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    Requerimiento de Glucosa

    Uploaded by

    Demian Legin
    This document provides instructions for quantitatively determining glucose concentration in serum or plasma samples using a liquid reagent and spectrophotometric method. Glucose is oxidized by glucose oxidase to produce hydrogen peroxide, which is detected using phenol and 4-aminophenazone in the presence of peroxidase to produce a colored compound. The intensity of color is directly proportional to the glucose concentration and is measured at 505nm. Normal glucose values are provided as well as storage conditions, potential interferences, and performance characteristics of the test.

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    © All Rights Reserved
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    Requerimiento de Glucosa

    Uploaded by

    Demian Legin
    57 views2 pages
    This document provides instructions for quantitatively determining glucose concentration in serum or plasma samples using a liquid reagent and spectrophotometric method. Glucose is oxidized by glucose oxidase to produce hydrogen peroxide, which is detected using phenol and 4-aminophenazone in the presence of peroxidase to produce a colored compound. The intensity of color is directly proportional to the glucose concentration and is measured at 505nm. Normal glucose values are provided as well as storage conditions, potential interferences, and performance characteristics of the test.

    Original Description:

    Practica de glucosa en sangre

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    This document provides instructions for quantitatively determining glucose concentration in serum or plasma samples using a liquid reagent and spectrophotometric method. Glucose is oxidized by glucose oxidase to produce hydrogen peroxide, which is detected using phenol and 4-aminophenazone in the presence of peroxidase to produce a colored compound. The intensity of color is directly proportional to the glucose concentration and is measured at 505nm. Normal glucose values are provided as well as storage conditions, potential interferences, and performance characteristics of the test.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    57 views2 pages

    Requerimiento de Glucosa

    Uploaded by

    Demian Legin
    This document provides instructions for quantitatively determining glucose concentration in serum or plasma samples using a liquid reagent and spectrophotometric method. Glucose is oxidized by glucose oxidase to produce hydrogen peroxide, which is detected using phenol and 4-aminophenazone in the presence of peroxidase to produce a colored compound. The intensity of color is directly proportional to the glucose concentration and is measured at 505nm. Normal glucose values are provided as well as storage conditions, potential interferences, and performance characteristics of the test.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 2

    GLUCOSE -LQ

    Glucose
    GOD-POD. Liquid
    Quantitative determination of glucose
    IVD

    CALCULATIONS
    ( A ) Sample
    x 100 (Standard conc.) = mg/dL glucose in the sample
    ( A ) S tan dard

    Store at 2-8C
    PRINCIPLE OF THE METHOD
    Glucose oxidase (GOD) catalyses the oxidation of glucose to gluconic
    acid. The formed hydrogen peroxide (H 2O2), is detected by a
    chromogenic oxygen acceptor, phenol, 4 aminophenazone (4-AP)
    in the presence of peroxidase (POD):
    -D-Glucose + O2 + H2O
    H2O2 + Phenol + 4-AP

    GOD
    POD

    Gluconic acid + H2O2


    Quinone + H2O

    The intensity of the color formed is proportional to the glucose


    1,2
    concentration in the sample .

    CLINICAL SIGNIFICANCE
    Glucose is a major source of energy for most cells of the body;
    insulin facilitates glucose entry into the cells.
    Diabetes is a disease manifested by hyperglycemia; patients with
    diabetes demonstrate an inability to produce insulin1,5,6.
    Clinical diagnosis should not be made on a single test result; it
    should integrate clinical and other laboratory data.
    REAGENTS
    TRIS pH 7.4
    Phenol
    Glucose oxidase (GOD)
    Peroxidase (POD)
    4 Aminophenazone (4-AP)

