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  • Clinical Chemistry Laboratory Reviewer

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    Nathaniel Derige Andes
    151 просмотров4 страницы
    1) The document discusses various clinical chemistry tests including total cholesterol, HDL cholesterol, triglycerides, total protein, and albumin. 2) It provides information on the principle, reagents, standards, reference values, and procedures for each of these assays. 3) The assays use enzymatic and colorimetric methods to measure the levels of these analytes and provide information on liver and heart health.

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    CC LAB

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    1) The document discusses various clinical chemistry tests including total cholesterol, HDL cholesterol, triglycerides, total protein, and albumin. 2) It provides information on the principle, reagents, standards, reference values, and procedures for each of these assays. 3) The assays use enzymatic and colorimetric methods to measure the levels of these analytes and provide information on liver and heart health.

    Авторское право:

    © All Rights Reserved
    Скачайте в формате DOCX, PDF, TXT или читайте онлайн в Scribd
    Отметить как неприемлемый контент
    151 просмотров4 страницы

    Clinical Chemistry Laboratory Reviewer

    Загружено:

    Nathaniel Derige Andes
    1) The document discusses various clinical chemistry tests including total cholesterol, HDL cholesterol, triglycerides, total protein, and albumin. 2) It provides information on the principle, reagents, standards, reference values, and procedures for each of these assays. 3) The assays use enzymatic and colorimetric methods to measure the levels of these analytes and provide information on liver and heart health.

    Авторское право:

    © All Rights Reserved
    Скачайте в формате DOCX, PDF, TXT или читайте онлайн в Scribd
    Отметить как неприемлемый контент
    из 4

    CLINICAL CHEMSTRY 2 H2O2 + 4-aminophenazone

    LABORATORY REVIEWER + phenol quinoneimine +


    4H2O
    EXERCISE 8: TOTAL CHOLESTEROL
    DISCUSSION:
    ASSAY REQUIREMENT
    Cholesterol
    major lipid in body Wavelength: 500 nm
    precursor of steroid
    Temperature: 20-25 C
    hormones
    usually associated with Measurement: Reagent Blank (0.01)
    coronary heart disease
    Unsaturated steroid alcohol Incubate 10 mins at 20-25 C
    2/3 in esterified form CALCULATION:
    transported by lipoprotein
    such as HDL, LDL, VLDL
    REAGENT:
    Phosphate Buffer
    4-Aminophenazole
    EXERCISE 9: HDL-C
    Phenol
    Peroxidase DISCUSSION:
    Cholesterol Oxidase
    Cholesterol esterase High Density Lipoprotein
    (HDL)
    STANDARD: 200 mg/dL or 5.17 good cholesterol
    mmol/L removes excess cholesterol
    from cells
    REFERENCE VALUE: <200 mg/dL or
    smallest and most dense
    <5.2
    lipoprotein
    PRINCIPLE comprises of both lipid and
    protein
    Determined by Enzymatic Apolipoprotein A-1 (apo A-1)
    Hydrolysis and Oxidation used a as index of the
    Uses cholesterol esterase, amount of HDL
    cholesterol oxidase, peroxidase larger the lipoprotein, the more
    Quinoneimine formed from H2O2 lipid is present relative to
    + 4-aminophenazone in protein lighter the density
    presence of (Phenol and
    peroxidase) REAGENT:
    ENZYME

    Cholesterolester + H2O Goods Buffer (6.6)


