Enzymes p400 en

ALP CP
ABX Pentra
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Mono-reagent application
2008/10/10
A93A00012Q EN
A11A01626
26 ml
6.5 ml
HORIBA ABX
BP 7290
34184 Montpellier- cedex 4 - France
S.A.S. au capital de 41.700.000 - RCS Montpellier 328 031 042 - SIRET 328 031 042 000 42 - APE 332 B Latest version documents on www.horiba-abx.com
ABX Pentra ALP CP (Mono-reagent application)
Ref.: A11A01626
Volume R1: 26 ml
Volume R2: 6.5 ml
Diagnostic reagent for quantitative in-vitro
determination of Alkaline Phosphatase (ALP) in
serum or plasma.
Clinical Interest (1,2)
Alkaline phosphatase (ALP), an hydrolytic enzyme acting optimally at
alkaline pH, exists in blood in numerous distinct forms which originate
mainly from bone and liver, but also from other tissues as kidney,
placenta, intestine, testes, thymus, lung and tumors. Physiological
increases are found during bone growth in childhood and in
pregnancy, while pathological increases are largely associated with
hepatobiliary and bone diseases. In hepatobiliary disease they
indicate obstruction of the bile ducts as in cholestasis caused by gall
stones, tumors or inflammation. Elevated activities are also observed
in infectious hepatitis. In bone diseases elevated ALP activities
originate from increased osteoblastic activity as in Pagets disease,
osteomalacia (rickets), bone metastases and hyperparathyroidism.
Method
Kinetic photometric test, according to the International Federation of
Clinical Chemistry (IFCC).
(ALP = Alkaline Phosphatase)
Reagents
ABX Pentra ALP CP is ready-to-use.
ABX Pentra ALP CP should be used according to this reagent notice.
HORIBA ABX cannot guarantee its performance if used otherwise.
Handling
Transfer the complete volume of Reagent R2 into the Reagent 1 container.
Recap the containers and homogenize the mixture by gentle inversion.
Pour the volume of the solution required for a daily workload into a
15, 10 or 4 mL reagent vial and place it in position 1 of one of the
available areas. Please use one of the following:
a 15 ml reagent vial
a 10 ml reagent vial + a specific adapter
a 4 ml reagent vial + a specific adapter
Reagent 1: 2-Amino-2-methyl-1-propanol pH 10.4 440 mmol/l
Magnesium sulphate 2.0 mmol/l
Zinc sulphate 1.25 mmol/l
HEDTA 2.5 mmol/l
Sodium azide < 1 g/l
Reagent 2: p-Nitrophenylphosphate 80 mmol/l
p-Nitrophenylphosphate + H
2
O Phosphate + p-Nitrophenol
ALP
Place the reagent rack in the refrigerated reagent compartment of the
ABX Pentra 400.
Important note: The residual amount of reagent should be discarded at
the end of day.
Calibrator
For calibration, use:
ABX Pentra MultiCal, Ref. A11A01652 (not included)
10 x 3 ml (lyophilisate)
Control
For internal quality control, use:
ABX Pentra N Control, Ref. A11A01653 (not included)
ABX Pentra P Control, Ref. A11A01654 (not included)
Each control should be assayed daily and/or after each calibration.
The frequency of controls and the confidence intervals should
correspond to laboratory guidelines and country-specific directives.
The results must be within the range of the defined confidence limits.
Each laboratory should establish a procedure to follow if the results
exceed these confidence limits.
ABX Pentra ALP CP
Reagent 1 + Reagent 2
ALP CP
ABX Pentra
Materials required but not provided
Automated clinical chemistry analyser
Standard laboratory equipment.
Specimen
Serum.
heparin plasma.
Loss of activity within 2 - 3 days at 15 - 25 C < 10 %.
Reference range (4,5)
Adults:
Children:
Storage and Stability
Reagents, in unopened cassettes, are stable up to the expiry date on
the label if stored at 2-8 C protected from light and contamination is
avoided.
Stability after opening : refer to the paragraph Performance on ABX
Pentra 400.
Do not freeze the reagents.
Assay Procedure
Test instructions for other automated systems than ABX Pentra 400 are
available on request.
Waste Management
1. Please refer to local legal requirements.
2. This reagent contains sodium azide (0.95 g/l) as a preservative. As
sodium azide may react with lead or copper to form explosive metal
azides, this reagent should be disposed of by flushing with copious
amounts of water.
General Precautions
1. This reagent is for professional in-vitro diagnostic use only.
2. Do not swallow! Avoid contact with skin and mucous membranes.
3. During reaction p-nitrophenol is produced which is poisonous
when inhaled, swallowed or absorbed through skin. If the reaction
mixture comes in contact with skin or mucous membranes wash
copiously with water.
4. Take the necessary precautions for the use of laboratory reagents.
5. The reagent cassettes are disposable and should be disposed of in
accordance with the local legal requirements.
6. Please refer to the MSDS associated with the reagent.
Performance on ABX Pentra 400
The performance data listed below have been obtained on the ABX
Pentra 400 analyser.
Number of tests: 125 tests
Reagent Stability:
Use fresh reagent each day. Discard the remaining reagent in container
after use. Once open, the cassette is stable for 29 days.
Sample volume: 4 l/test
Detection limit:
The detection limit is determined according to the Valtec protocol (6)
and equals 8 U/l.
Accuracy and Precision:
Repeatability (within-run precision)
3 specimens of low, medium and high concentration and 2 controls are
tested 20 times according to the recommendations found in the Valtec
protocol (6).
Reproducibility (run-to-run precision)
2 specimens of medium and high levels and 2 controls are tested in
duplicate for 20 days (2 series per day) according to the
recommendations found in the NCCLS, EP5-A protocol (7).
Stability : 7 days at 4 - 8 C
2 months at - 20 C
37 C
Women [U/l] 35 - 104
Men [U/l] 40 - 129
37 C
1 day [U/l] < 250
2 to 5 days [U/l] < 231
6 days to 6 months [U/l] < 449
7 months to 1 year [U/l] < 462
1 to 3 years [U/l] < 281
4 to 6 years [U/l] < 269
7 to 12 years [U/l] < 300
13 to 17 years (female) [U/l] < 187
13 to 17 years (male) [U/l] < 390
Mean value U/l CV %
Normal control 98 1,26
Pathological control 278 0,80
Specimen 1 36 4,01
Specimen 2 63 2,11
Specimen 3 467 0,60
Mean value U/l CV %
Normal control 96 2,75
Pathological control 270 2,52
Specimen 1 32 6,00
Specimen 2 474 2,15
ALP CP
ABX Pentra
Linearity and Measuring Range:
The reagent linearity is determined according to the recommendations
found in the NCCLS, EP6-P protocol (8).
Low linearity: 8 U/l
High linearity: 1825 U/l, with automatic post-dilution: 7300 U/l.
Correlation:
95 patient samples are correlated with a commercial reagent taken as
reference according to the recommendations found in the NCCLS, EP9-
A2 protocol (9).
The equation for the allometric line obtained is:
y = x with a correlation coefficient r
2
= 0.9936.
Interferences:
Calibration stability:
The calibration stability is 6 hours.
Note: A recalibration is recommended when reagent lots change, and
when quality control results fall outside the range established.
Application release
a
: 9.xx
Warning
It is the users responsibility to verify that this document is applicable
to the reagent used.
