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    Calibration of Micro - Pipette

    Uploaded by

    mdasifkhaleel
    This document provides instructions for calibrating micro-pipettes using the gravimetric method. It describes the types of micro-pipettes, requirements, apparatus needed including balances, recommended test volumes, calibration procedures for single and multi-channel pipettes, and how to convert mass measurements to volume using correction factors. The calibration procedure involves precisely measuring water volumes delivered from the pipette using an analytical balance and calculating the mean volume from multiple measurements.

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    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd

    Calibration of Micro - Pipette

    Uploaded by

    mdasifkhaleel
    287 views17 pages
    This document provides instructions for calibrating micro-pipettes using the gravimetric method. It describes the types of micro-pipettes, requirements, apparatus needed including balances, recommended test volumes, calibration procedures for single and multi-channel pipettes, and how to convert mass measurements to volume using correction factors. The calibration procedure involves precisely measuring water volumes delivered from the pipette using an analytical balance and calculating the mean volume from multiple measurements.

    Original Description:

    Pipette Clinic calibration of Micro Pipette

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    © © All Rights Reserved

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    This document provides instructions for calibrating micro-pipettes using the gravimetric method. It describes the types of micro-pipettes, requirements, apparatus needed including balances, recommended test volumes, calibration procedures for single and multi-channel pipettes, and how to convert mass measurements to volume using correction factors. The calibration procedure involves precisely measuring water volumes delivered from the pipette using an analytical balance and calculating the mean volume from multiple measurements.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    287 views17 pages

    Calibration of Micro - Pipette

    Uploaded by

    mdasifkhaleel
    This document provides instructions for calibrating micro-pipettes using the gravimetric method. It describes the types of micro-pipettes, requirements, apparatus needed including balances, recommended test volumes, calibration procedures for single and multi-channel pipettes, and how to convert mass measurements to volume using correction factors. The calibration procedure involves precisely measuring water volumes delivered from the pipette using an analytical balance and calculating the mean volume from multiple measurements.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    of 17

    Bangladesh BEST

    Programme

    Calibration of Micro-pipette
    By
    Gravimetric Method

    Nihal Gunasekara
    Sri Lanka

    Bangladesh BEST
    Programme

    Types of Micro-pipettes
    Single Channel
    Multi- Channel

    Bangladesh BEST
    Programme

    Requirements

    Unit of volume
    l
    Ref. temperature
    To deliver
    calibrated to
    transfer the stated
    another container.

    Cubic millimeter :
    20 C
    Apparatus is
    accurately
    volume to

    Bangladesh BEST
    Programme

    Apparatus Needed
    Balance
    Thermometer
    Hygrometer
    Barometer

    Distilled or deionized water


    Weighing vessel

    Bangladesh BEST
    Programme

    Recommended Apparatus Balance


    Selected Volume
    (V)

    1 l V

    Resolution Std.
    of
    Deviation
    Balance
    (mg)
    (mg)

    10 l

    0.002
    0.001

    10 l < V

    100

    0.01
    0.02

    100 l < V

    1000

    0.1
    0.2

    1 ml < V

    10 ml

    0.1
    0.2

    Bangladeshi BEST
    Programme

    Recommended
    Apparatus/Liquids

    Apparatus/Liqui
    d
    Thermometer

    0.2 C

    Hygrometer

    10%

    Barometer

    0.5 kPa

    Calibration
    Liquid

    Uncertainty

    Distilled /de-ionized
    water

    Bangladesh BEST
    Programme

    Conditions for the Test


    Temperature
    Maintained at 20 0.5 C and
    M
    during
    calibration 0. 2 C stability
    Humidity
    Maintained > 50 %
    Prefer to maintain at
    70 10 %
    Stabilization
    Volumetric instruments and
    test water
    1 to 2 hours
    Evaporation
    Special precautions below 50 l
    Test cycle

    Less than 60 s

    Bangladesh BEST
    Programme

    Test Volume

    Fixed Volume M
    Variable Volume
    volumes
    Preferably
    nominal volume
    Measurements

