0104498631190COINV3.

CHED2
Cholinesterase/Dibucaine Gen.2 Enzymes
Order information
Analyzer(s) on which cobasc pack(s) can be used
COBASINTEGRA 400 plus
04498631 190 Cholinesterase/Dibucaine Gen.2 (150 tests*) System-ID 0768456
COBAS INTEGRA 800
10759350 190 Calibrator f.a.s. (12 3 mL) System-ID 0737186
12149435 122 Precinorm U plus (10 3 mL) System-ID 0779997
12149443 122 Precipath U plus (10 3 mL) System-ID 0780006
10171743 122 Precinorm U (20 5 mL) System-ID 0779970
10171735 122 Precinorm U (4 5 mL) System-ID 0779970
10171778 122 Precipath U (20 5 mL) System-ID 0779989
10171760 122 Precipath U (4 5 mL) System-ID 0779989
05117003 190 PreciControl ClinChem Multi 1 (20 5 mL) System-ID 0774693
05947626 190 PreciControl ClinChem Multi 1 (4 5 mL) System-ID 0774693
05117216 190 PreciControl ClinChem Multi 2 (20 5 mL) System-ID 0774707
05947774 190 PreciControl ClinChem Multi 2 (4 5 mL) System-ID 0774707
*150tests per cassette with or without dibucaine or 75determinations of the dibucaine number

English CHE

System information butyrylthiocholine+H2O thiocholine+butyrate

Test CHET2, testID0019 (uninhibited activity of cholinesterase)
Test CHED2, testID0020 (inhibited activity of cholinesterase) thiocholine+potassium hexacyanoferrate(III)
Intended use dithiobis(choline)+potassium hexacyanoferrate(II)
In vitro test for the quantitative determination of the uninhibited and inhibited To determine the dibucaine number(DN), cholinesterase activity is
activity of cholinesterase (EC3.1.1.8; acylcholine acylhydrolase) in serum measured with and without enzyme inhibitor dibucaine. The dibucaine
and plasma using dibucaine as inhibitor number is calculated as follows:
Summary1,2,3,4,5
Cholinesterase (pseudocholinesterase or cholinesteraseII) is found in the CHE activity with inhibitor
DN=100 100
liver, pancreas, heart, serum and in the white matter of the brain. This CHE activity without inhibitor
enzyme must not be confused with acetylcholinesterase from erythrocytes
(EC3.1.1.7), which is also referred to as cholinesteraseI. Reagents - working solutions
The biological function of cholinesterase is unknown. Serum cholinesterase R1 Pyrophosphate buffer: 92mmol/L, pH7.7; potassium
serves as an indicator of possible insecticide poisoning. It is measured as hexacyanoferrate: 2.4mmol/L
an index of liver function. In pre-operative screening, cholinesterase is used
to detect patients with atypical forms of the enzyme and hence avoid R2 Dibucaine: 2.6mmol/L, pH6.3
prolonged apnea caused by slow elimination of muscle relaxants. SR GOOD's buffer: 10mmol/L, pH4.0; butyrylthiocholine: 46mmol/L;
At least 6different genetic variants of serum cholinesterase have been stabilizers
described (A, F, J, K, S, andU). The normal, most common phenotype is
designatedU. The atypical forms A, F, and S are described by high R1 is in positionA, R2 is in positionB and SR is in positionC.
resistance to dibucaine inhibition(A), increased resistance to fluoride Precautions and warnings
inhibition (F), and absence of catalytic activity(S). The homozygous forms
AA or FF are found in only 0.3 to 0.5% of the caucasian population. Pay attention to all precautions and warnings listed in
Section1/Introduction of this Method Manual.
The genotypes most susceptible to prolonged anesthesia and/or apnea
after succinyldicholine administration are AA, AS, and SS (severe risk); AF, Reagent handling
FS, and FF (moderate risk); and to some extent, UA (usually only during Ready for use
pregnancy). In such cases succinyldicholine anesthesia should be avoided.
Storage and stability
Measurements of total serum cholinesterase activity as well as
determination of dibucaine number and fluoride number are needed to Shelf life at 28C See expiration date on
characterize cholinesterase variants fully. cobasc pack label
The dibucaine number indicates the percentage inhibition of enzyme activity
in the presence of a standard concentration of dibucaine. COBASINTEGRA400 plus system
Depressed cholinesterase levels are found in cases of intoxication with On-board in use at 1015C 4weeks
organophosphorus compounds and in hepatitis, cirrhosis, myocardial COBASINTEGRA800 system
infarction, acute infections and atypical phenotypes of the enzyme.
This assay is based on the method published by Schmidt Eetal in 1992.5 On-board in use at 8C 4weeks
Test principle5,6 Specimen collection and preparation
Method with butyrylthiocholine5 using dibucaine as inhibitor.6 For specimen collection and preparation only use suitable tubes or
Cholinesterase catalyzes the hydrolysis of butyrylthiocholine to thiocholine collection containers.
and butyrate. Thiocholine instantaneously reduces the yellow Only the specimens listed below were tested and found acceptable:
hexacyanoferrate(III) to the almost colorless hexacyanoferrate(II). This Serum: Collect serum using standard sampling tubes.
decrease in color can be measured at wavelengths between 405 and Plasma: Liheparin, K2EDTA or K3EDTA plasma.
415nm. Do not use citrate and fluoride plasma.
The sample types listed were tested with a selection of sample collection
tubes that were commercially available at the time of testing, i.e. not all

