- Information
- AI Chat
AST & ALT Stanbio - notes
Clinical Chemistry 2 (MDT 3122L)
Our Lady of Fatima University
Recommended for you
Preview text
S STANBID Material Required But Not Provided Expected Values LABCRATORY An EKF Diagnostics Company CE Spectrophotometer capable of absorbance reading at 340 nm and 1 cm Normal Range: lightpath. Constant temperature block or bath, or temperature This range should serve only as a guideline. It is recommended that controlled cuvette well. Accurate pipetting devices, Test tubes and each laboratory establish its own range of expected values, since Interval timer differences exist between instruments, laboratories, and local Specimen Collection and Storage populations. serum is the specimen of choice. yet EDTA treated Performance Characteristics Stanbio plasma or heparinized plasma can be used. Whenever possible Comparison: A group of 62 sera ranging in AST activity from 463 specimens should be separated and analyzed on the day of collection. was assayed the described AST method and a similar Procedure No. 2920 Store serum in stoppered tubes. The enzyme in serum is reportedly commercially available AST reagent. Comparison of the results yielded stable for a minimum of 7 days at a correlation coefficient of 0 and the regression equation was y For the Kinetic Quantitative Determination of Interfering Substances: Hemolysis must be avoided as the 0 0. (Comparison studies were performed according to in Serum for Manual Automated Procedures concentration of AST in red cells is roughly 10 times that of serum. NCCLS Tentative Guideline, Summary and Principle Bilirubin levels up to 40 and triglyceride levels up to Precision: precision was established 20 assays on three different levels of commercial serum controls. Total Precision values Aspartate aminotransferase (AST) is one of several enzymes that catalyze show no interference in this test. Certain drugs and other substances the exchange of amino and OXO groups between acids and are also known to affect AST values. were obtained assaying the 3 commercial controls for 5 consecutive days. acids. It is widely distributed throughout body tissues with Procedure Serum 1 Serum 2 Serum 3 significant amounts in the heart and liver.¹ Lesser amounts are found in 1. Prepare AST Working Reagent according to instructions. Mean AST 25 51 116 skeletal muscle, kidneys, pancreas, spleen, lungs and brain. Injury to these 2. Zero spectropholometer at 340 nm with distilled water. Std. Deviation 0 1 0 tissues results in release of the AST enzyme to the general circulation. In 3. For each sample and control, add 1 mL Working Reagent to C. 3 3 0 myocardial infarction, serum AST may begin to rise within hours after cuvette or test tube and warm to for 3 minutes. Total Precision Serum 1 Serum 2 Serum 3 onset, peak within two days and return to normal the fourth or fifth day 4a. Add 100 uL (0 mL) serum to its respective tube and mix gently. of post infarction. An increase in serum AST is also found with hepatitis, 2 Mean AST 26 49 115 Running this method (1:10 ratio) will yield results linear to 300 or Std. Deviation 1 0 0 liver necrosis, cirrhosis and liver metastasis. 4b. Add 50 UL (0 mL) serum to its respective tube and mix gently. C. 4 1 0 Karmen4 first reported a kinetic method for measuring AST activity in Running this method (1:20 ratio) will yield results linear to 600 Precision studies were performed according to NCCLS Tentative serum. Subsequently, the method has been modified and optimized 5. Read and record absorbance at 1 minute. Continue incubating at Guideline, Bergmeyer et The Stanbio assay procedure for AST measurement is and record absorbance again at 2 and 3 minutes. Rate should Linearity: Linear to 600 at for running method 1:20 ratio and similar to the method recommended the International Federation of be constant. Clinical Chemistry The enzymatic reactions involved in the assay 300 running method at 1:10 ratio. Performed according to NCCLS 6. Determine the average absorbance per minute multiply Guideline procedure are as follows: factor 1768 (1:10 ratio) or 3376 (1:20 ratio) for results in Sensitivity: Based on an instrument resolution of A 0, the AST NOTE: If cuvette is not temperature controlled, incubate samples at Oxalacetate method presented shows a sensitivity of 1 between readings. Find Symbol Glossary at MDH Automated Procedure Oxalacetate NADH NAD References Special adaptations for automated analyzers are available contacting Malate dehydrogenase 1. Wilkinson JH. Principles and Practice for Diagnosis