    92 mmol/L
    0.3 mmol/L
    15000 U/L
    1000 U/L
    2.6 mmol/L

    GLUCOSE CAL Glucose aqueous primary standard 100 mg/dL


    PREPARATION
    Reagent and calibrator provided are ready to use.
    STORAGE AND STABILITY
    All the components of the kit are stable until the expiration date on
    the label when stored tightly closed at 2-8C, protected from light
    and contaminations prevented during their use.
    Do not use reagents over the expiration date.
    SIGNS OF REAGENT DETERIORATION:
    - Presence of particles and turbidity.
    - Blank absorbance (A) at 505 nm
    0.32.
    ADDITIONAL EQUIPMENT
    - Spectrophotometer or colorimeter measuring at 505 nm.
    - Matched cuvettes 1.0 cm light path.
    - General laboratory equipment.
    SAMPLES
    Serum or plasma, free of hemolysis 1:
    Serum should be removed from the clot as quickly as possible.
    Stability of the sample: Glucose in serum or plasma is stable at 28 for 3 days.
    PROCEDURE
    1. Assay conditions:
    Wavelength: . . . . . . . . . . . . . .. . . . . 505 nm (490-550)
    Cuvette: . . . . . . . . . . . . . . . . . . . . .. . . . 1 cm light path
    Temperature. . . . . . . . . . . . . . . . . . . . .. 37C / 15-25C
    2. Adjust the instrument to zero with distilled water.
    3. Pipette into a cuvette:
    Standard
    Blank
    Sample
    R (mL)
    1.0
    1.0
    1.0
    (Note 1,2)
    -10
    -Standard
    ( L)
    --10
    Sample ( L)
    4.
    5.

    Mix and incubate for 10 min at 37C or 20 min at room


    temperature (15-25C).
    Read the absorbance (A) of the samples and standard,
    against the Blank. The colour is stable for at least 30
    minutes.

    BSIS46-I

    20/01/12

    Conversion factor: mg/dL x 0.0555= mmol/L.


    QUALITY CONTROL
    Control sera are recommended to monitor the performance of assay
    procedures: SPINTROL H Normal and Pathologic (Ref. 1002120 and
    1002210).
    If control values are found outside the defined range, check the
    instrument, reagent and calibration for problems.
    Each laboratory should establish its own Quality Control scheme and
    corrective actions if controls do not meet the acceptable tolerances.
    REFERENCE VALUES1
    Serum or plasma:
    60 110 mg/dL
    3.33 6.10 mmol/L
    These values are for orientation purpose; each laboratory should
    establish its own reference range.
    PERFORMANCE CHARACTERISTICS
    Measuring range: From detection limit 1 mg/dL to linearity limit 500
    g/dL.
    If the concentration is greater than linearity limit dilute 1/2 the sample
    with ClNa 9 g/L and multiply the result by 2.
    Precision:
    Intra-assay (n=20)
    Inter-assay (n=20)
    Mean (mg/dL)
    94.9
    238
    98.6
    246
    SD
    1.99
    4.11
    3.04
    5.00
    CV (%)
    2.10
    1.73
    3.09
    2.03
    Sensitivity: 1 mg/dL = 0.0035 (A).
    Accuracy: Results obtained using SPINREACT reagents (y) did not
    show systematic differences when compared with other commercial
    reagent (x).
    The results obtained using 50 samples were the following:
    Correlation coefficient (r): 0.9929.
    Regression equation: y= 0.9901x + 1.0515.

    The results of the performance characteristics depend on the analyzer


    used.
    INTERFERENCES
    1

    Hemoglobin up to 19 g/L and bilirubin up to 100 mg/L, do not interfere .


    A list of drugs and other interfering substances with glucose determination
    3,4
    has been reported by Young et. al .
    NOTES
    1. GLUCOSE CAL: Proceed carefully with this product because due its
    nature it can get contamined easily.
    2. Calibration with the aqueous standard may cause a systematic error in
    automatic procedures. In these cases, it is recommended to use a
    serum Calibrator.
    3. Use clean disposable pipette tips for its dispensation.
    4. SPINREACT has instruction sheets for several automatic
    analyzers. Instructions for many of them are available on request.

    BIBLIOGRAPHY
    1.
    2.
    3.
    4.
    5.
    6.

    Kaplan L.A. Glucose. Kaplan A et al. Clin Chem The C.V. Mosby Co. St Louis.
    Toronto. Princeton 1984; 1032-1036.
    Trinder P. Ann Clin Biochem 1969; 6 24-33.
    Young DS. Effects of drugs on Clinical Lab. Tests, 4th ed AACC Press, 1995.
    Young DS. Effects of disease on Clinical Lab. Tests, 4th ed AACC 2001.
    Burtis A et al. Tietz Textbook of Clinical Chemistry, 3rd ed AACC 1999.
    Tietz N W et al. Clinical Guide to Laboratory Tests, 3rd ed AACC 1995.

