K factor, calibration updates and
calculating results are also discussed.
Roche Diagnostics
C-10 COBI CD · Version 1.0
cobas c 311 analyzer 6 Photometric calibration
Calibration checks
Calibration checks
For each photometric application, the following checks that automatically verify the
reliability of calibrations are available. If a check value lies outside the configured
check limits, an alarm is issued. This section briefly explains the calibration checks
and the associated alarms.
The limits of the calibration checks are configured under Utility > Application >
Calib.
SD limit When calibrating nonlinear or multipoint linear tests, the instrument performs the
following check:
For each calibrator, an absorbance value is calculated from the given concentration
and the current calibration curve. This calculated absorbance is compared to the
measured absorbance. If the difference of the two exceeds the SD limit value, an SD.E
alarm is issued. The SD limit value is defined in the SD Limit box (in Abs × 104). An
SD limit value of 999.9 denotes the check will be omitted.
In case an SD.E alarm occurs, measurement is still possible and the calibration curve
is updated. However, trace the cause of the alarm before you proceed to sample
measurement. The SD value is printed out together with the result of calibration.
Roche Diagnostics
COBI CD · Version 1.0 C-11
6 Photometric calibration cobas c 311 analyzer
Calibration checks
Duplicate limit All photometric calibrators are run in duplicate. The duplicate check calculates the
% error and the absolute absorbance error (difference) between these duplicate
measurements. The obtained check values are compared to the % error limit and the
absorbance error limit. The % error limit is defined in the first Duplicate Limit box.
The absorbance error limit is defined in the second Duplicate Limit box (Abs.). The
corresponding check values DE% and DEAbs. are calculated as follows:
Abs2 – Abs1
DE % = --------------------------------------------- ⋅ 100 and DE Abs. = Abs2 – Abs1 ,
( Abs2 + Abs1 ) ⁄ 2
where Abs1 and Abs2 denote the two absorbance readings, taken for each calibrator
(duplicate readings).
If both the % error and the absorbance error are out of range, a Dup.E alarm is issued
indicating a failed calibration. The calibration curve of the affected test is not
updated.
e For more details see also Duplicate limit check (Dup.E) on page D-9.
Sensitivity limit Sensitivity, here, refers to the ratio of an absorbance difference to a concentration
difference. It is calculated from the measured absorbance values and given
concentration values of the blank calibrator ( S1 ) and the span calibrator ( S N ):
Abs ( S N ) – Abs ( S 1 ) ⁄ Conc ( S N ) – Conc ( S 1 )
The sensitivity obtained in a calibration must lie within certain limits: The lower limit
is defined in the first Sensitivity Limit box. The upper limit is defined in the second
Sensitivity Limit box. If the obtained sensitivity is not within these limits, a Sens.E
alarm is issued indicating a failed calibration. The calibration curve of the affected test
is not updated.
S1 Abs. limit This check sets an upper and lower absorbance limit for the blank calibrator, Std (1).
If the absorbance for Std (1) falls outside these limits, the analyzer issues a S1A.E
alarm indicating an erroneous calibration. The calibration curve of the affected test is
not updated. An S1 Abs. Limit minimum of -32000 and maximum of 32000 denotes
the check will be omitted.
For linear calibrations, the reagent blank is simply the y-intercept of the calibration curve.
For all nonlinear calibration types, the reagent blank is the predicted absorbance for an
analyte concentration of zero.
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cobas c 311 analyzer 6 Photometric calibration
Calibration checks
Std. check If any of the data alarms listed below occurs in a calibration, an Std.E alarm is issued.
The calibration curve of the affected test is not updated. Choose Alarm (global
button) to verify which data alarm has occurred.
Updated and non-updated The table below shows the data output when data alarm is issued during a calibration.
calibration data If the working curve is not updated, take necessary measures and perform
recalibration. Recalibration may also be required depending on the cause of an alarm
even if the working curve is updated.
Data alarm Working curve Saving on hard disk Display on Alarm screen
SD.E Updated Yes Provided
Dup.E Not updated No Not provided
Sens.E Not updated No Provided
S1A.E Not updated No Not provided
Std.E Not updated No Provided
Table C-4 Data output in case of data alarm during calibration
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COBI CD · Version 1.0 C-13
6 Photometric calibration cobas c 311 analyzer
Calibration overview
Calibration overview
Calibration types The term calibration refers to the determination of a valid relation between the
measured signal [absorbance or (for rate assays) a rate of change in absorbance] and
the concentration of the analyte of interest. The graphical representation of such a
signal/concentration relation is the calibration curve also referred to as working
curve.
The analyzer uses different types of mathematical models to describe this relation.
These math models are referred to as calibration types.
Calibration methods Up to six calibrators—abbreviated Std (1), Std (2)... Std (6)—can be used for a full
calibration. However, not all of these need to be used in every update of a calibration.
Select one of four different calibration methods to define which calibrators are to be
used.
Calibration update types For calibration methods where only one calibrator is remeasured (Blank and Span),
there are three possibilities how the calibration curve is corrected. This choice is made
by setting the calibration update type.
e For more information, see:
Calibration types on page C-15
Calibration methods on page C-16
Calibration update types on page C-19
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C-14 COBI CD · Version 1.0
cobas c 311 analyzer 6 Photometric calibration
Calibration overview
Calibration types
Linear calibrations are used for tests when the absorbance readings plotted against
calibrator concentrations lie on a straight line. If a linear calibration is based on two
calibrator measurements, it is termed linear two-point calibration. If it is based on
more than two calibrators, it is termed linear multipoint calibration.
Nonlinear calibrations are used for tests whose absorbances at different
concentrations form a nonlinear but reproducible plot. At least three and a maximum
of six calibrators are required for calibration. Available Calibration types are RCM,
RCM2T1, and RCM2 T2.
