1.0 OBJECTIVE
To lay down the procedure for operation & calibration
of micropipette.
2.0
SCOPE
This SOP shall be applicable for operation & calibration of micropipette in Chemistry Lab.
3.0
RESPONSIBILITY
3.1
Sr. Microbiologist and above shall be responsible for the preparation and execution
of this SOP
3.2
Head QC & QA shall
be responsible for the checking and approval of this SOP.
4.0
ACCOUNTABILITY
Head QC
5.0
PROCEDURE
5.1
Operation
5.1.1 Hold the micropipette in one hand and with the
other hands turn the volume adjustment knob until the volume indicator displays
the desired volume.
5.1.2 Hold the micropipette so that the tip ejector the button is facing the user and the thumb is free to depress the push button.
5.1.3
Select and open a box containing the correct
size tips to be used.
5.1.4 Place the tip on the shaft of the pipette. Press
tip firmly using slight twisting action to ensure airtight sealing.
5.1.5 Hold the pipette in the nearly vertical position.
Using the thumb, depress the pushbutton to the first stop.
5.1.6
When aspirating fluid
the micropipette tip should normally be immersed to a depth of 2-3 mm.
5.1.7
Make sure the reagent solution does not touch
the micropipette shaft.
5.1.8
With the pipette tip immersed in the reagent the solution, draw the reagent into the tip by slowly raising the thumb and
releasing the pushbutton.
5.1.9
Remove the tip from the reagent solution by
lifting the micropipette.
5.1.10 Visually
examine the tip to make sure it contains reagent solution and there are no air
bubbles present.
5.1.11 Move
the micropipette towards the transfer container which is to receive the
measured reagent.
5.1.12 Ensure that the micropipette blowout action is fully
activated, while dispensing only.
5.1.13 Ensure that the volume is still set at the required position.
5.1.14 Press down the push button
slowly to the first stop and wait until the liquid stops flowing.
5.1.15 Press
down the push button to the second stop until the tip is completely empty.
5.1.16 With
the thumb still holding down the depressed push button, lift the micropipette and tip out of the transfer
container
5.1.17 With
the micropipette and tip out of the transfer container, slowly raise the thumb
to release the pushbutton.
5.1.18 Discard
the plastic tip.
5.1.19 Place
the pipette back into the respective stands after use.
5.2 Calibration
5.2.1
Check and ensure that the analytical balance is
calibrated before use.
5.2.2 Take distilled water in the beaker and adjust
the temperature of water to 25°C ±1°C
to be used for calibration.
5.2.3
Keep the previously dried and cooled empty
beaker in the analytical balance and tare the weight of the beaker
5.2.4 Fix the tip and deliver a fixed volume of
distilled water into the tarred beaker using the pipette to be calibrated. Note
down the weight and tare the weight of
the beaker.
5.2.5 Repeat the procedure using 10 deliveries in the
same beaker and note down the weight during each delivery.
5.2.6
Ensure that the temperature is maintained at
25°C ± 1°C during the repeated measurement.
5.2.7
Working range and the calibration points are,
|
Working Range (mL) |
Calibrate at volume (mL) |
|
20 – 200 |
20, 100 and 200 |
|
100 – 1000 |
100, 500 and 1000 |
5.2.8 After the completion of calibration, calibration the label shall be affixed into the respective micropipette.
5.2.9
Record the calibration detail in respective
format
5.2.10 Calibration of micropipette should
be done every 3 months with weight variation.
5.3 Calculations
For Calibration
5.3.1 Volume Calculation
Mean
volume = Mean of 10 weighing x
1000
0.99602
(Density of water at 25°c is 0.99602g/ml)
5.3.2 The
accuracy of the pipette is calculated as follow,
%
Accuracy = Mean volume – nominal
volume x 100
Nominal Volume
5.3.3 The precision of the pipette
is calculated as follow,
%
Precision = Standard Deviation x 100
Mean
Volume
5.4
Handling, Care, And Maintenance
5.4.1
Rack the micropipette when not in use.
5.4.2
Do
not lay down the pipette when a filled pipette tip is attached
5.4.3
Avoid dropping the micropipette or allowing
contact with dirt or grease.
5.4.4 Avoid exposing the unit to extreme temperature
changes, humidity, and dust (Operating temperatures are from15°C to 40°C).
5.4.5
Do not allow any liquid to enter into the pipette.
5.4.6
Do
not clean the pipette with acetone or aggressive solutions.
6.0
REFERENCES
Not Applicable
7.0
ANNEXURE
|
Annexure No. |
Title of the Annexure |
Format No. |
|
Annexure - I |
Calibration Record of Micropipette |
|
8.0
ABBREVIATIONS :
SOP : Standards Operating Procedure
QC :
Quality Control
HOD :
Head of the Department
QA : Quality Assurance
QCD : Quality Control Department
QAD : Quality Assurance Department
EQP : Equipment
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