BIOLABO

www.biolabo.fr CK-NAC I F C C Single Vial

MANUFACTURER:
BIOLABO SAS, Reagent for quantitative determination of Creatine Kinase activity
Les Hautes Rives [EC 2.7.3.2] in human serum.
02160, Maizy, France
REF 92207 R1 20 x 3 mL R2 1 x 60 mL
REF 92307 R1 8 x 20 mL R2 8 x 20 mL

TECHNICAL SUPPORT AND ORDERS

Tel : (33) 03 23 25 15 50
Fax: (33) 03 23 256 256
| IN VITRO DIAGNOSTIC USE
[email protected]

CLINICAL SIGNIFICANCE (1) REAGENTS COMPOSITION (7)

Creatine kinase (CK) is found mainly in cardiac, cerebral and skeletal R1 SUBSTRATES-ENZYMES SUBS CK Attention
muscle tissues. So, any dammage or disease of these tissues Creatine Phosphate 30 mmol/L
(myocardial infarction, acute cerebrovascular disease, muscular D-Glucose 20 mmol/L
dystrophy) will result in an increased level of CK activity in serum. There N-Acetyl-L-cystein 20 mmol/L
are 3 isoenzymatic forms of CK separable by electrophoresis or column
AMP 5 mmol/L
chromatography. Diagnosis specificity may be refined by the
ADP 2 mmol/L
determination of the level of each isoforms in serum (variable in
NADP 2 mmol/L
conjunction with tissue of origin).
Following myocardial infarction, CK activity begins to rise within 4 to 6 AP5A 10 µmol/L
hours, peaks between 18 and 30 hours and returns to normal by the G-6-PDH (Glucose-6-phosphate dehydrogenase) > 2500 UI/L
third day. HK (Hexokinase ) > 3000 UI/L
Before reconstitution:
Skin Irrit. 2 : H315 – Causes skin irritation
PRINCIPLE (4) (5) (6) STOT SE 3 : H335 – May cause respiratory irritation
Enzymatic method described by Oliver and modified by Rosalki and Acute Tox. 4: H302+H312+H332 – Harmful if swallowed, in contact with skin, if inhaled
Eye Irrit. 2: H319 - Causes serious eye irritation
later by Szasz.
P280: Wear protective gloves/protective clothing/eye protection/face protection, P264: Wash
1) Creatine phosphate + ADP CK Creatine + ATP hands thoroughly after handling, P302+P352 : IF ON SKIN: Wash with soap and water,
P304+P340 : IF INHALED: Remove victim to fresh air and keep at rest in a position comfortable
2) D-Glucose + ATP HK ADP + G-6-Phosphate for breathing, P305+351+338 : IF IN EYES: Rinse continuously with water for several minutes.
Remove contact lenses if present and easy to do – continue rinsing, P501: Dispose of
contents/container in accordance with dangerous waste regulations. Classification due to:
G6-PDH 6-Phosphogluconate Creatine phosphate 25 - < 50%.
3) G-6-Phosphate + NADP+
+ NADPH + H+
R2 BUFFER BUF CK Danger
The increase in absorbance, proportional to CK activity in the specimen,
is measured at 340 nm. Imidazole Acetate, pH 6,7 100 mmol/L
EDTA Na2 2 mmol/L
SAFETY CAUTIONS Magnésium Acetate 10 mmol/L
BIOLABO reagents are designated for professional, in vitro diagnostic Surfactant, stabilizer
use (do not pipette with mouth). Repro. 1B : H360 - May damage fertility or the unborn child
· Refer to current Material Safety Data Sheet available on request or P201: Obtain special instructions before use, P202: Do not handle until all safety precautions
on www.biolabo.fr have been read and understood, P308+P313: IF exposed or concerned: Get medical
advice/attention, P405: Store locked up, P501: Dispose of contents/container in accordance
· Verify the integrity of the contents before use. with dangerous waste regulations. Classification due to Imidazole < 1%.
· Waste disposal: Respect legislation in force in the country. For more details, refer to Safety Data Sheet (MSDS)
All specimens or reagents of biological origin should be handled as Once reconstituted: Working Reagent (R1+R2) is classified as contents of vial R2 (Buffer)
potentially infectious. Respect legislation in force in the country.

REAGENTS PREPARATION STABILITY AND STORAGE

· Substrate-Enzymes (R1): Stored away from light, well caped in the original vial at 2-8°C,
reagents are stable when stored and used as described in the
Use a non-sharp instrument to remove aluminium cap from the vial. insert:
● Buffer (R2): Ready to use
Unopened:
● Working reagent:
· Until expiry date stated on the label of the kit.
REF 92307: Transfer promptly the contents of R1 into R2
Once opened:
REF 92207: Transfer promptly 3 mL of R2 into R1. · Working reagent (R1+R2) is stable for 1 month
Mix gently and wait for complete dissolution. · Discard any reagent if cloudy or if reagent blank is > 0.500 at 340 nm.
· Don’t use working reagent after expiry date.
LIMITS (1) (3)
Discard any hemolysed specimen (some enzymes and intermediate SPECIMEN COLLECTION AND HANDLING (1) (2)
products released by erythrocytes interfere with the reaction).
Unhemolysed serum. Avoid anticoagulants such as heparin, EDTA,
For a more comprehensive review of factors affecting this assay refer to
citrate or fluoride. Protect from light and store in an airtight container to
the publication of Young D.S
prevent loss of CO2. Adjunction of thiols is not necessary.
MATERIAL REQUIRED BUT NOT PROVIDED CK activity is stable in serum for:
· 4 to 8 h at room temperature, 1 to 2 days at 2-8°C, 1 month at –20°c.
1. Basic medical analysis laboratory equipment.
2. Thermostated Spectrophotometer or Biochemistry Analyzer