Enzymology. Year Book AST catalyzes the transfer of the amino group aspartate to Customer Service Department. Medical Publishers, 1976 to yield oxalacetate and glutamate. The oxalacetate formed in the first Quality Control: Stanbio 1 Level 1 Control Serum, Cat. No. 2. Kachmar JR: Enzymes. In Fundamentals of Clinical Chemistry, NW Tietz. Editor, reaction is then reacted with NADH in the presence of malate and Stanbio 2 Level 2 Control Serum, Cat. No. Saunders, Philadelphia, 1976. p 674. dehydrogenase (MDH) to form NAD. AST activity is determined 86 are recommended for each run. Other commercially available 3. Sacks HJ. Lanchantin GF: An elevation of serum transaminase in the jaundice state. Am J Clin Pathol 33:97, 1960 measuring the rate of oxidation of NADH at 340 nm. Lactate controls with AST values assayed this method are also suitable. AST 4. Karmen A: A note on the spectrophotometric assay of dehydrogenase is included in the reagent to convert endogenous pyruvate activity determined in these materials, this procedure should fall transaminase in human blood. J Clin Invest 34:131, 1955 in the sample to lactate during the lag phase prior to measurement. within the ranges stated for the controls. Two levels of controls should 5. Bergmeyer HU, Scheibe P. Wahlefeld AW: Optimization of methods for aspartate be analyzed with each run. aminotransferase and alanine aminotransferase. Clin Chem 24: 58, 1978. Reagent AST Buffer (R1), Ref. No. 2921 Calibration: AST activity is based on the extinction 6. Expert Panel of Enzymes of the International Federation of Clinical Chemistry: Part 3. Revised IFCC method for aspartate aminotransferase. Clin Chem 24:720, Composition: 240 of NADH at 340 nm (see section). The instrument 1978, MDH 600 calibration guidelines should be followed to calibrate 7. Demetriou JA et al. In Clinical Chemistry Principles and Technics 2nd ed. RJ LDH 600 your analyzer. Assaying the AST contents of a control serum with known Henry et al. Eds. Harper Row, Hagerstown MD, 1974, p 873. AST values can be used to assure instrument calibration has been 8. International Federation of Clinical Chemistry. Provisional Recommendations on Tris Buffer, pH 7 80 performed correctly. IFCC Methods for the Measurement of Catalytic Concentrations of Enzymes. Clin AST Enzyme (R2), Ref. No. 2922 Chem 23: 887, 1977. Results Composition: 12 9. Young D. Effects of drugs on clinical laboratory tests. AACC Press, Washington NADH (Disodium salt) 0 Values are derived based on the micromolar extinction D., 1990 Precautions: The reagents are for Vitro Diagnostic Rx only. of NADH at 340 nm (0). Units per liter of 10 JB. Clinical Diagnosis and Management Laboratory Methods, 17th ed. activity is that amount of enzyme which oxidizes one WB Saunders Co., 1984. p 1437. Normal precautions exercised in handling laboratory reagents should be of NADH per minute. 11 Laboratory Data followed. The reagents contain sodium azide which may be toxic if STANBIO LABORATORY, LP DISCLAIMS ALL EXPRESS AND IMPLIED WARRANTIES OF THE ingested. Sodium azide may also react with lead and copper plumbing to (1:10 sample to reagent ratio) MERCHANTABILITY AND FITNESS PERTAINING TO THIS PRODUCT WHICH ARE NOT form highly explosive metal azides. Refer to Safety Data Sheet for any Absorptivity) X (Tot Vol Sam Vol) EXPRESSLY DETAILED IN THIS PACKAGING INFORMATION OR A WRITTEN AGREEMENT updated risk, hazard or safety information. 0) X BETWEEN THE BUYER AND SELLER OF THIS PRODUCT. X 1768 STANBIO LABORATORY LP MAINTAINS THAT THIS PRODUCT CONFORMS TO THE Reagent Preparation: Buffer and Enzyme liquid reagents are supplied INFORMATION CONTAINED IN THIS INSERT. PURCHASER MUST DETERMINE THE SUITABILITY Prepare Working Reagent in the ratio of 5 parts Buffer (R1) to (1:20 sample to reagent ratio) OF THE PRODUCT FOR THEIR PARTICULAR USE. USE ONLY IN ACCORDANCE WITH 1 part Enzyme (R2) (i., 25 mL Buffer and 5 mL Enzyme). 