Biochemistry Handbook For Mindray [PDF]

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MINDRAY Clinical Chemistry Solutions



PRODUCT LIST A. Biochemistry Reagents .................................................................................................................................. 4 1. Hepatic .................................................................................................................................................... 4 1.1 ALT (Alanine Aminotransferase) ..................................................................................................... 4 1.2 AST (Aspartate Aminotransferase) ................................................................................................. 5 1.3 GGT (gamma-Glutamyltransferase) ............................................................................................... 7 1.4 ALP (Alkaline Phosphatase) ........................................................................................................... 8 1.5 ALB (Albumin) .............................................................................................................................. 10 1.6 TP (Total Protein) ......................................................................................................................... 11 1.7 Bil-D (Bilirubin Direct, VOX Method) ............................................................................................ 13 1.8 Bil-D (Bilirubin Direct, DSA Method) ............................................................................................. 14 1.9 Bil-T (Bilirubin Total, VOX Method) ............................................................................................... 15 1.10 Bil-T (Bilirubin Total, DSA Method) ............................................................................................. 17 1.11 PA (Prealbumin).......................................................................................................................... 18 1.12 TBA (Total Bile Acids) ................................................................................................................. 19 1.13 CHE (Cholinesterase) ................................................................................................................ 21 2. Renal ..................................................................................................................................................... 22 2.1 Crea (Creatinine, Sarcosine Oxidase Method) ............................................................................. 22 2.2 Crea (Creatinine, Modified Jaffé Method)..................................................................................... 24 2.3 Urea/BUN (Blood Urea Nitrogen) ................................................................................................. 25 2.4 UA (Uric Acid) ............................................................................................................................... 27 2.5 CO2 (Carbon dioxide) ................................................................................................................... 29



3. Cardiac .................................................................................................................................................. 30 3.1 CK (Creatine Kinase) ................................................................................................................... 30 3.2 CK-MB (Creatine Kinase-MB Isoenzyme) .................................................................................... 32 3.3 a-HBDH (α-Hydroxybutyrate dehydrogenase) ............................................................................. 34 3.4 LDH (Lactate Dehydrogenase)..................................................................................................... 35 3.5 HCY (Homocysteine) ................................................................................................................... 36 4. Lipids ..................................................................................................................................................... 38 4.1 Apo A1 (Apolipoprotein A1) .......................................................................................................... 38 4.2 Apo B (Apolipoprotein B) .............................................................................................................. 39 4.3 Lp(a) (Lipoprotein(a)) ................................................................................................................... 41 4.4 HDL-C (High Density Lipoprotein - Cholesterol)........................................................................... 42 4.5 LDL-C (Low Density Lipoprotein - Cholesterol) ............................................................................ 44 4.6 TC (Total Cholesterol) .................................................................................................................. 45 4.7 TG (Triglycerides)......................................................................................................................... 47 5. Diabetes ................................................................................................................................................ 48 5.1 Glu (Glucose, HK method) ........................................................................................................... 48 5.2 Glu (Glucose, GOD-POD Method) ............................................................................................... 50 5.3 HbA1c (Hemoglobin A1c) ............................................................................................................. 51 5.4 FUN (Fructosamine) ..................................................................................................................... 53 6. Pancreatitis ........................................................................................................................................... 55 6.1 α-AMY (alpha-Amylase) ............................................................................................................... 55 6.2 LIP (Lipase) .................................................................................................................................. 57 7. Inorganic ions ........................................................................................................................................ 58 7.1 Ca (Calcium) ................................................................................................................................ 58 7.2 Mg (Magnesium) .......................................................................................................................... 60 7.3 P (Phosphorus) ............................................................................................................................ 61 7.4 Fe (Iron) ....................................................................................................................................... 62 8. Immune ................................................................................................................................................. 64 8.1 Ig A (Immunoglobulin A) ............................................................................................................... 64 8.2 Ig G (Immunoglobulin G) .............................................................................................................. 66 8.3 Ig M (Immunoglobulin M) ............................................................................................................. 67 8.4 C3 (Complement C3) ................................................................................................................... 69 8.5 C4 (Complement C4) ................................................................................................................... 70 8.6 CRP (C-reactive Protein).............................................................................................................. 72 8.7 RF (Rheumatoid Factor) .............................................................................................................. 73 8.8 ASO (Antibodies Against Streptolysin O)...................................................................................... 74 B. Calibrator & Quality Control ......................................................................................................................... 76 1. Calibrator ............................................................................................................................................... 76 1.1 Multi Sera Calibrator .................................................................................................................... 76 1.2 Lipids Calibrator ........................................................................................................................... 77 1.3 Specific Proteins Calibrator .......................................................................................................... 78 1.4 CK-MB Calibrator ......................................................................................................................... 79 1.5 Lipoprotein(a) Calibrator .............................................................................................................. 80 1.6 Prealbumin Calibrator .................................................................................................................. 80



2. Quality Control ...................................................................................................................................... 81 2.1 Multi Control Sera N ..................................................................................................................... 81 2.2 Multi Control Sera P ..................................................................................................................... 82 2.3 Lipids Control N............................................................................................................................ 83 2.4 Lipids Control P ............................................................................................................................ 84 2.5 HDL&LDL Cholesterol Control P .................................................................................................. 84 2.6 Specific Proteins Control N .......................................................................................................... 85 2.7 Specific Proteins Control P ........................................................................................................... 86 2.8 CK-MB Control N ......................................................................................................................... 86 2.9 CK-MB Control P.......................................................................................................................... 87 2.10 Lipoprotein(a) Control N&P ........................................................................................................ 87 2.11 Prealbumin Control N&P ............................................................................................................ 88



A. Biochemistry Reagents 1. Hepatic 1.1 ALT (Alanine Aminotransferase) Order Information Cat. No.



Package size



ALT0102



R1 4×35 mL + R2 2×18 mL



ALT0103



R1 6×40 mL + R2 2×32 mL



ALT0104



R1 6×60 mL + R2 3×32 mL



ALT0105



R1 2×250 mL + R2 1×125 mL



Clinical significance Alanine aminotransferase (EC 2.6.1.2, ALT), formerly called Glutamic Pyruvic Transaminase (GPT), is one of liver-specific enzymes. It can catalyze the interconversion of amino acids and α-ketoacids by transfer of amino groups. Elevated ALT levels can indicate myocardial infarction, muscular dystrophy, especially in hepatobiliary diseases. Measurement of ALT is often used in diagnosis and monitoring treatment of liver diseases and heart diseases. The AST/ALT ratio is often used for differential diagnosis in liver diseases: if the AST/ALT ratio < 1, it indicates mild liver damage; otherwise it is associated with severe, often chronic liver diseases. Method UV-assay according to IFCC (International Federation of Clinical Chemistry and Laboratory Medicine) without pyridoxal phosphate activation. Reaction Principle α-oxoglutarate + L-alanine pyruvate + NADH + H+



LDH



ALT



L-glutamate + pyruvate L-lactate + NAD+



Alanine aminotransferase catalyzes the reversible transamination of L-alanine and α-oxoglutarate to pyruvate and L-glutamate. The pyruvate is then reduced to lactate in the presence of lactate dehydrogenase (LDH) with the concurrent oxidation of reduced β-nicotinamide adenine dinucleotide (NADH) to β-nicotinamide adenine dinucleotide (NAD). This change in absorbance is directly proportional to the activity of ALT in the sample. Reagent Components and Concentrations



R1



R2



TRIS buffer



150 mmol/L



L-Alanine



750 mmol/L



LDH



≥1200 U/L



α-oxoglutarate



90 mmol/L



NADH



0.9 mmol/L



Storage and Stability Stable up to expiry date indicated on the label, when stored unopened at 2℃－8℃ and protected from light. Once opened, the reagent is stable for 28 days when refrigerated on the analyzer or refrigerator. Contamination of the reagents must be avoided. Do not freeze the reagents. Assay procedure Blank



Sample



Reagent 1



1000 μL



1000 μL



Dist water



100 μL



－



Sample



－



100 μL



250 μL



250 μL



Mix, incubate for 5 min, then add: Reagent 2



Mix thoroughly, read the absorbance after 1 min and monitor time. Read the absorbance again for additional 3 min. ΔA/min = [ΔA/min sample]- [ΔA/min blank]



Reference Intervals Sample Type



Conventional Units



S.I.Units



Male



≤45 U/L



≤0.75 μkat/L



Female



≤34 U/L



≤0.57 μkat/L



Sample Type



Conventional Units



S.I.Units



Serum / Plasma



4－500 U/L



0.07－8.33 μkat/L



Reportable Range



If the value of sample exceeds 500 U/L, the sample should be diluted with 9 g/L NaCl solution (e.g. 1+ 9) and repeat the assay using this dilution, the result should be multiplied by 10.