IVD REF LOT ®

Manufacturer Use by In vitro diagnostic Temperature limitation Catalogue number See insert Batch number Store away from light sufficient for dilute with

H2O
Demineralized water Biological hazard

Made in France Latest revision : www.biolabo.fr Revision : 11/02/2019

CALIBRATION QUALITY CONTROL
· REF 95015 BIOLABO Multicalibrator (traceable to internal Masterlot) · REF 95010 BIOLABO EXATROL-N Level I
· REF 95011 BIOLABO EXATROL-P Level II
The calibration frequency depends on proper instrument functions and
· External quality control program.
on the preservation of reagent
It is recommended to control in the following cases:
· At least once a run.
EXPECTED VALUES (1) (2)
· At least once within 24 hours.
CK activity in serum is influenced by age, sexe, origin, corpulence, · When changing vial of reagent.
physic activity and other less known genetic factors. · After maintenance operations on the instrument.
If control is out of range, apply following actions:
IU/L (37°C) µKat/L 1. Repeat the test with the same control.
Newborn and paediatric 145-1578 (moy 382) [2,47-26,85] 2. If control is still out of range, prepare a fresh control serum and
Alter caesarian births 2-3 x adult repeat the test.
4 days 3 x adult 3. If control is still out of range, verify analysis parameters:
wavelength, temperature, specimen/reagent ratio, time counting,
6 weeks-12 years Adult values
calibration factor.
Adult Men > 19 years 20-200 UI/L [0,34-3,40] 4. If control is still out of range, use a new vial of reagent and
reassay.
Adult Women > 19 years 20-180 UI/L [0,34-3,06] 5. If control is still out of range, please contact BIOLABO technical
support or your local Agent.
Heterogeneity of serum CK activities have been described among
racial groups ; 97.5th percentiles were respectively as follows:
PROCEDURE
Population IU/L
Detailed Kenza 240TX procedure is available on request
African-American Men 520
Wavelength: 340 nm
Caucasian Men 370
Temperature: 37°C
African-American Women 290
Let stand reagent and specimen at room temperature.
Caucasian Women 145
Automated Manual
Normal Range can be assay and Instrument dependent. analyzer procedure
Each laboratory should establish its own normal ranges for the
population that it serves. Reagent 240 µL 1000 µL

PERFORMANCES at 37°C on KENZA 240TX Calibrator/Control or Specimen 12 µL 50 µL

Linearity Range: between 12 and 800 IU/L Mix. Start a timer and record initial absorbance at 340 nm after 2 minutes. Record
Detection limit: approx. 4 IU/L the absorbance again every minute during 3 minutes.
Precision:
Calculate absorbance change per minute (DAbs/min).
Within-run Low Normal High Between run Low Normal High
N = 20 level level level N = 20 level level level Notes:
Mean (IU/L) 63 123.4 556.4 Mean (IU/L) 65.8 131.2 583.4 1- Performances and stability data’s have been validated on KENZA
240TX and KENZA 450TX.
S.D. IU/L 0.7 1.4 3.0 S.D. IU/L 2.2 3.8 14.4
2-With Manual Procedure on Spectrophotometer and on other
C.V. % 1.1 1.1 0.5 C.V. % 3.3 2.9 2.5
automated analyzers, performances and stability should be validated
Comparison studies with commercially available reagent: by user.
Realised on human specimens (n=100) between 12 and 1063 IU/L 3- Applications proposal are available on request
y = 0,9745 x + 4.6059 r = 0.9995
Analytical sensitivity: approx. 0.0035 abs/min for 10 IU/L CALCULATION
Interferences:
Calculate the result as follows:
Total bilirubin Negative interference from 247 µmol/L
Direct bilirubin No interference up to 427 µmol/L With seric multicalibrator:
Ascorbic acid No interference up to 2500 mg/dL (∆Abs/min) Assay
CK Activity= x Calibrator Concentration
Glucose No interference up to 1060 mg/dL
(∆Abs/min) Calibrator
Turbidity Positive interference from 0.337 abs
Haemoglobin Positive interference from 257 µmol/L With theoretical factor :
Other substances may interfere (see § Limits)
Activity (IU/L) = ΔAbs/min x Factor
On-board stability: Working Reagent is stable 21 days
Calibration Frequency: 14 days VR x 1000
Make a new calibration when changing reagent batch, if quality control Factor = ———————
results are found out of the established range and after maintenance 6.22 x VE x P
operations With:
VR = Total reactional volume (mL)
VE = Specimen volume (mL)
REFERENCES
6.22 = Molar extinction coefficient for NADPH at 340 nm
(1) TIETZ N.W. Text book of clinical chemistry, 3rd Ed. C.A. Burtis, E.R. P = Pathlength (cm).
Ashwood, W.B. Saunders (1999) p. 657-666, 728, 1185-1190.
(2) Clinical Guide to Laboratory Test, 4th Ed., N.W. TIETZ (2006) p. 306-309 Example, with manual Procedure,
(3) p .3-185 to 3-190. (Path length 1 cm, 37°C, 340 nm):
(4) Oliver I.T., Biochem J., 61, (1955), p.116-122.
(5) Rosalki S.B., J. Lab. Clin. Med., 69, (1967), p.696-705.
(6) Szasz G., Gruber W., and Bernt E., Clin. Chem., 22 (1976), p.650-656. IU/L = (DAbs/min) x 3333
(7) Horder M and al, Approved IFCC recommendation on methods for the
measurement of catalytic concentration of enzymes. Part 7. IFCC method UI/L
µKat/L =
for creatine kinase [EC 2. 7. 3. 2]. Eur J. clin. Chem. Clin. Biochem., 29, 60
p435-456 (1991).

Made in France Latest revision : www.biolabo.fr Revision : 11/02/2019