0) X LABELING INSTRUCTIONS. Reagent Storage and Stability: Reagents are stable until the X 3376 Stanbio Laboratory 1261 North Main Street . Boerne, TX 78006 USA expiration date on their respective labels, when properly stored at Limitations Ph: (830) Fax (830) and protected from light. Reagents should appear clear and colorless. If the is greater than 0, dilute 1 part sample with 9 parts Discard if either appears cloudy or contains particulate matter. The Working isotonic saline and Multiply the result 10. AST values for RBR.2920.CE.EN Last Revision: Procedure No. 2920CE Reagent is stable for 4 weeks at or 5 days at room temperature neonatal patients have not been established with this procedure. The Working Reagent should be discarded if the initial absorbance Grossly icteric or turbid specimen may require the use of a sample read against distilled water at 340 nm is below 0. blank. S STANBI EKF Diagnostics Company S STANBID Material Required But Not Provided Expected Values LABCRATORY An EKF Diagnostics Company CE Spectrophotometer capable of absorbance reading at 340 nm and 1 cm Normal Range: lightpath. Constant temperature block or bath, or temperature This range should serve only as a guideline. It is recommended that controlled cuvette well. Accurate pipetting devices, Test tubes and each laboratory establish its own range of expected values, since Interval timer differences exist between instruments, laboratories, and local Specimen Collection and Storage populations. serum is the specimen of choice. yet EDTA treated Performance Characteristics Stanbio plasma or heparinized plasma can be used. Whenever possible Comparison: A group of 62 sera ranging in AST activity from 463 specimens should be separated and analyzed on the day of collection. was assayed the described AST method and a similar Procedure No. 2920 Store serum in stoppered tubes. The enzyme in serum is reportedly commercially available AST reagent. Comparison of the results yielded stable for a minimum of 7 days at a correlation coefficient of 0 and the regression equation was y For the Kinetic Quantitative Determination of Interfering Substances: Hemolysis must be avoided as the 0 0. (Comparison studies were performed according to in Serum for Manual Automated Procedures concentration of AST in red cells is roughly 10 times that of serum. NCCLS Tentative Guideline, Summary and Principle Bilirubin levels up to 40 and triglyceride levels up to Precision: precision was established 20 assays on three different levels of commercial serum controls. Total Precision values Aspartate aminotransferase (AST) is one of several enzymes that catalyze show no interference in this test. Certain drugs and other substances the exchange of amino and OXO groups between acids and are also known to affect AST values. were obtained assaying the 3 commercial controls for 5 consecutive days. acids. It is widely distributed throughout body tissues with Procedure Serum 1 Serum 2 Serum 3 significant amounts in the heart and liver.¹ Lesser amounts are found in 1. Prepare AST Working Reagent according to instructions. Mean AST 25 51 116 skeletal muscle, kidneys, pancreas, spleen, lungs and brain. Injury to these 2. Zero spectropholometer at 340 nm with distilled water. Std. Deviation 0 1 0 tissues results in release of the AST enzyme to the general circulation. In 3. For each sample and control, add 1 mL Working Reagent to C. 3 3 0 myocardial infarction, serum AST may begin to rise within hours after cuvette or test tube and warm to for 3 minutes. Total Precision Serum 1 Serum 2 Serum 3 onset, peak within two days and return to normal the fourth or fifth day 4a. Add 100 uL (0 mL) serum to its respective tube and mix gently. of post infarction. An increase in serum AST is also found with hepatitis, 2 Mean AST 26 49 115 Running this method (1:10 ratio) will yield results linear to 300 or Std. Deviation 1 0 0 liver necrosis, cirrhosis and liver metastasis. 4b. Add 50 UL (0 mL) serum to its respective tube and mix gently. C. 4 1 0 Karmen4 first reported a kinetic method for measuring AST activity in Running this method (1:20 ratio) will yield results linear to 600 Precision studies were performed according to NCCLS Tentative serum. Subsequently, the method has been modified and optimized 5. Read and record absorbance at 1 minute. Continue incubating at Guideline, Bergmeyer et The Stanbio assay procedure for AST measurement is and record absorbance again at 2 and 3 minutes. Rate should Linearity: Linear to 600 at for running method 1:20 ratio and similar to the method recommended the International Federation of be constant. Clinical Chemistry The enzymatic reactions involved in the assay 300 running method at 1:10 ratio. Performed according to NCCLS 6. Determine the average absorbance per minute multiply Guideline procedure are as follows: factor 1768 (1:10 ratio) or 3376 (1:20 ratio) for results in Sensitivity: Based on an instrument resolution of A 0, the AST NOTE: If cuvette is not temperature controlled, incubate samples at Oxalacetate method presented shows a sensitivity of 1 between readings. Find Symbol Glossary at MDH Automated Procedure Oxalacetate NADH NAD References Special adaptations for