    PACKAGING
    R: 2 x 50 mL, CAL: 1 x 2 mL

    Ref: 41010

    R: 2 x 100 mL, CAL: 1 x 2 mL

    Ref. 41012
    Ref. 41011
    Ref. 41013

    Cont.
    .

    R: 2 x 250 mL, CAL: 1 x 5 mL


    R: 1 x 1000 mL, CAL: 1 x 5 mL

    SPINREACT,S.A./S.A.U Ctra.Santa Coloma, 7 E-17176 SANT ESTEVE DE BAS (GI) SPAIN


    Tel. +34 972 69 08 00 Fax +34 972 69 00 99. e-mail: spinreact@spinreact.com

    GLUCOSE -LQ

    Glucosa
    GOD-POD. Lquido
    Determinacin cuantitativa de glucosa
    CLCULOS

    IVD

    ( A ) Muestra
    x 100 (Conc. Patrn) = mg/dL de glucosa en la muestra
    ( A ) Patrn

    Conservar a 2-8C
    PRINCIPIO DEL MTODO
    La glucosa oxidasa (GOD) cataliza la oxidacin de glucosa a
    cido glucnico. El perxido de hidrgeno (H2 O2) producido se
    detecta mediante un aceptor cromognico de oxigeno, fenol, 4
    aminofenazona (4-AF), en presencia de la peroxidasa (POD):
    -D-Glucosa + O2 + H2O

    GOD

    cido glucnico + H2O2

    POD

    H2O2 + Fenol + 4-AF


    Quinona + H2 O
    La intensidad del color formado es proporcional a la
    concentracin de glucosa presente en la muestra ensayada1,2.
    SIGNIFICADO CLNICO
    La glucosa es la mayor fuente de energa para las clulas del
    organismo; la insulina facilita la entrada de glucosa en las clulas.
    La diabetes mellitus es una enfermedad que se manifiesta por
    una hiperglucmia, causada por un dficit de insulina1,5,6.
    El diagnostico clnico debe realizarse teniendo en cuenta todos
    los datos clnicos y de laboratorio.
    REACTIVOS

    GLUCOSE CAL

    TRIS pH 7,4
    92 mmol/L
    Fenol
    0,3 mmol/L
    Glucosa oxidasa (GOD)
    15000 U/L
    Peroxidasa (POD)
    1000 U/L
    4 - Aminofenazona (4-AF)
    2,6 mmol/L
    Patrn primario acuoso de Glucosa 100 mg/dL

    PREPARACIN
    El reactivo y el calibrador estn listos para su uso.
    CONSERVACIN Y ESTABILIDAD
    Todos los componentes del kit son estables, hasta la fecha de
    caducidad indicada en la etiqueta del vial, cuando se mantienen
    los viales bien cerrados a 2-8C, protegidos de la luz y se evita la
    contaminacin durante su uso.
    No usar reactivos fuera de la fecha indicada.
    Indicadores de deterioro de los reactivos:
    - Presencia de partculas y turbidez.
    - Absorbancias (A) del Blanco a 505 nm 0,32.
    MATERIAL ADICIONAL
    - Espectrofotmetro o analizador para lecturas a 505 nm.
    - Cubetas de 1,0 cm de paso de luz.
    - Equipamiento habitual de laboratorio.

    Factor de conversin: mg/dL x 0,0555= mmol/L.


    CONTROL DE CALIDAD
    Es conveniente analizar junto con las muestras sueros control valorados:
    SPINTROL H Normal y Patolgico (Ref. 1002120 y 1002210)
    Si los valores hallados se encuentran fuera del rango de tolerancia, se debe
    revisar los instrumentos, los reactivos y la calibracin.
    Cada laboratorio debe disponer su propio Control de Calidad y establecer
    correcciones en el caso de que los controles no cumplan con las
    tolerancias.