1.2 AST (Aspartate Aminotransferase) Order Information Cat. No.



Package size



AST0102



R1 4×35 mL + R2 2×18 mL



AST0103



R1 6×40 mL + R2 2×32 mL



AST0104



R1 6×60 mL + R2 3×32 mL



AST0105



R1 2×250 mL + R2 1×125 mL



Clinical significance



Aspartate aminotransferase (EC 2.6.1.1, AST)，formerly called Glutamic Oxalacetic Transaminase (GOT), is present in both cytoplasm and mitochondria of cells，belonging to the transaminase family, which catalyze the conversion of amino acids and α-oxoglutarate by transfer of amino groups. AST is commonly found in various human tissues. The heart muscle is found to have the most activity of the enzyme, secondly in the brain, liver, gastric mucosa, skeletal muscle and kidneys. The serum AST present low activity in the healthy human body, but when these tissues injury or damage, AST is released into blood and results in high blood AST activity. Measurement of AST in serum and plasma is mainly used for the diagnosis of heart muscle damages, liver damages and skeletal muscle diseases as well as for monitoring the treatment. The AST/ALT ratio is often used for differential diagnosis in liver diseases. While the ratio < 1, it indicates mild liver damage, otherwise it is associated with severe, often chronic liver diseases. Method UV-assay according to IFCC (International Federation of Clinical Chemistry and Laboratory Medicine) without pyridoxal phosphate activation. Reaction Principle L-aspartate + α-oxoglutarate oxaloacetate + NADH + H+



AST



oxaloacetate + L-glutamate L-malate + NAD+



MDH



(MDH－Malate dehydrogenase，EC1.1.1.37) In the assay reaction, the AST catalyzes the reversible transamination of L-aspartate and α-oxoglutarate to oxaloacetate and L-glutamate. The oxaloacetate is then reduced to malate in the presence of malate dehydrogenase with NADH being oxidized to NAD+. The rate of the photometrically determined NADH decrease is directly proportional to the rate of formation of oxaloacetate and thus the AST activity. Reagents Components and Concentrations



R 1：



R 2：



TRIS buffer



100 mmol/L



L-aspartate



300 mmol/L



LDH



≥900 U/L



MDH



≥600 U/L



α-oxoglutarate



60 mmol/L



NADH



0.9 mmol/L



Storage and Stability Stable up to expiry date indicated on the label, when stored unopened at 2℃－8℃ and protected from light. Once opened, the reagent is stable for 28 days when refrigerated on the analyzer or refrigerator. Contamination of the reagents must be avoided. Do not freeze the reagents. Assay procedure



Reagent 1



Blank



Sample



1000 μL



1000 μL



Dist water



100 μL



－



Sample



－



100 μL



250 μL



250 μL



Mix, incubate for 5 min, then add: Reagent 2



Mix thoroughly, read the absorbance after 1 min and monitor time. Read the absorbance again for additional 3 min. ΔA/min = [ΔA/min sample]- [ΔA/min blank]



Reference Intervals Sample Type



Conventional Units



S.I. Units



Male



≤35 U/L



≤0.58 μkat/L



Female



≤31U/L



≤0.52 μkat/L



Reportable Range Sample Type



Conventional Units



S.I. Units



Serum / Plasma



4－350 U/L



0.07－5.83 μkat/L



If the value of sample exceeds 350 U/L, the sample should be diluted with 9 g/L NaCl solution (e.g. 1+ 9) and repeat the assay using this dilution, the result should be multiplied by 10.



1.3 GGT (gamma-Glutamyltransferase) Order Information Cat. No.



Package size



GGT0102



R1 4×35 mL + R2 2×18 mL



GGT0103



R1 6×40 mL + R2 2×32 mL



GGT0104



R1 6×60 mL + R2 3×32 mL



GGT0105



R1 2×250 mL + R2 1×125 mL



Clinical significance Gamma-Glutamyltransferase (EC 2.3.2.2, GGT) is a transferase, widely distributed in tissues, particularly in the liver, pancreas, kidney and spleen. The measurement of GGT is often used in the diagnosis and monitoring of hepatobiliary diseases. Increased GGT activity can indicate the damage of hepatobiliary tissue. GGT test is also a sensitive screening test for occult alcoholism. Method UV-assay for the quantitative determination of gamma-glutamyltransferase (GGT) according to Szasz. Reaction Principle L-γ-glutamyl-3-carboxy-4-nitroanilide + glycyl-glycine



GGT



L-γ-glutamyl- glycyl-glycine



+ 5-amino-2-nitrobenzoate Gamma-glutamyltransferase transfers the gamma-glutamyl group of gamma-glutamyl-3-carboxy-4-nitroanilide to glycyl-glycine with the production of p-nitroaniline. The amount of 5-amino-2-nitrobenzoate results in the elevated absorbance which is directly



proportional to the activity of GGT in the sample. Reagent Components and Concentrations R1 R2



TRIS buffer



100 mmol/L



Glycyl-glycine



150 mmol/L



L-γ-glutamyl-3-carboxy-4-nitroanilide



20 mmol/L



Storage and Stability Stable up to expiry date indicated on the label, when stored unopened at 2℃－8℃ and protected from light. Once opened, the reagent is stable for 30 days when refrigerated on the analyzer or refrigerator. Contamination of the reagents must be avoided. Do not freeze the reagents. Assay procedure Blank



Sample



Reagent 1



1000 μL



1000 μL



Dist water



100 μL



－



Sample



－



100 μL



250 μL



250 μL



Mix, incubate for 1 min, then add: Reagent 2



Mix thoroughly, read the absorbance after 1 min and monitor time. Read the absorbance again for additional 3 min. ΔA/min = [ΔA/min sample]- [ΔA/min blank]



Reference Intervals Sample Type



Conventional Units



S.I.Units



Male



7KU/L



Ascorbic acid oxidase



2KU/L



Catalase



>100KU/L



ESPMT



0.47mM



Creatininase



>400KU/L



Peroxidase



>50KU/L



4-aminoantipyrine



2.95 mmol/L



Storage and stability Stable up to expiry date indicated on the label, when stored unopened at 2-8℃ and protected from light. Once opened, the reagents are stable for 30 days when refrigerated on the analyzer or refrigerator. Contamination of the reagents must be avoided. Do not freeze the reagents. Assay procedure Blank



Sample



Reagent 1



1800 μL



1800 μL



Dist. water



60 μL



－



Sample



－



60 μL



Mix, incubate for 3 min. at 37℃, then add:



600 μL



Reagent 2



600 μL



Mix thoroughly, incubate at 37℃ for 5 min, and then read the absorbance change value. ΔA = [ΔA sample]- [ΔA blank]



Reference Intervals Sample Type Serum/Plasma



Conventional Units



S.I. Units



Male



0.8-1.3 mg/dL



70-115 μmol/L



Female



0.5-0.9 mg/dL



44-80 μmol/L



Reportable Range Sample Type



Conventional Units



S.I. Units



Serum / Plasma



0.11-102 mg/dL



10-9000 μmol/L



If the value of sample exceeds 9000 μmol/L, the sample should be diluted with 9 g/L NaCl solution (e.g. 1+ 9) and rerun; the result should be multiplied by 10.



2.2 Crea (Creatinine, Modified Jaffé Method) Order Information Cat. No.



Package size



CRE0102



R1 3×35 mL + R2 3×35 mL



CRE0103



R1 4×40 mL + R2 4×40 mL



CRE0104



R1 4×45 mL + R2 4×45 mL



CRE0105



R1 2×250 mL + R2 2×250 mL



Clinical significance Creatinine is synthesized at a constant rate form creatine phosphate during muscle contractions. Since the excretion of creatinine in healthy individuals is independent of diet and it is constantly produced, the clearance ratio of creatinine is one of the most sensitive indexes for glomerular filtration rate (GFR) detecting. Many renal diseases such as glomerular nephritis, nephropathy syndrome, and serious renal failure will lead to elevated levels of creatinine in serum. It is a practical method to measure the creatinine level together with the urea level to distinguish the reason of azotemia. Method Modified Jaffé method Reaction Principle -



Creatinine + Picric acid



OH Creatinine-Picric acid complex



At an alkaline solution, creatinine combines with picric acid to form an orange-red colored complex. The absorbency increase is directly proportional to the concentration of creatinine. Reagents Components and concentration R1：



Sodium hydroxide



0.38 mol/L



Picric acid



R2：



15 mmol/L



Storage and stability Up to expiration date indicated on the label, when stored unopened at 2-8℃and protected from light. Once opened, the reagents are stable for 21 days when refrigerated on the analyzer or refrigerator. Contamination of the reagents must be avoided. Do not freeze the reagents. Assay procedure Blank



Sample



Reagent 1



180 μL



180 μL



Dist. water



18 μL



－



Sample



－



18 μL



180 μL



180 μL



Mix, incubate for 1 min. at 37℃ , then add: Reagent 2



Mix thoroughly, incubate at 37℃ for 30 s and then read the absorbance change value over a further 2 min. ΔA = [ΔA sample]- [ΔA blank]



Reference Intervals Sample Type Serum / Plasma



S.I. Units Men Women Men



Urine



Women



50 years: 72–127 µmol/L 58–96 µmol/L 800–2000 mg/24 hours 600–1800 mg/24 hours



Reportable Range Sample Type



S.I. Units



Serum / Plasma / Urine



9–2420 μmol/L



If the value of sample exceeds 2420 μmol/L, the sample should be diluted with 9 g/L NaCl solution (e.g. 1+ 9) and rerun; the result should be multiplied by 10.