automated analyzers are available contacting Malate dehydrogenase 1. Wilkinson JH. Principles and Practice for Diagnosis Enzymology. Year Book AST catalyzes the transfer of the amino group aspartate to Customer Service Department. Medical Publishers, 1976 to yield oxalacetate and glutamate. The oxalacetate formed in the first Quality Control: Stanbio 1 Level 1 Control Serum, Cat. No. 2. Kachmar JR: Enzymes. In Fundamentals of Clinical Chemistry, NW Tietz. Editor, reaction is then reacted with NADH in the presence of malate and Stanbio 2 Level 2 Control Serum, Cat. No. Saunders, Philadelphia, 1976. p 674. dehydrogenase (MDH) to form NAD. AST activity is determined 86 are recommended for each run. Other commercially available 3. Sacks HJ. Lanchantin GF: An elevation of serum transaminase in the jaundice state. Am J Clin Pathol 33:97, 1960 measuring the rate of oxidation of NADH at 340 nm. Lactate controls with AST values assayed this method are also suitable. AST 4. Karmen A: A note on the spectrophotometric assay of dehydrogenase is included in the reagent to convert endogenous pyruvate activity determined in these materials, this procedure should fall transaminase in human blood. J Clin Invest 34:131, 1955 in the sample to lactate during the lag phase prior to measurement. within the ranges stated for the controls. Two levels of controls should 5. Bergmeyer HU, Scheibe P. Wahlefeld AW: Optimization of methods for aspartate be analyzed with each run. aminotransferase and alanine aminotransferase. Clin Chem 24: 58, 1978. Reagent AST Buffer (R1), Ref. No. 2921 Calibration: AST activity is based on the extinction 6. Expert Panel of Enzymes of the International Federation of Clinical Chemistry: Part 3. Revised IFCC method for aspartate aminotransferase. Clin Chem 24:720, Composition: 240 of NADH at 340 nm (see section). The instrument 1978, MDH 600 calibration guidelines should be followed to calibrate 7. Demetriou JA et al. In Clinical Chemistry Principles and Technics 2nd ed. RJ LDH 600 your analyzer. Assaying the AST contents of a control serum with known Henry et al. Eds. Harper Row, Hagerstown MD, 1974, p 873. AST values can be used to assure instrument calibration has been 8. International Federation of Clinical Chemistry. Provisional Recommendations on Tris Buffer, pH 7 80 performed correctly. IFCC Methods for the Measurement of Catalytic Concentrations of Enzymes. Clin AST Enzyme (R2), Ref. No. 2922 Chem 23: 887, 1977. Results Composition: 12 9. Young D. Effects of drugs on clinical laboratory tests. AACC Press, Washington NADH (Disodium salt) 0 Values are derived based on the micromolar extinction D., 1990 Precautions: The reagents are for Vitro Diagnostic Rx only. of NADH at 340 nm (0). Units per liter of 10 JB. Clinical Diagnosis and Management Laboratory Methods, 17th ed. activity is that amount of enzyme which oxidizes one WB Saunders Co., 1984. p 1437. Normal precautions exercised in handling laboratory reagents should be of NADH per minute. 11 Laboratory Data followed. The reagents contain sodium azide which may be toxic if STANBIO LABORATORY, LP DISCLAIMS ALL EXPRESS AND IMPLIED WARRANTIES OF THE ingested. Sodium azide may also react with lead and copper plumbing to (1:10 sample to reagent ratio) MERCHANTABILITY AND FITNESS PERTAINING TO THIS PRODUCT WHICH ARE NOT form highly explosive metal azides. Refer to Safety Data Sheet for any Absorptivity) X (Tot Vol Sam Vol) EXPRESSLY DETAILED IN THIS PACKAGING INFORMATION OR A WRITTEN AGREEMENT updated risk, hazard or safety information. 0) X BETWEEN THE BUYER AND SELLER OF THIS PRODUCT. X 1768 STANBIO LABORATORY LP MAINTAINS THAT THIS PRODUCT CONFORMS TO THE Reagent Preparation: Buffer and Enzyme liquid reagents are supplied INFORMATION CONTAINED IN THIS INSERT. PURCHASER MUST DETERMINE THE SUITABILITY Prepare Working Reagent in the ratio of 5 parts Buffer (R1) to (1:20 sample to reagent ratio) OF THE PRODUCT FOR THEIR PARTICULAR USE. USE ONLY IN ACCORDANCE WITH 1 part Enzyme (R2) (i., 25 mL Buffer and 5 mL Enzyme). 