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    Total Protein Spinreact

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    ariyan khan
    The Biuret method is a colorimetric technique for determining total protein concentration in plasma or serum. It works by forming a blue complex between protein in an alkaline solution and copper salts. The intensity of the blue color is proportional to the protein level. The method is linear up to 15g/dl and involves mixing the sample with Biuret reagent and measuring absorbance after 15 minutes to calculate the protein concentration compared to a standard. Normal total protein levels in serum are 6.7-8.7 g/dl for adults and 5.2-9.1 g/dl for newborns.

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    Total Protein Spinreact

    Uploaded by

    ariyan khan
    288 views1 page
    The Biuret method is a colorimetric technique for determining total protein concentration in plasma or serum. It works by forming a blue complex between protein in an alkaline solution and copper salts. The intensity of the blue color is proportional to the protein level. The method is linear up to 15g/dl and involves mixing the sample with Biuret reagent and measuring absorbance after 15 minutes to calculate the protein concentration compared to a standard. Normal total protein levels in serum are 6.7-8.7 g/dl for adults and 5.2-9.1 g/dl for newborns.

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    this is my important document. my dmlt 2nd year practical

    Original Title

    TOTAL_PROTEIN_SPINREACT

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    The Biuret method is a colorimetric technique for determining total protein concentration in plasma or serum. It works by forming a blue complex between protein in an alkaline solution and copper salts. The intensity of the blue color is proportional to the protein level. The method is linear up to 15g/dl and involves mixing the sample with Biuret reagent and measuring absorbance after 15 minutes to calculate the protein concentration compared to a standard. Normal total protein levels in serum are 6.7-8.7 g/dl for adults and 5.2-9.1 g/dl for newborns.

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    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    288 views1 page

    Total Protein Spinreact

    Uploaded by

    ariyan khan
    The Biuret method is a colorimetric technique for determining total protein concentration in plasma or serum. It works by forming a blue complex between protein in an alkaline solution and copper salts. The intensity of the blue color is proportional to the protein level. The method is linear up to 15g/dl and involves mixing the sample with Biuret reagent and measuring absorbance after 15 minutes to calculate the protein concentration compared to a standard. Normal total protein levels in serum are 6.7-8.7 g/dl for adults and 5.2-9.1 g/dl for newborns.

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    © All Rights Reserved
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    of 1

    TOTAL PROTEIN

    BIURET METHOD

    Principle: Linearity:
    Colorimetric determination of the total protein based on The method is linear up to 15g/dl or 150g/l.
    the principle of Biuret reaction (copper salts in an alkaline If the protein concentration is greater than 15 g/dl in the
    medium). Protein in plasma or serum forms a blue serum or plasma, dilute sample 1:2 with saline solution
    coloured complex when treated with cupric icons in and repeat test. Multiply result by 2.
    alkaline solution. The intensity of the blue colour is
    proportional to the protein concentration.
    Normal Values:
    Reagent Concentration:
    Biuret Reagent:
    Sodium iodide 100 mmol/l Serum, Adults: 6.7-8.7 g/dl (67-87 g/l)
    Potassium sodium tartrate 15 mmol/l Newborn: 5.2-9.1 g/dl (52-91g/l)
    Copper sulphate 5 mmol/l Children: 5.4-8.7 g/dl (54-87 g/l)
    Potassium Iodide 15 mmol/l
    Notes:
    Standard:
    The sample blank for clear, colourless sera is equivalent to
    Albumin Bovine Fraktion V 7 g/dl
    0.2 g/dl and is therefore negligible. A sample blank must
    be determined for haemolytic and liperimic sera by
    Preparation and Stability pipetting 20µl serum to 1000µl physiological saline and
    Reagent and Standard are ready to use. measurement against distilled water. The absorbance of
    The reagents are stable up to the stated expiry date when the sample blank has to be subtracted from the absorbance
    stored at +2 to +8 ºC. of the sample.
    Patients who have received intravenous amounts of
    Samples: polydextrans yield values which are too high to be
    measured by the Biuret method. In such cases the protein
    Serum, heparinised or EDTA-plasma.
    concentration should be measured using the Kjeldal
    Stability in serum at +2 to +8 ºC is up to 1 month and at
    method.
    +15 to +25ºC is up to 1 week.

    Procedure: Quality Control:


    Wavelength: Hg 540nm (530-570nm) For accuracy and reproducibility control:-
    Assayed Multi-Sera Normal and Elevated.
    Temperature: +25ºC
    For reproducibility control:-
    Cuvette: 1cm light path
    Multi-Sera Loaw, Normal and Elevated.
    Zero adjustment: Reagent blank
    Each series needs one reagent
    blank only Presentation:
    TOP0350 2 x 125ml, 250 tests
    Blank Standard Sample Biuret Reagent 2x125ml
    Reagent/R1 1000µl 1000µl 1000µl Standard 1x5ml
    Standard/R4 --- 25µl ---
    Serum or Plasma --- --- 25µl Safety precautions:
    For in vitro diagnostic use only. Do not pipette by mouth.
    Mix. Incubate for 15 minutes at 30-37C. Wait for 5 Exercise the normal precautions required for handling
    minutes at room temperature. Measure absorbance of laboratory reagents.
    sample and standard against reagent blank.
    Health and Safety data sheets are available on request.

    Literature:
    Calculation: Weichselbaum, T. E., Amer. Clin. Path., 16, 40 – 48
    By Standard: (1946)
    ? A sample Josephson, B. and Gyllenswärd, A. Scand. J. Clin. Lab.
    -------------- x standard conc (g/dl. = Protein in g/dl Invest., 9, 29 (1975).
    ? A standard Henry R.J et al. Anal. Chem.92 1491 (1957)
    Peters T.J. Clin. Chem. 14 1147 (1968)
    Standard concentration: 7g/dl

    TOP0350.V1
    09/12/02

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