2.3 Urea/BUN (Blood Urea Nitrogen) Order Information Cat. No.



Package size



URE0102



R1 4×35 mL + R2 2×18 mL



URE0103



R1 6×40 mL + R2 2×32 mL



URE0104



R1 6×60 mL + R2 3×32 mL



URE0105



R1 2×250 mL + R2 1×125 mL



Clinical significance Urea is the final products of the protein and aminophenol catabolism. Adult produces 16 g urea everyday. Diseases associated with elevated levels of urea in blood are referred to as uremia or azotemia. Parallel determination of urea and creatinine is used to distinguish the reason of azotemia. Prerenal azotemia may cause by starvation, pyrexia, dehydration, increased protein catabolism, cortisol treatment or decreased renal perfusion (e.g. serious heart failure, lack of water), while creatinine level remains within the reference ranges. Postrenal azotemia may cause by the obstruction of the urinary tract, in this regard, both urea and creatinine levels rise, but urea is in a higher extent. Method Urease-glutamate Dehydrogenase, UV method Reaction Principle



Urease



2NH4+ + CO3 2-



Urea + 2H2O



α-Oxoglutarate + NH4+ + NADH



GLDH



L-Glutamate + NAD+ + H2O



Urea is hydrolyzed by urease, and one of the products, ammonia, helps to turn NADH to NAD+ with the catalysis of GLDH. The absorbency decrease is directly proportional to the concentration of urea. Reagents Components and concentrations



R1:



R2:



Tris buffer



120 mmol/L



ADP



750 mmol/L



Urease



≥40 KU/L



GLDH



≥0.4 KU/L



NADH



1.2 mmol/L



α-Oxoglutarate



25 mmol/L



Storage and stability Up to expiration date indicated on the label, when stored unopened at 2-8℃ and protected from light. Once opened, the reagents are stable for 21 days when refrigerated on the analyzer or refrigerator. Contamination of the reagents must be avoided. Do not freeze the reagents. The standard is stable, even after opening, up to the stated expiration date when stored at 2-8℃. Assay procedure Blank



Sample



Reagent 1



1000 μL



1000 μL



Dist. water



10 μL



－



Sample



－



10 μL



250 μL



250 μL



Mix, incubate for 2 min. at 37℃. , then add: Reagent 2



Mix thoroughly, incubate at 37℃ for 30 s and then read the absorbance change value over a further 1-2 min. ΔA = [ΔA sample]- [ΔA blank]



Reference Intervals Sample Type



S.I. Units



Serum / Plasma



Adult



2.8-7.2 mmol/L



Urine



Morning Urine



141-494 mmol/L



Reportable Range Sample Type



S.I. Units



Serum / Plasma / Urine



1-40 mmol/L



If the value of sample exceeds 40 mmol/L, the sample should be diluted with 9 g/L NaCl solution (e.g. 1+ 3) and rerun; the result should be multiplied by 4.



2.4 UA (Uric Acid) Order Information Cat. No.



Package size



UA0102



R1 4×40 mL + R2 2×20 mL



UA0103



R1 6×40 mL + R2 2×32 mL



UA0104



R1 6×60 mL + R2 3×32 mL



UA0105



R1 2×250 mL + R2 1×125 mL



Clinical significance Uric acid is synthesized in liver and excreted via kidney, and it is the final products of the purine metabolism. The most common complication of hyperuricemia is the formation of urate crystals, which is called tophus, around the joints. Further causes of elevated blood concentrations of uric acid are renal function disease, starvation, drug abuse, toxicosis, malignant tumour, and increased alcohol and incretion disorders. Reasons of Hypouricemia are hereditary metabolic disorders, renal diseases, severe hepatic diseases and drug effects. Method Uricase-Peroxidase (Uricase-POD) method Reaction Principle



Ascorbic acid + O2 Uric acid + H2O + O2 TOOS + 4-AAP + 2H2O2 + H+



Ascorbate oxidase dehydro-ascorbic acid + H2O Uricase Allantoin + CO2 + H2O2 POD Quinoneimine + 4H2O



By using ascorbic oxidase to eliminate the interference of ascorbic acid, the uric acid is catalyzed to produce H2O2, which oxidize the 4-AAP to yield a colored dye of quinoneimine. The absorbency decrease is directly proportional to the concentration of uric acid.



Reagents Components and concentrations



R1:



R2:



Phosphate buffer



70 mmol/L



Peroxidase



5000 U/L



Ascorbate oxidase



3000 U/L



TOOS



0.72 mmol/L



Phosphate buffer



70 mmol/L



Peroxidase



10000 U/L



4-Aminoantipyrine



1.7 mmol/L



Uricase



750 U/L



Storage and stability Up to expiration date indicated on the label, when stored unopened at 2-8℃ and protected from light. Once opened, the reagents are stable for 14 days when refrigerated on the analyzer or refrigerator. Contamination of the reagents must be avoided. Do not freeze the reagents. Assay procedure Blank



Sample



Reagent 1



1200 μL



1200 μL



Dist. water



25 μL



－



Sample



－



25 μL



Mix, incubate for 5 min. at 37 ℃, and read the blank absorbance, then add: 300 μL



Reagent 2



300 μL



Mix thoroughly at 37 ℃, and read the absorbance again 4-5 min. later. ΔA = [ΔA sample]- [ΔA blank]



Reference Intervals Sample Type Serum / Plasma Urine



Conventional Units



S.I. Units



Men



3.6-8.2 mg/dL



214-488 μmol/L



Women



2.3-6.1 mg/dL



137-363 μmol/L



Normal diet



2.8 U/mL



Storage and stability Stable up to expiry date indicated on the label, when stored unopened at 2-8℃ and protected from light. Once opened, the reagents are stable for 30 days when refrigerated on the analyzer or refrigerator. Contamination of the reagents must be avoided. Do not freeze the reagents. Assay procedure Blank



Sample



Reagent 1



1000 μL



1000 μL



Dist. water



50 μL



－



Sample



－



50 μL



250 μL



250 μL



Mix, incubate for 3 min. at 37℃, then add: Reagent 2



Mix thoroughly, incubate at 37℃ for 3 min., and then read the absorbance change value within 1-3 min. ΔA/min = [ΔA/min sample]- [ΔA/min blank]



Reference Intervals Sample Type



Conventional Units



S.I. Units



Serum



≤ 24 U/L



≤ 0.4 μkat/L



Reportable Range Sample Type



Conventional Units



S.I. Units



Serum



5-600 U/L



0.08-10.00 μkat/L



If the value of sample exceeds 600 U/L, the sample should be diluted with 9 g/L NaCl solution (e.g. 1+ 9) and rerun; the result should be multiplied by 10.



3.3 a-HBDH (α-Hydroxybutyrate dehydrogenase) Order Information Cat. No.