Reference
1. Thomas L. Clinical Laboratory Diagnostics. 1
st
ed. Frankfurt: TH-
Books Verlagsgesellschaft; 1998. p. 36-46.
2. Moss DW, Henderson AR. Clinical enzymology. In: Burtis CA,
Ashwood ER, editors. Tietz Textbook of Clinical Chemistry. 3
rd
ed.
Philadelphia: W.B Saunders Company; 1999. p. 617-721.
3. Tietz NW, Rinker D, Shaw LM. IFCC method for alkaline
phosphatase. J. Clin. Chem. Clin. Biochem. 1983; 21:731-748.
4. Abitch K, El-Samalouti V, Junge W, Kroll M, Luthe H, Treskes M et
al.. Multicenter evaluation of new GGT and ALP reagents with new
reference standardization and determination of 37C reference
intervals. Clin Chem Lab Med 2001; 39, Special Supplement pp S
346.
5. Tietz NW, Shuey DF. Reference intervals for Alkaline Phosphatase
Activity Determined by IFCC and AACC Reference Methods. Clin
Chem 1986; 32: 1593-1594.
6. Vassault A., Grafmeyer D. Naudin C. et al., Protocole de validation
de techniques (document B), Ann. Biol. Clin., 1986, 44, 686-745.
7. Evaluation of Precision Performance of Clinical Chemistry Devices,
Approved Guideline, NCCLS document EP5-A, Vol. 19, No. 2,
february 1999.
8. Evaluation of the Linearity of Quantitative Analytical Methods,
Proposed Guideline, NCCLS document EP6-P, Vol. 6, No. 18,
september 1986.
9. Method Comparison and Bias Estimation Using Patient Samples,
Approved Guideline, 2
nd
ed., NCCLS document EP9-A2, Vol. 22, No.
19, 2002.
Haemoglobin: No significant influence is observed up to 59 mol/l
Triglycerides: No significant influence is observed up to 7 mmol/l
Total Bilirubin: No significant influence is observed up to 502 mol/l
Direct Bilirubin: No significant influence is observed up to 371 mol/l
a. Modification from index P to Q: new application release.
ALP CP
ABX Pentra
ALP CP
ABX Pentra
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Use in reagent rack
2007/06/08
A93A00012N EN
A11A01626
26 ml
6.5 ml
HORIBA ABX
BP 7290
ABX Pentra ALP CP (Use in reagent rack)
Ref.: A11A01626
Volume R1: 26 ml
Volume R2: 6.5 ml
determination of Alkaline Phosphatase (ALP)
in serum or plasma.
Alkaline phosphatase (ALP), an hydrolytic enzyme acting optimally at
alkaline pH, exists in blood in numerous distinct forms which originate
mainly from bone and liver, but also from other tissues as kidney,
placenta, intestine, testes, thymus, lung and tumors. Physiological
increases are found during bone growth in childhood and in
pregnancy, while pathological increases are largely associated with
hepatobiliary and bone diseases. In hepatobiliary disease they
indicate obstruction of the bile ducts as in cholestasis caused by gall
stones, tumors or inflammation. Elevated activities are also observed
in infectious hepatitis. In bone diseases elevated ALP activities
originate from increased osteoblastic activity as in Pagets disease,
osteomalacia (rickets), bone metastases and hyperparathyroidism.
Method
Kinetic photometric test, according to the International Federation of
Clinical Chemistry (IFCC).
(ALP = Alkaline Phosphatase)
Reagents
ABX Pentra ALP CP is ready-to-use.
ABX Pentra ALP CP should be used according to this reagent notice.
Handling
Transfer the required volume of Reagent 1 in a container 15, 10 or 4
ml.
Transfer the required volume of Reagent 2 in a container 10 or 4 ml.
Reagent 1 and Reagent 2 should be placed on the same reagent rack
sector A, B or C (see diagram below, sector A is taken as an example).
Reagent 1: 2-Amino-2-methyl-1-propanol pH 10.4 440 mmol/l
Magnesium sulphate 2.0 mmol/l
Zinc sulphate 1.25 mmol/l
HEDTA 2.5 mmol/l
Reagent 2: p-Nitrophenylphosphate 80 mmol/l
p-Nitrophenylphosphate + H
2
O Phosphate + p-Nitrophenol
ALP
Place Reagent 1 in position 1 of one available sector using either:
Reagent container 15 ml
Reagent container 10 ml + its specific adaptor
Place Reagent 2 in position 2 of same selected sector using either:
Place the reagent rack in the refrigerated ABX Pentra 400 reagent
compartment.
Calibrator
Control
ABX Pentra ALP CP
Reagent 2
ABX Pentra ALP CP
Reagent 1
ALP CP
ABX Pentra
Specimen
Serum.
heparin plasma.
Loss of activity within 2 - 3 days at 15 - 25 C < 10 %.
Adults:
Children:
avoided.
Pentra 400.
Assay Procedure
Waste Management
amounts of water.
General Precautions
2. Do not swallow! Avoid contact with skin and mucous membranes.
3. During reaction p-nitrophenol is produced which is poisonous
when inhaled, swallowed or absorbed through skin. If the reaction
mixture comes in contact with skin or mucous membranes wash
copiously with water.
Reagent Stability:
Use fresh reagent each day. Discard the remaining reagent in container
after use. Once open, the cassette is stable for 29 days.
Detection limit:
and equals 6 U/l.
protocol (6).
Stability : 7 days at 4 - 8 C
2 months at - 20 C
37 C
Women [U/l] 35 - 104
Men [U/l] 40 - 129
37 C
1 day [U/l] < 250
2 to 5 days [U/l] < 231
6 days to 6 months [U/l] < 449
7 months to 1 year [U/l] < 462
1 to 3 years [U/l] < 281
4 to 6 years [U/l] < 269
7 to 12 years [U/l] < 300
13 to 17 years (female) [U/l] < 187
13 to 17 years (male) [U/l] < 390
Mean value U/l CV %
Normal control 90.79 1.27
Pathological control 252.68 0.62
Specimen 1 28.05 3.98
Specimen 2 54.88 2.42
Specimen 3 430.87 0.84
ALP CP
ABX Pentra
Correlation:
A2 protocol (9).
Y = 0.99 x - 1.22 with a correlation coefficient r
2
= 0.99.
Interferences:
The calibration stability is 7 hours.
Application release
a
: 7.xx
Warning
Reference
st
ed. Frankfurt: TH-
Books Verlagsgesellschaft; 1998. p. 36-46.
rd
ed.
3. Tietz NW, Rinker D, Shaw LM. IFCC method for alkaline
phosphatase. J. Clin. Chem. Clin. Biochem. 1983; 21:731-748.
4. Abitch K, El-Samalouti V, Junge W, Kroll M, Luthe H, Treskes M et al..
Multicenter evaluation of new GGT and ALP reagents with new
reference standardization and determination of 37C reference
intervals. Clin Chem Lab Med 2001; 39, Special Supplement pp S 346.
5. Tietz NW, Shuey DF. Reference intervals for Alkaline Phosphatase
Activity Determined by IFCC and AACC Reference Methods. Clin
Chem 1986; 32: 1593-1594.
february 1999.
september 1986.
nd
19, 2002.