0) X LABELING INSTRUCTIONS. Reagent Storage and Stability: Reagents are stable until the X 3376 Stanbio Laboratory 1261 North Main Street . Boerne, TX 78006 USA expiration date on their respective labels, when properly stored at Limitations Ph: (830) Fax (830) and protected from light. Reagents should appear clear and colorless. If the is greater than 0, dilute 1 part sample with 9 parts Discard if either appears cloudy or contains particulate matter. The Working isotonic saline and Multiply the result 10. AST values for RBR.2920.CE.EN Last Revision: Procedure No. 2920CE Reagent is stable for 4 weeks at or 5 days at room temperature neonatal patients have not been established with this procedure. The Working Reagent should be discarded if the initial absorbance Grossly icteric or turbid specimen may require the use of a sample read against distilled water at 340 nm is below 0. blank. S STANBI EKF Diagnostics Company CE Material Required But Not Provided differences exist between instruments, laboratories, and local STANBIO Spectrophotometer capable of absorbance reading at 340 nm and 1 cm populations. S lightpath. Constant temperature block or bath, or temperature Performance Characteristics LABORATORY controlled cuvette well. Accurate pipetting devices, Test tubes and Comparison: A group of 82 sera ranging in ALT activity from 3 682 An EKF Diagnostics Company Interval timer was assayed the described ALT method and a similar Specimen Collection and Storage commercially available ALT reagent. Comparison of the results yielded a Stanbio serum is the specimen of choice. Whenever possible correlation coefficient of and the regression equation was y Procedure No. 2930 specimens should be separated and analyzed on the day of collection. 0 0. (Comparison studies were performed according to Store serum in stoppered tubes. About ALT is lost 3. days at NCCLS Tentative Guideline, For the Kinetic Quantitative Determination of and in 1 day at Precision: precision was established 20 assays on three in Serum for Manual Automated Interfering Substances: Hemolysis must be avoided as the different levels of commercial serum controls. Total Precision values Procedures concentration of ALT in red cells is roughly 5 times that of serum.² were obtained assaying the 3 commercial controls for 5 consecutive Bilirubin levels up to 40 and triglyceride levels up to days. Summary and Principle show no interference in this test Certain drugs and other substances The enzyme alanine aminotransferase is widely reported in a variety of are also known to affect ALT values. 6 Serum 1 Serum 2 Serum 3 tissue sources. The major source of ALT is of hepatic origin and has led to Mean ALT 22 45 102 Procedure the application of ALT determinations to the study of hepatic diseases. Std. Deviation 1 1 1 1. Prepare ALT Working Reagent according to instructions. Elevated serum levels are found in hepatitis, cirrhosis, and obstructive C. 6 2 1 2. Zero spectrophotometer at 340 nm with distilled water. jaundice. Levels of ALT are only slightly elevated in patients following a Total Precision 3. For each sample and control, add 1 mL Working Reagent to myocardial infarction. Serum 1 Serum 2 Serum 3 cuvette or test tube and warm to for 3 minutes. UV methods for ALT determination were first developed Wroblewski and Mean ALT 23 43 99 4a. Add 100 uL (0 mL) serum to its respective tube and mix gently. LaDue in 1956. The method was based on the oxidation of NADH Std. Deviation 0 0 0,9 Running this method (1:10 ratio) will yield results linear to 300 or lactate dehydrogenase (LDH). In 1980, the International Federation of C. 3 1 0,9 4b. Add 50 uL (0 mL) serum to its respective tube and mix gently. Clinical Chemistry recommended a reference procedure for the Precision studies were performed according to NCCLS Tentative Running this method (1:20 ratio) will yield results linear to 600 measurements of ALT based on the Wroblewski and LaDue procedure. Guideline, 5. Read and record absorbance at 1 minute. Continue incubating at The Stanbio ALT reagent is based on a modified formulation of the IFCC2 Linearity: Linear to 600 at running method 1:20 ratio and 300 and record absorbance again at 2 and 3 minutes. Rate should and Bergmeyer methods. running method at 1:10 ratio. Performed according to NCCLS be constant. Guideline ALT Oxoglutarate Pyruvate 6. Determine the average absorbance per minute multiply Sensitivity: Based on an instrument resolution of A 0, the factor 1768 (1:10 ratio) or 3376 (1:20 ratio) for results in LDH NOTE: If cuvette is not temperature controlled, incubate samples at method presented shows a sensitivity of 2 Pyruvate NADH H2O between readings. Find Symbol Glossary at Quality Control: Stanbio 1, Level 1Control Serum, Cat. No. References Alanine aminotransferase (ALT) catalyzes the transfer of the amino group from alanine to to form glutamate and pyruvate. The and Stanbio 2. Level 2 Control Serum, Cat. No. 1. Henry, J.: Clinical Diagnosis and Management Laboratory pyruvate formed is then reduced to lactate in the presence of LDH, with are recommended for