WVR TSL Pipetting and Calibration En

42
Pipetting & Calibration Norway – June 2008

description

pipette

Transcript of WVR TSL Pipetting and Calibration En

  • Pipetting & CalibrationNorway June 2008

  • 2Pipettors calibration & Service

  • History Engaged in metrology and calibration

    2th company in the world with ISO 17025-accreditation

    Over 10 years of know-how

    75.000 pipette-calibrations a year.

    Customer focused Provide choice

    Onsite

    ISO 8655 or customer driven

    Specifications

    Quality focused

    ISO 17025 accreditated

    SOPs based

    Known uncertainty budget

    Skilled technicians

    Pipettors calibration & Service

  • ISO 17025 - accreditation

  • Multilateral agreement

    Country Body Country Body

    Austria BMWA Lithuania LA

    Germany DKD Netherlands RvA

    Czech Republic CAI Norway NA

    Denmark DANAK Portugal IPQ

    Estonia EAK Romania RENAR

    Finland FINAS Slovakia SNAS

    France COFRAC Slovenia SA

    Belgium BKO Greece ESYD

    Spain ENAC Latvia LATAK

    Sweden SWEDAC Ireland INAB

    Switzerland SAS Italy SINAL

    United Kingdom UKAS Standard for all countries is ISO 17025

  • Customer made calibration / ISO 8655; Gravimetric method; Complete programme;

    As found calibration; Maintenance; Calibration; Adjustment; Repair

    Multichannels, measurement of all channels; Qualified technicians; Validated software; ISO 17025 calibration-certificates; Warranties responsibility; Due time reminder system.

    Pipettors calibration & Service

  • Pipettes performed 2007:

    59.812 pieces;

    Coverage: Europe

    Manufacturer: 113x

    Problem solved: 98,2%

    Rejection: 1.913x

    Turnaroundtime: 3,2 days

    Brand deviation

    40%

    18%16%

    7%

    6%1%

    3% 2%7%

    GilsonFinnpipetteEppendorfBiohitSocorexBrandRaininVWROthers

    Pipette-Calibration at Competence Center

  • 867 Headcounts Technicians: 48 Quality: 4 Dispatch / administr: 7 Logistic: 2 Sales: 2 HR: 1 Financial: 3

    Pipette Calibration-facilities VWR

    = VWR technicians

    = VWR partner

    = ISO 17025 accredited calibration-lab

  • Pipetting??

    Every lab-technician can do

    that!!!

  • - Principle pipettors

    - Liquid Handling group

    - Pipetting technics

    - Terminology

    - Calibration

    - Wrong Pipetting

    - Maintenance & Repair

    - Measure uncertainty

  • Accuracy and Precision

    Precision

    Precision is an agreement between replicate

    measurements. Precision is quantified by imprecision

    High precision, i.e. small imprecision means very little

    variation between repeated measurements

    Accuracy

    It is possible to be very consistent, but

    consistently wrong

    Inaccuracy is the numerical difference between the

    mean of a set of replicate measurements and the true

    value

  • Accuracy and Precision

    Accurate Precise Accurate Not PrecisePrecise Not Accurate Not Precise Not Accurate

  • Liquid Handling

    Glassware

    Pipettors, mechanical / electronical

    Bottle top dispenser

    Burette

    Automatic Transfer Pipette station

  • Basics of a pipettor

    Vertical movement Diameter

    Length of the piston-movement

  • Pipettor types

    Monochannel

    Multichannel

    Stepper

    Multistepper

  • Pipettor working principles

    Air displacement

    Air between liquid and piston

    Length and diameter of piston define the amount

    Watery liquids

    Sensitive for contamination

    Worse accuracy

    Cheap system

  • Pipettor working principles

    Positive displacement

    Contact between liquid and piston

    Length, diameter and tip of piston makes the amount

    Viscosity liquids

    Volatile liquids

    Non-Sensitive for contamination

    Good accuracy

    Expensive system

  • Monochannels, fix and variable volume Air- and positive displacement system

    Pipettor types

    Multichannels, fix and variable volume Air- displacement system

    Stepper, variable volume Air- and positive displacement system

    Multistepper, variable volume Air- displacement system

  • Forward technique

    Pipetting technique

    Reverse technique

    Whole blood technique

    Repetition technique

  • Forward Technique

    - Most common technique

    - Only for watery liquids

    - Manufacturer adjustable

    - VWR / TSL common technique

    Pipetting technique

    The worst pipetting technique

  • Reverse Technique

    - Best technique for viscosity liquids

    - Best technique for volatile liquids

    - Best technique for small volumes

    - Only with air displacement pipettor

    Pipetting technique

    Best technique for lab-technicians who cant pipetting

  • Reverse technique

    97.598

    98.599

    99.5100

    100.5101

    101.5

    1 2 3 4 5 6 7 8 9 10

    Subsequent pipettings

    l

    P rP d (4x25ul)