Mean value U/l CV %
Specimen 1 64.11 4.36
Specimen 2 190.44 2.66
a. Modification from index L to N: suppression of minor index.
ALP CP
ABX Pentra
ALT CP
ABX Pentra
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2007/06/08
A93A00022I EN
A11A01627
56 ml
14 ml
HORIBA ABX
BP 7290
ABX Pentra ALT CP
Ref.: A11A01627
Volume R1: 56 ml
Volume R2: 14 ml
determination of Alanine AminoTransferase (ALT) in
serum or plasma.
Alanine Aminotransferase (ALAT/ALT), formerly called Glutamic
Pyruvic Transaminase (GPT) and Aspartate Aminotransferase (ASAT/
AST), formerly called Glutamic Oxalacetic Transaminase (GOT) are the
most important representatives of a group of enzymes, the
aminotransferases or transaminases, which catalyze the conversion of
-keto acids into amino acids by transfer of amino groups.
As a liver specific enzyme ALT is only significantly elevated in
hepatobiliary diseases. Increased AST levels, however, can occur in
connection with damages of heart or skeletal muscle as well as of liver
parenchyma. Parallel measurement of ALT and AST is therefore applied
to distinguish liver from heart or skeletal muscle damages. The AST/
ALT ratio is used for differential diagnosis in liver diseases. While
ratios < 1 indicate mild liver damage, ratios > 1 are associated with
severe, often chronic liver diseases.
Method
Optimized UV-test according to IFCC (International Federation of
Clinical Chemistry) modified method without pyridoxal phosphate.
(ALT = Alanine Aminotransferase, LDH = Lactate Dehydrogenase)
Reagents
ABX Pentra ALT CP is ready-to-use.
ABX Pentra ALT CP should be used according to this reagent notice.
Handling
Remove both caps of the cassette, place in the refrigerated ABX Pentra
400 reagent compartment.
If present, remove foam by using a plastic pipette.
Reagent 1: TRIS pH 7.5 140 mmol/l
L-Alanine 709 mmol/l
LDH (lactate dehydrogenase) 1700 U/l
Reagent 2: 2-Oxoglutarate 85 mmol/l
NADH 1.09 mmol/l
L-Alanine + 2-Oxoglutarate L-Glutamate + Pyruvate
ALT
Pyruvate + NADH + H
+
D-Lactate + NAD
+
LDH
Calibrator
Control
Specimen
Serum.
heparin Plasma or EDTA Plasma.
Loss of activity within 3 days:
at 2 - 8 C: < 10 %
at 15 - 25 C : < 17 %
Stability at -20 C: at least 3 months
ALT CP
ABX Pentra
Reference range (4)
avoided.
Pentra 400.
Assay Procedure
Waste Management
amounts of water.
General Precautions
2. Do not swallow. Avoid contact with skin and mucous membranes.
On board Reagent Stability:
Once opened, the reagent cassette placed in the refrigerated ABX
Pentra 400 compartment is stable for 42 days.
Detection limit:
and equals 4 U/l.
protocol (5).
2 specimens of low and high levels and 2 controls are tested in
Correlation:
A2 protocol (8).
Y = x + 5 with a correlation coefficient r
2
= 0.9964.
Interferences:
The reagent is calibrated on Day 0. The calibration stability is checked
by testing 2 control specimens.
The calibration stability is at least 8 days.
Application release
a
: 3.xx
Warning
37 C
Women: 34 U/l
Men: 45 U/l
Mean value U/l CV %
Normal control 39.73 1
Specimen 1 17.43 3.07
Specimen 2 28.44 2.28
Specimen 3 127.88 0.59
Mean value U/l CV %
Specimen 1 31.49 6
Specimen 2 87.6 2.48
Haemoglobin: No significant influence is observed up to 195 mol/l.
Triglycerides: No significant influence is observed up to 5 mmol/l.
Total Bilirubin: No significant influence is observed up to 450 mol/l.
a. Modification from index H to I: suppression of minor index.
ALT CP
ABX Pentra
Reference
1. Thomas L. Alanine aminotransferase (ALT), Aspartate
aminotransferase (AST). In: Thomas L, editor. Clinical Laboratory
Diagnostics. 1
st
ed. Frankfurt: TH-Books Verlagsgesellschaft; 1998.
p. 55-65.
rd
ed.
3. Bergmeyer HU, Horder M, Rej R. International Federation of
Clinical Chemistry (IFCC) Scientific Committee, Analytical section:
approved recommendation (1985) on IFCC methods for the
measurement of catalytic concentration of enzymes. Part 3. IFCC
method for alanine aminotransferase (L-alanine:2-oxoglutarate
aminotransferase, EC 2.6.1.2). J. Clin. Chem. Clin. Biochem. 1986;
24:481-495.
4. IFCC Primary Reference Procedures for the Measurement of
Catalytic Activity Concentrations of Enzymes at 37C; Part 4 ; Clin
Chem Lab Med 2002; 40(7) : 718-724.
february 1999.
september 1986.
nd
19, 2002.
ALT CP
ABX Pentra
Amylase CP
ABX Pentra
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2008/10/10
A93A00042M EN
A11A01628
26 ml
6.5 ml
HORIBA ABX
BP 7290
ABX Pentra Amylase CP
Ref.: A11A01628
Volume R1: 26 ml
Volume R2: 6.5 ml
determination of -Amylase in serum,
plasma and urine.
-Amylases are hydrolytic enzymes which break down starch into
maltose. In the human body -amylases originate from various organs:
the pancreatic amylase is produced by the pancreas and released into the
intestinal tract, the salivary amylase is synthesized in the salivary glands
and secreted into saliva. The amylase present in the blood is eliminated
through the kidney and excreted into the urine. Therefore, elevation of
serum activity is reflected in a rise of urinary amylase activity.
Measurement of -amylase in serum and urine is mainly used for the
diagnosis of pancreatic disorders as well as for detecting the
development of complications. In acute pancreatitis the blood amylase
activity increases within few hours after onset of abdominal pain,
peaks after approx. 12 hours and returns to values within the reference
range at the latest after 5 days. The specificity of -amylase for
pancreatic disorders is not very high as elevated levels are measured
also in various non-pancreatic diseases, e.g. parotitis and renal
insufficiency. Therefore, for confirmation of an acute pancreatitis
measurement of lipase should be additionally performed.
Method
Enzymatic photometric test, in which the substrate 4,6-ethylidene-
(G7)-p-nitrophenyl-(G1)--D-maltoheptaoside (EPS-G7) is cleaved by
-amylases into various fragments. These are further hydrolyzed in a
second step by -glucosidase producing glucose and p-nitrophenol.
The increase in absorbance represents the total (pancreatic and
salivary) amylase activity in the sample (3,4).
(PNP = p-Nitrophenol, G =Glucose)
Reagents
ABX Pentra Amylase CP is ready-to-use.
Reagent 1: Goods buffer pH 7.1 0.1 mol/l
NaCl 62.5 mmol/l
MgCl2 12.5 mmol/l
-Glucosidase 2.5 kU/l
Reagent 2: Goods buffer pH 7.1 0.1 mol/l
EPS-G7 8.5 mmol/l
5 EPS-G7+5 H
2
O
2 EthylideneG5 + 2G2PNP + 2 Ethylidene-G4 +
2G3PNP + Ethylidene-G3 + G4PNP
-Amylase
5 PNP + 14 G 2 G2PNP + 2 G3PNP + G4PNP + 14 H
2
O
-Glucosidase
ABX Pentra Amylase CP should be used according to this reagent
notice. HORIBA ABX cannot guarantee its performance if used
otherwise.