each run. Other commercially available Methods, W. Saunders and Co., Philadelphia, PA, simultaneous oxidation of reduced nicotinamide adenine dinucleotide controls with ALT values assayed this method are also suitable. ALT (1974) (NADH). The rate of decrease in absorbance at 340 nm is directly activity determined in these materials, this procedure should fall 2. Wroblewski, F. and LaDue, J. Proc. Soc. Exper. Biol. and Med. proportional to ALT activity. within the ranges stated for the controls. Two levels of controls should 91:569 (1956) be analyzed with each run. Reagent 3. International Federation of Clinical Chemistry. Provisional Calibration: ALT activity is based on the extinction Concentrations of Enzymes. Clin Chem 23: 887, 1977. ALT Buffer (R1), Ref. No. 2931 of NADH at 340 nm (see section). The instrument 4. Bergmeyer HU, Scheibe P, Wahlefeld AW: Optimization of methods Composition: 500 calibration guidelines should be followed to calibrate for aspartate aminotransferase and alanine aminotransferase. Clin LDH 1200 your analyzer. Assaying the ALT contents of a control serum with known Chem 24: 58, 1978. Tris Buffer, pH 7 100 ALT values can be used to assure instrument calibration has been 5. Bergmeyer, H. Principles of Enzymatic Analysis. Verlag Chemic, performed correctly. 1978. ALT Enzyme (R2), Ref. No. 2932 Results 6. Young Effects of drugs on clinical laboratory tests. AACC Press, Composition: 2 Oxoglutarate 15 Values are derived based on the micromolar extinction Washington D., 1990 NADH (Disodium salt) 0 of NADH at 340 nm (0). Units per liter of 7. Henry JB. Clinical Diagnosis and Management Laboratory activity is that amount of enzyme which oxidizes one of Methods, 17th ed. WB Saunders Co., 1984, p 1437. NADH per minute. 8. Stanbio Laboratory Data Precautions: The reagents are for Vitro Diagnostic Rx only. STANBIO LABORATORY, LP DISCLAIMS ALL EXPRESS AND IMPLIED WARRANTIES OF THE MERCHANTABILITY Normal precautions exercised in handling laboratory reagents should be (1:10 sample to reagent ratio) AND FITNESS PERTAINING TO THIS PRODUCT WHICH AR NOT EXPRESSLY DETAILED IN THIS PACKAGING Absorptivity) X (Tot Vol Sam Vol) INFORMATION OR A WRITTEN AGREEMENT BETWEEN THE BUYER AND SELLER OF THIS PRODUCT followed. The reagents contain sodium azide which may be toxic if ingested. Sodium azide may also react with lead and copper plumbing to 1 0) x STANBIO LABORATORY LP MAINTAINS THAT THIS PRODUCT CONFORMS TO THE INFORMATION CONTAINED IN THIS INSERT. PURCHASER MUST DETERMINE THE SUITABILITY OF THE PRODUCT FOR THEIR PARTICULAR form highly explosive metal azides. Refer to Safety Data Sheet for any x 1768 USE USE ONLY IN ACCORDANCE WITHLABELING INSTRUCTIONS updated risk, hazard or safety information. (1:20 sample to reagent ratio) Manufactured : Reagent Preparation: Buffer and Enzyme liquid reagents are supplied 0) x Stanbio Laboratory 1261 North Main Street Boerne, TX 78006 USA X 3376 Ph: (830) Fax (830) Prepare Working Reagent in the ratio of 5 parts Buffer (R1) to 1 part Enzyme (R2), (i., 25 mL Buffer and 5 mL Enzyme). Limitations Reagent Storage and Stability: Reagents are stable until the If the is greater than 0, dilute 1 part sample with 9 parts RBR.2930.CE.EN Last Revision: Procedure No. 2930CE expiration date on their respective labels, when properly stored at isotonic saline and Multiply the result 10. ALT values for and protected from light. Reagents should appear clear and colorless. neonatal patients have not been established with this procedure. Discard if either appears cloudy or contains particulate matter. The Working Grossly icteric or turbid specimen may require the use of a sample Reagent is stable for 4 weeks at or 5 days at room temperature blank. The Working Reagent should be discarded if the initial absorbance, Expected Values8 read against distilled water at 340 nm is below 0. Normal Range: 3 35 This range should serve only as a guideline. It is recommended that each laboratory establish its own range of expected values, since S STANBID LABORATORY An EKF Diagnostics Company CE Material Required But Not Provided differences exist between instruments, laboratories, and local STANBIO Spectrophotometer capable of absorbance reading at 340 nm and 1 cm populations. S lightpath. Constant temperature block or bath, or temperature Performance Characteristics LABORATORY controlled cuvette well. Accurate pipetting devices, Test tubes and Comparison: A group of 82 sera ranging in ALT activity from 3 682 An EKF Diagnostics Company Interval timer was assayed the described ALT method and a similar Specimen Collection and Storage commercially available ALT reagent. Comparison of the results yielded a Stanbio serum is the specimen of choice. Whenever possible correlation coefficient of and the regression equation was y Procedure No. 2930 specimens should be separated and analyzed on the day of collection. 