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Cholinesterase/Dibucaine Gen.2 Enzymes

available tubes of all manufacturers were tested. Sample collection systems Pipetting parameters
from various manufacturers may contain differing materials which could
affect the test results in some cases. When processing samples in primary CHET2 Diluent (H2O)
tubes (sample collection systems), follow the instructions of the tube R1 120L
manufacturer.
Sample 2L 5L
Centrifuge samples containing precipitates before performing the assay.
SR 24L
Stability:4,7,8 6hours at 1525C
Total volume 151L
7days at 28C
1year at 20C CHED2 Diluent (H2O)
R1 100L
Materials provided
See Reagents working solutions section for reagents. R2 20L
Assay Sample 2L 5L
For optimum performance of the assay follow the directions given in this SR 24L
document for the analyzer concerned. Refer to the appropriate operators
manual for analyzerspecific assay instructions. Total volume 151L

Application for total and inhibited cholinesterase in serum and plasma Ratio definition for dibucaine number
COBASINTEGRA400plus test definition Abbreviated ratio name CHE2R (0030)
Measuring mode Absorbance Equation 100-(CHED2/CHET2)100
Abs. calculation mode Kinetic Use the predefined profile (CHE2P; 0050) for simultaneous order entry of
CHET2 and CHED2 tests from the same sample. The ratio for dibucaine
Reaction mode CHET2: R1SSR number will automatically be calculated after result output of both tests.
CHED2: R1/R2SSR Calibration
Reaction direction Decrease CHET2
Wavelength A/B 409/659nm Calibrator Calibrator f.a.s.
Calc. first/last CHET2: 43/52 Use deionized water as zero calibrator.
CHED2: 43/52
Calibration mode Linear regression
Unit U/L Calibration replicate Duplicate recommended
Pipetting parameters Calibration interval Each lot
CHET2 Diluent (H2O) Traceability: This test is standardized against a reference method using a
manual application of the butyrylthiocholine/ hexacyanoferrate(III) method
R1 120L on a photometer and the published molar absorptivity of
Sample 2L 5L hexacyanoferrate(III).
SR 24L CHED2
Total volume 151L Calibrator Use deionized water as zero calibrator.