Package size



HBD0102



R1 4×35 mL + R2 2×18 mL



HBD0103



R1 6×40 mL + R2 2×32 mL



HBD0104



R1 4×45 mL + R2 4×12 mL



HBD0105



R1 2×250 mL + R2 1×125 mL



Clinical significance α-Hydroxybutyrate dehydrogenase is an isoenzyme of lactate dehydrogenase. The level of α-HBDH is higher than other isoenzymes of LDH in heart muscle tissue, so it is somewhat more sensitive and more specific in the diagnosis of myocardial infarction. The HBDH/LDH ratio can be used for differentiation liver disease from heart disease. A decreased ratio indicates liver diseases, on the contrary an increased ratio means myocardial infarction. Method UV-assay according to German Society of Clinical Chemistry (DGKC). Reaction Principle +



α-Oxobutyrate + NADH + H



α-HBDH α-Hydroxybutyric acid + NAD+



By the catalysis of α-HBDH, α-Oxobutyrate is deoxidated into α-Hydroxybutyric acid, at the same time NADH is oxidized into NAD+. The absorbency decrease is directly proportional to the activity of α-HBDH. Reagents Components and concentrations R1： R2：



Tris buffer



100 mmol/L



α-Oxobutyrate



4.4 mmol/L



Tris buffer



50 mmol/L



NADH



1.27 mmol/L



Storage and stability Up to expiration date, when stored unopened at 2-8℃and protected from light. Once opened, the reagents are stable for 28 days when refrigerated on the analyzer or refrigerator. Contamination of the reagents must be avoided. Do not freeze the reagents. Assay procedure Blank



Sample



Reagent 1



240 μL



240 μL



Dist. water



10 μL



－



Sample



－



10 μL



Mix, incubate at 37℃ for 3 min., then add: 60 μL



Reagent 2



60 μL



Mix thoroughly, incubate 37℃ for 1-2 min., and then read the absorbance change value within 2-3 min. ΔA/min = [ΔA/min sample]- [ΔA/min blank]



Reference Intervals Sample Type Serum / Plasma



Adult



Conventional Units



S.I. Units



72-182 U/L.



1.2-3.0 μkat/L



Reportable Range Sample Type



Conventional Units



S.I. Units



Serum / Plasma



10-1000 U/L



0.17-16.67 μkat/L



If the value of sample exceeds 1000 U/L, the sample should be diluted with 9 g/L NaCl solution (e.g. 1+ 5) and rerun; the result should be multiplied by 6.



3.4 LDH (Lactate Dehydrogenase) Order Information Cat. No.



Package size



LDH0102



R1 4×35 mL + R2 2×18 mL



LDH0103



R1 6×40 mL + R2 2×32 mL



LDH0104



R1 4×45 mL + R2 4×12 mL



LDH0105



R1 2×250 mL + R2 1×125 mL



Clinical significance Lactate dehydrogenase (EC 1.1.1.27, LDH) is a tetramer composed of two different subunits. It has five isoenzymes. LDH is widely distributed in tissues, particularly in the liver, muscles, kidneys and heart. Increased LDH activities can indicate a variety of pathological conditions, such as myocardial infarction, liver diseases, blood diseases and cancers. Because of the lack of organ specificity of LDH, parallel measurement of its isoenzymes or other enzymes such as ALP, ALT and AST is necessary for differential diagnosis. Method UV-assay according to IFCC (International Federation of Clinical Chemistry and Laboratory Medicine). Reaction Principle L-lactate + NAD+



pyruvate + NADH + H+



LDH



Lactate Dehydrogenase catalyzes the conversion of L-lactate to pyruvate in the presence of lactate dehydrogenase (LDH). In the process, β-nicotinamide adenine dinucleotide (NAD) is deoxidized to NADH. This change in absorbance is directly proportional to the activity of LDH in the sample.



Reagent Components and Concentrations R1



TRIS buffer



50 mmol/L



L-Lactate



5 mmol/L



+



R2



NAD



7.0 mmol/L



Storage and Stability Stable up to expiry date indicated on the label, when stored unopened at 2℃－8℃ and protected from light. Once opened, the reagent is stable for 30 days when refrigerated on the analyzer or refrigerator. Contamination of the reagents must be avoided. Do not freeze the reagents. Assay procedure Blank



Sample



Reagent 1



2400 μL



2400 μL



Dist water



100 μL



－



Sample



－



100 μL



600 μL



600 μL



Mix, incubate for 2－3 min, then add: Reagent 2



Mix thoroughly, read the absorbance after 1 min and monitor time. Read the absorbance again for additional 3 min. ΔA/min = [ΔA/min sample]- [ΔA/min blank]



Reference Intervals Sample Type



Conventional Units



S.I.Units



Men



< 248 U/L



< 4.13 μkat/L



Women



< 247 U/L



< 4.12 μkat/L



Reportable Range Sample Type



Conventional Units



S.I.Units



Serum / Plasma



4－1000 U/L



0.07－16.67 μkat/L



If the value of sample exceeds 1000 U/L, the sample should be diluted with 9 g/L NaCl solution (e.g. 1+ 9) and repeat the assay using this dilution, the result should be multiplied by 10.



3.5 HCY (Homocysteine) Order Information Cat. No. HCY0102 HCY0103 HCY0104



Package size R1a 6×10mL + R1b 6 + R1c 6 + R2 1×11mL + Calibrator 1×1mL + Quality control 2×1mL R1a 6×10mL + R1b 6 + R1c 6 + R2 1×11mL + Calibrator 1×1mL + Quality control 2×1mL R1a 12×10mL + R1b 12 + R1c 12 + R2 2×11mL + Calibrator 1×1.5mL + Quality control 2×1.5mL



Clinical significance Homocysteine (Hcy) is a thiol-containing amino acid produced by the intracellular demethylation of methionine. Total homocysteine (tHcy) represents the sum of all forms of Hcy including forms of oxidized, protein bound and free. Elevated level of tHcy has emerged as an important risk factor in the assessment of cardiovascular disease. Excess Hcy in the blood stream may cause injures to arterial vessels due to its irritant nature, and result in inflammation and plaque formation, which may eventually cause blockage of blood flow to the heart. Elevated tHcy levels are resulted from four major causes including: a) genetic deficiencies in enzymes involved in Hcy metabolism such as cystathionine beta-synthase (CBS), methionine synthase (MS), and methylenetetrahydrofolate reductase (MTHFR); b) nutritional deficiency in B vitamins such as B6, B12 and folate; c) renal failure for effective amino acid clearance, and d) drug interactions such as nitric oxide, methotrexate and phenytoin that interfere with Hcy metabolisms. Elevated levels of tHcy are also linked with Alzheimer’s disease and Osteoporosis. Guidelines for tHcy determination in clinical laboratories have recently been established. Method Enzymatic Assay Method Reaction Principle Oxidated HCY＋Reducing reagent Deoxidizd HCY



HCY enzyme



Deoxidizd HCY Sulfureted hydrogen



Sulfureted hydrogen＋ Chromogen ＋ oxidant



Methyl blue complex



Reagents Components and concentration R1a：



Tris



20 mmol/L



R1b：



HCY enzyme



/



R1c：



Reducer



/



R2:



Oxidant



2.8g/L



Calibrator



Concentration see label



Quality control



Concentration see label



Storage and stability Up to expiration date indicated on the label, when stored unopened at 2-8℃ and protected from light. Once opened, the R2 reagent is stable for 28 days when refrigerated on the analyzer or refrigerator, Once mixed the R1 working reagent can stable for 7 days. Contamination of the reagents must be avoided. Do not freeze the reagents. Once opened, the calibrator and control are stable for 28 days at 2–8℃, do not freeze. Assay procedure Blank



Sample



Reagent 1



200 μL



200 μL



Dist. water



20 μL



－



Sample



－



20 μL



Mix, incubate for 5 min. at 37 ℃, and read the blank absorbance, then add:



30 μL



Reagent 2



30 μL



Mix thoroughly at 37 ℃, and read the absorbance again 5 min. later. ΔA = [ΔA sample]- [ΔA blank]



Reference Intervals Sample Type



S.I. Units



Serum/plasma



5～15 μ mol/L



Reportable Range Sample Type



S.I. Units



Serum/plasma



1.0～40.0 μ mol/L



If the value of sample exceeds 40.0 μ mol/L, the sample should be diluted with 9 g/L NaCl solution (e.g. 1+ 9) and the result should be multiplied by 10.



4. Lipids 4.1 Apo A1 (Apolipoprotein A1) Order Information Cat. No.



Package size



APA0102



R1 1×35 mL + R2 1×12 mL



APA0103



R1 4×40 mL + R2 2×28 mL



APA0104



R1 3×45 mL + R2 3×15 mL



APA0105



R1 1×240 mL + R2 1×80 mL



Clinical significance ApolipoproteinA1 is the major protein component of high-density lipoprotein, which takes charge of transferring excess cholesterol to the liver, so it is a protective factor of atherosclerosis. ApoA1 can foreshow the early coronary heart disease; furthermore evaluate the function of lowering fat therapy. Decrease of ApoA1 level exists in inherited hypolipoproteinemia, cholestasis, sepsis and atherosclerosis. And increase exists in liver disease, pregnancy and estrogen treatment. Method Turbidimetry Method Reaction Principle Anti-human ApoA1 antibody + ApoA1



Immunocomplex (agglutination)



Determination of the concentration of ApoA1 through photometric measurement of immunocomplex between antibodies of ApoA1 and ApoA1 present in the sample, the absorbency increase is directly proportional to the concentration of ApoA1.



