St FX- Chemistry

Variable Volume Pipet

Calibration

Revision: A
Date: Aug15,2011

Page: 1 of 3

Standard Operating Procedure

Pipet Calibration
1.0 Purpose
This procedure describes how to calibrate and use a volumetric pipet. Pipets are
designed for transferring known volumes of liquids from one container to another.
Pipets have to be tested regularly since their accuracy may be affected (i.e. by
aggressive chemicals or by the frequency of cleaning). The pipet should be
calibrated and the calibration should fall within the intended volume.
2.0 Method Summary
A known volume of deionized water is transferred to a weighing flask with a clean
variable volume pipet, and weighed. Using the density of water, the volume of
water actually pipetted is determined. The amount actually delivered, compared to
the volume marked on the pipet, indicates the accuracy of the pipet. Repeated
weighings give a measure of the precision of the pipet.
3.0 When it should be performed:
Calibration should be performed at least every 3 months .
4.0 Equipment
Pipet
Pipet tip
Beaker
Analytical balance (0.0000 Calibrated See SOP Balance Calibration)
Erlenmeyer flask
Thermometer
Distilled or deionized water -degassed or at equilibrium with air will be used. The
liquid must be at room temperature.
5.0. Procedure
1. Weight the erlenmeyer flask, record.
2. Place appropriate tip firmly on the end of the pipet.
3. Press the top button (plunger knob) down to the first stop.

St FX- Chemistry
Variable Volume Pipet
Calibration

Revision: A
Date: Aug15,2011

Page: 2 of 3

4. Keep the tip of the pipet below the surface of the water when drawing liquid into
the pipet. Pull the liquid into the tip by releasing the top button slowly. Make
certain there are no air bubbles in the liquid or foam at the surface.
5. With draw the tip from the sample solution. Do not wipe the tip.
6. While holding the pipet in a vertical position, observe the meniscus of the liquid
on the tip opening. If there is a leak in the pipette, a drip will form at the tip
opening.
7. Place the pipet tip against the side wall of the weighed Erlenmeyer . Smoothly
depress the plunger knob to the first sop, pause; then depress the knob to the
second stop, to allow the water to drain. Rotate to remove any droplets adhering
to the tip.
8. With the knob still held in its lowest position, slowly withdraw the tip while
sliding it against the wall of the receiving vessel.
9. Return the knob to the release position. Do not allow the knob to SNAP back.
10. Weigh the Erlenmeyer flask and determine the amount of water transferred by
difference.
11. Transfer three aliquots (at a minimum) and weigh to determine precision in
volume measurement. Calculate the volume.
12. Complete the Test Report for the Pipet (Appendix 1) . If the average pipet
volume average falls within the intended volume confidence interval, the pipet is
calibrated and can be used.
Notes: Smooth depression and release of the plunger knob will give the most
consistent result .
Volume errors can also occur when pipetting liquids with a high vapor pressure,
where the density or viscosity deviates considerably from the value of water.

References: Eppendorf SOP - Standard Operating Procedure for pipettes See Eppendorf/
Nichiryo manual specific for the model calibrated.

Revision: A
Date: Aug15,2011

St FX- Chemistry
Variable Volume Pipet
Calibration

Page: 3 of 3

APPENDIX 1:
Test Report for the Pipet
Pipet # :

_________________________

Brand :

_________________________

Calibration :

Fixed Volume

Variable Volume

Repeater Volume
Volume Delivered:
_________________________ L
Error limits
_______________________ L
Damages :
None
Type of damage

_________________________

___________________________________________
Test Conditions :

Test temperature : ____________________________ C

Balance # :________________

Evaluation :

Mean volume : _____________ L

Test passes (within error limits)

Test not passed (not within error limits)

Date : ________________________

Examiner : ________________________