    Pipetting Viscous Liquids

    (Glycerol)

  • Whole blood Technique

    - Accuracy for pipetting whole blood

    - Best technique for measurement of or in whole

    blood (erythrocytes)

    Pipetting technique

  • Terminology

    - Calibration- Specifications

    - Adjustment

    - Validation

    - Traceability

    - Nominal volume

    - Dead volume

  • Calibration methods

    - Gravimetrical

    - Titrimetrical

    - Photometrical

    - Radio-active

  • Traceability chain

    Primary standard

    National standard

    Reference standard

    Working standard

    Pipettor

  • Gravimetric method

    Given a certain mass of water with a known specific gravity, its volume

    can then be predicted.

    - Only official, described method- Most common calibration method

    - Method is several times accreditated

    - Uncertainty is known

    - Expensive method

    - Time consumable

  • ISO 8655 gravimetric method

    Given a certain mass of water with a known specific gravity, its volume

    can then be predicted.

    Test room: Stable environment. Constant 0,5C and humidity > 50%

    Evaporation: < 50l precautions

    Test volumes: Nominal volume / 50% of nominal volume / the lower limit of the

    volume range or 10% of the nominal volume (whichever is the greater)

    Number of measurements: 10 per test volume

    Test liquid: Deionized water grade 3 as in ISO 3696, degassed

    During calibration: measurement of aquadest with 0,2 C accuracy

    Multichannel: all channels!!!!!

  • ISO 8655 gravimetric method

    Given a certain mass of water with a known specific gravity, its volume

    can then be predicted.

    Weighing equipment:

    1 l < V < 10 l: Microscale / resolution 0,001 mg.

    10 l < V < 100 l: Semi-microscale / resolution 0,01 mg.

    100 l < V < 1.000 l: Analytical scale / resolution 0,1 mg.

    1 ml < V < 10 ml: Analytical scale / resolution 0,1 mg.

    10 ml < V < 200 ml: Scale / resolution 1 mg.

  • ISO 8655 gravimetric method

    Given a certain mass of water with a known specific gravity, its volume

    can then be predicted.

    Weighing vessel: Never plastics!!!!

    Thermometer: Uncertainty of < 0,2 C

    Hygrometer: Uncertainty of < 10%

    Barometer: Uncertainty of < 0,5 kPa

    Certified weights!!!!

    Most important: Technicians who are trained in pipetting

  • ISO 8655 procedure

    Preparation

    As found-calibration + certificate

    Preventive maintenance

    Calibration

    Meet specifications?

    Adjustment / Repair

    As left-calibration + certificate

  • Pipette service

    Preventative Maintenance:

    Disassemble pipette;Inspect and clean exterior and interior parts;Clean Piston & tipholder & seals;Check Spring tension;Grease/oil;Reassemble Pipette.

    Functional testLeakage test

  • Pipette service

    Calibration

    Check temperature and humidity;Pre-wet tips by pipetting 4 times;Calibrate pipette by 10 measures on lowest and highest volume;Pipette-technique is Forward Technique;Meeting specifications for accuracy and precision;Check temperature;Adjustment or repair if necessary;Recalibration;ISO 17025 - Calibration certificateCalibration sticker with Cal. Date and recal.-date

  • Specifications

    - Manufacturer

    - Customer; validation!

    - ISO 8655 (DIN 12650)

    You are responsible, not the manufacturer or auditor!

    Depends on the asked accuracy in the analytical test

  • Calibration certificate

  • Calibration

    Stable testroom

    Scales

    Instruments

    Traceability

    Acclimatisation

    Trained technicians

    Small volume !!!

  • Temperature

    Biohit Proline 200 - 1000 l

    900

    920

    940

    960

    980

    1000

    1020

    1040

    5.2 22.2 28.1

    Water temperature (C)

    T

    e

    s

    t

    v

    o

    l

    u

    m

    e

    (

    l

    )

    Pipettor at 28C

    Pipettor at 22C

    Pipettor at 5C

  • Common failures at pipetting

    No pre-rinse of pipettortip

    Use of other pipetting-technique as at calibration

    No wipping of pipettortip

    Multi-use of pipettortip

    Uneven movement of push button

    Angle at aspirate and deliver

    Temperature-difference instrument and materials

    Poorly seated pipettips

    Keep the pipettor vertical

    Wrong pipettor for the amount of liquid

    Wrong pipettor for the sort of liquid

  • Pipettors are precision-instruments!!!

    Use them in that way!!!!

  • Questions, please.