CHED2 Diluent (H2O) Calibration mode Blank

R1 100L Calibration interval Each lot

R2 20L Note
Only CHET2 has to be calibrated with Calibratorf.a.s. For CHED2
Sample 2L 5L calibration always use the calibration factor obtained in the CHET2
SR 24L calibration. A blank calibration is necessary for CHED2. Use the following
procedure:
Total volume 151L COBASINTEGRA400 plus system
COBASINTEGRA800 test definition 1. Calibrate CHET2 test using Calibrator f.a.s.
Measuring mode Absorbance 2. Find the calibration factor(F) for CHET2 test in menu
Results>Calibration (click on the CHET2).
Abs. calculation mode Kinetic
3. In menu Configuration>Definitions>Tests choose CHED2(0020).
Reaction mode CHET2: R1SSR Then in Laboratory enter the calibration factor in the field Correlation
Factors.
CHED2: R1R2SSR
4. Calibrate (Blank) the CHED2 test. Use deionized water as CHED2
Reaction direction Decrease calibrator (SystemID0768596).
Wavelength A/B 409/659nm COBASINTEGRA800 system
Calc. first/last CHET2: 60/75 1. Calibrate CHET2 test using Calibrator f.a.s.
CHED2: 60/75 2. Find the calibration factor(F) for CHET2 test in window Detail of
Calibration. (Click on the CHET2 calibration result with detail mouse
Unit U/L button (middle button) to obtain window Detail of Calibration.)

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Cholinesterase/Dibucaine Gen.2 Enzymes