In addition, there are two calibration types whose calibration curves are piecewise
defined interpolation functions: Spline and Line Graph. The following table provides
an overview of all available calibration types.
page C-22
Conc. (x)
x c page C-25
1 + ⎛⎝--- ⎞⎠
b Conc. (x)
page C-29
Conc. (x)
K factor
A K factor is used in the calculation of sample results. Any test requiring more than
just a blank during calibration will have its K factor calculated via the measured
absorbances of the blank calibrator Std (1) and the other calibrator(s).
e For more details, see K factor calculation on page C-19.
A fixed K factor is used for some tests and is derived at the time of system installation.
The respective tests have only their blank (baseline) values updated during
calibration.
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COBI CD · Version 1.0 C-15
6 Photometric calibration cobas c 311 analyzer
Calibration overview
Calibration methods
In the following sections, we explain these calibration methods and show how the
calibration curve parameters are updated. The following parameters are used
throughout:
Definition of parameters S1Abs Calibration curve parameter displayed in the S1 Abs. column of the
Calibration Result window(a) and on Working Information window(b)
K Calibration curve parameter displayed in the K column
' A diacritical mark (’) denotes an updated parameter. For example, B ' is
the new B parameter of the calibration curve after the calibration update.
(a) To display this window, choose Calibration > Status > Calibration Result.
(b) To display this window, choose Calibration > Status > Calibration Result > Working Information.
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cobas c 311 analyzer 6 Photometric calibration
Calibration overview
Blank calibration
A Blank calibration is a one-point calibration. Tests are calibrated with the Std(1)
calibrator only, and the signal correction is a simple proportional adjustment. The
calculation method for various applicable calibration types are listed below.
S1Abs
Linear S1Abs ' = s b K ' = ---------------- ⋅ K
sb
sb
RCM S1Abs ' = s b Previous value Previous value B ' = ---------------- ⋅ B
S1Abs
sb sb
RCM2T1 S1Abs ' = -------- ⋅ S1Abs K ' = -------- ⋅ K Previous value Previous value
)
)
sb sb
sb sb
RCM2T2 S1Abs ' = ------------------------- ⋅ S1Abs K ' = ------------------------- ⋅ K Previous value
S1Abs + K S1Abs + K
s b
curve for Std (1) calibrator s b = S1Abs + K ⋅ sinh B ⁄ ( 1 + B 2 )
)
Span calibration
A Span calibration is a one-point calibration. Tests are calibrated with only one
calibrator, and this has to be a standard solution other than Std (1). The signal
correction is a simple proportional adjustment.
The calibrator that corresponds to the Span point (entered on Utility > Application >
Calib.) is measured and the previously measured calibration curve is corrected for
each applicable calibration type as listed below.
sN sN
Linear S1Abs ' = --------- ⋅ S1Abs K ' = --------- ⋅ K
sN
)
sN
sN sN
RCM S1Abs ' = --------- ⋅ S1Abs Previous value Previous value B ' = --------- ⋅ B
)
sN sN
sN sN
RCM2T1 S1Abs ' = --------- ⋅ S1Abs K ' = --------- ⋅ K Previous value Previous value
)
sN sN
sN sN
RCM2T2 S1Abs ' = ------------------------- ⋅ S1Abs K ' = ------------------------- ⋅ K Previous value
S1Abs + K S1Abs + K
s N
given concentration value of Std (N)
Roche Diagnostics
COBI CD · Version 1.0 C-17
6 Photometric calibration cobas c 311 analyzer
Calibration overview
)
The calculated signal value S N is obtained simply by insertion of the given
concentration value x of Std (N) into the function of the calibration curve. For a
Linear calibration, for example, s N = S1Abs + ( 1 ⁄ K ) ⋅ x . Likewise for an RCM
)
calibration,
= S1Abs – B- + B .
)
Equation C-4 s N ------------------------
⎛ x---⎞ A
1+⎝ ⎠
K
2 Point calibration
Tests are calibrated using Std (1) calibrator and one calibrator Std (N) with N > 1. For
this calibration update, the signal correction is linear: s ' = q + p ⋅ s .
The number of the second calibrator Std (N) is displayed in the Span box on Utility >
Application > Calib. The calculation method depends on the calibration type as listed
below.
x A
RCM S1Abs' = s b + ( s b – p ⋅ S1Abs ) ⎛⎝--- ⎞⎠ Previous value Previous value B' = p ⋅ B
K
sinh B
RCM2T1 S1Abs' = s b – p ⋅ K --------------- K' = p ⋅ K Previous value Previous value
1+B
)
p N
s b
Std (1) calibrator
Signal value calculated from the (non-updated) calibration curve for the
)
s N
given concentration value of Std (N)
Full calibration
Tests are calibrated using all calibrators specified on Utility > Application > Others.
After this calibration, all parameters of the calibration curve are updated. The
parameters of a test’s calibration curve are displayed on the Calibration Result
window (choose Calibration > Status > Calibration Result). Parameters of linear
calibration curves are updated by linear regression, and nonlinear calibration curves
are updated using a nonlinear regression algorithm.
Applicable calibration types are Linear multipoint (with more than two calibrators),
RCM, RCM2T1, RCM2T2, Spline, and Line Graph.
Roche Diagnostics
C-18 COBI CD · Version 1.0
cobas c 311 analyzer 6 Photometric calibration
Calibration overview
If you are updating a calibration using either a Blank update or a Span update you
may select how the calibration is updated from Utility > Application > Calib. using
the Update Type box.
The following calibration types can be updated by either the Blank or Span method
and may use this update feature: RCM, RCM2T1, RCM2T2, Spline, and Line Graph.
The calibrator used is either Std (1), for a Blank calibration, or it is defined in the
Span box on Utility > Application > Calib. > Calibration Type area.
There are three update types available: None, Difference, and Ratio.
None If None is chosen as the entry in the Update Type box, then neither Difference nor
Ratio calibration update values are applied. The calibration update occurs as
described for either a blank or span calibration, depending on the calibration type
chosen.
Difference The absorbance difference to the previous calibration is measured for the one defined
calibrator only. This difference is then added to each of the test’s standard absorbance
values. This moves the calibration curve up or down, maintaining its original slope.