Handling
Remove both caps of the cassette. If present, remove foam by using a
plastic pipette.
Position the respective protective cap, ref. GBM0969 on R1 and Ref.
GBM0970 on R2 and place in the refrigerated ABX Pentra 400 reagent
compartment.
Calibrator
Control
ABX Pentra Urine Control L/H, Ref. A11A01674 (not included)
1 x 10 ml + 1 x 10 ml
Amylase CP
ABX Pentra
ABX Pentra Clean-Chem CP, Ref. A11A01755, 30 ml
Specimen
Serum.
heparin Plasma.
Urine.
Reference range (4)
the label if stored at 2 - 8 C, protected from light and contamination
is avoided.
Pentra 400.
Assay Procedure
Waste Management
amounts of water.
General Precautions
2. Saliva and skin contain -amylase therefore never pipette reagents
by mouth and avoid skin contact with the reagents.
Serum, Plasma
Sample volume: 4.0 l/test
Detection limit:
and equals 9 U/l.
protocol (5).
3 specimens of low, medium and high levels and 2 controls are tested
in duplicate for 20 days (2 series per day) according to the
Correlation:
A2 protocol (8).
Y = 0.96 x + 2.46 with a correlation coefficient r
2
= 0.9955.
Stability in:
Serum/Plasma: 7 days at 20 - 25 C
7 days at 4 - 8 C
1 year at - 20 C
Urine: 2 days at 20 - 25 C
10 days at 4 - 8 C
3 weeks at - 20 C
Women Men
Serum/plasma < 100 U/l < 100 U/l
Urine < 447 U/l < 491 U/l
Mean value U/l CV %
Specimen 2 151.8 1.23
Specimen 3 383.6 1.32
Mean value U/l CV %
Specimen 1 52.80 5.29
Specimen 2 148.70 4.14
Specimen 3 389.60 2.50
Amylase CP
ABX Pentra
Interferences:
Application release
a
: 10.xx
Urine
Once opened, the reagent cassette placed in the refregerated ABX
Detection limit:
and equals 4.92 U/l.
protocol (5).
2 controls are tested in duplicate for 20 days (2 series per day)
according to the recommendations found in the NCCLS, EP5-A protocol
(6).
Low linearity: 4.92 U/l
Correlation:
A2 protocol (8).
2
= 0.99.
Interferences:
Application release: 7.xx
Warning
Reference
1. Lorentz K. -Amylase. In : Thomas L, editor. Clinical laboratory
diagnostics. 1
st
ed. Frankfurt: TH-Book Verlagsgesellschaft ; 1998.
p.192-202.
2. Moss DW, Henderson AR. Digestive enzymes of pancreatic origin.
In: Burtis CA, Ashwood ER, editors. Tietz Textbook of Clinical
Chemistry. 3
rd
ed. Philadelphia: W.B Saunders Company; 1999.
p.689-698.
3. Kruse-JarresJD,KaiserC,Hafkenscheid JC, Hohenwallner W, SteinW.,
Bohner J et al. Evaluation of a new alpha-amylase assay using 4,6-
ethylidene-(G7)-1-4-nitrophenyl-(G1)-alpha,D-maltoheptaoside as
substrate. J. Clin. Chem. Biochem. 1989; 27:103 - 113.
4. Junge W, Wortmann W, Wilke B, Waldenstroem J et al.
Development and evaluation of assays for determination of total
and pancreatic amylase at 37C according to the principle
recommended by the IFCC. Clin Biochem 2001; 34:607-15.
Approved Guideline, NCCLS document EP5-sA, Vol. 19, No. 2,
february 1999.
a.Modification from index L to M: New application release.
Mean value U/l CV %
Specimen 1 86.35 1.65
Specimen 2 157.73 0.63
Specimen 3 286.80 0.63
Mean value U/l CV %
Amylase CP
ABX Pentra
september 1986.
nd
19, 2002.
AST CP
ABX Pentra
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2007/06/08
A93A00032I EN
A11A01629
56 ml
14 ml
HORIBA ABX
BP 7290
ABX Pentra AST CP
Ref.: A11A01629
Volume R1: 56 ml
Volume R2: 14 ml
determination of Aspartate AminoTransferase (AST)
in serum or plasma.
Aspartate Aminotransferase (ASAT/AST), formerly called Glutamic
Oxalacetic Transaminase (GOT) and Alanine Aminotransferase (ALAT/
ALT), formerly called Glutamic Pyruvic Transaminase (GPT) are the
most important representatives of a group of enzymes, the
aminotransferases or transaminases, which catalyze the conversion of
-keto acids into amino acids by transfer of amino groups.
As a liver specific enzyme ALT is only significantly elevated in
hepatobiliary diseases. Increased AST levels, however, can occur in
connection with damages of heart or skeletal muscle as well as of liver
parenchyma. Parallel measurement of ALT and AST is therefore applied
to distinguish liver from heart or skeletal muscle damages. The AST/
ALT ratio is used for differential diagnosis in liver diseases. While
ratios < 1 indicate mild liver damage, ratios > 1 are associated with
severe, often chronic liver diseases.
Method
Optimized UV-test according to IFCC (International Federation of
Clinical Chemistry) modified method without pyridoxal phosphate.
(AST = Aspartate Aminotransferase, MDH = Malate Dehydrogenase)
Reagents
ABX Pentra AST CP is ready-to-use.
ABX Pentra AST CP should be used according to this reagent notice.
Handling
L-Aspartate 340 mmol/l
MDH (malate dehydrogenase) 900 U/l
LDH (lactate dehydrogenase) 900 U/l
Reagent 2: 2-Oxoglutarate 85 mmol/l
NADH 1.09 mmol/l
L-Aspartate + 2-Oxoglutarate L-Glutamate + 2-Oxalacetate
AST
Oxalacetate + NADH + H
+
L-Malate + NAD
+
MDH
Calibrator
Control
Specimen
Serum.
Heparin Plasma or EDTA Plasma.
Loss of activity within 3 days:
at 2 - 8 C : < 8 %
at 15 - 25 C: < 10 %
Stability at -20 C: at least 3 months
AST CP
ABX Pentra
Reference range (4)
is avoided.
Pentra 400. Do not freeze the reagents.
Assay Procedure
Waste Management
amounts of water.
General Precautions
Detection limit:
and equals 4 U/l.
protocol (5).
Correlation:
A2 (8).
2
= 0.9977 .
Interferences:
Application release
a
: 4.xx
Warning
37 C
Women: 31 U/l
Men: 35 U/l
Mean value U/l CV %
Specimen 3 145.4 1.08
Mean value U/l CV %
Specimen 2 348.0 4.97
Triglycerides: No significant influence is observed up to 4.6 mmol/l
a. Modification from index H to I: suppression of minor index.
AST CP
ABX Pentra
Reference
1. Thomas L. Alanine aminotransferase (ALT), Aspartate
aminotransferase (AST). In: Thomas L, editor. Clinical Laboratory
Diagnostics. 1
st
ed. Frankfurt: TH-Books Verlagsgesellschaft; 1998.
p. 55-65.
rd
ed.