Reagents Components and concentration R1： R2：



Phosphate buffer



100 mmol/L



PEG



1.15mmol/L



Reportable Range Sample Type



Conventional Units



Serum



0.05-6 mmol/L



If the value of sample exceeds 6 mmol/L, the sample should be diluted with 9 g/L NaCl solution (e.g. 1+ 9) and rerun; the result should be multiplied by 10.



4.5 LDL-C (Low Density Lipoprotein - Cholesterol) Order Information Cat. No.



Package size



LDL0102



R1 1×40 mL + R2 1×14 mL



LDL0103



R1 4×40 mL + R2 2×28 mL



LDL0104



R1 4×60 mL + R2 2×42 mL



LDL0105



R1 3×250 mL + R2 1×250 mL



Clinical significance LDL-Cholesterol is directly related to the risk of developing coronary heart disease. A low HDL/LDL-Cholesterol ratio is directly related to the risk of developing coronary artery disease. Elevated LDL-Cholesterol is the primary target of cholesterol-lowering therapy. Method Direct method Reaction Principle （1） HDL,VLDL, Chylomicrons 2H2O2 （2） LDL



Cholestenone + H2O2



Catalase



2H2O + O2



CHE + CHO



Cholestenone + H2O2



H2O2 + TOOS + 4-aminoantipyrin



POD



Quinonimine



The System monitors the change in absorbance at 600 nm. This change in absorbance is directly proportional to the concentration of cholesterol in the sample and is used by the System to calculate and express the LDL-cholesterol concentration. Reagents Components and Concentrations



R 1：



R 2：



Good’s buffer



50 mmol/L



Cholesterol esterase



600 U/L



Cholesterol oxidase



500 U/L



Catalase



600 KU/L



TOOS



2 mmol/L



Good’s buffer



50 mmol/L



4-aminoantipyrine



4 mmol/L



Peroxidase



4 U/mL



Storage and stability Stable up to expiry date indicated on the label, when stored unopened at 2-8℃ and protected from light. Once opened, the reagents are stable for 30 days when refrigerated on the analyzer or refrigerator.



Contamination of the reagents must be avoided. Do not freeze the reagents.



Assay procedure Blank



Sample



Reagent 1



900 μL



900 μL



Dist. water



12 μL



－



Sample



－



12 μL



300 μL



300 μL



Mix, incubate for 5 min. at 37℃, then add: Reagent 2



Mix thoroughly, incubate at 37℃ for 5 min., and then read the absorbance change value. ΔA = [ΔA sample]- [ΔA blank]



Reference Intervals Sample Type



Conventional Units



Serum



0-4.11 mmol/L



Reportable Range Sample Type



Conventional Units



Serum



0.05-25.8 mmol/L



If the value of sample exceeds 25.8 mmol/L, the sample should be diluted with 9 g/L NaCl solution (e.g. 1+ 9) and rerun; the result should be multiplied by 10.



4.6 TC (Total Cholesterol) Order Information Cat. No.



Package size



TC0102



R 4×40 mL



TC0103



R 6×40 mL



TC0104



R 6×60 mL



TC0105



R 4×250 mL



Clinical significance Cholesterol is a main component of cell membranes and lipoprotein and it is the precursor for steroid hormones and bile acids synthesizing. Cholesterol is transported in plasma by low-density lipoprotein. The level of the individual’s total cholesterol is used in screening early atherosclerosis and monitoring the clinical effect of drugs or low-fat diet. Method Cholesterol oxidase- Peroxidase (CHOD-POD) method Reaction Principle CHE



Cholesterol ester + H2O



Cholesterol + Fatty acid CHO



Cholesterol + O2



4-Cholestenone + H2O2 POD



2H2O2 + 4-Aminoantipyrine + Phenol



Quinoneimine + 4H2O



By the catalysis of CHE and CHO, Cholesterol ester is catalyzed to yield H2O2, which oxidates 4- Aminoantipyrine with phenol to form a colored dye of quinoneimine. The absorbency increase is directly proportional to the concentration of cholesterol. Reagents Components and concentrations



R：



Phosphate buffer



100 mmol/L



Phenol



5 mmol/L



4-Aminoantipyrine



0.3 mmol/L



Cholesterol esterase



>150 KU/L



Cholesterol oxidase



>100 KU/L



Peroxidase



5 KU/L



Storage and stability Up to expiration date indicated on the label, when stored unopened at 2-8℃ and protected from light. Once opened, the reagents are stable for 28 days when refrigerated on the analyzer or refrigerator. Contamination of the reagents must be avoided. Do not freeze the reagents. Assay procedure Blank



Sample



R



1000 μL



1000 μL



Dist. water



10 μL



－



Sample



－



10 μL



Mix thoroughly at 37℃, and read the absorbance 10 min. later. ΔA = [ΔA sample]- [ΔA blank]



Reference Intervals Sample Type



S.I. Units



Serum / Plasma



≤5.2 mmol/l



Reportable Range Sample Type



Conventional Units



S.I. Units



Serum / Plasma



3.85-769.23 mg/dL



0.1-20.0 mmol/L



If the value of sample exceeds 20.0 mmol/L, the sample should be diluted with 9 g/L NaCl solution (e.g. 1+ 1) and rerun; the result should be multiplied by 2.



4.7 TG (Triglycerides) Order Information Cat. No.



Package size



TG0102



R 4×40 mL



TG0103



R 6×40 mL



TG0104



R 6×60 mL



TG0105



R 4×250 mL



Clinical significance Triglycerides are the most abundant naturally lipids. It consists of three fatty acids and one glycerol, and is transported in plasma combing with apolipoproteins. Measurement of triglycerides is used for detecting early atherosclerotic risks, classing hyperlipoproteinemia and monitoring the clinical effect of drugs or low-fat diet. High triglyceride levels often lead to liver or kidneys disease, diabetes and pancreas disease. Method Glycerokinase Peroxidase- Peroxidase Method Reaction Principle



Lipase



Triglycerides + 3H2O



Glycerol + fatty acid GK Glycerol-3-phosphate + ADP



Glycerol + ATP



GPO Dihydroxyacetone Phosphate + H2O2



Glycerol-3-phosphate + O2 H2O2 + 4-Aminoantipyrine + 4-Chlorophenol



POD Quinoneimine + HCl + H2O



Through a sequence of enzymatic catalysis steps by lipase, GK and GPD, triglycerides is catalyzed to yield H2O2, which oxidize 4Aminoantipyrinel to yield a colored dye of quinoneimine. The absorbency increase is directly proportional to the concentration of triglycerides. Reagents Components and concentrations Phosphate buffer



50 mmol/L



4-Chlorophenol



5 mmol/L



ATP



2 mmol/L



2+



R：



Mg



4.5 mmol/L



Glycerokinase



≥0.4 U/mL



Peroxidase



≥0.5 U/mL



Lipoprotein lipase



≥1.3 U/mL



4-Aminoantipyrine Glycerol-3-phosphate-oxidase



0.25 mmol/L ≥1.5 U/mL



Storage and Stability Up to expiration date indicated on the label, when stored unopened at 2-8℃ and protected from light.



Once opened, the reagents are stable for 21 days when refrigerated on the analyzer or refrigerator. Contamination of the reagents must be avoided. Do not freeze the reagents. Assay procedure Blank



Sample



R



1000 μL



1000 μL



Dist. water



10 μL



－



Sample



－



10 μL



Mix thoroughly at 37 ℃, and read the absorbance 10 min. later. ΔA = [ΔA sample]- [ΔA blank]



Reference Intervals Sample Type



S.I. Units



Serum / Plasma



≤ 2.3 mmol/L



Reportable Range Sample Type



S.I. Units



Serum / Plasma



0.1-12.5 mmol/L



If the value of sample exceeds 12.5 mmol/L, the sample should be diluted with 9 g/L NaCl solution (e.g. 1+ 1) and rerun; the result should be multiplied by 2.



