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Insight V-CHEM General Health Profile

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Insight V-CHEM General Health Profile 【Product Name】 General Health Profile 【Packing Specification】 1 Disc / Sample 【Instrument】 See the InSight V-CHEM chemistry analyser Operator’s Manual for complete information on use of the analyser. 【Intended Use】 The General Health Profile used with the InSight V-CHEM chemistry analyser is intended to be used for the in vitro quantitative determination of total Protein (TP), albumin (ALB), total bilirubin (TBIL), alanine aminotransferase (ALT), blood urea (BUN), creatinine (CRE), amylase (AMY), creatinekinase (CK), calcium (Ca2+), phosphorus (P), alkaline Phosphatase (ALP), glucose (GLU), and total cholesterol (CHOL)in heparinised whole blood, heparinised plasma, or serum in a clinical laboratory setting or point of care location. The General Health Profile and the InSight V-CHEM chemistry analyser comprise an in vitro diagnostic system that aids the physician in the following disorders: liver and gall bladder diseases, urinary system diseases, carbohydrate metabolism disorders, lipid metabolism disorders, cardiovascular disease and pancreatic diseases. 【Test Principles】 This product, which is based on spectrophotometry, is used to quantitatively determine the concentration or activity of the 13 biochemical indicators in the sample. The test principles are as follows: (1) Total Protein (TP) The total protein method is a Biuret reaction, the protein solution is treated with cupric [Cu(II)] ions in a strong alkaline medium. The Cu(II) ions react with peptide bonds between the carbonyl oxygen and amide nitrogen atoms to form a coloured Cu-protein complex. The amount of total protein present in the sample is directly proportional to the absorbance of the Cu-protein complex. The total protein test is an endpoint reaction and the absorbance is measured as the difference in absorbance between 550 nm and 800 nm. - Total Protein + Cu(II) ⎯ OH⎯⎯ →Cu-Protein Complex (2) Albumin (ALB) Bromcresol green (BCG), when bound with albumin, changes from a yellow to green colour. The absorbance maximum changes with the colour shift. BCG + Albumin ⎯ pH4⎯⎯.2→Albumin Complex Bound albumin is proportional to the concentration of albumin in the sample. This is an endpoint reaction that is measured as the difference in absorbance between 630 nm and 700 nm. (3) Total Bilirubin (TBIL) In the enzyme procedure, bilirubin is oxidised by bilirubin oxidase (BOD) into biliverdin. Bilirubin is quantitated as the difference in absorbance between 450 nm and 546 nm. The initial absorbance of this endpoint reaction is determined from the bilirubin blank cuvette and the final absorbance is obtained from the bilirubin test cuvette. The amount of bilirubin in the sample is proportional to the difference between the initial and final absorbance measurements. BOD Bilirubin + O2 ⎯ ⎯⎯ → Biliverdin + H2O (4) Alanine Aminotransferase (ALT) ALT catalyses the transfer of an amino group from L-alanine to ɑ-ketoglutarate to form L-glutamate and pyruvate. Lactate dehydrogenase catalyses the conversion of pyruvate to lactate. Concomitantly, NADH is oxidised to NAD+, as illustrated in the following reaction scheme. L-Alanine + ɑ-Ketoglutarate ⎯ ALT⎯⎯ →L-Glutamate + Pyruvate Pyruvate + NADH + H+ ⎯ LDH⎯⎯ → Lactate + NAD+ The rate of change of the absorbance difference between 340 nm and 405 nm is due to the conversion of NADH to NAD+ and is directly proportional to the amount of ALT present in the sample. (5) Urea(BUN) In the coupled-enzyme reaction, urease hydrolyses urea into ammonia and carbon dioxide. Upon combining ammonia with 2-oxoglutarate and reduced nicotinamide adenine dinucleotide (NADH), the enzyme glutamate dehydrogenase (GLDH) oxidizes NADH to NAD+. Urease BUN + H2O ⎯ ⎯⎯ → NH3 + CO2 GLDH + NH3 + 2-Oxoglutarate + NADH ⎯ ⎯⎯ → L-Glutamate + H2O + NAD The rate of change of the absorbance difference between 340 nm and 405 nm is caused by the conversion of NADH to NAD+ and is directly proportional to the amount of urea present in the sample. (6) Creatinine (CRE) In the coupled enzyme reactions, creatinineamidohydrolase (CAH) hydrolyses creatinine to creatine. A second enzyme, creatineamidinohydrolase (CRH), catalyses the formation of sarcosine from creatine. Sarcosine oxidase (SAO) causes the oxidation of sarcosine to glycine, formaldehyde and hydrogen peroxide (H2O2). Peroxidase (POD) catalyses the reaction among the hydrogen peroxide, N-Ethyl-N-(2- hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium salt (FDAOS) and 4-aminoantipyrine (4-AAP) into a red quinoneimine dye. Potassium ferrocyanide and ascorbate oxidase are added to the reaction mixture to minimise the potential interference of bilirubin and ascorbic acid, respectively. CAH Creatinine + H2O ⎯ ⎯⎯ →Creatine CRH Creatine + H2O ⎯ ⎯⎯ →Sarcosine + Urea