3. Bergmeyer HU, Horder M, Rej R. International Federation of
Clinical Chemistry (IFCC) Scientific Committee, Analytical section:
approved recommendation (1985) on IFCC methods for the
measurement of catalytic concentration of enzymes. Part 2. IFCC
method for aspartate aminotransferase (L-aspartate: 2-
oxoglutarate aminotransferase, EC 2.6.1.1). J. Clin. Chem. Clin.
Biochem. 1986; 24:497-510.
Chem Lab Med 2002; 40(7) : 725-733.
february 1999.
september 1986.
nd
19, 2002.
AST CP
ABX Pentra
CK- MB RTU
ABX Pentra
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A93A00062L EN
A11A01643
1 x 20 ml
1 x 5 ml
HORIBA ABX
BP 7290
ABX Pentra CK-MB RTU
Ref. : A11A01643
Volume R1: 1 x 20 ml
Volume R2: 1 x 5 ml
determination of CK-MB in serum.
Creatine kinase (CK) is an enzyme which consists of isoenzymes mainly
of the muscle (CK-M) and the brain (CK-B). CK exists in serum in
dimeric forms as CK-MM, CK-MB, CK-BB and as macro-enzymes.
Measurement of CK-MB is a quite specific test for detection of cardiac
muscle damage and is therefore used for diagnosis and monitoring of
myocardial infarction.
Method
Optimized UV test according to DGKC (German Society of Clinical
Chemistry) and IFCC (International Federation of Clinical Chemistry)
for CK with inhibition of CK-M isoenzymes by polyclonal antibodies.
CK-MB consists of the subunits CK-M and CK-B.
Specific antibodies against CK-M inhibits the complete CK-MM activity
(main part of the total CK activity) and the CK-M subunit of CK-MB.
Only CK-B activity is measured, which is half of the CK-MB activity.
Reagents
ABX Pentra CK-MB RTU is ready-to-use.
ABX Pentra CK-MB RTU should be used according to this reagent notice.
Reagent 1:
Imidazole pH 6.7 62.5 mmol/l
Glucose 27.5 mmol/l
N-Acetyl cysteine (NAC) 27.5 mmol/l
Magnesium acetate 13.8 mmol/l
EDTA-Na2 2.1 mmol/l
NADP 2.75 mmol/l
Hexokinase (HK) 3 kU/l
Polyclonal antibodies (sheep) against
human CK-M; inhibiting capacity
2500 U/l
Reagent 2:
Imidazole 160 mmol/l
Creatine phosphate 30 mmol/l
EDTA-Na2 2.1 mmol/l
ADP 11 mmol/l
AMP 28 mmol/l
Diadenosine pentaphosphate 56 mol/l
Glucose-6-phosphate dehydrogenase (G6P-DH) 7.5 kU/l
Handling
Reagent 1 and Reagent 2 should be placed on the same reagent rack
sector A, B or C (see diagram below, sector A is taken as an example).
Place Reagent 1 in position 1 of one available sector using either:
Place Reagent 2 in position 2 of same selected sector using either:
Reagent container 4 ml + its specific adaptor.
Place the reagent rack in the refrigerated ABX Pentra 400 reagent
compartment.
Calibrator
N/A.
Control
ABX Pentra CK Control, Ref. A11A01786 (not included)
Reagent 2
Reagent 1
CK- MB RTU
ABX Pentra
Specimen
Serum.
Reference range (1)
Reagents, in unopened vials, are stable up to the expiry date on the
label if stored at 2 - 8 C, protected from light and contamination.
Once opened, ABX Pentra CK-MB RTU is stable 28 days at 2-8C.
Assay Procedure
Waste Management
amounts of water.
General Precautions
4. The reagent vials should be discarded after use.
Detection limit:
and equals 5.24 U/l.
3 specimens of low, medium and high concentration and 1 control are
protocol (5).
2 specimens of medium and high levels and 1 control are tested in
Correlation:
A2 protocol (8).
2
= 0.98.
Interferences:
Application release
a
: 8.xx
Warning
Reference
1. Stein W. Creatine kinase (total activity), creatine kinase
isoenzymes and variants. In: Thomas L, ed. Clinical laboratory
diagnostics.Frankfurt: TH-Books Verlagsgesellschaft;1998.p.71-80.
Loss of activity:
after 24 h at 2 - 8 C < 10 %
after 1 h at 15 - 25 C < 10 %
Stability: at -20 C 4 weeks (in the dark)
37 C
< 24 U/l
Mean value U/l CV %
Specimen 1 50.36 2.81
Specimen 2 179.85 1.45
Specimen 3 282.23 0.87
Mean value U/l CV %
Specimen 1 46.8 6.5
Haemoglobin: Positive interference starting from 56 mol/l
a.Modification from index K to L: new application release.
CK- MB RTU
ABX Pentra
rd
ed.
3. Wrzburg U, Hennrich N, Orth HD, Lang H. Quantitative
determination of creatine kinase isoenzyme catalytic
concentrations in serum using immunological methods. J. Clin.
Chem. Clin. Biochem. 1977; 15:131-137.
4. Recommendations of the German Society for Clinical Chemistry.
Standardization of methods for the estimation of enzyme activities
in biological fluids: Standard method for the determination of
creatine kinase activity. J. Clin. Chem. Clin. Biochem. 1977;
15:255-260.
february 1999.
september 1986.
nd
19, 2002.
CK- MB RTU
ABX Pentra
CK NAC CP
ABX Pentra
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A93A00052K EN
A11A01632
26 ml
6.5 ml
HORIBA ABX
BP 7290
ABX Pentra CK NAC CP
Ref.: A11A01632
Volume R1: 26 ml
Volume R2: 6.5 ml
determination of Creatine Kinase (CK) in
serum or plasma.
Creatine kinase (CK) is an enzyme which consists of isoenzymes mainly
of the muscle (CK-M) and the brain (CK-B). CK exists in serum in
dimeric form as CK-MM, CK-MB, CK-BB and as macroenzyme. Elevated
CK values are observed in cardiac muscle damages and in skeletal
muscle diseases. Measurement of CK is used especially in conjunction
with CK-MB for diagnosis and monitoring of myocardial infarction.
Method
Optimized UV-test according to DGKC (German Society of Clinical
Chemistry ) and IFCC (International Federation of Clinical Chemistry).
(CK = Creatine Kinase, HK = Hexokinase, G6P-DH = Glucose-6-phosphate
dehydrogenase)
Reagents
ABX Pentra CK NAC CP is ready-to-use.
Reagent 1:
Imidazole pH 6.7 62.5 mmol/l
Glucose 27.5 mmol/l
N-Acetylcysteine (NAC) 27.5 mmol/l
Magnesium acetate 13.8 mmol/l
EDTA-Na
2
2.1 mmol/l
NADP 2.75 mmol/l
Hexokinase (HK) 3 kU/l
Reagent 2:
Imidazole 160 mml/l
Creatine phosphate 170 mmol/l
EDTA-Na
2
2.1 mmol/l
ADP 11 mmol/l
AMP 28 mmol/l
Diadenosine pentaphosphate 56 mol/l
Glucose-6-phosphate dehydrogenase(G6P-DH) 7.5 kU/l
CK
Creatine + ATP Creatine phosphate+ ADP
Glucose + ATP
HK
Glucose-6-phosphate+ADP
Glucose-6-phosphate+NADP
+
G6P-DH
Gluconate-6-phosphate+NADPH+H
+
ABX Pentra CK NAC CP should be used according to this reagent
otherwise.