0 0. (Comparison studies were performed according to Store serum in stoppered tubes. About ALT is lost 3. days at NCCLS Tentative Guideline, For the Kinetic Quantitative Determination of and in 1 day at Precision: precision was established 20 assays on three in Serum for Manual Automated Interfering Substances: Hemolysis must be avoided as the different levels of commercial serum controls. Total Precision values Procedures concentration of ALT in red cells is roughly 5 times that of serum.² were obtained assaying the 3 commercial controls for 5 consecutive Bilirubin levels up to 40 and triglyceride levels up to days. Summary and Principle show no interference in this test Certain drugs and other substances The enzyme alanine aminotransferase is widely reported in a variety of are also known to affect ALT values. 6 Serum 1 Serum 2 Serum 3 tissue sources. The major source of ALT is of hepatic origin and has led to Mean ALT 22 45 102 Procedure the application of ALT determinations to the study of hepatic diseases. Std. Deviation 1 1 1 1. Prepare ALT Working Reagent according to instructions. Elevated serum levels are found in hepatitis, cirrhosis, and obstructive C. 6 2 1 2. Zero spectrophotometer at 340 nm with distilled water. jaundice. Levels of ALT are only slightly elevated in patients following a Total Precision 3. For each sample and control, add 1 mL Working Reagent to myocardial infarction. Serum 1 Serum 2 Serum 3 cuvette or test tube and warm to for 3 minutes. UV methods for ALT determination were first developed Wroblewski and Mean ALT 23 43 99 4a. Add 100 uL (0 mL) serum to its respective tube and mix gently. LaDue in 1956. The method was based on the oxidation of NADH Std. Deviation 0 0 0,9 Running this method (1:10 ratio) will yield results linear to 300 or lactate dehydrogenase (LDH). In 1980, the International Federation of C. 3 1 0,9 4b. Add 50 uL (0 mL) serum to its respective tube and mix gently. Clinical Chemistry recommended a reference procedure for the Precision studies were performed according to NCCLS Tentative Running this method (1:20 ratio) will yield results linear to 600 measurements of ALT based on the Wroblewski and LaDue procedure. Guideline, 5. Read and record absorbance at 1 minute. Continue incubating at The Stanbio ALT reagent is based on a modified formulation of the IFCC2 Linearity: Linear to 600 at running method 1:20 ratio and 300 and record absorbance again at 2 and 3 minutes. Rate should and Bergmeyer methods. running method at 1:10 ratio. Performed according to NCCLS be constant. Guideline ALT Oxoglutarate Pyruvate 6. Determine the average absorbance per minute multiply Sensitivity: Based on an instrument resolution of A 0, the factor 1768 (1:10 ratio) or 3376 (1:20 ratio) for results in LDH NOTE: If cuvette is not temperature controlled, incubate samples at method presented shows a sensitivity of 2 Pyruvate NADH H2O between readings. Find Symbol Glossary at Quality Control: Stanbio 1, Level 1Control Serum, Cat. No. References Alanine aminotransferase (ALT) catalyzes the transfer of the amino group from alanine to to form glutamate and pyruvate. The and Stanbio 2. Level 2 Control Serum, Cat. No. 1. Henry, J.: Clinical Diagnosis and Management Laboratory pyruvate formed is then reduced to lactate in the presence of LDH, with are recommended for each run. Other commercially available Methods, W. Saunders and Co., Philadelphia, PA, simultaneous oxidation of reduced nicotinamide adenine dinucleotide controls with ALT values assayed this method are also suitable. ALT (1974) (NADH). The rate of decrease in absorbance at 340 nm is directly activity determined in these materials, this procedure should fall 2. Wroblewski, F. and LaDue, J. Proc. Soc. Exper. Biol. and Med. proportional to ALT activity. within the ranges stated for the controls. Two levels of controls should 91:569 (1956) be analyzed with each run. Reagent 3. International Federation of Clinical Chemistry. Provisional Calibration: ALT activity is based on the extinction Concentrations of Enzymes. Clin Chem 23: 887, 1977. ALT Buffer (R1), Ref. No. 2931 of NADH at 340 nm (see section). The instrument 4. Bergmeyer HU, Scheibe P, Wahlefeld AW: Optimization of methods Composition: 500 calibration guidelines should be followed to calibrate for aspartate aminotransferase and alanine aminotransferase. Clin LDH 1200 your analyzer. Assaying the ALT contents of a control serum with known Chem 24: 58, 1978. Tris Buffer, pH 7 100 ALT values can be used to assure instrument calibration has been 5. Bergmeyer, H. Principles of Enzymatic Analysis. Verlag Chemic, performed correctly. 