SAO Sarcosine + H2O + O2 ⎯ ⎯⎯ → Glycine + Formaldehyde + H2O2 POD H2O2 + FDAOS + 4-AAP ⎯ ⎯⎯ →Red Quinoneimine Dye + H2O Two cuvettes are used to determine the concentration of creatinine in the sample. Endogenous creatine is measured in the blank cuvette, which is subtracted from the combined endogenous creatine and the creatine formed from the enzyme reactions in the test cuvette. Once the endogenous creatine is eliminated from the calculations, the concentration of creatinine is proportional to the intensity of the red colour produced. The endpoint reaction is measured as the difference in absorbance between 550 nm and 700 nm. (7) Amylase (AMY) In the coupled-enzyme reaction, amylase hydrolyses Ethylidene-nitrophenol-maltgglucoside into smaller molecules malt oligomeric glycoside. ɑ-glucosidase (ɑ-GLU) catalyses oligomeric glycoside to produce p-nitrophenolc and glucose. The absorbance of this rate reaction is measured at 405 nm. The production is directly proportional to the amylase activity in the sample. AMY E-4-NP-G7+ H2O ⎯ ⎯⎯ →E-G5 + G2-4-NP+E-G4 + G3-4-NP+E-G3+G4-4-NP -GLU G2-4-NP+G3-4-NP+G4-4-NP+H2O ⎯ ⎯ ⎯ →4-NP+GLU (8) Glucose (GLU) The reaction of glucose with adenosine triphosphate (ATP)catalysed by hexokinase (HK), produces glucose-6-phosphate (G-6-P) and adenosine diphosphate (ADP). Glucose-6-phosphate dehydrogenase (G-6-PD) catalyses the reaction of G-6-P into 6-phosphogluconate and the reduction of nicotinamide adenine dinucleotide phosphate (NADP) to NADPH. HK Glucose + ATP ⎯ ⎯ →Glucose-6-Phosphate + ADP G-6-P + NADP ⎯ G⎯-6-⎯PD→6-Phosphogluconate + NADPH + H+ The absorbance is measured dichromatically at 340 nm and 405 nm. The production of NADPH is directly proportional to the amount of glucose present in the sample. (9) Total Cholesterol (CHOL) The reaction of CHOL is an enzymatic end-point method that uses cholesterol esterase (CE) and Cholesterol oxidase (COD). Peroxidase (POD) catalyses the reaction among the hydrogen peroxide, N- Ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methylanilineodium salt (TOOS) and 4-aminoantipyrine (4-AAP) into a red quinoneimine dye. CE Cholesterol Esters + H2O ⎯ ⎯ → Cholesterol + Fatty Acids COD Cholesterol +O2 ⎯ ⎯⎯ →Cholestenone+ H2O2 POD H2O2+4-AAP+TOOS ⎯ ⎯⎯ →Quinoneimine dye +H2O (10) Calcium (Ca2+) Calcium in the patient sample binds with Arsenazo III to form a calcium-dye complex. Ca2+ + Arsenazo III ⎯⎯ → Ca2+-Arsenazo III Complex It is an endpoint reaction. The amount of total calcium in the sample is proportional to the absorbance. (11) Phosphorus (P) The reaction of inorganic phosphorus with inosine, catalysed by purine nucleoside phosphorylase (PNP), produces hypoxanthine and ribose phosphate. The hypoxanthine is then oxidised to uric acid (UA) catalysed by Xanthine oxidase (XOD). Peroxidase (POD) catalyses the reaction among the hydrogen peroxide, N-Ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methylaniline odium salt (TOOS) and 4- aminoantipyrine (4-AAP) into a red quinoneimine dye. P + Inosine ⎯ PNP⎯⎯ → Hypoxanthine + Ribose Phosphate XOD Hypoxanthine + O2 +H2O ⎯ ⎯⎯ → UA + H2O2 H2O2 + 4-AAP + TOOS Quinoneimine dye + H2O (12) Creatine Kinase (CK) Creatine kinase catalyses the formation of creatine and adenosine triphosphate (ATP) from creatine phosphate and adenosine diphosphate (ADP). With hexokinase (HK) as a catalyst, ATP reacts with D- glucose to form ADP and D-glucose-6-phosphate (G-6-P), which is reacted with nicotinamide adenine dinucleotide phosphate (NADP) in the presence of glucose-6-phosphate dehydrogenase (G-6-PD) to produce 6-Phosphogluconate (6-PG) and NADPH. The formation of NADPH is measured as a change in absorbance at 340 nm relative to 405 nm. This absorbance change is directly proportional to creatine kinase activity in the sample. CK Creatine phosphate + ADP ⎯ ⎯ →Creatine + ATP ATP + D-⎯glucose⎯HK→ ADP + G-6-P G-6-PD G-6-P + NADP ⎯ ⎯ ⎯ → 6-Phosphogluconate + NADPH (13) Alkaline Phosphatase (ALP) Under the catalysis of ALP, the Phosphoric acid on nitrobenzene (4-NNP) was turned into Para nitro phenol (4-NP).4-NP shows a yellow colour in alkaline solution. At the wavelength of 405nm, the ALP activity can be calculated by monitoring the absorbance change rate. ALP 4-NNP ⎯ ⎯⎯ → Acyl phosphate + 4-NP 【Principle of Operation】 Refer to the InSight V-CHEM chemistry analyser for the Principles and Limitations of the Procedure. 【Description of Reagents】 ⎯ POD⎯⎯ → Each General Health Profile contains dry test-specific reagent beads. Each sealing pouch contains a reagent disc and a pack of desiccant. A one-time use disc is for only one sample. Each sealing pouch has a unique code printed on the surface. The ingredient of each General Health Profile are as follows (after redissolution): Component Quantity Total protein assay reagent 13.5μL Albumin assay reagent 13.5μL Total Bilirubin assay reagent 13.5μL Alanine
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