STANDARD OPERATING PROCEDURE Operation and Calibration of Micropipette
STANDARD OPERATING PROCEDURE |
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Department: Quality Control
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SOP No.: QC/MB/xxxx
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Operation and Calibration of Micropipette
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Effective
Date:
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Review
Date:
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Page
No.: 1 of 5
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1.0
OBJECTIVE:
1.1
To lay down a procedure for Operation and Calibration of Micropipettes.
2.0
SCOPE:
2.1
This SOP is applicable to different ranges of Micropipettes
used in Microbiology laboratory at Pharma Private Limited.
3.0
RESPONSIBILITY:
3.1
Microbiologist – Responsible for operation and calibration of the
instrument.
3.2
Section In-charge – Responsible
for reviewing the procedure.
3.3
QC Head – Responsible for implementing the procedure.
3.4
QA Head – Overall compliance.
4.0
PROCEDURE:
4.1
Operating procedure of Micropipettes:
4.1.1
Take the suitable range micropipette.
4.1.2
Set the desired volume as per requirement by turning the volume knob.
4.1.3
Attach a new sterilized tip. Press the shaft into the tip with only
sufficient force to make a good seal.
4.1.4
Press the plunger button to the first stop and place the tip into the
sample to the proper depth.
4.1.5
Relax the thumb pressure on the plunger. The set volume will aspirate
into the micro tip.
4.1.6
When the plunger bottom is pressed completely, the aspirated volume will
be dispensed.
4.1.7
Press tip ejector to release the tip after use and discard.
4.1.8
Care must be taken to avoid finger contact to micropipette tip.
4.1.9
As a precision instrument, micropipettes are advised to store or hang
under LAF.
4.2
Calibration procedure of
micropipettes:
4.2.1
All the different ranges of micropipettes are calibrated as per the below
table.
Pipette ranges
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10
µL – 100 µL
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100
µL - 1000 µL
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1000
µL – 10,000 µL
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Set Volume 1
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10µL
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100µL
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1000
µL
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Set Volume 2
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25µL
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250µL
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5000
µL
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Set Volume 3
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50µL
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500µL
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10,000
µL
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Set Volume 4
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100µL
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1000µL
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4.2.2
Take 100 mL of purified water in a clean container and stabilize the
water to 25 ± 1.0ºC.
4.2.3
Take the micropipette and adjust the required set volume for calibration
and operate as per the procedure mentioned above (4.1.3 to 4.1.8).
4.2.4
Collect the dispensed volume into a pre-weighed container (W1)
and weigh the container along with dispensed water (W2).
4.2.5
Calculate the weight of the dispensed volume (W2- W1).
4.2.6
For each set volume repeat the procedure for three times and ensure
that each observed weight is within the acceptance criteria.
4.2.7
Follow the above procedure to all ranges of different set volume
micropipettes.
4.2.8
For all the fixed volume pipettes calibrate only with the volume intended
by using the procedure as mentioned above.
4.3
Calculation:
4.3.1
Consider that the density of 1 mL of water = 1000 mg at 25 ± 1.0ºC
(0.997).
Weight of Empty beaker /
petridish: W1
Weight of beaker / petridish + x µL water: W2
W2
- W1
Observed weight = -------------- X 1000
0.997
4.3.2
Record the calibration data in Annexure I (Micropipette calibration
record).
4.4
Acceptance Criteria: The weight of the dispensed volume shall be within the ± 2% range of
volume.
S.No.
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Volume (µL)
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Acceptance criteria
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Lower limits (mg)
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Upper limits (mg)
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1.
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10
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± 2%
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9.8
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10.2
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2.
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25
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± 2%
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24.5
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25.5
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3.
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50
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± 2%
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49
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51
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4.
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100
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± 2%
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98
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102
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5.
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250
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± 2%
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245
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255
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6.
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500
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± 2%
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490
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510
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7.
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1000
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± 2%
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980
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1020
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8.
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5000
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± 2%
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4960
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5040
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9.
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10,000
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± 2%
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9940
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10060
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4.5
Frequency of calibration: Monthly once ± 3 days or after any maintenance work.
4.6
If micropipette is failed to meet an acceptance criteria of calibration,
then out of calibration shall be investigated as per current version of SOP
QA/xx/xxx.
5.0
ABBREVIATIONS:
QC: Quality Control
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QA: Quality Assurance
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SOP: Standard
Operating Procedure
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LAF: Laminar Air Flow
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6.0
ANNEXURES :
6.1
Annexure I - Micropipette Calibration Report: Current Version of QC/MB/xxxx.
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