SOP for Calibration of Micropipettes

Calibration frequency:

  • 6 Months

Calibration Procedure for Micropipette:

  • Gravimetric method is used for calibration of Micro-pipette.
  • Attach the selected tip to the Micropipettes.
  • Calibration should be performed in Controlled room temperature (20-25°C) and vibration-free environments.
  • Tare the balance pan, place an empty dried vial and record the weight.
  1. For micropipette (10 – 100 µL)
    • Aspirate and dispense 10 µL of Milli- Q Water into previously weighed vial and record the weight.
    • Follow the above procedure for 50 µL and 100 µL with Milli- Q Water.
    • Repeat the same procedure for five times.
    • Record the observation.         
  2. For micropipette (100 – 1000 µL)     
    1. Aspirate and dispense 100 µL of Milli- Q Water into the previously weighed vial and record the weight.
    2. Follow the above procedure for 500 µL and 1000 µL with Milli- Q Water.
    3. Repeat the above procedure for five times.
    4. Record the observation.         
  3. For micropipette (500 – 5000 µL)     
    • Aspirate and dispense 500 µL of Milli- Q Water into the previously weighed vial and record the weight.
    • Follow the above procedure for 2000 µL and 5000 µL with Milli- Q Water.
    • Repeat the above procedure for five times.
    • Record the observation.         
Calibration of Micropipette

Calculate the Actual dispense volume for Micropipette as follows:             

The density of water (g/mL) against temperature is as mentioned below.

Chart for density of water (g/ml) against temperature (°c)   

Temp. (°C)0.00.10.20.30.40.50.60.70.80.9
00.9998410.9998470.9998540.9998600.9998660.9998720.9998780.9998840.9998890.999895
10.9999000.9999050.9999090.9999140.9999180.9999230.9999270.9999300.9999340.999938
20.9999410.9999440.9999470.9999500.9999530.9999550.9999580.9999600.9999620.999964
30.9999650.9999670.9999680.9999690.9999700.9999710.9999720.9999720.9999730.999973
40.9999730.9999730.9999730.9999720.9999720.9999720.9999700.9999690.9999680.999966
50.9999650.9999630.9999610.9999590.9999570.9999550.9999520.9999500.9999470.999944
60.9999410.9999380.9999350.9999310.9999270.9999240.9999200.9999160.9999110.999907
70.9999020.9998980.9998930.9998880.9998830.9998770.9998720.9998660.9998610.999855
80.9998490.9998430.9998370.9998300.9998240.9998170.9998100.9998030.9997960.999789
90.9997810.9997740.9997660.9997580.9997510.9997420.9997340.9997260.9997170.999709
100.9997000.9996910.9996820.9996730.9996640.9996540.9996450.9996350.9996250.999615
110.9996050.9995950.9995850.9995740.9995640.9995530.9995420.9995310.9995200.999509
120.9994980.9994860.9994750.9994630.9994510.9994390.9994270.9994150.9994020.999390
130.9993770.9993640.9993520.9993390.9993260.9993120.9992990.9992850.9992720.999258
140.9992440.9992300.9992160.9992020.9991880.9991730.9991590.9991440.9991290.999114
150.9990990.9990840.9990690.9990540.9990380.9990230.9990070.9989910.9989750.998959
160.9989430.9989260.9989100.9988930.9988770.9988600.9988430.9988260.9988090.998792
170.9987740.9987570.9987390.9987220.9987040.9986860.9986680.9986500.9986320.998613
180.9985950.9985760.9985580.9985390.9985200.9985010.9984820.9984630.9984440.998424
190.9984050.9983850.9983650.9983450.9983250.9983050.9982850.9982650.9982440.998224
200.9982030.9981830.9981620.9981410.9981200.9980990.9980780.9980560.9980350.998013
210.9979920.9979700.9979480.9979260.9979040.9978820.9978600.9978370.9978150.997792
220.9977700.9977470.9977240.9977010.9976780.9976550.9976320.9976080.9975850.997561
230.9975380.9975140.9974900.9974660.9974420.9974180.9973940.9973690.9973450.997320
240.9972960.9972710.9972460.9972210.9971960.9971710.9971460.9971200.9970950.997069
250.9970440.9970180.9969920.9969670.9969410.9969140.9968880.9968620.9968360.996809
260.9967830.9967560.9967290.9967030.9966760.9966490.9966210.9965940.9965670.996540
270.9965120.9964850.9964570.9964290.9964010.9963730.9963450.9963170.9962890.996261
280.9962320.9962040.9961750.9961470.9961180.9960890.9960600.9960310.9960020.995973
290.9959440.9959140.9958850.9958550.9958260.9957960.9957660.9957360.9957060.995676
300.9956460.9956160.9955860.9955550.9955250.9954940.9954640.9954330.9954020.995371

Acceptance Criteria for Micropipette:

1. %RSD not more than 2.0.

2. % Accuracy is between 98 to 102.

Calculation:

Volume Calibration

  • Balance ID: _______________        Next Calibration due date: _______________      
  • Thermometer ID: ______________   Next Calibration due date: _______________   
  • Temperature: ______________         Density of water: _________________

Actual dispensed volume (mL) = Weight of water dispensed (mg) / Density of water (g/mL) at RT

% Accuracy =  Mean value of actual dispensed volume (µL) × 100 /Set volume (µL)

Observation Table:

Micropipette ID and Volume Capacity: _______________

Serial No. of micropipette:_______________

Set No.Weight of vial after Dispensing (mg)Weight of vial before Dispensing (mg)Weight of water dispensed (mg)Actual dispensed volume (mL)
1    
2    
3    
4    
5    
Mean  NA
SD  NA
%RSD  NA
%Accuracy  NA

What is the Use of Micropipette ?

A micropipette is a very essential laboratory Analytical instrument used to accurately and precisely transfer volumes of liquid in the microliter range mainly in Dissolution and GC operation where, volume accuracy make a difference.

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Reference:

https://www.eppendorf.com/my-en/eShop-Products/Manual-Liquid-Handling/Manual-Pipetting-Dispensing/Eppendorf-Research-plus-p-PF-534798

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