3. In menu Configuration>Definitions>Tests choose CHED2(0020). In very rare cases, gammopathy, in particular type IgM (Waldenstrms
Then in Modify>Laboratory enter the calibration factor in the field macroglobulinemia), may cause unreliable results.12
Correlation Factor of window Laboratory Parameters of Basic Test For diagnostic purposes, the results should always be assessed in
CHED2. conjunction with the patients medical history, clinical examination and other
4. Calibrate (Blank) the CHED2 test. Enter0 for both high and low findings.
calibrator values. ACTION REQUIRED
Quality control Special Wash Programming: The use of special wash steps is mandatory
when certain test combinations are run together on COBASINTEGRA
Reference range PrecinormU, PrecinormUplus or analyzers. Refer to the CLEAN Method Sheet for further instructions and for
PreciControlClinChemMulti1 the latest version of the Extra wash cycle list.
Where required, special wash/carry-over evasion programming must
Pathological range PrecipathU, PrecipathUplus or be implemented prior to reporting results with this test.
PreciControlClinChemMulti2
Limits and ranges
Control interval 24hours recommended Measuring range
Control sequence User defined CHET2: 20014000U/L (3.34234kat/L)
Control after calibration Recommended CHED2: 2007000U/L(3.34117kat/L)
Determine samples having higher activities via the rerun function. Dilution
For quality control, use control materials as listed in the Order information of samples via the rerun function is a 1:2 dilution. Results from samples
section. In addition, other suitable control material can be used. diluted using the rerun function are automatically multiplied by a factor of2.
The control intervals and limits should be adapted to each laboratorys Lower limits of measurement
individual requirements. Values obtained should fall within the defined
limits. Each laboratory should establish corrective measures to be taken if Lower detection limit of the test:
values fall outside the defined limits. CHET2: 200U/L (3.34kat/L)
Follow the applicable government regulations and local guidelines for CHED2: 250U/L (4.16kat/L)
quality control. The lower detection limit represents the lowest measurable analyte level
Calculation that can be distinguished from zero. It is calculated as the value lying
CHET2 3standard deviations above that of a zero sample (zero sample+3SD,
repeatability, n=21).
COBASINTEGRAanalyzers automatically calculate the analyte activity of
each sample. For more details, please refer to Data Analysis in the Online Expected values*,13
Help (COBASINTEGRA400plus/800analyzers). Children, men, women (aged 40years or more):
CHED2 532012920U/L (89215kat/L)
The cholinesterase activity of each sample is determined by multiplying the Women aged 1639years, not pregnant, not taking hormonal
measured rates (A/min) with the conversion factor. For more details, contraceptives:
please refer to Data Analysis in the Online Help 426011250U/L (71187kat/L)
(COBASINTEGRA400plus/800analyzers). Women aged 1841years, pregnant or taking contraceptives:
Dibucaine number 36509120U/L (61152kat/L)
For calculation of the dibucaine number, please refer to Section Test *Calculated with a temperature conversion factor of 1.52 (2537C)14
principle and Ratio Definition for Dibucaine Number in this method sheet.
Dibucaine Number
Conversion factor: U/L0.0167=kat/L
Limitations - interference Dibucaine Number for the cholinesterase variantsU (usual) and A
Criterion: Recovery within10% of initial value. (atypical):
CHET2 Normal individuals UU Dibucaine Number 73
Icterus:9 No significant interference up to an Iindex of 60 for conjugated Heterozygous individuals UA Dibucaine Number 5772
and unconjugated bilirubin (approximate conjugated and unconjugated
bilirubin concentration: 1026mol/L or 60mg/dL). Homozygous individuals AA Dibucaine Number 50
Hemolysis:9 No significant interference up to an Hindex of 350 DN results below 57 should be considered to indicate a potentially high risk
(approximate hemoglobin concentration: 217mol/L or 350mg/dL). of succinyldicholine sensitivity.15
Lipemia:9 No significant interference up to an Lindex of 1000. There is poor Each laboratory should investigate the transferability of the expected values
correlation between the Lindex (corresponds to turbidity) and triglycerides to its own patient population and if necessary determine its own reference
concentration. ranges.
CHED2 Specific performance data
Icterus:9 No significant interference up to an Iindex of 60 for conjugated Representative performance data on the COBASINTEGRA analyzers are
bilirubin and 50 for unconjugated bilirubin (approximate conjugated bilirubin given below. Results obtained in individual laboratories may differ.
concentration: 1026mol/L or 60mg/dL; approximate unconjugated
bilirubin concentration: 855mol/L or 50mg/dL). Precision
Precision was determined using human samples and controls in an internal
Hemolysis:9 No significant interference up to an Hindex of 150 protocol with repeatability (n=21) and intermediate precision (1aliquot per
(approximate hemoglobin concentration: 93mol/L or 150mg/dL). run, 1run per day, 21days). The following results were obtained:
Lipemia:9 No significant interference up to an Lindex of 400. There is poor
correlation between the Lindex (corresponds to turbidity) and triglycerides CHET2 Repeatability Intermediate precision
concentration.
Sample Mean CV Mean CV
CHET2/CHED2 U/L (kat/L) % U/L (kat/L) %
Drugs: No interference was found at therapeutic concentrations using
common drug panels.10,11 Human serum 6374 (106) 0.5 6675 (111) 1.4
Anticoagulants: Citrate and fluoride inhibit the reaction and must not be PrecinormU 6263 (105) 0.6 6213 (104) 1.1
used. PrecipathU 6015 (100) 0.6 5964 (100) 0.9

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Cholinesterase/Dibucaine Gen.2 Enzymes