Ratio The test’s absorbance value is measured with the one defined calibrator only. The
ratio of this value to the previous yields an adjustment factor. Each of the test’s
standard absorbance values is then multiplied by this factor. This adjusts the slope of
the calibration curve, maintaining its original y-intercept.
K factor calculation
This section shows how K factors are calculated from absorbance and concentration
values for tests that are based on linear two-point calibration curves. Two examples
are given: One for an endpoint assay and one for a rate assay.
After a successful calibration, an updated S1 Abs. value is shown on both the Working
Information window and the first column of the S1 on the Calibration Monitor
report.
The absorbance value (or rate of change in absorbance) of the second calibrator is
printed in the first column under S2 on the Calibration Monitor report.
Roche Diagnostics
COBI CD · Version 1.0 C-19
6 Photometric calibration cobas c 311 analyzer
Calibration overview
These new values are used to calculate the K factor. When displayed on the Working
Information window, the K factor is automatically rounded and multiplied by the
correct power of 10, according to the decimal placement of the Std (1) concentration.
A glucose test is calibrated with water as Std (1) and a second calibrator, Std (2), with
a glucose concentration of 10.8 mmol/L. Mean values of the measured absorbance
values are 0.0036 for Std (1) and 0.8739 for Std (2). The K factor is calculated as
follows:
Equation C-6 K = ( 10,8 – 0,00 ) ⁄ ( 0,8739 – 0,0036 )
K = 10,8 ⁄ 0,8703 = 12,41
This K factor can now be used to calculate results from test absorbances.
e For more information on calculation of endpoint assay results, see:
Example of a 2 Point End assay on page B-12
Calculation of concentration on page B-15
An AST (aspartate aminotransferase) test is calibrated with water as Std (1) and a
second calibrator with a concentration of 94.2 U/L. Mean values of the measured
rates of change in absorbance are v b = – 0,0006 for Std (1) and v x = – 0,01575 for the
second calibrator. The K factor is calculated as follows:
Equation C-8 K = ( 94,2 – 0,0 ) ⁄ [ – 0,0486 – ( – 0,0006 ) ]
K = 94,2 ⁄ ( – 0,0480 ) = – 1962,5
This K factor can now be used to calculate results from test absorbance rates.
e For more information on calculation of rate assay results, see:
Example of a Rate A assay on page B-17
Result calculation on page B-19
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cobas c 311 analyzer 6 Photometric calibration
Calibration overview
Introduction to weighting
It is possible to apply a weighting function during the curve fitting process that favors
those calibrator points with a lower absorbance (or rate of change in absorbance).
This may result in a more accurate curve fit in that particular concentration range.
∑ [ Ai – f (Ci ) ]
2
Equation C-9 → min
i=1
n
Equation C-10 ∑ { wi [ Ai – f (Ci ) ] }2 → min
i=1
Weighting factors
The weighting factor is inversely related to the absorbance of the calibrator, so that
those calibrator points with a lower absorbance will have a larger weighting factor.
o If an entry of 1 is made in the Weight field, then the weighting factor for calibrator
point i is: w i = 1 ⁄ [ g(Ai ) ] , where g(Ai ) is a function of the absorbance (or rate of
change in absorbance) of calibrator i .
o If an entry of 2 is made in the Weight field, then the weighting factor for calibrator
point i is: w i = 1 ⁄ [ g(Ai ) ]2 .
Roche Diagnostics
COBI CD · Version 1.0 C-21
6 Photometric calibration cobas c 311 analyzer
Linear calibration
Linear calibration
Water is commonly used as a zero or blank calibrator. For Linear 2 point calibration,
the absorbance of water and a second calibrator is measured. These two points are
used to establish a linear plot, and its slope is used in the calculation of subsequent
control and patient results.
Parameters on Utility > Application > Calib.:
A S2
Absorbance
Ax
Ab
Cb Cx C S2 Concentration
When C b = 0
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cobas c 311 analyzer 6 Photometric calibration
Linear calibration
A S2
Absorbance
Ax
Ab
Cb Cx C S2 Concentration
When C b ≠ 0
The math model for a Linear calibration is the equation for a straight line
y = a + b ⋅ x , where a is the y-intercept and b is the slope. For our purpose, we
interpret this equation’s variables in as follows:
Slope The slope of a straight line can be derived either by the formula b = ( ∆y ) ⁄ ( ∆x ) (when
two points are used) or by the least squares method (when multiple points are used).
For the first case, comparison with Figure C-4 shows that ∆y = A S2 – A b and
∆x = C S2 – C b .
The formula for the slope can then be solved to b = ( A S2 – A b ) ⁄ ( C S2 – C b ) . This
equation shows that b is equal to the reciprocal K factor defined earlier.
Therefore, b = 1 ⁄ K .
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COBI CD · Version 1.0 C-23
6 Photometric calibration cobas c 311 analyzer
Linear calibration
y-intercept Comparison with Figure C-4 shows that the y-intercept a = A b – ( b ⋅ C b ) , where A b
is the absorbance and C b the concentration value for Std (1)/blank calibrator.
With slope and y-intercept thus determined, it is now possible to solve the equation
y = a + b ⋅ x to x , to calculate the analyte concentration in a patient sample C x :
C x = [K ( A x – A b ) + C b ]
Two additional constants are applied to this formula to correct the result for
systematic bias deriving from the instrument. The final formula for calculation of the
concentration is
Equation C-13 C x = [ K ( A x – A b ) + C b ] ⋅ IFA + IF B
K K factor
Ax Sample absorbance value
Assay types
Linear 2 Point calibration can be used with the following assay types:
o 1 Point assay
o 2 Point End assay
o 2 Point Rate assay
o Rate A assay
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cobas c 311 analyzer 6 Photometric calibration
RCM calibration
RCM calibration
The RCM calibration applies a working curve in which the absorbance increases or
decreases in a nonlinear manner as the concentration increases.