    VALORES DE REFERENCIA1
    Suero o plasma:
    60 110 mg/dL
    3,33 6,10 mmol/L
    Estos valores son orientativos. Es recomendable que cada laboratorio
    establezca sus propios valores de referencia.
    CARACTERSTICAS DEL MTODO
    Rango de medida: Desde el limite de deteccin 1 mg/dL hasta el limite de
    linealidad 500 mg/dL.
    Si la concentracin de la muestra es superior al limite de linealidad, diluir
    1/2 con ClNa 9 g/L y multiplicar el resultado final por 2.
    Precisin:
    Intraserie (n=20)
    Interserie (n=20)
    Media (mg/dL)
    94,9
    238
    98,6
    246
    SD
    1,99
    4,11
    3,04
    5,00
    CV (%)
    2,10
    1,73
    3,09
    2,03
    Sensibilidad analtica: 1 mg/dL = 0,0035 (A).
    Exactitud: Los reactivos SPINREACT (y) no muestran diferencias
    sistemticas significativas cuando se comparan con otros reactivos
    comerciales (x).
    Los resultados obtenidos con 50 muestras fueron los siguientes:
    Coeficiente de regresin (r): 0,9929.
    Ecuacin de la recta de regresin: y= 0,9901x + 1,0515.
    Las caractersticas del mtodo pueden variar segn el analizador utilizado.

    INTERFERENCIAS
    No se han observado interferencias con hemoglobina hasta 19 g/L y
    bilirrubina hasta 100 mg/L1.
    Se han descrito varias drogas y otras substancias que interfieren en la
    determinacin de la glucosa3,4.

    MUESTRAS
    Suero o plasma, libre de hemlisis1.
    El suero debe separarse lo antes posible del cogulo.
    Estabilidad de la muestra: La glucosa en suero o plasma es
    estable 3 das a 2-8C.

    NOTAS
    1. GLUCOSE CAL: Debido a la naturaleza del producto, es aconsejable
    tratarlo con sumo cuidado ya que se puede contaminar con facilidad.
    2. La calibracin con el Patrn acuoso puede dar lugar a errores
    sistemticos en mtodos automticos. En este caso, se recomienda
    utilizar calibradores sricos.
    3. Usar puntas de pipeta desechables limpias para su dispensacin.
    4. SPINREACT dispone de instrucciones detalladas para la
    aplicacin de este reactivo en distintos analizadores.

    PROCEDIMIENTO

    BIBLIOGRAFA

    1.

    Condiciones del ensayo:


    Longitud de onda: . . . . . . . . . . . . . . . 505 nm (490-550)
    Cubeta:. . . . . . . . . . . . . . . . . . . . . . . .. .1 cm paso de luz
    Temperatura. . . . . . . . . . . . . . . . . . . . . . . 37C / 15-25C
    2. Ajustar el espectrofotmetro a cero frente a agua destilada.
    3. Pipetear en una cubeta:
    Blanco
    Patrn
    Muestra
    R (mL)
    1,0
    1,0
    1,0
    -10
    -Patrn (Nota1,2) ( L)
    --10
    Muestra ( L)

    1.
    2.
    3.
    4.
    5.
    6.

    Kaplan L.A. Glucose. Kaplan A et al. Clin Chem The C.V. Mosby Co. St Louis.
    Toronto. Princeton 1984; 1032-1036.
    Trinder P. Ann Clin Biochem 1969; 6: 24-33.
    Young DS. Effects of drugs on Clinical Lab. Tests, 4th ed AACC Press, 1995.
    Young DS. Effects of disease on Clinical Lab. Tests, 4th ed AACC 2001.
    Burtis A et al. Tietz Textbook of Clinical Chemistry, 3rd ed AACC 1999.
    Tietz N W et al. Clinical Guide to Laboratory Tests, 3rd ed AACC 1995.

    PRESENTACIN

    5.

    Mezclar e incubar 10 minutos a 37C 20 min a


    temperatura ambiente (15-25C).
    Leer la absorbancia (A) del patrn y la muestra, frente al
    Blanco de reactivo. El color es estable como mnimo 30
    minutos.

    BSIS46-E

    20/01/12

    R: 2 x 100 mL, CAL: 1 x 2 mL

    Ref. 41012
    Ref. 41011

    4.

    R: 2 x 50 mL, CAL: 1 x 2 mL

    Ref: 41010

    Ref. 41013

    Cont.
    .

    R: 2 x 250 mL, CAL: 1 x 5 mL


    R: 1 x 1000 mL, CAL: 1 x 5 mL

    SPINREACT,S.A./S.A.U Ctra.Santa Coloma, 7 E-17176 SANT ESTEVE DE BAS (GI) SPAIN


    Tel. +34 972 69 08 00 Fax +34 972 69 00 99. e-mail: spinreact@spinreact.com

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