CHED2 Repeatability Intermediate precision 13 den Blaauwen DH, Poppe WA, Tritschler W. Cholinesterase (EC
3.1.1.8) mit Butyrylthiocholiniodid als Substrat:Referenzwerte in
Sample Mean CV Mean CV Abhngigkeit von Alter und Geschlecht unter Bercksichtigung
U/L (kat/L) % U/L (kat/L) % hormonaler Einflsse und Schwangerschaft. J Clin Chem Clin Biochem
Human serum 1533 (25.6) 1.2 1634 (27.2) 2.3 1983;21:381-386.
PrecinormU 1539 (25.7) 1.5 1524 (25.5) 2.6 14 Zawta B, Klein G, Bablok W. Temperature Conversion in Clinical
Enzymology? Klin Lab 1994;40:33-42.
PrecipathU 1439 (24.0) 1.7 1461 (24.4) 3.0 15 Weber F, Klein G, Kytzia H-J. Changed Reference values for Dibucaine
Method comparison for CHET2 Numbers with the new Roche CHE liquid Test, Data on File at Roche.
CHE2 values for human serum samples obtained on a 16 Bablok W, Passing H, Bender R, et al. A general regression procedure
COBASINTEGRA800 analyzer(y) were compared with those determined for method transformation. Application of linear regression procedures
using the corresponding reagent on a Roche/Hitachi917 analyzer(x). for method comparison studies in clinical chemistry, Part III.
JClinChemClinBiochem 1988 Nov;26(11):783-790.
Sample size (n)=57
A point (period/stop) is always used in this Method Sheet as the decimal
Passing/Bablok16 Linear regression separator to mark the border between the integral and the fractional parts of
y=1.03x+54 y=1.03x+83 a decimal numeral. Separators for thousands are not used.
=0.979 r=1.00 Symbols
Roche Diagnostics uses the following symbols and signs in addition to
SD (md95)=96.8 Sy.x=37.6 those listed in the ISO 152231 standard.
The activities of the samples were between 1159U/L and 13932U/L
(19.4233kat/L). Contents of kit
Method comparison for CHED2 Volume after reconstitution or mixing
A method comparison based on human sera between CHET2(x) and
CHED2(y) on COBASINTEGRA800analyzer gave the following dibucaine COBAS, COBASINTEGRA, COBASC, PRECICONTROL, PRECINORM, and PRECIPATH are trademarks of
Roche.
inhibition and correlation (U/L):
All other product names and trademarks are the property of their respective owners.
Sample size (n) 56 Significant additions or changes are indicated by a change bar in the margin.
2014, Roche Diagnostics
Corr. coefficient (r) 0.996
Passing/Bablok16 y=0.248x- 19
References
Roche Diagnostics GmbH, SandhoferStrasse116, D-68305 Mannheim
1 Moss DW, Henderson AR, Kachmar JF. Enzymes. In: Tietz NW, ed. www.roche.com
Fundamentals of Clinical Chemistry, 3rd ed. Philadelphia, PA: WB
Saunders 1987;346-421.
2 Evans RT. Cholinesterase phenotyping: clinical aspects and laboratory
applications. CRC Crit Rev Clin Lab Sci 1986;23:35-64.
3 George PM, Joyce SL, Abernethy MH. Screening for plasma
cholinesterase deficiency: an automated succinylcholine based assay.
Clin Biochem 1988;21:159-162.
4 Tietz NW, ed. Clinical Guide to Laboratory Tests, 3rd ed. Philadelphia
PA: WB Saunders Company 1995;132-133.
5 Schmidt E, Gerhardt W, Henkel E, et al. Proposal of Standard Methods
for the Determination of Enzyme Catalytic Concentrations in Serum and
Plasma at 37 C. Eur J Clin Chem Clin Biochem 1992;30:163-170.
6 Garry PJ. Serum cholinesterase variants: examination of differential
inhibitors, salts, and buffers used to measure enzyme activity Clin
Chem 1971;17:183-191.
7 Use of Anticoagulants in Diagnostic Laboratory Investigations. WHO
Publication WHO/DIL/LAB/99.1 Rev. 2. Jan. 2002.
8 Huizenga JR, van der Belt K, Gips CH. The Effect of Storage at
Different Temperatures on Cholinesterase Activity in Human Serum. J
Clin Chem Clin Biochem 1985;24:283-385.
9 Glick MR, Ryder KW, Jackson SA. Graphical Comparisons of
Interferences in Clinical Chemistry Instrumentation.
ClinChem1986;32:470-475.
10 Breuer J. Report on the Symposium Drug effects in Clinical Chemistry
Methods. Eur J Clin Chem Clin Biochem 1996;34:385-386.
11 Sonntag O, Scholer A. Drug interference in clinical chemistry:
recommendation of drugs and their concentrations to be used in drug
interference studies. Ann Clin Biochem 2001;38:376-385.
12 Bakker AJ, Mcke M. Gammopathy interference in clinical chemistry
assays: mechanisms, detection and prevention.
ClinChemLabMed2007;45(9):1240-1243.

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