Parameters on Utility > Application > Calib.:
AN
AX
Absorbance
A S3
A S2
Ab
Cb C S2 C S3 CX CN
Concentration
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COBI CD · Version 1.0 C-25
6 Photometric calibration cobas c 311 analyzer
RCM calibration
RCM calculation
The math model for the RCM calibration curve approximation is shown below:
a–d -+d
Equation C-14 A = --------------------
C c
1 + ⎛⎝--- ⎞⎠
b
The above calibration curve parameters correspond to the values on the Working
Information window as follows (to display this window, select Calibration > Status >
Calibration Result > Working Information):
S1 Abs. column displays parameter a .
K column displays parameter b .
A column displays parameter c .
B column displays parameter d .
The formula for sample concentration calculation is shown below:
Equation C-15 C x = ( C + C b ) ⋅ IFA + IF B with
a – Ax 1 ⁄ c
C = b ⋅ ⎛ ---------------⎞
⎝ Ax – d ⎠
Assay types
Nonlinear RCM calibration can be used with the following assay types:
o 1 Point assay
o 2 Point End assay
o 2 Point Rate assay
o Rate A assay
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cobas c 311 analyzer 6 Photometric calibration
RCM2T1 calibration
RCM2T1 calibration
The RCM2T1 calibration applies a working curve in which the absorbance increases
in a nonlinear manner as the concentration increases.
Parameters on Utility > Application > Calib.:
AN
AX
Absorbance
A S3
A S2
Ab
Cb C S2 C S3 CX CN
Concentration
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COBI CD · Version 1.0 C-27
6 Photometric calibration cobas c 311 analyzer
RCM2T1 calibration
RCM2T1 calculation
The math model for the RCM2T1 calibration curve approximation is shown below:
sinh z
Equation C-16 A = a + b ⋅ -------------2- with z = c ⋅ C + d
1 +z
The above calibration curve parameters correspond to the values on the Working
Information window as follows (to display this window, select Calibration > Status >
Calibration Result > Working Information):
S1 Abs. column displays parameter a .
K column displays parameter b .
A column displays parameter c .
B column displays parameter d .
The model function for RCM2T1 (Equation C-16) cannot be inverted analytically.
However, the iteration series z n + 1 = arc sinh [ y ⋅ ( 1 + z n2) ] can be used to solve the
equation y = sinh z ⁄ ( 1 + z 2) . Thus, the formula for the sample concentration is as
follows:
Equation C-17 C x = ( C + C b ) ⋅ IFA + IF B where C is calculated by iteration.
Assay types
Nonlinear RCM2T1 calibration can be used with the following assay types:
o 1 Point assay
o 2 Point End assay
o 2 Point Rate assay
o Rate A assay
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cobas c 311 analyzer 6 Photometric calibration
RCM2T2 calibration
RCM2T2 calibration
The RCM2T2 calibration applies a working curve in which the absorbance decreases
in a nonlinear manner as the concentration increases.
Parameters on Utility > Application > Calib.:
Ab
Absorbance
A S2
AX
A S3
AN
Cb C S2 C X C S3 CN
Concentration
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COBI CD · Version 1.0 C-29
6 Photometric calibration cobas c 311 analyzer
RCM2T2 calibration
RCM2T2 calculation
The math model for the RCM2T2 calibration curve approximation is shown below:
Equation C-18 A = a + r ⋅ ( 1 + s ⋅C ) – 2
The above calibration curve parameters correspond to the values on the Working
Information window as follows (to display this window, select Calibration > Status >
Calibration Result > Working Information):
S1 Abs. column displays parameter a .
K column displays parameter r .
A column displays parameter s .
The formula for sample concentration calculation is shown below:
Equation C-19 C x = ( C + C b ) ⋅ IFA + IF B where
C = --- ⋅ ⎛ --------------
- – 1⎞⎠
1 r
s ⎝ Ax – a
Assay types
Nonlinear RCM2T2 calibration can be used with the following assay types:
o 1 Point assay
o 2 Point End assay
o 2 Point Rate assay
o Rate A assay
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cobas c 311 analyzer 6 Photometric calibration
Spline calibration
Spline calibration
When this calibration type is applied, the ranges between the data points of the
measured calibrators are approximated by third degree polynomials so that a smooth
calibration curve is obtained.
Parameters on Utility > Application > Calib.:
AN
A N-1
Absorbance
AX
A S3
A S2
Ab
Cb C S2 C S3 CX C N-1 CN
Concentration
C S2 , C S3 , ... C N Concentration value for Std (2), Std (3), ...Std (N)
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COBI CD · Version 1.0 C-31
6 Photometric calibration cobas c 311 analyzer
Spline calibration
Spline calculation
In the math model of a spline calibration, data points of calibrators are taken as
supporting points for the determination of interpolating functions. The simplest
interpolating functions are used in linear interpolation, where adjacent data points
are connected by a straight line. This method is used for Line Graph calibrations.
e See Line Graph calibration on page C-33.
In contrast to the angular polygon of a Line Graph calibration, a smooth curve will
result when using piecewise polynomials of a higher degree for the interpolation. The
routine applied for Spline calibrations determines a smooth cubic spline
approximation using third degree polynomials.
Assay types
Nonlinear spline calibration can be used with the following assay types:
o 1 Point assay
o 2 Point End assay
o 2 Point Rate assay
o Rate A assay
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cobas c 311 analyzer 6 Photometric calibration
Line Graph calibration
When this calibration type is applied, the ranges between the data points of the
measured calibrators are approximated by linear interpolation. An angular polygon is
obtained as calibration curve.
Parameters on Utility > Application > Calib.
AN
A N-1
A S4
Absorbance
Ax
A S3
A S2
Ab
Cb C S2 C S3 C x C S4 C N-1 CN
Concentration
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COBI CD · Version 1.0 C-33
6 Photometric calibration cobas c 311 analyzer
Line Graph calibration
The math model for Line Graph calibration curve approximation is shown below:
C N – C N-1
Equation C-20 K N -1 = ------------------------
A N – A N-1
KN Calibration factor for the interval between C N-1 and C N , or A N-1 and
-1
A N , respectively
AN Absorbance of Std (N)
For a calibration based upon N standard solutions—Std (1) to Std (N)—there are
N - 1 calibration curve intervals. The sample absorbance value (or rate of change in
absorbance for rate assays) A x determines which of the calibration curve intervals
and which of the calibration factors is relevant for the calculation of C x . If A x lies
between A N-1 and A N the relevant calibration factor is KN -1 .