5. Diabetes 5.1 Glu (Glucose, HK method) Order Information Cat. No.



Package size



GLU0202



R1 4×36 mL + R2 2×34 mL



GLU0203



R1 4×40 mL + R2 2×40 mL



GLU0204



R1 6×45 mL + R2 3×45 mL



GLU0205



R1 2×250 mL + R2 1×250 mL



Clinical significance Carbohydrates supply the body energy with glucose, which is the most important monosaccharide in blood, and it is an indispensable energy supplier for cellular function. Measuring blood glucose is used for the diagnosis of carbohydrate metabolism disorders and monitoring of treatment in diabetes mellitus, neonatal hypoglycemia, idiopathic hypoglycemia, pharmic hypoglycemia and insulinoma. Method HK method



Reaction Principle



HK glucose-6-phosphate + ADP



Glucose + ATP



Hexokinase catalyzes the phosphorylation of glucose to glucose-6-phosphate by ATP. Glucose-6-phosphate + NAD+



G6P-DH



6-phosphogluconate + NADH + H+



Glucose-6-phosphate dehydrogenase oxidizes glucose-6-phosphate in the presence of NADP to gluconate-6-phosphate. No other carbohydrate is oxidized. The rate of NADPH formation during the reaction is directly proportional to the glucose concentration and can be measured photometrically. Reagents Components and concentration R1：



R2：



Phosphate buffer



100 mmol/L



G-6-PDH



20 KU/L



ATP



10 mmol/L



Phosphate buffer



100 mmol/L



HK



1 KU/L +



NAD



0.5 mmol/L



Storage and stability Up to expiration date indicated on the label, when stored unopened at 2-8℃ and protected from light. Once opened, the reagents are stable for 28 days when refrigerated on the analyzer or refrigerator. Contamination of the reagents must be avoided. Do not freeze the reagents. The standard is stable, even after opening, up to the stated expiration date when stored at 2-8℃. Assay procedure Blank



Sample



Reagent 1



2500 μL



2500 μL



Dist. water



30 μL



－



Sample



－



30 μL



Mix, incubate at 37 ℃ for 3 min., and read the blank absorbance, then add: 1250 μL



Reagent 2



1250 μL



Mix thoroughly 37 ℃, and read the absorbance again 5 min. later. ΔA = [ΔA sample]- [ΔA blank]



Reference Intervals Sample Type



Conventional Units



S.I. Units



Capillary vessel whole blood



70-100 mg/dL



3.9-5.5 mmol/L



60-100 mg/dL



3.5-5.5 mmol/L



70-115 mg/dL



3.9-6.4 mmol/L



Vein whole blood Vein plasma



Adult



Reportable Range Sample Type



S.I. Units



Serum / Plasma



0.6-33.0 mmol/L



If the value of sample exceeds 33.0 mmol/L, the sample should be diluted with 9 g/L NaCl solution (e.g. 1+1) and rerun; the result should be multiplied by 2.



5.2 Glu (Glucose, GOD-POD Method) Order Information Cat. No.



Package size



GLU0102



R1 4×40 mL + R2 2×20 mL



GLU0103



R1 4×40 mL + R2 2×20 mL



GLU0104



R1 6×60 mL + R2 3×32 mL



GLU0105



R1 2×250 mL + R2 1×125 mL



Clinical significance Carbohydrates supply the body energy with glucose, which is the most important monosaccharide in blood, and it is an indispensable energy supplier for cellular function. Measuring blood glucose is used for the diagnosis of carbohydrate metabolism disorders and monitoring of treatment in diabetes mellitus, neonatal hypoglycemia, idiopathic hypoglycemia, pharmic hypoglycemia and insulinoma. Method Glucose oxidase-Peroxidase (GOD-POD) method Reaction Principle D-Gluconic acid + H2O2



D-Glucose + O2



2H2O2 + 4-AAP + p-Hydroxybenzoic acid sodium + H3O+



GOD POD



Quinoneimine + 5H2O



By the catalysis of GOD, Glucose is oxidated to yield H2O2, and then at the present of POD, H2O2 oxidates 4-Aminoantipyrine with p-Hydroxybenzoic acid sodium to form a colored dye of quinoneimine. The absorbency increase is directly proportional to the concentration of glucose. Reagents Components and concentration R1：



R2：



Phosphate buffer



100 mmol/L



Ascorbate oxidase



4700 U/L



Glucose oxidase



4000 U/L



Phosphate buffer



100 mmol/L



Peroxidase



6700 U/L



4-Aminoantipyrine



0.7 mmol/L



p-Hydroxybenzoic acid sodium



1.3 mmol/L



Storage and stability Up to expiration date indicated on the label, when stored unopened at 2-8℃ and protected from light. Once opened, the reagents are stable for 28 days when refrigerated on the analyzer or refrigerator. Contamination of the reagents must be avoided. Do not freeze the reagents. Assay procedure Blank



Sample



Reagent 1



240 μL



240 μL



Dist. water



3 μL



－



Sample



－



3 μL



Mix, incubate at 37 ℃ for 5 min., and read the blank absorbance, then add: 60 μL



Reagent 2



60 μL



Mix thoroughly 37 ℃, and read the absorbance again 5-10 min. later. ΔA = [ΔA sample]- [ΔA blank]



Reference Intervals Sample Type



Conventional Units



S.I. Units



Capillary vessel whole blood



70-100 mg/dL



3.9-5.5 mmol/L



60-100 mg/dL



3.5-5.5 mmol/L



70-115 mg/dL



3.9-6.4 mmol/L



Vein whole blood Vein plasma



Adult



Reportable Range Sample Type



S.I. Units



Serum / Plasma



0.3-35 mmol/L



If the value of sample exceeds 35 mmol/L, the sample should be diluted with 9 g/L NaCl solution (e.g. 1+1) and rerun; the result should be multiplied by 2.



5.3 HbA1c (Hemoglobin A1c) Order Information Cat. No. HBA0102 HBA0103 HBA0104



Clinical significance



Package size R1 2×30mL+R2 1×12mL+Calibrator 2×1mL + Quality control 2×1mL Pretreatment Solution 1×150ml R1 2×40mL+R2 1×15mL+Calibrator 2×1mL + Quality control 2×1mL Pretreatment Solution 1×200ml R1 4×40mL+R2 2×15mL+Calibrator 2×1mL + Quality control 2×1mL Pretreatment Solution 2×150ml



Hemoglobin (Hb) consists of four protein chains with four heme portions, and is the red-pigmented protein located in the erythrocytes. Its main function is the transport of oxygen and carbon dioxide in blood. Each Hb molecule is able to bind four oxygen molecules. Hb consists of a variety of subfractions and derivatives. Among this heterogeneous group of hemoglobins HbA1c is one of the glycated hemoglobins, a subfraction formed by the attachment of various sugars to the Hb molecule. HbA1c is formed in two steps by the non-enzymatic reaction of glucose with the N-terminal amino group of the β-chain of normal adult Hb (HbA). The first step is reversible and yields labile HbA1c. This slowly rearranges in the second reaction step to yield stable HbA1c. In the erythrocytes, the relative amount of HbA converted to stable HbA1c increases with the average concentration of glucose in the blood. The conversion to stable HbA1c is limited by the erythrocyte’s life span of approximately 100 to 120 days. As a result, HbA1c reflects the average blood glucose level during the preceding 2 to 3 months. HbA1c is thus suitable to monitor long-term blood glucose control in individuals with diabetes mellitus. More recent glucose levels have a greater influence on the HbA1c level. The approximate relationship between HbA1c and mean blood glucose value during the preceding 2 to 3 months has been analyzed by several studies. Method Enzymatic Assay Method Reaction Principle In the first reaction, the concentration of hemoglobin is measured at absorbance of fixed wavelength, and simultaneously the fructosyl dipeptides are generated from the N-terminus amino groups of the beta-chain of HbA1c by the reaction of protease. In the second reaction, the reaction of Fructosyl peptide oxidase(FPOX) with fructosyl dipeptides, generated hydroperoxide allows 10-(carboxymethylaminocarbonyl)-3,7-bis(dimethylamino) phenothiazine sodium salt to develop a color in the presence of peroxidase. The change in absorbance is measured for HbA1c determination. The combined assay results for hemoglobin and HbA1c are used by the system to calculate and express HbA1c(%). Reagents Components and concentration R1： R2： Pretreatment



Tris buffer



2.7 mmol/L



Peroxidase



1500 U/L



Fructosyl peptide oxidase



1500 U/L



Hemolysin



5 g/L



Solution Calibrator



Concentration see label



Quality control



Concentration see label



Storage and stability Up to expiration date indicated on the label, when stored unopened at 2-8℃ and protected from light. Once opened, the reagents are stable for 28 days when refrigerated on the analyzer or refrigerator. Contamination of the reagents must be avoided. Do not freeze the reagents. Once dissolved, the calibrator and control are stable for 15 days at 2–8℃,do not freeze. Assay procedure Pretreatment Procedure of Whole Blood Prior to testing, whole blood samples should be centrifuged at 2000 rpm for 5 min. Get 25uL blood corpuscle from the blood corpuscle deposition into a sample cup or an Eppen-dorf microfuge tube.