1978. ALT Enzyme (R2), Ref. No. 2932 Results 6. Young Effects of drugs on clinical laboratory tests. AACC Press, Composition: 2 Oxoglutarate 15 Values are derived based on the micromolar extinction Washington D., 1990 NADH (Disodium salt) 0 of NADH at 340 nm (0). Units per liter of 7. Henry JB. Clinical Diagnosis and Management Laboratory activity is that amount of enzyme which oxidizes one of Methods, 17th ed. WB Saunders Co., 1984, p 1437. NADH per minute. 8. Stanbio Laboratory Data Precautions: The reagents are for Vitro Diagnostic Rx only. STANBIO LABORATORY, LP DISCLAIMS ALL EXPRESS AND IMPLIED WARRANTIES OF THE MERCHANTABILITY Normal precautions exercised in handling laboratory reagents should be (1:10 sample to reagent ratio) AND FITNESS PERTAINING TO THIS PRODUCT WHICH AR NOT EXPRESSLY DETAILED IN THIS PACKAGING Absorptivity) X (Tot Vol Sam Vol) INFORMATION OR A WRITTEN AGREEMENT BETWEEN THE BUYER AND SELLER OF THIS PRODUCT followed. The reagents contain sodium azide which may be toxic if ingested. Sodium azide may also react with lead and copper plumbing to 1 0) x STANBIO LABORATORY LP MAINTAINS THAT THIS PRODUCT CONFORMS TO THE INFORMATION CONTAINED IN THIS INSERT. PURCHASER MUST DETERMINE THE SUITABILITY OF THE PRODUCT FOR THEIR PARTICULAR form highly explosive metal azides. Refer to Safety Data Sheet for any x 1768 USE USE ONLY IN ACCORDANCE WITHLABELING INSTRUCTIONS updated risk, hazard or safety information. (1:20 sample to reagent ratio) Manufactured : Reagent Preparation: Buffer and Enzyme liquid reagents are supplied 0) x Stanbio Laboratory 1261 North Main Street Boerne, TX 78006 USA X 3376 Ph: (830) Fax (830) Prepare Working Reagent in the ratio of 5 parts Buffer (R1) to 1 part Enzyme (R2), (i., 25 mL Buffer and 5 mL Enzyme). Limitations Reagent Storage and Stability: Reagents are stable until the If the is greater than 0, dilute 1 part sample with 9 parts RBR.2930.CE.EN Last Revision: Procedure No. 2930CE expiration date on their respective labels, when properly stored at isotonic saline and Multiply the result 10. ALT values for and protected from light. Reagents should appear clear and colorless. neonatal patients have not been established with this procedure. Discard if either appears cloudy or contains particulate matter. The Working Grossly icteric or turbid specimen may require the use of a sample Reagent is stable for 4 weeks at or 5 days at room temperature blank. The Working Reagent should be discarded if the initial absorbance, Expected Values8 read against distilled water at 340 nm is below 0. Normal Range: 3 35 This range should serve only as a guideline. It is recommended that each laboratory establish its own range of expected values, since S STANBID LABORATORY An EKF Diagnostics Company PROCEDURE Method M 340nm: I 6 3 1 mL Working Reagent (h) M. IV 6 103 X 0 SV at for 3 mins. Retinetion colfficies 1 x No Add 100ml soun (1:10 ratio 300VL) 0 0 N W. t SAMPLE F Lincoly x1768 X.P. 1 mL t 0 Incubate at for i mn. Ref Range: 13 0 m mL (100wl) 10 106 009 Read obs. at 340 nm vs. H2O blank ALT Communication Mading for the next 2 mins. AST 00622 X 0 1 003 1 mih. intowal in between readings Limitations. NOTE: Incubate in between rading: D. AHOS Mr. media 10 datema Calculate for the knear Vdlve: 300 Dilute the sample wing 1:10 Name of Patient Ratio isotinic saline 1:10 1pat sample 19 pats diluent Andyot 1 the result the dilution factor 10 PROCEDURE Method M 340nm: I 6 3 1 mL Working Reagent (h) M. IV 6 103 X 0 SV at for 3 mins. Retinetion colfficies 1 x No Add 100ml soun (1:10 ratio 300VL) 0 0 N W. t SAMPLE F Lincoly x1768 X.P. 1 mL t 0 Incubate at for i mn. Ref Range: 13 0 m mL (100wl) 10 106 009 Read obs. at 340 nm vs. H2O blank ALT Communication Mading for the next 2 mins. AST 00622 X 0 1 003 1 mih. intowal in between readings Limitations. NOTE: Incubate in between rading: D. AHOS Mr. media 10 datema Calculate for the knear Vdlve: 300 Dilute the sample wing 1:10 Name of Patient Ratio isotinic saline 1:10 1pat sample 19 pats diluent Andyot 1 the result the dilution factor 10
AST & ALT Stanbio - notes
Course: Clinical Chemistry 2 (MDT 3122L)
University: Our Lady of Fatima University