Assay types
Nonlinear Line Graph calibration can be used with the following assay types:
o 1 Point assay
o 2 Point End assay
o 2 Point Rate assay
o Rate A
Roche Diagnostics
C-34 COBI CD · Version 1.0
Calculating data alarms D
This chapter provides you with an overview of how some important data alarms are
calculated by the cobas c 311 analyzer.
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7 Calculating data alarms cobas c 311 analyzer
Table of contents
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D-4 COBI CD · Version 1.0
cobas c 311 analyzer 7 Calculating data alarms
Introduction
Introduction
Several methods are used by the analyzer to ensure that final results are valid. Data
alarms appear on the results printout to indicate possible data errors and are sent to
Host. Some of these also activate the audible alarm and initiate the display of the
alarm indicator on the global Alarm button.
Prozone effect
Antigen readdition The analyzer may perform a check for the prozone effect by adding a dilution of the
antigen as an additional reagent (R2 or R3). If the reaction continues in the same
direction (increasing or decreasing absorbance) as in the initial reaction, then an
excess of reagent (antibody) still exists—prozone effect is not occurring. If the
reaction proceeds in the opposite direction, after additional antigen is added, then
prozone effect is occurring and the result is invalid. The corresponding data alarm is
printed on the patient report.
The antigen readdition method is applied when two prozone measuring points are
defined on Utility > Application > Analyze ( [ pmp1 ] [ pmp2 ] [ 0 ] [ 0 ] ).
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7 Calculating data alarms cobas c 311 analyzer
Prozone effect
R3
Absorbance
Apmp 2
R2
S, R1
Apmp 1
C1 C2 C3
S Pipetting of sample
(a) See Chapter 3 Photometric principles: Cell Blank Measurement report on page B-21.
Reaction rate method The reaction rate method verifies the existence of an excess of reagent (antibody) not
by repeated addition of antigen but by observation of the reaction rate in the course
of the reaction. This method is applied when four prozone measuring points are
defined on Utility > Application > Analyze ( [ pmp1 ] [ pmp2 ] [ pmp 3 ] [ pmp4 ] ).
v (pmp 3,pmp 4)
∆A (pmp 4,pmp 3)
v (pmp 1,pmp 2)
Absorbance
S, R1
C1 C2 C3
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cobas c 311 analyzer 7 Calculating data alarms
Linearity verification (>Lin)
To verify the linearity of a rate reaction (rate of change in absorbance), the percentage
of nonlinearity is calculated. This values must be less than the Linearity Limit, defined
on the Utility > System > Application. If the calculated value is above the limit, a >Lin
data alarm is issued. If any absorbance reading taken during the programmed interval
exceeds the Abs. Limit parameter, that absorbance reading is excluded from the least
squares rate calculation.
Depending on the number of measure points of an application, the analyzer
calculates the nonlinearity in one of the following ways:
o If there are less than 5 measure points, linearity check is not performed.
o If there are 5 to 13 measure points, two times three points are used in the
calculation.
o If there are 14 or more measure points, two times six points are used in the
calculation.
∆v
f
Absorbance
S, R1 R2/R3 vx
∆v
i
Time
mp 1 mp 2
S Pipetting of sample
R1, R2/R3 Pipetting of reagent at R1 timing, and at R2 or R3 timing
mp 1 First photometric measure point
vf Rate of change in absorbance calculated for the final five measure points
The percentage of nonlinearity is the difference between the slope of the initial part of
the curve and the slope of the final part of the curve scaled to the overall slope. An
alarm is issued, if [ ( v i – v f ) ⁄ ( v x ) ] ⋅ 100 > LL 1 , where LL 1 is the value of the first box in
the Linearity Limit line on Utility > Application > Analyze.
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7 Calculating data alarms cobas c 311 analyzer
Linearity verification (>Lin)
14 or more measure points In principle, the percentage of nonlinearity for 14 or more measure points is
calculated in the same way as for 5 to 13 measure points. The only difference is that v i
and v f are calculated on the basis of the initial and final six measure points,
respectively.
An alarm is issued, if [ ( v i – v f ) ⁄ ( v x ) ] ⋅ 100 > LL 2 , where LL 2 is the value of the second
box in the Linearity Limit line on Utility > Application > Analyze.
∆v f
Absorbance
S, R1
R2/R3 vx
∆v i
Time
mp 1 mp 2
S Pipetting of sample
R1, R2/R3 Pipetting of reagent at R1 timing, and at R2 or R3 timing
mp 1 First photometric measure point
Additional conditions for the To the right of the Linearity Limit field on the Utility > System > Application there
linearity check are four boxes:
Linearity Limit [ limit 5-13 mp ]% [ limit ≥ 14 mp]% [ condition 1 ] [ condition 2 ]
o The first two boxes indicate the linearity limits (in Abs × 104/min) for 5 to 13 and
14 or more measure points, respectively.
o The third and forth boxes define additional conditions for the linearity check.
The entry in the third box defines a minimum rate of change in absorbance
(allover slope in Abs × 104/min) for v x . If the measured rate falls below this
minimum, the linearity check is neglected.—That is, the third box defines the
variable T for the following condition:
–4
O If vx < T ×10 , linearity check is not performed.
The entry in the fourth box defines a minimum difference between v i and v f (in
Abs × 104/min). If the measured difference vi – vf falls below this minimum, the
linearity check is neglected.—That is, the fourth box defines the variable D for the
following condition:
–4
O If vi – vf < D ×10 , linearity check is not performed.