Add 500 μL of Pretreatment Solution into sample cup or an Eppen-dorf microfuge tube. Close the test tube and lysis the blood by shaking vigorously. Then mix the solution by either swirling gently or by using a vibration mixer. The hemolysate can be used as working samples after 5 minute. 1). Hb Blank



Sample



Reagent 1



180 μL



180 μL



Dist. water



12 μL



－



Sample



－



12 μL



Mix, incubate for 5 min. at 37 ℃, and read the blank absorbance A ΔA = [ΔA sample]- [ΔA blank] 2). HbA1c Blank



Sample



Reagent 1



180 μL



180 μL



Dist. water



12 μL



－



Sample



－



12 μL



Mix, incubate for 5 min. at 37 ℃, and read the blank absorbance, then add: 60 μL



Reagent 2



60 μL



Mix thoroughly at 37 ℃, and read the absorbance again 5 min later. ΔA = [ΔA sample]- [ΔA blank]



Reference Intervals Sample Type Who blood



S.I. Units According to IFCC



2.9-4.2%



According to NGSP/DCCT



4.8-5.9%



According to JCCLS



4.3-5.8%



Reportable Range Sample Type



S.I. Units



Who blood



3-16 %



If the value of sample exceeds 16%, the sample should be diluted with Pretreatment Solution (e.g. 1+ 1) and rerun.



5.4 FUN (Fructosamine) Order Information Cat. No.



Package size



FUN0102



R1 2×30 mL+R2 1×15 mL+Calibrator 1×1.5 mL+Control 1×1 mL



FUN0103



R1 4×40 mL+R2 2×20 mL+Calibrator 1×1.5 mL+Control 1×1 mL



FUN0104



R1 4×40 mL+R2 2×20 mL+Calibrator 1×1.5 mL+Control 1×1 mL



Clinical significance Fructosamine is a time-averaged indicator of blood glucose levels and is used to assess the glycemic status of diabetics. The concentration of glycated proteins such as glycohemoglobin, glycoalbumin or glycated total protein is generally recognized to be



valuable in evaluating the glycemic status of diabetic patients. Method Colorimetric Assay. Reaction Principle



fructosamine + NBT



colored complex



This colorimetric assay is based on the ability of ketoamines to reduce nitrotetrazolium-blue (NBT) to formazan in an alkaline solution. The rate of formation of formazan is directly proportional to the concentration of fructosamine. Uricase serves to eliminate uric acid interference and detergent eliminates matrix effects. The rate of reaction is measured photometrically at 546 nm. Reagents Components and concentrations



R1:



R2:



Carbonate buffer



100 mmol/L



Uricase



≥400 U/L



Sodium cholate



3.0 mmol/L



Surfactant



appropriate



Phosphate buffer



20 mmol/L



NBT



3.0 mmol/L



Calibrator:



1-Deoxy-1-morpholino-D-Fructose



Control:



Lyophilized control based on human serum



Storage and stability Up to expiration date indicated on the label, when stored unopened at 2-8℃ and protected from light. Once opened, the reagents are stable for 30 days when refrigerated on the analyzer or refrigerator. Once opened, the calibrator are stable for 30 days at 2-8℃, do not freeze. Once dissolved, the control is stable for 30 days at -20℃ (when frozen once) Contamination of the reagents must be avoided. Do not freeze the reagents. Assay procedure Blank



Sample



Reagent 1



200 μL



200 μL



Dist. water



20 μL



－



Sample



－



20 μL



Mix, incubate for 5 min. at 37 ℃, and read the blank absorbance, then add: Reagent 2



50 μL



Mix thoroughly at 37 ℃ 4-5 min later, and read the absorbance again 1-2 min. later. ΔA/min = [ΔA/min sample]- [ΔA/min blank]



Reference Intervals ≤286 µmol/L



50 μL



Reportable Range Sample Type



S.I. Units



Serum / Heparin plasma



5-1000 μmol/L



If the value of sample exceeds 1000 μmol/L, the sample should be diluted with 9 g/L NaCl solution (e.g. 1+3) and rerun; the result should be multiplied by 4.



6. Pancreatitis 6.1 α-AMY (alpha-Amylase) Order Information Cat. No.



Package size



AMY0102



R1 1×38 mL + R2 1×10 mL



AMY0103



R1 4×20 mL + R2 2×10 mL



AMY0104



R1 4×45 mL + R2 4×12 mL



AMY0105



R1 2×250 mL + R2 1×125 mL



Clinical significance α-Amylases originate from various organs and are mainly produced by the pancreas (P-type) and the salivary glands (S-type). α-Amylases catalyze the hydrolytic degradation of polymeric carbohydrates, such as amylose, amylopectin and glycogen by cleaving 1, 4-α-glucosidic bonds into various fragments. 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. Because of its small molecular weight, the blood amylase is eliminated through the kidney and excreted into urine. Elevation of urine amylase activity reflects the rise of serum amylase activity. Measurement of α-amylase in serum and urine is mainly used for the diagnosis of pancreatic disorders, as well as for detecting the development of complications. Normally the α-amylases present low activity, in acute pancreatitis the blood amylase activity increases within 8-12 hours after onset of abdominal pain, peaks after approx. 12-24 hours. The blood amylase activity returns to normal values at the latest after 2-5 days, but the high urine amylase level will still last for 5-7 days. However, various nonpancreatic diseases, e.g. parotitis, renal insufficiency or pulmonary inflammation, can also increase amylase levels. To confirm pancreatic disorders, additional pancreas specific enzyme, lipase or pancreatic-α-amylase, is recommended to be determined at the same time. Method This method is in accordance with the continuous monitoring of recommendations on the IFCC (International Federation of Clinical Chemistry). Reaction Principle



α-amylase 5Ethylidene-G7-PNP + 5H2O



2Ethylidene-G5 + 2G2-PNP + 2Ethylidene-G4



+ 2G3-PNP + Ethylidene-G3 + G4-PNP



α-glucosidase 2G2-PNP + 2G3-PNP + G4-PNP + 14H2O



5PNP + 14Glucose



(PNP－p-nitrophenol; G－α-glucose) In the assay reaction, the substrate 4, 6- ethylidene-(G7)-1, 4-nitrophenyl-(G1) –α, D-maltoheptaoside (EPS-G7) is cleaved by α-amylases and subsequent hydrolysis of all the degradation products to p-nitrophenol with the aid of α-glucosidase (100% chromophore liberation). The increase in absorbance of the p-nitrophenol formed at 405 nm is directly proportional to the activity of α-amylases. Reagents Components and Concentrations R 1：



R 2：



TRIS buffer



50 mmol/L



Magnesium sulphate



10 mmol/L



α-Glucosidase



4500 U/L



TRIS buffer



50 mmol/L



E-pNP-G7



5.5 mmol/L



Storage and Stability Stable up to expiry date indicated on the label, when stored unopened at 2℃－8℃ and protected from light. Once opened, the reagent is stable for 30 days when refrigerated on the analyzer or refrigerator. Contamination of the reagents must be avoided. Do not freeze the reagents. Assay procedure Blank



Sample



Reagent 1



200 μL



200 μL



Dist water



5 μL



－



Sample



－



5 μL



50 μL



50 μL



Mix, incubate for 5 min, then add: Reagent 2



Mix thoroughly, read the absorbance after 1 min and monitor time. Read the absorbance again for additional 2 min. ΔA/min = [ΔA/min sample]- [ΔA/min blank]



Reference Intervals Sample Type



Conventional Units



S.I. Units



Serum (Male/ Female)



≤100 U/L



≤1.67 μkat/L



Spontaneous urine



≤1000 U/L



≤16.67 μkat/L



Collected urine



≤900 U/24h



≤15.00 μkat/24h



Reportable Range SAMPLE TYPE



CONVENTIONAL UNITS



S.I. UNITS



Serum / Plasma /Urine



5－1500 U/L



0.08－25 μkat/L



If the value of sample exceeds 1500 U/L, the sample should be diluted with 9 g/L NaCl solution (e.g. 1+ 9) and repeat the assay using this dilution, the result should be multiplied by 10.



