CHOLINESTERASE

(DGKC 94 Substrate:Butyrylthiocholine )

Cat.No
117 081
117 082

Package Size
(Hit I) 5 x 25 mL
5 x 5 mL
(Hit II) 4 x 50 mL
2 x 20 mL

PRINCIPLE
The enzyme cholinesterase splits butyrylthiocholine
into butyrate and thiocholine. Thiocholine reacts
with yellow potassium hexacyanoferrate(III) to the
colorless Fe(II)-form.
The decrease of O.D. measured at 405 nm is
proportional to the activity of cholinesterase in the
sample.

REAGENTS
Reagent composition (Concentrations in the
test) :
Reagent R1:
Phosphate buffer(pH 7.6)
Potassium hexacyanoferrate(III)
mmol/L
Reagent R2(Lyophilisate) :
S-Butyrylthiocholineiodide
mmol/L

75.0 mmol/L
2.0

15.0

Stability
The sealed reagents are stable up to the indicated
expiry date if stored at 2 - 8C.

Preparation and Stability of Working

Reagents
Reagent R1 is ready to use.
Reagent R2 has to be dissolved with the
indicated volume of dist. water.
Reaction Mixture (Monoreagent) :
Mix Reagent 1 (5 parts) with
Reagent 2 (1 part).
Stability

at 2 - 8C: 4 weeks
at 18 - 22C: 5 days

SAMPLE MATERIAL
Serum, Plasma(EDTA/Heparine)
Stability:

8 days
at
2 - 8 C
3 months at
- 20 C
(Freeze only once!)

REFERENCE VALUES:
Female:
3.93-10.80kU/l (65..5-180kat/l)
Male:
4.62-11.50kU/l (77.0-192kat/l)
Note: Reference values depend - beside other factors on age structure and ethnic composition of population.For
orientation purposes the following reference ranges,which
were obtained from a middle european population, can
be taken.

ASSAY PROCEDURE
Wavelength : 405 nm Hg
Light path:
1 cm
Temperature : 37C

R1 / R2 Procedure (with Start-Reagent)

Pipette into Cuvettes
Reagent R1
1000 l
Sample
20 l
Mix well , incubate for abt. 5 min
Reagent R2
200 l
Mix well, incubate for 90 sec, then read initial
absorbance A1 and at the same time start stop watch.
Repeat measurement after exactly 1, 2 and 3 min

Monoreagent Procedure ( Sample-Start)

Pipette into Cuvettes
Monoreagent
1000 l
Sample
20 l
Mix well, incubate for 90 sec, then read initial
absorbance A1 and at the same time start stop watch.
Repeat measurement after exactly 1, 2 and 3 min

CALCULATION
For procedure with Start-Reagent:

A/min x 98.7 = kU/l

A/min x 1645 = kat/l
For procedure with Monoreagent:

A/min x 79.5 = kU/l

A/min x 1325 = kat/l
CALIBRATION AND QUALITY CONTROL
For the calibration of automated analyzers Greiner
Multicalibrator Unical-M is recommended, for quality
control use Greiner normal and abnormal Controlsera Unitrol-I and Unitrol-II .

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Greiner Diagnostic GmbH - Unter Gereuth 10 D-79353 Bahlingen Germany

www.greiner-diagnostic.com

PERFORMANCE DATA (37C)

LITERATURE

-Linearity
Up to CHE activity of 33kU/l (550kat/l).
If the enzyme activity exceeds limit,dilute the sample
1+3 with physiological saline(0.9%) and repeat the
assay. The result must be multiplied by 4.

1. Lothar Thomas, Labor und Diagnose, TH-Books

Verlagsgesellschaft mbH, Frankfurt/Main, 2000,
5.Auflage, Seite 67 ff.
2. Schmidt et al., Eur.J.Clin.Chem.Clin.Biochem.
1992, 30, 165.
SYMBOLS USED

- Sensitivity
The low detection limit is equal to 10 U/L.
- Precision
Within-run reproducibility
Within
series
n = 11
Sample 1
Sample 2
Sample 3

For in vitro diagnostic medical use

Mean
value
[kU/L]

Standarddeviation
[kU/L]

CV
[%]

6405
5755
4523

43,9
73,0
42,2

0,686
1,27
0,933

Between-run reproducibility
Day to day
n = 11
Sample 1
Sample 2
Sample 3

Batch Code

Use by

Mean
value
[U/L]

Standarddeviation
[U/L]

CV
[%]

6357
5735
4518

103,6
67,9
87,9

1,63
1,18
1,93

Temperature limitation

- Correlation
A comparative study has been performed between
Greiner method and another commercial reagent on
30 human serum samples. The parameters of linear
regression are as follows:
Linear regression:
y 1,004 x - 189 U/l; r= 0,999 .

INTERFERENCES
-

Ascorbic Acid: no interference up to 100 mg/dl

Bilirubin: no interference up to 40 mg/dl
Hemoglobin no interference up to 1000mg/dl
Triglycerides: no interference up to 2000 mg/dl

10/2007//rev03/2014

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Greiner Diagnostic GmbH - Unter Gereuth 10 D-79353 Bahlingen Germany