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cobas c 311 analyzer 7 Calculating data alarms
Sensitivity limit check (Sens.E)
An upper and lower sensitivity limit is designated on Utility > Application > Calib.
for each photometric application. These values relate to the minimum and maximum
absorbance changes which must be satisfied between the blank and span calibrators
during calibration. The sensitivity observed during calibration is calculated as
follows:
Abs ( S N ) – Abs ( S 1 )
Equation D-1 ----------------------------------------------------------
-
Conc ( S N ) – Conc ( S 1 )
where S N is the span calibrator and S 1 is the blank. If the sensitivity observed is not
within the sensitivity limits, a Sens.E alarm is issued indicating a failed calibration. All
calibrations that affect the factor setting for the test will be error checked against the
sensitivity limit calculated from the span calibrator.
A duplicate limit for calibrator acceptability is designated on Utility > Application >
Calib. The entry in the first Duplicate Limit text box defines the % error limit. The
entry in the second box defines the absorbance error limit. The corresponding check
values are calculated as follows:
Abs2 – Abs1
DE % = --------------------------------------------- ⋅ 100 ; DE Abs. = Abs2 – Abs1
( Abs2 + Abs1 ) ⁄ 2
DE % Relative duplicate error: Calculated value for the % error of a calibrator’s
absorbance readings (duplicate)
DE Abs. Absolute duplicate error
Abs1 , Abs2 Two absorbance readings, taken for each calibrator (duplicate readings)
All photometric calibrators are run in duplicate. If both the % error and the
absorbance error are out of range, a Dup.E alarm is issued indicating a failed
calibration.
The following flowchart describes how a decision is made to flag a calibration for
violating the duplication limit.
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7 Calculating data alarms cobas c 311 analyzer
Duplicate limit check (Dup.E)
Measure Abs1
and Abs2 for
calibrator Std(N)
Compute
DE % and DE Abs.
No
Is DE % < Yes
Continue
% error limit?
No
Set Dup.E
alarm
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cobas c 311 analyzer 7 Calculating data alarms
Technical limit check (>Test)
If a result does not fall into the concentration range specified by the upper and lower
Technical Limits on Utility > Application > Range, a data alarm >Test or <Test is
issued. >Test indicates results that exceed the upper limit. <Test indicates results
below the lower limit.
Technical Limit for diluted The Technical Limit for diluted samples is checked by multiplying the Technical Limit
samples entries on Utility > Application > Range by a concentration conversion coefficient (γ)
as below.
V ( S2 )1 ⋅ V ( S1 )1 ⁄ [ V ( S1 )1 + V (Dil ) 1 ]
Equation D-2 γt 1 < C1 < γt 2 , γ = -------------------------------------------------------------------------------------------
V ( S2 ) ⋅ V ( S1 ) ⁄ [ V ( S1 ) + V (Dil ) ]
V ( S1 ) Normal sample volume from a cup (or tube) to a cuvette when the
primary sample type is diluted.
V (Dil ) Normal diluent volume for the primary sample type.
V ( S2 ) Sample volume from cuvette to cuvette when the primary sample type is
diluted.
V ( S1 )1 Normal sample volume from a cup (or tube) to a cuvette when any
sample type, with the exception of the primary sample type, is diluted.
V (Dil ) 1 Normal diluent volume for any sample type with the exception of the
primary sample type.
V ( S2 )1 Normal sample volume from cuvette to cuvette when any sample type,
with the exception of the primary sample type, is diluted.
Primary is assigned to the sample type that is used during calibration. All other sample
types are corrected to the sample type used in calibration. When a sample is diluted,
V ( S1 )1 , V (Dil ) 1 and V ( S2 )1 refer to the currently chosen sample type.
When a sample is not diluted, diluent volumes and S2 volumes (cuvette to cuvette)
are zero. Therefore, γ in the equation above simplifies to the following:
V ( S1 )1
γ = -----------------
V ( S1 )
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7 Calculating data alarms cobas c 311 analyzer
Repeat limit check (>Rept)
The Repeat Limit is checked with the final concentration (printed concentration).
Relationships between parameters on Utility > Application > Range and check values
of the Technical Limit and Repeat Limit are shown below:
Check item on Utility > Application > Range Check value Check range
Technical Limit [ t1 ] - [ t2 ] Conc.1 [ t1 · γ ] - [ t2 · G ]
Repeat Limit [ a1 ] - [ a2 ] Conc.2 [ a1 ] - [ a2 ]
Table D-1 Parameters on Utility > Application > Range and check values
When the result (Conc.1) is less than the lower Technical Limit (t1), the result is
flagged with a <Test data alarm. When the result (Conc.1) is greater than the upper
Technical Limit (t2), the result is flagged with a >Test data alarm.
When the result (Conc.2) is less than the lower Repeat Limit (a1), the result is flagged
with a <Rept data alarm. When the result (Conc.2) is greater than the upper Repeat
Limit (a2), the result is flagged with a >Rept data alarm.
In rate assays, correct data cannot be obtained if the concentration or activity value is
beyond the quantitative range. For this reason, a check is performed with reference to
a set upper or lower absorbance limit. For rate assays with ascending absorbances, the
limit is an upper limit; for assays with descending absorbances, the limit is a lower
limit. The reaction limit value is displayed on Utility > Application > Analyze.
Figure D-6 Reaction limits for ascending and descending rate assays
A data alarm (>React) is issued if only 3 or less measure points remain in the within
the set absorbance limit. The alarm is not issued if there are 4 or more measure points
within the absorbance limit.
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cobas c 311 analyzer 7 Calculating data alarms
Reaction limit check (>React)
Absorbance
Reaction Limit
Time
Reaction Limit
Time
Reaction Limit
Time
Table D-2 Relationship between reaction limit check and photometric points
Automatic correction of Reaction The reaction limit check is performed with reference to the absorbance at the main
Limit absorbance wavelength. The analyzer automatically corrects the given reaction limit value by
adding absorbance due to sample turbidity, etc.:
Reaction limit absorbance after correction =
Input reaction limit absorbance
+ (L1 - Lb)
L1: Main wavelength absorbance of sample at photometric point 1
Lb: Main wavelength absorbance of reagent blank at photometric point 1
When L1 - Lb ≤ 0, the reaction limit absorbance is not corrected.