6.2 LIP (Lipase) Order Information Cat. No.



Package size



LIP0102



R1 : 1×35mL + R2: 1×9mL + Calibrator:1×3mL + Quality control 1×5mL



LIP0103



R1 : 1×40mL + R2: 1×10mL + Calibrator:1×3mL + Quality control: 1×5mL



LIP0104



R1 : 2×40mL + R2: 2×10mL + Calibrator:1×3mL + Quality control: 1×5mL



Clinical significance Lipases are glycoproteins with a molecular weight of 47000 daltons. They are defined as triglyceride hydrolases which catalyse the cleavage of triglycerides to diglycerides with subsequent formation of monoglycerides and fatty acids. Lipase enzymes are produced in the pancreas and also secreted in small amounts by the salivary glands as well as by gastric, pulmo-nary and intestinal mucosa. Determination of lipase is used for di-agnosis and treatment of diseases of the pancreas such as acute and chronic pancreatitis and obstruction of the pancreatic duct. Method Enzymatic Colorimetric Assay Method Reaction Principle The method for the determination of lipase is based on the cleavage of specific chromogenic lipase substrate 1,2-O-dilaurylrac-glycero-3-glutaric acid-(6’methyl-resorufin)-ester emulsified in stabilized micro-particles. In the presence of specific activators of pancreatic lipase as colipase, calcium ions and bile acids, the substrate is con-verted to 1,2-O-dilauryl-rac-glycerol and glutaric acid-6’-methylresorufinester which decomposes spontaneously to glutaric acid and methylresorufin. The increase of absorbance at 580 nm, due to methylresorufin formation, is proportional to the activity of lipase in the sample. Reagents Components and concentrations



R1



R2:



Tris



40 mmol/L



Desoxycholate



1.8 mmol/L



Taurodesoxycholate



7.2 mmol/L



Colipase



>1mg/L



Tartrate buffer,



15 mmol/L



Calcium chloride



0.13 mmol/L



Lipase Substrate



≥ 0.7 mmol/L



Standards



Activity see label



Quality control



Activity see label



Storage and stability Up to expiration date indicated on the label, when stored unopened at 2-8 ℃ and protected from light. Once opened, the reagents are stable for 28 days when refrigerated on the analyzer or refrigerator.



Contamination of the reagents must be avoided. Do not freeze the reagents. Once dissolved, the calibrator and control are stable for 30 days at -20℃(only freeze once). Assay procedure Blank



Sample



Reagent 1



200 μL



200 μL



Dist. water



2 μL



－



Sample



－



2 μL



40 μL



40 μL



Mix, incubate at 37℃ for 3 min., then add: Reagent 2



Mix thoroughly, incubate 37℃ for 2 min., and then read the absorbance change value within 2 min. ΔA/min = [ΔA/min sample]- [ΔA/min blank]



Reference Intervals Sample Type



S.I. Units



Serum/plasma



≤ 38U/L



Reportable Range Sample Type



S.I. Units



Serum/plasma



5～250U/L



If the value of sample exceeds 250U/L, the sample should be diluted with 9 g/L NaCl solution (e.g. 1+ 1) and the result should be multiplied by 2.



7. Inorganic ions 7.1 Ca (Calcium) Order Information Cat. No.



Package size



CA0102



R 4×40 mL



CA0103



R 6×40 mL



CA0104



R 4×45 mL



CA0105



R 4×250 mL



Clinical significance In plasma, calcium consists of three forms: free, conglutinated with proteins or complex with anions such as phosphate, bicarbonate and citrate. Calcium is an absolutely necessary cation for cell functions. For example: muscle contraction, bone mineralization, glycogen metabolism, blood concretion and nerve impulses conduction. Renal diseases, liver diseases, intestinal malabsorption, acute pancreas inflammation, vitamin D deficiency, adrenal cortical hormone therapy, diuretic treatment and hypoparathyroidism all may result in low levels of total calcium.



Hyperparathyroidism, hyperthyroidism, Addison’s disease, intussuscept vitamin D or vitamin A excessively, malignant diseases with metastases and sarcoidosis will lead to high levels of total calcium. Method Arsenazo Ⅲ method Reaction Principle pH=7 a blue colored complex



Calcium + Arsenazo Ⅲ



By using 8-hydroxyquinoline-5-sulfonic acid to eliminate the interference of magnesium, calcium ions combine with Arsenazo Ⅲ to produce a blue colored complex at a neutral solution. The absorbency increase is directly proportional to the concentration of calcium. Reagents Components and concentrations R：



Phosphate buffer



50 mmol/L



8-Hydroxyquinoline-5-sulfonic acid



5 mmol/L



Arsenazo Ⅲ



0.12 mmol/L



Storage and stability Up to expiration date indicated on the label, when stored unopened at 2-8℃ and protected from light. Once opened, the reagents are stable for 28 days when refrigerated on the analyzer or refrigerator. Contamination of the reagents must be avoided. Do not freeze the reagents. Assay procedure Blank



Sample



R



1000 μL



1000 μL



Dist. water



10 μL



－



Sample



－



10 μL



Mix thoroughly at 37 ℃, and read the absorbance 5 min. later. ΔA = [ΔA sample]- [ΔA blank]



Reference Intervals Sample Type



S.I. Units



Serum / Plasma



2.20-2.65 mmol/L



Urine



Male



< 250 mg/24 h (6.2 mmol/24 h)



Female



< 300 mg/24 h (7.5 mmol/24 h)



Reportable Range Sample Type



S.I. Units



Serum / Plasma / Urine



0.1-3.75 mmol/L



If the value of sample exceeds 3.75 mmol/L, the sample should be diluted with 9 g/L NaCl solution (e.g. 1+1) and rerun; the result should be multiplied by 2.



7.2 Mg (Magnesium) Order Information Cat. No.



Package size



MG0102



R 4×40 mL



MG0103



R 6×40 mL



MG0104



R 4×45 mL



MG0105



R 4×250 mL



Clinical significance Magnesium is one of the most abundant cations in the body, and plays an important role in cell respiration, glucose metabolism and transmembrane transport. Magnesium can activate more than 300 kinds of enzymes, the famous of which is Na+-K+-ATP enzyme. Hypomagnesemia may cause by defective gastrointestinal absorption, body fluid losses, diuretic therapy, aminoglucoside treatment, endocrinopathy and alcoholism, and the inherited disease is also an infrequent reason. Hypermagnesemia is found in acute and chronic renal failure, magnesium excess, dehydration and diabetic acidosis. Method Xylidyl blue method Reaction Principle Xylidyl blue + Magnesium



-



OH Xylidyl blue-Magnesium complex



By using the EGTA to eliminate the interference of calcium, magnesium ions combine with xylidyl blue to produce a xylidyl blue -magnesium complex at an alkaline solution. The absorbency increase is directly proportional to the concentration of magnesium. Reagents Components and concentrations



R：



Ethanolamine



49 mmol/L



EGTA



0.13 mmol/L



Xylidyl blue



0.09 mmol/L



Surfactant



＜2% (m/v)



Storage and stability Up to expiration date indicated on the label, when stored unopened at 2-8℃ and protected from light. Once opened, the reagents are stable for 10 days when refrigerated on the analyzer or refrigerator. Contamination of the reagents must be avoided. Do not freeze the reagents. Assay procedure Blank



Sample



R



1000 μL



1000 μL



Dist. water



10 μL



－



Sample



－



10 μL



Mix thoroughly at 37℃, and read the absorbance 5 min. later. ΔA = [ΔA sample]- [ΔA blank]



Reference Intervals Sample Type



Serum / Plasma



Urine



S.I. Units Neonates



0.48–1.05 mmol/l



Children



0.60–0.95 mmol/l



Women



0.77–1.03 mmol/l



Men



0.73–1.06 mmol/l



Male



3–5 mmol/24 h



Reportable Range Sample Type



Conventional Units



S.I. Units



Serum / Plasma / Urine



0.04-4.16 mg/dL



0.04–2.05 mmol/L



If the value of sample exceeds 2.05 mmol/L, the sample should be diluted with 9 g/L NaCl solution (e.g. 1+ 1) and rerun; the result should be multiplied by 2.



7.3 P (Phosphorus) Order Information Cat. No.



Package size



P0102



R 4×40 mL



P0103



R 6×40 mL



P0104



R 4×45 mL



P0105



R 4×250 mL



Clinical significance PO43+ is the main anion in the cell, and its metabolism is closely related with Ca2+. In the body, the main form of Phosphorus is calcium phosphate salt, which is the inorganic substance of the bones. The others take part in the glucose metabolism, constitution of phospholipids, phosphoproteins, nucleic acids and nucleic protein. Phosphorus plays an important role in energy transfer, muscle contraction and nerve conduction. Increased values occur in renal failure, hypoparathyroidism, pseudo hypoparathyroidism, acute metabolically acid toxicosis and calcium phosphate loss of bones and cells. Decreased values occur in malabsorption, hyperparathyroidism, alcoholism and vitamin D deficiency. Method Phosphomolybdate Method Reaction Principle



phosphomolybdate complex



Ammonium molybdate + Sulphuric acid + Phosphate



Ammonium molybdate combines with phosphate in present of sulphuric acid to produce a phosphomolybdate complex. The absorbency increase is directly proportional to the concentration of phosphate. Reagents Components and concentrations R：



Ammonium molybdate



0.3 mmol/L



Sulphuric acid



0.5 mol/L



Surfactant