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    Chloride, Bicarbonate and Lactate

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    Ella Ortega
    This document provides information on determining chloride, bicarbonate, and lactate levels in clinical chemistry. It discusses specimen considerations for each analyte and lists the most common methods for determination. For chloride, blood, urine, and sweat can be used as specimens. Ion-selective electrodes and titration methods are described for analysis. Bicarbonate is the major buffer in blood and total CO2 is indicative of levels. Enzymatic and ISE methods are used for measurement. Lactate is a product of anaerobic metabolism and measurements are useful for critically ill patients. Specimens must be separated and analyzed quickly to avoid glycolysis. The enzymatic method uses lactate oxidase to produce a colored complex

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    Chloride, Bicarbonate and Lactate

    Uploaded by

    Ella Ortega
    35 views25 pages
    This document provides information on determining chloride, bicarbonate, and lactate levels in clinical chemistry. It discusses specimen considerations for each analyte and lists the most common methods for determination. For chloride, blood, urine, and sweat can be used as specimens. Ion-selective electrodes and titration methods are described for analysis. Bicarbonate is the major buffer in blood and total CO2 is indicative of levels. Enzymatic and ISE methods are used for measurement. Lactate is a product of anaerobic metabolism and measurements are useful for critically ill patients. Specimens must be separated and analyzed quickly to avoid glycolysis. The enzymatic method uses lactate oxidase to produce a colored complex

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    This document provides information on determining chloride, bicarbonate, and lactate levels in clinical chemistry. It discusses specimen considerations for each analyte and lists the most common methods for determination. For chloride, blood, urine, and sweat can be used as specimens. Ion-selective electrodes and titration methods are described for analysis. Bicarbonate is the major buffer in blood and total CO2 is indicative of levels. Enzymatic and ISE methods are used for measurement. Lactate is a product of anaerobic metabolism and measurements are useful for critically ill patients. Specimens must be separated and analyzed quickly to avoid glycolysis. The enzymatic method uses lactate oxidase to produce a colored complex

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    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    35 views25 pages

    Chloride, Bicarbonate and Lactate

    Uploaded by

    Ella Ortega
    This document provides information on determining chloride, bicarbonate, and lactate levels in clinical chemistry. It discusses specimen considerations for each analyte and lists the most common methods for determination. For chloride, blood, urine, and sweat can be used as specimens. Ion-selective electrodes and titration methods are described for analysis. Bicarbonate is the major buffer in blood and total CO2 is indicative of levels. Enzymatic and ISE methods are used for measurement. Lactate is a product of anaerobic metabolism and measurements are useful for critically ill patients. Specimens must be separated and analyzed quickly to avoid glycolysis. The enzymatic method uses lactate oxidase to produce a colored complex

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    of 25

    CLINICAL CHEMISTRY 2 LABORATORY

    CHLORIDE, BICARBONATE
    AND LACTATE
    PROF. ANGELA DELA PEÑA
    OBJECTIVES:
    By the end of this laboratory lecture the students are able to:

    Determine the different specimen considerations of


    chloride, bicarbonate and lactate.

    Identify the different methods to be used for its determination.

    Determine the reference range.


    CHLORIDE
    CHLORIDE
    - major extracellular anion.

    FUNCTIONS:
    Involved in maintaining osmolality,
    blood volume, and electric neutrality
    Chloride shifts secondarily to a
    movement of sodium or bicarbonate
    ions.
    DETERMINATION OF CHLORIDE

    SPECIMEN:
    • Blood
    • Urine
    • Sweat
    SPECIMEN
    CONSIDERATION

    BLOOD
    Lithium Heparin is the anticoagulant of choice.
    Serum or Plasma may be used.
    Hemolysis does not cause a significant change in serum
    or plasma values.
    With marked hemolysis, Chloride levels may be
    decreased as a result of dilutional effect.
    Whole blood samples may be used with some analyzers .
    SPECIMEN
    CONSIDERATION
    URINE
    • 24-HOUR URINE COLLECTION
    - The specimen of choice for urine chloride analyses.

    SWEAT
    - Analysis of chloride ion concentration in sweat is used to confirm the diagnosis
    of Cystic Fibrosis.
    - Chloride concentration in sweat is best determined by coulometric titration.
    METHODS:
    ION- SELECTIVE ELECTRODES (ISE)
    - most commonly used method.
    Principle: An ion-exchange membrane (tri-n-octylpropylammonium decanol) is used
    to selectively bind chloride ions.
    METHODS:

    AMPEROMETRIC-COULOMETRIC TITRATION
    - a method using coulometric generation of Silver ions which combine with
    chloride ions to quantitate Chloride concentrations.
    - Endpoint: The surge of free silver ions.
    - COTLOVE CHLORIDOMETER- instrument that uses this principle.
    METHODS:

    MERCURIMETRIC TITRATION ( SCHALES AND SCHALES)


    - PFF preparation: uses tungstic acid as precipitating agent of protein.
    - Principle: PFF is titrated with standard solution of mercuric ions (mercuric nitrate) to form
    a soluble compound of mercuric chloride which does not dissociate to mercuric
    ions.

    Excess mercuric + s -diphenylc a rba zone = Blue- violet color


    ions (indic ator dye)
    METHODS:

    MERCURIC THIOCYANATE (WHITEHORN TITRATION) METHOD


    - Specimen is mixed with a solution of mercuric thiocyanate.
    - Mercuric chloride – final product

    Free thiocyanate Ferric ions Reddish- brown


    + =
    (from Ferric nitrate) color
    REFERENCE RANGE:
    BICARBONATE
    BICARBONATE
    - S econd mos t abundant anion in the ECF
    Function: Major component of the
    buffering system in the blood.

    Total CO2 measurement is indicative of


    Bicarbonate measurement (Serum
    bicarbonate accounts for more than 90%
    of serum total CO2 at physiologic ph.)
    DETERMINATION OF TOTAL CO2
    Specimen
    • Venous serum or heparinized plasma
    • Anaerobic collection
    - If the sample is left uncapped before analysis,
    CO2 escapes (decrease by 6 mmol/L per hour)
    METHODS:

    ION- SELECTIVE ELECTRODES (ISE)


    - Uses an acid reagent to convert all the forms of CO2 to CO2 gas and
    is measured by a pCO2 electrode (Severinghaus Electrode).
    METHODS:
    ENZYMATIC METHOD
    - Alkalinizes the sample to convert all forms of CO2 to HCO3-
    LACTATE
    LACTATE
    - is a by product of an emergency mechanism that
    produces a small amount of ATP when oxygen delivery is
    severely diminished.

    Pyruvate is the normal end product of glucose


    metabolism. The conversion from pyruvate to
    lactate is activated when a deficiency of oxygen
    leads to an accumulation of excess NADH.

    Other function:
    A precursor for producing glucose.
    Clinical Applications
    Clinical Applications Measurements of blood lactate are useful
    for metabolic monitoring in critically ill patients, for indicating
    the severity of the illness, and for objectively determining
    patient prognosis.
    LACTATE
    SPECIMEN HANDLING
    • Tourniquet should not be used
    • After sample collection, anaerobic glycolysis will occur.
    • Heparinized blood must be delivered on ice and the plasma
    must be separated quickly.
    • Fluoride and iodoacetate inhibits glycolysis but the specific
    method directions must be consulted.
    LACTATE
    • Enzymatic Method
    Uses lactate oxidase to produce pyruvate and hydrogen peroxide.

    Addition of chromogen and catalyzed by peroxidase to form a colored complex.


    Reference Books:

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