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Reaction limit check (>React)
Roche Diagnostics
D-14 COBI CD · Version 1.0
Quality control E
Applying QC rules
This chapter provides you with an overview of the application of quality control rules
by the cobas c 311 analyzer. The multi-rule Shewhart-type method using the
Westgard algorithm is described as well as possible alarms generated.
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Table of contents
Roche Diagnostics
E-4 COBI CD · Version 1.0
cobas c 311 analyzer 8 Applying QC rules
Introduction
Introduction
If selected in the software, the analyzer can utilize the Realtime QC to evaluate QC by
a multi-rule Shewhart-type method using the Westgard algorithm. For each test, this
algorithm applies a set of rules selected on QC > Individual > Realtime QC > Rules.
Any combination of rules may be specified. A pair of controls for each test being
processed is compared against a known standard deviation (SD) and mean. If one or
both of the controls fail a rule, the analyzer continues applying the testing criteria for
all selected rules. When at least one rule violation is found, the appropriate data alarm
for that rule is issued, and both the graph on the screen and the QC results on
Workplace > Data Review are flagged. The QC alarm for the last rule violated is
issued. The following is an explanation of each QC rule, using a display example
where appropriate.
All data alarms are listed in the Data alarms chapter of the Operator’s Manual.
No
1-2SD Inside control range
Yes
No
No No No No No No No No
1-2.5SD 1-3SD 2-2SA R-4SD 2-2SW 4-1SA 4-1SW 10XA 10XW
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8 Applying QC rules cobas c 311 analyzer
Rule 1: 1-2SD
Rule 1: 1-2SD
1-2SD represents the control rule where one control result exceeds limits defined as
the mean ± 2SD. Each control sample pair X and Y is compared against its respective
expected mean and standard deviation. If both X and Y are within the mean ± 2SD,
the QC results are accepted. No alarm is issued and no additional rules are checked
for this control pair. If either X or Y results are outside the mean ± 2SD, the test fails,
but no alarm is issued. The next selected rule is then applied and tested.
1-2.5SD symbolizes the control rule violated when one control result exceeds the limit
defined as the mean ± 2.5SD.
If tighter QC restrictions are desired, this rule may be selected in place of Rule 2: 1-
3SD. If both the 1-3SD and 1-2.5SD rules are selected in the QC > Individual >
Realtime QC > Select Rules window and a set of controls fails both rules, the Q3SD
alarm is issued. The deviation of a single sample is compared against 2.5 SD for each
control. If either control X or Y is outside the mean ± 2.5 SD, the rule is violated. A
Q2.5SD data alarm is issued for an indeterminate QC error, and a “ ” displays on
the Yoden plot
. 1-2.5SD
Y
2.5SD
cross-hatched area?
Q2.5SD
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cobas c 311 analyzer 8 Applying QC rules
Rule 3: 1-3SD (Q3SD alarm)
1-3SD is the control rule violated when a single control X or Y result exceeds the limit
defined as the mean ± 3SD. Each control sample X and Y is compared against its
respective expected mean and standard deviation. If both X and Y are within the
mean ± 3SD, the QC results are accepted.
If either X or Y results are outside the mean ± 3SD, a Q3SD alarm is issued indicating
an indeterminate QC error has occurred. A “ ” displays on the screen in the
appropriate part.
1-3SD
Y
3SD
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8 Applying QC rules cobas c 311 analyzer
Rule 4: 2-2SA (S2-2Sa alarm)
The results of one assay on each control are evaluated (a total of two control results
are tested).
The results of the most recent control pair of X and Y are compared against standard
deviations. If both X and Y deviate outside ± 2SD and both are either above or below
the mean, the rule is violated. A S2-2Sa data alarm is issued indicating a systematic
QC error has occurred. A “ ” displays on the Yoden plot.
The S2-2Sa alarm is issued when the two results (both X and Y in this case) are
outside the ± 2SD limit, across control materials. This is a systematic violation.
2-2SD
Y
2SD
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cobas c 311 analyzer 8 Applying QC rules
Rule 5: R-4SD (R4SD alarm)
R-4SD is the control rule in which there is a range or a difference between the control
materials that exceeds 4SD, as would be the case if the X control exceeded the -2SD
limit and the Y control exceeded the +2SD limit.
The run size specified when R-4SD is selected on the QC > Individual > Realtime
QC > Select Rules window determines the number of consecutive control X and Y
samples tested. The maximum deviations of X minus the minimum deviations of Y,
and the maximum deviations of Y minus the minimum deviations of X are
computed. If either of these differences is greater than 4SD, the rule is violated. A
R4SD data alarm is issued for a random QC error, and a “ ” displays on the Yoden
plot.
R-4SD
Y
2SD
cross-hatched area? (n is
entered in the screen)
R4SD
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COBI CD · Version 1.0 E-9
8 Applying QC rules cobas c 311 analyzer
Rule 6: 2-2SW (S2-2Sw alarm)
The results of the two most recent assays of each control are evaluated. A total of four
control results are tested. If either or both X and Y results deviate outside ± 2SD, the
rule is violated. A S2-2Sw data alarm is issued indicating a systematic QC error has
occurred. A ” ” displays on the Yoden plot.
The S2-2Sw alarm is issued when two consecutive control results are outside of the
2SD limit, within a control material. This is a systematic violation.
2-2SD
Y Y
-2SD 2SD
2SD
X X
-2SD
Are X n and X n – 1 or
S2-2Sw
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E-10 COBI CD · Version 1.0
cobas c 311 analyzer 8 Applying QC rules
Rule 7: 4-1SA (S4-1Sa alarm)
4-1SA is the control rule violated when four consecutive control results exceed the
same limit, either mean + 1SD or mean - 1SD. The S4-1Sa alarm is issued when three
control results are outside the ± 1SD limit and one control result is outside the ± 2SD
limit across control materials. This is a systematic alarm.