Handling
Calibrator
Control
ABX Pentra CK Control, Ref. A11A01786 (not included)
CK NAC CP
ABX Pentra
Specimen
Serum.
Heparin Plasma or EDTA Plasma.
is avoided.
Pentra 400.
Assay Procedure
Waste Management
amounts of water.
General Precautions
Detection limit:
and equals 8 U/l.
protocol (6).
Correlation:
A2 protocol (9).
2
= 0.994 .
Interferences:
Stability: 1 week at 2 - 8 C
1 day at 15 - 25 C
Stability at - 20 C: 4 weeks (in the dark)
in [U/l] 37 C
Adults:
Women 145
Men 171
Children:
Umbilical cord blood 175 - 402
Newborns 468 - 1200
< 5 days 195 - 700
< 6 months 41 - 330
> 6 months 24 - 229
Mean value U/l CV %
Normal control 165 1.20
Pathological control 474 0.92
Specimen 2 115 1.14
Specimen 3 347 0.79
Mean value U/l CV %
Specimen 2 311 2.61
Haemoglobin: No significant influence is observed up to 55 mol/l.
Direct Bilirubin: No significant influence is observed up to 100 mol/l.
CK NAC CP
ABX Pentra
Application release
a
: 4.xx
Warning
Reference
1. Stein W. Creatine kinase (total activity), creatine kinase
isoenzymes and variants. In: Thomas L, ed. Clinical laboratory
diagnostics.Frankfurt: TH-Books Verlagsgesellschaft;1998.p.71-80.
rd
ed.
Chem Lab Med 2002; 40(6) : 635-642.
4. Recommendations of the German Society for Clinical Chemistry.
Standardization of methods for the estimation of enzyme activities
in biological fluids: Standard method for the determination of
creatine kinase activity. J Clin Chem Clin Biochem 1977;15:255-
260.
5. Lorentz K, Rhle G, Siekmann L. Introduction of new standard
methods 1994 for the determination of catalytic enzyme
concentrations at 37 C. DG Klinische Chemie Mitteilungen
1995;26:290-293.
february 1999.
september 1986.
nd
19, 2002.
a. Modification from index J to K: suppression of minor index.
CK NAC CP
ABX Pentra
GGT CP
ABX Pentra
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A93A00072H EN
A11A01630
56 ml
14 ml
HORIBA ABX
BP 7290
ABX Pentra GGT CP
Ref.: A11A01630
Volume R1: 56 ml
Volume R2: 14 ml
determination of Gamma-GlutamylTransferase
(GGT) in serum or plasma.
Clinical Interest (1)
Gamma-glutamyltransferase (Gamma-GT or GGT), also called gamma-
glutamyltranspeptidase, is an enzyme present in liver and bile duct
which is the most sensitive indicator of hepatobiliary diseases.
Because of a high negative predictive value for these diseases the
measurement of gamma-GT is widely used to rule out an hepatic or
biliary origin. Together with other enzymes such as alanine
aminotransferase (ALAT), aspartate aminotransferase (ASAT) and
cholinesterase gamma-GT is a valuable tool for the differential
diagnosis in liver diseases.
Method (2)
Kinetic photometric test according to Szasz modified (1974).
Gamma-GT catalyzes the transfer of glutamic acid to acceptors like
glycylglycine in this case.
This process releases 5-amino-2-nitrobenzoate, which can be
measured at 405 nm. The increase in absorbance at this wavelength is
directly related to the activity of gamma-GT.

Reagents
ABX Pentra GGT CP is ready-to-use.
ABX Pentra GGT CP should be used according to this reagent notice.
Handling
Calibrator
Glycylglycine 137 mmol/l
Reagent 2: L-Gamma-glutamyl-3-carboxy-4-nitroanilide 22 mmol/l
Gamma-GT
L-Gamma-glutamyl-3-carboxy-4-nitranilide + Glycylglycine
Gamma-glutamyl-glycylglycine + 5-Amino-2-nitrobenzoate
Control
Specimen
Serum.
EDTA Plasma.
Stability: at least 1 week between - 20 C and 25 C.
Reference range (3)
Reagents, in unopened cassettes, are stable up to the expiry date on the
label if stored at 2 - 8 C protected from light and contamination is avoided.
Pentra 400.
37 C
Women: 38 U/l
Men: 55 U/l
GGT CP
ABX Pentra
Assay Procedure
Waste Management
amounts of water.
General Precautions
Detection limit:
and equals 4 U/l.
protocol (4).
Correlation:
A2 protocol (7).
2
= 0.9984 .
Interferences:
Application release
a
: 3.xx
Warning
Reference
st
ed. Frankfurt: TH-
Books Verlagsgesellschaft; 1998. p.80-86.
2. Persijn JP, van der Silk W. A new method for the determination of
gamma-glutamyltransferase in serum. J. Clin. Chem. Clin. Biochem.
1976; 14:421-427.
Chem Lab Med 2002; 40(7) : 734-738.
february 1999.
Mean value U/l CV %
Specimen 1 47 3.37
Specimen 2 50 1.41
Specimen 3 394 0.82
Mean value U/l CV %
Specimen 1 43 5.75
Specimen 2 399 3.69
a. Modification from index G to H: suppression of minor index.
GGT CP
ABX Pentra
september 1986.
nd
19, 2002.
GGT CP
ABX Pentra
LDH CP
ABX Pentra
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A93A01192D EN
A11A01824
26 ml
6.5 ml
HORIBA ABX
BP 7290
S.A.S. au capital de 41.700.000 - RCS Montpellier 328 031 042 - SIRET 328 031 042 000 42 - APE 332 B Latest version documents on www.horiba-abx.comS.
ABX Pentra LDH CP
Ref.: A11A01824
Volume R1: 26 ml
Volume R2: 6.5 ml
determination of Lactate Dehydrogenase (LDH)
in serum or plasma.
Lactate dehydrogenase (LDH) is an enzyme, consisting of five different
isoenzymes that catalyze the interconversion of L-lactate and
pyruvate. LDH is present in the cytoplasm of all human tissues with
higher concentrations in liver, heart and skeletal muscle, and lower in
erythrocytes, pancreas, kidney and stomach. Increased LDH activities
are found in a variety of pathological conditions such as myocardial
infarction, liver diseases, blood diseases, cancer or muscle diseases.
However, because of the lack of organ specificity , determination of its
isoenzymes or other enzymes such as alkaline phosphatase or ALAT /
ASAT is necessary for differential diagnosis.
Method (3)
Optimized test according to German Society of Clinical Chemistry
(DGKC) .
(LDH = Lactate Dehydrogenase)
Reagents
ABX Pentra LDH CP is ready-to-use.
ABX Pentra LDH CP should be used according to this reagent notice.
Handling
Calibrator
Control
Reagent 1: Phosphate buffer, pH 7.5 64 mmol/l
Pyruvate 0.81 mmol/l
Reagent 2: Goods buffer, pH 9.6
NADH 1.05 mmol/l
LDH
Pyruvate + NADH + H
+
Lactate + NAD
+
Specimen
Serum.
Heparin plasma or EDTA plasma.
