METABELIAN SL(n; C) REPRESENTATIONS OF KNOT GROUPS, III: DEFORMATIONS
AST/GOT 4+1 SL - Manmancompany.com/en/wp-content/uploads/2014/10/AST-GOT-4+1-SL.pdf · AST/GOT 4+1...
2
AST/GOT 4+1 SL CLINICAL SIGNIFICANCE (1-4) Aspartate aminotransferase (AST) also known as glutamate oxa- loacetate transaminase (GOT) is a transaminase. AST catalyses the transfer of the aminogroup of L-aspartate to -ketoglutarate to give L-glutamate. AST is widely distributed in the body, but the highest levels are found in heart, liver, skeletal muscles and kid- neys. Damages to cells of these tissues induce AST increase in serum. In case of fulminant forms of hepatitis, especially viral hepatitis the enzyme level is markedly elevated. In case of myocardial in- farction, AST activity increases and reaches a peak after 18-24 hours. The activity falls back to normal after 4-5 days, provided no new infarct has occurred. The following pathological states are examples of disorders also resulting in an increase of enzyme activity : liver cell necrosis or injury of any cause (for example intake of alcohol, delirium tre- mens, and administration of drug induce moderate AST eleva- tion), alcoholic hepatitis, muscular dystrophy and gangrene, in- fectious mononucleosis, acute pancreatitis, heart affection as myocarditis or pericarditis, pulmonary emboli.... On the contrary, AST serum level can decrease in case of vitamin B6 deficiency. METHOD (5) IFCC method without pyridoxal phosphate (P-5’-P). Kinetic. UV. PRINCIPLE (5) Kinetic determination of the aspartate aminotransferase (AST) activity: AST L-Aspartate + -Ketoglutarate Oxaloacetate +L-Glutamate MDH Oxaloacetate + NADH + H + L-Malate + NAD + MDH = Malate dehydrogenase. REAGENTS COMPOSITION Reagent 1: R1 Tris buffer, pH 7.80 (30°C) 100 mmol/L L-Aspartate 330 mmol/L LDH B 2000 U/L MDH B 1000 U/L Reagent 2: R2 C-Ketoglutarate 78 mmol/L NADH 1.1 mmol/L MAERIAL REQUIRED BUT NOT PROVIDED CONT-0060 ELITROL I 10 × 5 mL CONT-0160 ELITROL II 10 × 5 mL PRECAUTIONS - The reagents contain less than 0.1 % sodium azide. Sodium azide can react with copper and lead plumbing to form explosive metal azides. If discharge in the canalisations, rinse with plenty of water. - Use clean or single use laboratory equipment only to avoid con- taminations. - High AST values may induce falsely low results due to the deple- tion of the substrate (total consumption of NADH before reading of the result). If an analyser is used, verify the presence of a dep- letion factor on the application. References: ASSL - 0410 2 x 62,5 mL ASSL - 0430 4 x 62,5 mL ASSL - 0510 5 x 125 mL WASTE MANAGEMEN Disposal of all Waste material should be in accordance with local and legal requirements. STABILITY OF REAGENTS To store at 2-8°C and protected from light. The reagents are stable until the expiry date stated on the label. On board stability : The stability is specific for each analyser (for Selectra refer to § PERFORMANCE DATA) PREPARATION AND STABILITY OF WORKING REAGENT - One-reagent procedure Mix 4 volumes of the reagent R1 with 1 volume of the reagent R2. Stability: 5 days at 20 - 25 °C 2 weeks at 2 - 8 °C - Two-reagent procedure The reagents are ready to use. SAMPLES (2,6) - Specimen Serum free from hemolysis. Lithium heparinized or EDTA plasma, free of hemolysis. - Storage Sera are stable 24 hours at room temperature, 7 days at 2-8°C and 3 months at -20°C. REFERENCE VALUES (2) Serum, plasma (37°C): < 40 U/L. Reference values for infants are higher than for adults. Note : It is recommended for each laboratory to establish and main- tain its own reference values. The data given here are only an indi- cation. PROCEDURE This reagent can be used on most analysers, semi automated ana- lysers and manual method. The applications are available on request. Wavelength: 340 nm Temperature: 37 °C Read against distilled water. - One-reagent procedure Working reagent : 200 μL Sample : 20 μL Mix and after a 50 second incubation, measure the change of ab- sorbance per minute (MA/min.) during 175 seconds. - Two-reagent procedure Reagent R1 : 200 μL Reagent R2 : 50 μL Mix, wait 25 seconds and add: Sample : 25 μL Mix and after a 50 second incubation, measure the change of ab- sorbance par minute (MA/min.) during 150 seconds. CALCULATION At 340 nm, with the one-reagent procedure and the two-reagent procedure for a 1 cm path light cuvette: Activity (U/L) = MN/min. x 1746 In vitro diagnostic reagent, for professional use only Kit composition: R1 2 x 50 mL + R2 1x 26 mL R1 4 x 50 mL + R2 2x 26 mL R1 5x 100 mL + R2 1 x 127 mL SEPPIM S.A.S. - Zone Industrielle - 61500 SEES FRANCE
Transcript of AST/GOT 4+1 SL - Manmancompany.com/en/wp-content/uploads/2014/10/AST-GOT-4+1-SL.pdf · AST/GOT 4+1...