The results of two consecutive assays of each control are evaluated (total four samples
tested). If insufficient data are available, the test is not performed. If all X and Y
results exceed ± 1SD, and either the current X or Y value exceeds ± 2SD, and all are on
the same side of the mean, then the rule is violated. A S4-1Sa data alarm is issued for a
systematic QC alarm, and a ” ” displays on the Yoden plot.
4-1SA
Y
1SD
Are X n , Y n , X n – 1 and
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COBI CD · Version 1.0 E-11
8 Applying QC rules cobas c 311 analyzer
Rule 8: 4-1SW (S4-1Sw alarm)
The results of four consecutive assays of each control are evaluated (total of eight
samples tested). If fewer than four samples are available, the test is not performed. If
all X or Y results exceed one standard deviation, and fall on the same side of the mean,
and the current X or Y value exceeds ± 2SD then the rule is violated. A S4-1Sw data
alarm is issued for a systematic QC alarm, and a “ ” displays on the Yoden plot.
The S4-1Sw alarm is issued when three consecutive control results are outside the ±
1SD limit and one control result is outside the ± 2SD limit within a control material.
This is a systematic violation.
4-1SW
Y Y
-1SD 1SD
1SD
X X
-1SD
S4-1Sw
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E-12 COBI CD · Version 1.0
cobas c 311 analyzer 8 Applying QC rules
Rule 9: 10XA (S10Xa alarm)
10X is the control rule where there are 10 consecutive control observations (5 pairs)
on the same side of the mean. The S10Xa alarm is issued when nine consecutive
control results are on the same side of the mean and one control is outside the ± 2SD
limit, across control materials. This is a systematic violation.
The results of five consecutive assays of each control are evaluated (total 10 samples
tested). If fewer than five samples are available for each control, the test is not
performed. The signs of all sample deviations for both controls are compared with
zero. If all are nonzero and have the same sign, and one of the current X and Y
samples exceeds 2SD, then the rule is violated. A S10Xa data processing alarm is
issued for a systematic QC alarm, and a “ ” displays on the Yoden plot.
10XA
Y
cross-hatched area?
S10Xa
Roche Diagnostics
COBI CD · Version 1.0 E-13
8 Applying QC rules cobas c 311 analyzer
Rule 10: 10XW (S10Xw alarm)
The results of 10 consecutive assays of each control are evaluated (total 20 samples
tested). If fewer than 10 samples are available for each control, the test is not
performed. All sample deviations are compared with zero. If all are nonzero and have
the same sign, and the current sample (X or Y) exceeds ± 2SD, the rule is violated. A
S10Xw data alarm is issued for a systematic QC alarm, and a “ ” displays on the
Yoden plot.
The S10Xw alarm is issued when nine consecutive control results are on the same side
of the mean and one control result is outside the ± 2SD limit within a control
material. This is a systematic violation.
10XW
Y Y
X X
Y n to Y n – 9 in the same
cross-hatched area?
S10Xw
Roche Diagnostics
E-14 COBI CD · Version 1.0
Index F
cobas c 311 analyzer Index
Index
A D
Absorbance limit, rate assays, D-12 Data alarm calculation
Antigen readdition, D-5 – absorbance limit, D-12
Approvals, 2 – calibrator duplicates, D-9
Assay principles – linearity, D-7
– ion selective electrode, B-3 – repeat limit, D-12
– photometric: See assay types. – sensitivity limit, D-9
– serum index, B-47 – substrate depletion, D-12
Assay types, photometric – technical limit, D-11
– overview, B-9 Document information, 2
– 1 Point, B-24 Duplicate limit, D-9
– 2 Point End, B-12
– 2 Point Rate, B-36 E
– Rate A, B-17
– Rate A with sample blank, B-33 Edition notice, 2
– summary, B-44 Electromotive force (EMF), B-5
EMF (electromotive force), B-5
B Endpoint assays
– 1 Point, B-24
Bichromatic measurement, A-5 – 2 Point End, B-27
Blank calibration, C-17
F
C
Full calibration, C-18
C. Value, ISE calibration, C-7
Calibration methods, photometric H
– 2 Point calibration, C-18
– Blank calibration, C-17 Hemolysis index, calculation, B-50
– Full calibration, C-18
– Span calibration, C-17 I
Calibration types, photometric, C-15
Calibration update types, photometric, C-19 Icterus index, calculation, B-50
Cell Blank Measurements report, B-21 Instrument approvals, 2
Concentration calculation Intended use, 2
– 1 Point assay, B-26 Internal standard calculation, C-6
– 2 Point End assay, B-28 ISE calibration
– 2 Point Rate assay, B-36 – internal standard, C-6
– Rate A assay, B-31 – reference electrode, C-8
Contact addresses, 3 – slope calculation, C-6
Copyrights, 2 ISE reference electrode, C-8
ISE, calculating concentrations, B-5
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COBI CD · Version 1.0 F-3
Index cobas c 311 analyzer
K Q
K factor QC rules
– calculation, C-19 – rule 1: 1-2SD, E-6
– definition, C-15 – rule 2: 1-2.5SD, E-6
– rule 3: 1-3SD, E-7
L – rule 4: 2-2SA, E-8
– rule 5: R-4SD, E-9
Line Graph calibration, C-33 – rule 6: 2-2SW, E-10
Linear 2 point calibration, C-22 – rule 7: 4-1SA, E-11
Linearity verification, D-7 – rule 9: 10XA, E-13
Lipemia index, calculation, B-50 – rule10: 10XAW, E-14
N R
Non-linear calibration Rate assays
– Line Graph, C-33 – 2 Point Rate, B-36
– RCM, C-25 – Rate A with sample blank, B-33
– RCM2T1, C-27 RCM calibration, C-25
– RCM2T2, C-29 RCM2T1 calibration, C-27
– Spline, C-31 RCM2T2 calibration, C-29
Reaction limit, D-12
O Reaction rate method, D-6
Realtime QC, E-5
One-point calibration Repeat limit, D-12
– ISE, C-7 Revision History, 2
– photometric test, C-17
Others tab, Utility > Application screen, B-23 S
Roche Diagnostics
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