    Cholesterol + Fatty Acid Sodium Chloride
    Cholesterol + O2 Cholesterol Esterase
    cholestene-3-one + H2O2 Cholesterol Oxidase
    POD Catalase
    Ascorbate oxidase Triglycerides
    Preservative contains 3 fatty acid
    attached to glycerol by ester
    SUBSTRATE
    bonds
    Goods Buffer (7.0) Fatty Acid can be saturated
    Peroxidase or unsaturated
    4-aminoantipyrin Saturated (Solid in RT,
    Sodium Azide animals)
    Detergents Unsaturated (Liquid in
    RT, plants)
    STANDARD: Check the bottle
    REAGENT:
    REFERENCE VALUE: >35.14 mg/dl or
    >0.91 mmol/L PIPES buffer (7.5)
    4-chlorophenol
    PRINCIPLE 4-aminoantipyrine
    Magnesium Ions
    Determined by Direct
    ATP
    homogenous assay
    Lipase
    elimination of chylomicrons,
    Peroxidase
    VLDL, LDL using Enzymatic
    Glycerol Kinase
    reaction with cholesterol
    Glycerol-3-Phosphate oxidase
    esterase, cholesterol oxidase,
    and catalase STANDARD: 200 mg/dL or 2.28
    surfactant is added together mmol/L
    with chromogen and enzyme
    peroxidase REFERENCE VALUE: <194 mg/dL or
    < 2.2 mmol/L

    ASSAY REQUIREMENT PRINCIPLE

    Wavelength: 593 nm major enzyme used in methods


    of measurement is Lipase
    Temperature: 37 C
    Triglyceride glycerol + fatty
    Measurement: Reagent Blank (0.01)
    substrate (0.25); Enzyme (0.75) acids
    Glycerol + ATP glycerol-3-
    Incubate 5 mins at 37 C phosphate +ADP
    Glycerol-3-phosphate + O2
    CALCULATION: dihydroxyacetone phosphate +
    H2O2
    H2O2 + 4-aminoantipyrine +4
    chlorophenol quinoneimine +
    HCL + H20
    ASSAY REQUIREMENT
    EXERCISE 10: Triglyceride
    Determination Wavelength: 500 nm
    DISCUSSION: Temperature: 20-25C
    Measurement: Reagent Blank (0.01) Wavelength: 520-580 nm
    Incubate 10 mins at 20-25C Temperature: 20-25C
    CALCULATION: Measurement: Reagent Blank (0.01)
    Incubate 10 mins at 20-25C standing
    time 30 mins
    CALCULATION:

    EXERCISE 11: Total Protein


    Determination EXERCISE 12: Albumin
    Determination
    DISCUSSION:
    DISCUSSION:
    Protein
    synthesized in liver Albumin
    significant in the assessment major protein found in
    of synthetic function of liver plasma
    Protein Deficiency: also significant in assessing
    Malabsorption and liver function
    Malnutrition Other function: oncotic
    Most requested: Total Protein function and pH
    and Albumin also binds in various
    substance such as thyroid
    REAGENT:
    hormones, bilirubin, iron,
    Sodium Hydroxide fatty acids
    Potassium Sodium Tartrate
    REAGENT:
    Copper Sulfate
    Potassium Iodide Citrate Buffer (4.2)
    Bromcresol green
    STANDARD: 80 g/L or 8 g/dL
    STANDARD: 40 g/L or 4 g/dL
    REFERENCE VALUE: 66-87 g/L or 6.6-
    8.7 g/dL REFERENCE VALUE: 38-51 g/L or 3.8-
    5.1 g/dL
    PRINCIPLE
    PRINCIPLE
    proteins comprises of several
    peptides bonds capacity of albumin to bind with
    Cupric ions- chelates peptide various
    bonds to for Violet colored Albumin binds with certain dyes
    compound in alkaline solution froms color complex measured
    Biurets Reaction- Absorbance spectrophotometrically
    of this is complex is proportional Bromcresol Green mixed with
    to the protein concentration in albumin in the sample pH is
    the sample maintained by citrate buffer
    Potassium Iodide (KI)- The absorbance of this complex
    formed is proportional to the
    ASSAY REQUIREMENT
    albumin concentration in Incubate 10 mins at 20-25C standing
    sample time 30 mins
    ASSAY REQUIREMENT CALCULATION:
    Wavelength: 520-580 nm
    Temperature: 20-25C
    Measurement: Reagent Blank (0.01)

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