AST/GOT 4+1 SL
CLINICAL SIGNIFICANCE (1-4) Aspartate aminotransferase (AST) also known as glutamate oxa-loacetate transaminase (GOT) is a transaminase. AST catalyses the transfer of the aminogroup of L-aspartate to α-ketoglutarate to give L-glutamate. AST is widely distributed in the body, but the highest levels are found in heart, liver, skeletal muscles and kid-neys. Damages to cells of these tissues induce AST increase in serum. In case of fulminant forms of hepatitis, especially viral hepatitis the enzyme level is markedly elevated. In case of myocardial in-farction, AST activity increases and reaches a peak after 18-24 hours. The activity falls back to normal after 4-5 days, provided no new infarct has occurred. The following pathological states are examples of disorders also resulting in an increase of enzyme activity : liver cell necrosis or injury of any cause (for example intake of alcohol, delirium tre-mens, and administration of drug induce moderate AST eleva-tion), alcoholic hepatitis, muscular dystrophy and gangrene, in-fectious mononucleosis, acute pancreatitis, heart affection as myocarditis or pericarditis, pulmonary emboli.... On the contrary, AST serum level can decrease in case of vitamin B6 deficiency.
METHOD (5) IFCC method without pyridoxal phosphate (P-5’-P). Kinetic. UV.
PRINCIPLE (5) Kinetic determination of the aspartate aminotransferase (AST) activity: AST L-Aspartate + α-Ketoglutarate Oxaloacetate +L-Glutamate
MDH Oxaloacetate + NADH + H+ L-Malate + NAD+
MDH = Malate dehydrogenase. REAGENTS COMPOSITION Reagent 1: R1 Tris buffer, pH 7.80 (30°C) 100 mmol/L L-Aspartate 330 mmol/L LDH ≥ 2000 U/L MDH ≥ 1000 U/L
Reagent 2: R2 C-Ketoglutarate 78 mmol/L NADH 1.1 mmol/L
�MAERIAL REQUIRED BUT NOT PROVIDED CONT-0060 ELITROL I 10 × 5 mL CONT-0160 ELITROL II 10 × 5 mL
�PRECAUTIONS - The reagents contain less than 0.1 % sodium azide. Sodium azide can react with copper and lead plumbing to form explosive metal azides. If discharge in the canalisations, rinse with plenty of water. - Use clean or single use laboratory equipment only to avoid con-taminations. - High AST values may induce falsely low results due to the deple-tion of the substrate (total consumption of NADH before reading of the result). If an analyser is used, verify the presence of a dep-letion factor on the application.
References: ASSL - 0410 2 x 62,5 mL ASSL - 0430 4 x 62,5 mL ASSL - 0510 5 x 125 mL
�WASTE MANAGEMEN Disposal of all Waste material should be in accordance with local and legal requirements.
STABILITY OF REAGENTS To store at 2-8°C and protected from light. The reagents are stable until the expiry date stated on the label.
On board stability:The stability is specific for each analyser (for Selectra refer to § PERFORMANCE DATA)
PREPARATION AND STABILITY OF WORKING REAGENT - One-reagent procedure Mix 4 volumes of the reagent R1 with 1 volume of the reagent R2.
Stability: 5 days at 20 - 25 °C 2 weeks at 2 - 8 °C
- Two-reagent procedure The reagents are ready to use.
�SAMPLES (2,6) - Specimen
Serum free from hemolysis. Lithium heparinized or EDTA plasma, free of hemolysis.
- Storage Sera are stable 24 hours at room temperature, 7 days at 2-8°C and 3 months at -20°C.
REFERENCE VALUES (2) Serum, plasma (37°C): < 40 U/L. Reference values for infants are higher than for adults.
Note: It is recommended for each laboratory to establish and main-tain its own reference values. The data given here are only an indi-cation.
PROCEDURE This reagent can be used on most analysers, semi automated ana-lysers and manual method. The applications are available on request.
Wavelength: 340 nm Temperature: 37 °C
Read against distilled water. - One-reagent procedure
Working reagent : 200 µLSample : 20 µLMix and after a 50 second incubation, measure the change of ab-sorbance per minute (∆A/min.) during 175 seconds.
- Two-reagent procedure
Reagent R1 : 200 µLReagent R2 : 50 µLMix, wait 25 seconds and add:
Sample : 25 µLMix and after a 50 second incubation, measure the change of ab-sorbance par minute (∆A/min.) during 150 seconds.
CALCULATION At 340 nm, with the one-reagent procedure and the two-reagent procedure for a 1 cm path light cuvette:
Activity (U/L) = ∆Α/min. x 1746
In vitro diagnostic reagent, for professional use only
Kit composition: R1 2 x 50 mL + R2 1 x 26 mL R1 4 x 50 mL + R2 2 x 26 mL R1 5 x 100 mL + R2 1 x 127 mL
SEPPIM S.A.S. - Zone Industrielle - 61500 SEES FRANCE
AST/GOT 4+1 SL
QUALITY CONTROL To ensure adequate quality, control sera such as ELITROL I (nor-mal control) and ELITROL II (abnormal control) are recommended.
