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PFSA-Pipette Calibration Worksheet & Guidelines

This document provides instructions for calibrating micropipettes using a gravimetric method. It specifies test conditions such as temperature, balance resolution requirements, and volume ranges to test. Calibration involves dispensing minimum and maximum volumes of water 10 times each and calculating the accuracy, precision, and errors to ensure they are within manufacturer specifications. If not, the pipette must be adjusted or taken out of service.

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4K views2 pages

PFSA-Pipette Calibration Worksheet & Guidelines

This document provides instructions for calibrating micropipettes using a gravimetric method. It specifies test conditions such as temperature, balance resolution requirements, and volume ranges to test. Calibration involves dispensing minimum and maximum volumes of water 10 times each and calculating the accuracy, precision, and errors to ensure they are within manufacturer specifications. If not, the pipette must be adjusted or taken out of service.

Uploaded by

sbspu
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
You are on page 1/ 2

Micropipettes Calibration Worksheet

Pipette Code: ________________ Volume Capacity (L): _____________ Manufacturer:


Balance used: ________________
Analyst:

Thermometer used: _____________ H2O Temperature: ____________

________________

Minimum Volume
Sr. No.

____________

Check Date:

(L)

Maximum Volume

Nominal
Actual
Nominal
Weight (mg)
Volume (L)
Volume(L) Volume (L)

____________

(L)

Actual
Weight (mg)
Volume(L)

Water
Density
(mg/L)

1
2
3
4
5
6
7
8
9
10
Average
Std. Deviation
Precision
Error (%)
Accuracy
Error (%)

Instructions regarding device requirements and test conditions

Water Density Table

For accurate calibration follow gravimetric method with recommended balance


Temperature (C) Density (mg/uL)
types and resolution value as follow:
15
0.999103
Volume range ( l)
Balance Type
Readable resolution (mg)
16
0.998946
1 - 10
Microscale
0.001
17
0.998778
>10 - 100
Semi-Microscale
0.01
18
0.998599
>100 - 1.000
Analytical Scale
0.1
19
0.998408
Use distilled or deionized water.
20
0.998207
Perform calibration in draught-free room at constant temperature of water, pipette
21
0.997996
and air between 15C to 30C (0.5C). The relative humidity must be above 50%.
Place water, pipettes and tips in calibration room at least 2 hours before the start of
22
0.997774
calibration to reach equilibrium with the test room conditions.
23
0.997542
A new tip should be pre-wetted 3 to 5 times to improve the accuracy.
24
0.997300
Always pipette water from a reservoir, do not take it back from the balance.
25
0.997048
Aspirate the volume to be tested slowly and evenly.
26
0.996787
Calibrate each pipette at the maximum volume and the minimum volume range.
30
0.995650
Do 10 pipetting with the minimum volume.
Do 10 pipetting with the maximum volume.
Always start the test with minimum volume first.
RESULTS
Calculate accuracy and precision of both series.
Pass
If the calculated results are within the limits specified by manufacturers
acceptable error limit, the pipette calibration is correct. If not, the pipette has to
Fail
be adjusted and checked again. If fail, place pipette out of service.
Check calibration at least once a year.

Out of service

Page 1 of 2

Evaluation of accuracy
The specified accuracy is the limit to the systematic error, which is the difference between the mean volume of actual
measurements and the true value of the volume set on the instrument. The systematic error (E) can be estimated as follows:
E
V0

systematic error
nominal volume
mean volume
The accuracy of a pipette can be expressed as a percentage of the nominal volume:

Evaluation of precision
The specified precision is the limit to the random error, which is the distribution of the measured values around a mean value. For
pipettes, precision refers to a within-series group of data, and therefore to repeatability. The random error is then quantified by
the standard deviation of measurements performed at a given volume setting under the same measuring conditions. The standard
deviation (SD or s) can be estimated as follows:

The precision of a pipette can also be expressed as a percentage of the mean volume. This is known as relative standard deviation
(RSD) or coefficient of variation (CV), and is estimated as follows:

Resources
1.
2.
3.
4.

Clinical and Laboratory Standards Institute (CLSI) HIS-A2. A quality system model for healthcare; approved guideline
second edition.
Johns Hopkins Hospital Clinical Laboratory Procedure Manual (1995). Eppendorf Pipette Calibration
Operating Instructions for Eppendorf Pipette. (ND). Brinkman Instruments Inc. N.Y.
Westgard, J.O., PhD. (2002). Basic QC Practices: 2nd Edition. Westgard QC Inc. Madison, WI.

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