Reference range (4)
Reagents, in unopened vials, are stable up to the expiry date on the
label if stored at 2 - 8 C protected from light and contamination is
avoided.
Assay Procedure
Loss of activity within 3 days at 2 - 8 C < 8 %
and at 15 - 25 C < 2 %
Stability at -20C: 6 weeks
Adults [U/l] : < 480
LDH CP
ABX Pentra
Waste Management
amounts of water.
General Precautions
On board Reagent Stability: 32 days
Detection limit:
and equals 10 U/l.
protocol (5).
Correlation:
A2 protocol (8).
2
= 0.991.
Interferences:
Application release
a
: 2.xx
Warning
Reference
1. Thomas L. Clinical laboratory diagnostics. 1
st
ed. Frankfurt: TH-
Books Verlagsgesellschaft; 1998. 89-94.
2. Moss DW, Henderson AR. Clinical enzymology In: Burtis CA,
rd
ed.
Philadelphia: W.B Saunders Company; 1999. 617-721.
3. Deutsche Gesellschaft fr Klinische Chemie. Empfehlungen der
deutschen Gesellschaft fr Klinische Chemie (DGCK).
Standardisierung von Methoden zur Bestimmung von
Enzymaktivitten in biologischen Flssigkeiten. (Recommendation
of the German Society of Clinical Chemistry. Standardization of
methods for measurement of enzymatic activities in biological
fluids.) Z. Klin. Chem. Klin. Biochem. 1972; 10:182-192.
4. Fischbach F., Zawta B., Age-dependent reference limits of several
enzymes in plasma at different measuring temperatures. Klin. Lab.
1992; 38:555-561.
Mean value U/l CV %
Specimen 1 147 1.46
Specimen 2 269 1.13
Specimen 3 681 0.56
Mean value U/l CV %
Specimen 1 272 4.38
Specimen 2 701 2.78
Haemoglobin: Do not use haemolysed samples.
Triglycerides: No significant influence is observed up to 7 mmol/l.
Direct Bilirubin: No significant influence is observed up to 900 mol/l.
a. Modification from index C to D: suppression of minor index.
LDH CP
ABX Pentra
february 1999.
september 1986.
nd
19, 2002.
LDH CP
ABX Pentra
LDH IFCC CP
ABX Pentra
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A93A01218D EN
A11A01871
26 ml
6.5 ml
HORIBA ABX
BP 7290
ABX Pentra LDH IFCC CP
Ref.: A11A01871
Volume R1: 26 ml
Volume R2: 6.5 ml
Intended use
determination of Lactate Dehydrogenase (LDH)
in serum or plasma.
Lactate dehydrogenase (LDH) is an enzyme, consisting of five different
isoenzymes that catalyze the interconversion of L-lactate and
pyruvate. LDH is present in the cytoplasm of all human tissues with
higher concentrations in liver, heart and skeletal muscle, and lower in
erythrocytes, pancreas, kidney and stomach. Increased LDH activities
are found in a variety of pathological conditions such as myocardial
infarction, liver diseases, blood diseases, cancer or muscle diseases.
However, because of the lack of organ specificity , determination of its
isoenzymes or other enzymes such as alkaline phosphatase or ALAT /
ASAT is necessary for differential diagnosis.
Method (3)
Optimized UV-test according to International Federation of Clinical
Chemistry and Laboratory Medecine (IFCC) .
(LDH = Lactate Dehydrogenase)
Reagents
ABX Pentra LDH IFCC CP is ready-to-use.
1g/l
ABX Pentra LDH IFCC CP should be used according to this reagent
otherwise.
Handling
Calibrator
Control
Reagent 1: N-Methyl-D-Glucamine 420 mmol/l
L-Lactate 65 mmol/l
Reagent 2: NAD
+
50 mmol/l
LDH
L-Lactate + NAD
+
Pyruvate + NADH + H
+
Automated clinical chemistry analyser: ABX PENTRA 400
Calibrator: ABX Pentra Multical, Ref. A11A01652
Controls: ABX Pentra N Control, Ref. A11A01653, and
ABX Pentra P Control, Ref. A11A01654
Specimen (12)
Serum.
Heparin-lithium plasma.
Reference range (4)
Each laboratory should establish its own reference ranges. The values
given here are used as guidelines only.
Stability:
7 days at 20 - 25 C
4 days at 2 - 8C
6 weeks at -20C
Women [U/l] at 37C: < 247
Men [U/l] at 37C: < 248
LDH IFCC CP
ABX Pentra
the label if stored at 2 - 8 C and contamination is avoided.
Reagent 2 must be protected from light.
Do not freeze the reagents!
Stability after opening: refer to the paragraph "Performance on ABX
Pentra 400".
Assay Procedure
available on request (not available in the USA).
Waste Management
amounts of water.
General Precautions
On board Reagent Stability (refrigerated area):
If the ABX Pentra LDH IFCC CP cassette is left on board the instrument
at all times, the cassette is stable for 31 days.
Detection limit:
The detection limit is determined according to CLSI (NCCLS), EP17-A
protocol (13) and equals 11 U/l.
protocol (5).
Reproducibility (total precision)
3 specimens of low, medium and high levels and 2 controls are tested
in duplicate for 20 days (2 series per day) according to the
recommendations found in the CLSI (NCCLS), EP5-A protocol (6).
Measuring Range:
The assay confirmed a measuring range from 11 U/l to 800 U/l, with
an automatic post-dilution up to 2400 U/l.
The reagent linearity has been assessed up to 800 U/l according to the
recommendations found in the CLSI (NCCLS), EP6-A protocol (7).
Correlation:
128 patient samples (serum) are correlated with a commercial reagent
taken as reference according to the recommendations found in the
CLSI (NCCLS), EP9-A2 protocol (8). Values ranged from 15 U/l to 791
U/l.
The equation for the allometric line obtained using Passing-Bablock
regression procedure (9) is:
2
= 0.9959.
Interferences:
Other limitations are given by Young as a list of drugs and preanalytical
variables known to affect this methodology (10,11).
Application release
a
: 5.xx
Mean value U/l CV %
Control specimen 1 197.2 2.66
Specimen 1 113.4 2.58
Specimen 2 256.5 1.67
Specimen 3 519.5 1.05
Mean value U/l CV %
Specimen 1 157.2 3.31
Specimen 2 316.0 2.67
Specimen 3 914.5 2.33
Haemoglobin: Do not use hemolysed samples.
Triglycerides: No significant influence is observed up to 612.5 mg/dl
(7 mmol/l).
(as Intralipid, representative of lipemia).
Total Bilirubin: No significant influence is observed up to 29.3 mg/dl
(500 mol/l).
Direct Bilirubin: No significant influence is observed up to 29.3 mg/dl
(500 mol/l).
a. Modification from index C to D: new application release.
LDH IFCC CP
ABX Pentra
Warning
Reference
1. Thomas L. Clinical laboratory diagnostics. 1
st
ed. Frankfurt: TH-
Books Verlagsgesellschaft; 1998. 89-94.
2. Moss DW, Henderson AR. Clinical enzymology In: Burtis CA,
rd
ed.
Philadelphia: W.B Saunders Company; 1999. 617-721.
3. Deutsche Gesellschaft fr Klinische Chemie. Empfehlungen der
deutschen Gesellschaft fr Klinische Chemie (DGCK).