PERFORMANCE DATA at 37°C for Selectra - Analytical range The reagent is linear from 10 to 250 U/L.
- Detection limit (7) Determined according to SFBC protocol, the detection limit is equal to 2 U/L.
- Analytical Sensitivity The average variation of the analytical signal is 0.57 m∆A/min per U/L of AST for a light path of 1 cm.
-Precision Within–run reproducibility
One-reagent procedure Two-reagent procedure n Mean (U/L) CV% n Mean (U/L) CV%
SH 1 20 18 3.9 19 18 4.5 SH 2 20 49 3.0 20 50 4.0 SH 3 20 230 0.8 20 231 1.2
Between-run reproducibility
One-reagent procedure Two-reagent procedure n Mean (U/L) CV% n Mean (U/L) CV%
SH 1 80 18 4.8 80 18 7.1 SH 2 79 46 4.0 78 46 6.3 SH 3 80 222 2.9 80 221 2.8
- Correlation A comparative study has been performed between ELITech me-thod and another commercial reagent (IFCC method without pyri-doxal phosphate) on 86 human sera samples. The sample concentrations ranged from 5 to 250 U/L.
The parameters of linear regressions are as follows:
One-reagent procedure Two-reagent procedure Correlation coefficient (r) 0.996 0.999
Linear regression y = 0.97 x + 0.7 U/L y = 0.94 x + 0.7 U/L
- Interferences (7, 8) According to SFBC recommendations, studies have been per-formed to determine the level of interference from different com-pounds:
Glucose: No significant interference up to 500 mg/dL (5 g/L, 28 mmol/L).
Ascorbic acid: No significant interference up to 40 mg/dL (400 mg/L, 2.3 mmol/L).
Pyruvate: No significant interference up to 2 mg/dL (20 mg/L, 0.23 mmol/L).
UnconjugatedBilirubin: Negative bias from 17.5 mg/dL (175 mg/L, 300 µmol/L).
ConjugatedBilirubin: Negative bias from 11.5 mg/dL (115 mg/L, 200 µmol/L).
Turbidity: No significant interference up to 600 mg/dL Triglyceride equivalent (6 g/L, 6.9 mmol/L).
Methyl-dopa: No significant interference up to 5 mg/dL (50 mg/L, 6.9 µmol/L).
Other compounds may interfere. (9-11)
Note: Hemolysed sera should not be used since significant hemo-lysis may increase AST concentration because of high levels of AST in erythrocytes. �- On board stability (no refrigered) On board stability: 5 days for one-reagent procedure and 14 days for two-reagent procedure (capped vials and stored at 2-8°C during the night). �BIBLIOGRAPHY 1. Henderson, A.R., Moss, D.W., Enzymes, Tietz Fundamentals of Clinical Chemistry, 5th Ed., Burtis, C.A. & Ashwood, E.R. (W.B. Saunders eds. Philadelphia USA), (2001), 352. 2. Tietz, N.W., Clinical guide to laboratory tests, 3rd Ed., (W.B. Saunders eds. Philadelphia USA), (1995), 76. 3. Scherwin, J.E, Liver function. Clinical Chemistry : Theory, Analysis, Correlation, 4th Ed., Kaplan, L.A, Pesce, A.J., Kaz-mierc-zak, S.C., (Mosby Inc. eds St Louis USA), (2003), 492 and appendix. 4. Ward, M.K., Cockayne, S., Enzymology. Clinical Chemistry : Concepts and Application, Anderson, S.C., Cockayne, S. (W.B. Saunders eds. Philadelphia USA), (1993), 238. 5. Schumann, G., et al. Clin Chem Lab Med., (2002), 40, 725. 6. Guder,W,G., et al.,Use of antycoagulans in diagnostic labora-tory investigation and stability of blood, plasma, plasma and serum samples.(2002).WHO/DIL/LAB/99.1 Rev.2 7. Vassault A., et al., Ann. Biol. Clin., (1986), 44, 686. 8. Young, D.S., Effects of preanalytical variables on clinical la-boratory tests, 2nd edition, AACC Press (1997). 9. Vassault A., et al., Ann. Biol. Clin., (1999), 57, 685 10. Young D.S., Effects of drugs on clinical laboratory tests, 4th edition, AACC Press (1995). 11. Berth, M. & Delanghe, J. Protein precipitation as a possible important pitfall in the clinical chemistry analysis of blood sam-ples containing monoclonal immunoglobulins: 2 case reports and a review of literature, Acta Clin Belg., (2004), 59, 263.
SYMBOLS USED ON LABELS
In vitro diagnostic reagent, for professional use only
Revised Version: PIT-ASSL4/1-10 10/2010
SEPPIM S.A.S. - Zone Industrielle - 61500 SEES FRANCE