    Nominal Volume
    At least three

    10%, 50 % and

    10 at each volume

    Bangladesh BEST
    Programme

    Calibration Procedure- Single


    Channel
    Place the test liquid in the weighing
    vessel to a depth of at least up to
    3mm and record the temperature,
    barometric pressure & humidity
    Fit the selected disposable tip to the
    micro pipette barrel
    Fill the tip with test liquid and expel to
    waste five time to reach a humidity
    equilibrium in the dead air volume
    Place the weighing vessel with its added
    water on the balance pan

    Bangladesh BEST
    Programme

    Calibration Procedure- Single


    Channel
    Prior to any calibration test make sure
    that all instruments reached the
    ambient temperature in the laboratory
    During the calibration maximum
    temperature variation in the
    laboratory should not exceed 0.2 oC.
    Be sure that all the apparatus use for
    calibration and the distilled water are
    at room temperature.

    Bangladesh BEST
    Programme

    Calibration Procedure
    Replace the disposable tip of the piston pipette
    Fill the piston pipette with test liquid, immersing its
    delivery orifice 2 to 3 mm below the surface of water
    Release the operating button slowly and draw the
    pipette vertically against the side wall of container .
    Expel the water to waste to pre-wet the tip and refill
    the piston pipette as above.
    Record the mass m0 of the weighing vessel or tare the
    balance to zero
    Deliver the contents of the pipette into the weighing
    vessel by touching pipette tip against the container
    wall just above liquid surface at an angle 30 to 45
    Record the mass m1 of the weighing vessel and
    determine the quantity delivered
    Repeat the above cycle for ten measurements

    Bangladesh BEST
    Programme

    Calibration Procedure- MultiChannel


    Procedure is similar to single
    channel micro pipette
    During the calibration each channel
    should be considered as a single
    channel

    Bangladesh BEST
    Programme

    Conversion of Mass into Volume

    vi mi Z
    The Z correction factor is a function
    of test temperature and air
    pressure

    13

    Bangladesh BEST
    Programme
    Correction
    Factors for Distilled

    Water

    (Z values in micro-litres per milligram )


    Temp.
    Air Pressure kPa
    C

    80

    85

    90

    95

    100

    101.3

    105

    17
    17.5

    1.0021 1.0021 1.0022 1.0022 1.0023 1.0023 1.0023


    1.0022 1.0022 1.0023 1.0023 1.0024 1.0024 1.0024

    18.0
    18.5

    1.0022 1.0023 1.0023 1.0024 1.0025 1.0025 1.0025


    1.0023 1.0024 1.0024 1.0025 1.0025 1.0026 1.0026

    19.0
    19.5

    1.0024 1.0025 1.0025 1.0026 1.0026 1.0027 1.0027


    1.0025 1.0026 1.0026 1.0027 1.0027 1.0028 1.0028

    20.0
    20.5

    1.0026 1.0027 1.0027 1.0028 1.0028 1.0029 1.0029


    1.0027 1.0028 1.0028 1.0029 1.0029 1.0030 1.0030

    21.0
    21.5

    1.0028 1.0029 1.0029 1.0030 1.0031 1.0031 1.0031


    1.0030 1,0030 1.0031 1.0031 1.0032 1.0032 1.0032

    22.0
    22.5

    1.0031 1.0031 1.0032 1.0032 1.0033 1.0033 1.0033


    1.0032 1.0032 1.0033 1.0033 1.0034 1.0034 1.0034

    Bangladesh BEST
    Programme

    Mean Volume
    1 10
    V Vi
    10 i 1

    15

    Bangladesh BEST
    Programme

    Data Recording
    Use spread sheet for data recording
    Record Temperature, Humidity,
    Barometric pressure
    Record final data in a data recording
    sheet
    Review from a third party
    Prepare the final calibration
    certificate

    Bangladesh BEST
    Programme

    Thank you

    17

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