Standardisierung von Methoden zur Bestimmung von
Enzymaktivitten in biologischen Flssigkeiten. (Recommendation
of the German Society of Clinical Chemistry. Standardization of
methods for measurement of enzymatic activities in biological
fluids.) Z. Klin. Chem. Klin. Biochem. 1972; 10:182-192.
4. Schumann G., Bonora R., Ceriotti F., Frard G. et al., IFCC primary
reference procedure for the measurement of catalytic activity
concentrations of enzymes at 37C. Part 3: Reference procedure for
the measurement of catalytic concentration of lactate
dehydrogenase. Clin. Chem. Lab. Med., 2002, 40: 643-648.
february 1999.
september 1986.
nd
19, 2002.
9. Passing H., Bablock W. A new biometrical procedure for testing the
equality of measurements from two different analytical methods.
J. Clin. Chem. Clin. Biochem. 1983; 21: 709-20.
10. Young D.S., Effects of Drugs on Clinical Laboratory Tests, 4
th
Edition, Washington, DC, AACC Press, 1995, 3: 143-163.
11. Young D.S., Effects of Preanalytical Variables on Clinical
Laboratory Tests, 2
nd
Edition, Washington, DC, AACC Press, 1997,
3: 120-132.
12. Guder W.G., Zawta B., The Quality of Diagnostics Samples. Samples:
From the Patient to the Laboratory. 1
st
ed. Guder W.G., Narayanan
S., Zawta B. (WHILEY -VCH, darmstadt, Germany), (2001), 24.
13. Protocols for determination of limits of detection and limits of
quantitation, Approved Guideline, CLSI (NCCLS) document EP17-A,
Vol. 24, No. 34, 2004.
LDH IFCC CP
ABX Pentra
Lipase CP
ABX Pentra
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A93A00092I EN
A11A01631
24 ml
7 ml
HORIBA ABX
BP 7290
ABX Pentra Lipase CP
Ref.: A11A01631
Volume R1: 24 ml
Volume R2: 7 ml
determination of Lipase in serum or plasma.
Lipases are enzymes which hydrolyze glycerol esters of long fatty
acids. The enzyme and its cofactor colipase is produced in the
pancreas, lipase being also secreted in small amounts by the salivary
glands as well as by gastric, pulmonary and intestinal mucosa. Bile
acids and colipase form micellar complexes with the lipids and bind
lipase on the substrate / water interface. Determination of lipase is
used for investigation of pancreatic disorders. In acute pancreatitis
the lipase concentrations rise to 2-50 fold the upper reference limit
within 4-8 hours after begin of abdominal pain peaking at 24 hours
and decreasing within 8 to 14 days. Elevated lipase values can also be
observed in chronic pancreatitis and obstruction of the pancreatic
duct.
Method
Enzymatic colorimetric test.
A synthetically produced lipase substrate (1,2-o-dilauryl-rac-glycero-
3-glutaric acid-(6-methylresorufin) ester) is added to a microemulsion
which is specifically split by lipase in the presence of colipase and bile
acids. The combination of lipase and bile acids make this specific and
reliable for pancreatic lipase without any reaction due to lipolytic
enzymes or esterases. The reagent composition has been thoroughly
optimised so there is no serum matrix effects. The generated
methylresorufin-ester is spontaneously degraded to methylresorufin.
The absorbance by this red dye is directly proportional to the lipase
activity in the sample.
Lipase catalyses the reaction:
1,2-o-Dilauryl-rac-glycero-3-glutaric acid(6-methylresorufin) ester
1,2-o-Dilauryl-rac-glycerin + Glutaric acid-(6-methylresorufin)-ester
Lipase / Colipase
Glutaric acid-(6-methylresorufin)-ester
Glutaric acid + Methylresorufin
Spontaneous degradation
Reagents
ABX Pentra Lipase CP is ready-to-use.
ABX Pentra Lipase CP should be used according to this reagent
otherwise.
Handling
a
Remove both caps of the cassette. If present, remove foam by using a
plastic pipette.
Position the respective protective cap, ref. GBM0969 on R1 and Ref.
GBM0970 on R2 and place in the refrigerated ABX Pentra 400 reagent
compartment.
Calibrator
Reagent 1: Goods Buffer pH 8.0 50 mmol/l
Taurodesoxycholate 4.3 mmol/l
Desoxycholate 3.25 mmol/l
Calcium chloride 15 mmol/l
Colipase 1.2 mg/l
Detergent
Preservative
Reagent 2: Tartrate Buffer pH 4.0 7.5 mmol/l
Taurodesoxycholate 17 mmol/l
Color Substrate 0.65 mmol/l
Coemulgator
Stabilizer
Preservative
a. Modification from index H to I: new handling.
Lipase CP
ABX Pentra
Control
ABX Pentra Clean-Chem CP, Ref. A11A01755, 30 ml
Specimen
Serum.
Heparin Plasma.
Reference range
< 60 U/l.
the label if stored at 2 - 8 C protected from light and contamination
is avoided.
Pentra 400.
Do not freeze the reagent.
Assay Procedure
Waste Management
Please refer to local legal requirements.
General Precautions
2. As many other clinical reagents contain lipase, avoid carry over.
On board Reagent Stability
a
:
Pentra 400 compartment is stable 40 days.
Detection limit:
and equals 4.6 U/l.
protocol (8).
Correlation:
A2 protocol (11).
2
= 0.985.
Stability: 24 hours at 15 - 25 C
5 days at 2 - 8 C
1 year at -20 C
a. Modification from index H to I: new on board reagent stability.
Mean value U/l CV %
Specimen 3 213.5 1.71
Mean value U/l CV %
Specimen 2 165.2 4.82
Lipase CP
ABX Pentra
Interferences:
Calibration stability
a
:
Application release
b
: 7.xx
Warning
Reference
1. Lorentz K. Lipase. In: Thomas L, editor. Clinical laboratory diagnostics.
1
st
ed. Frankfurt: TH-Books Verlagsgesellschaft; 1998. p. 95-97.
2. Moss DW, Henderson AR. Digestive enzymes of pancreatic origin.
In: Burtis CA, Ashwood ER, editors. Tietz Textbook of Clinical
Chemistry. 3
rd
ed. Philadelphia: W.B Saunders Company; 1999. p.
689-708.
3. Tietz N, Shuey DF. Lipase in serum - the elusive enzyme: an
overview. Clin. Chem. 1993; 39:746-756.
4. Lott J, Patel ST, Sawhney AK, Kazmierczak SC, Love JE. Assays of
serum lipase: analytical and clinical considerations. Clin. Chem.
1986; 32:1290-1302.
5. Leybold A, Junge W. Importance of colipase for the measurement
of serum lipase activity. Adv. Clin. Enzymol. 1986; 4:60-67.
6. Borgstrm B. The action of bile salts and other detergents on
pancreatic lipase and the interaction with colipase. Biochimica et
Biophysika Acta 1977; 488:381-91.
7. Gargouri Y, Julien R, Bois A, Verger R, Sarda L. Studies on the
detergent inhibition of pancreatic lipase activity. J. of Lipid.
Research 1983; 24:1336-42.
february 1999.
september 1986.
nd
19, 2002.
a. Modification from index H to I: modification of calibration stability.
b. Modification from index G to H: suppression of minor index.
Lipase CP
ABX Pentra
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