UNIVERSITY OF MINES AND TECHNOLOGY(UMaT),TARKWA

SCHOOL OF PETROLEUM STUDIES

DEPARTMENT OF CHEMICAL AND PETROCHEMICAL ENGINEERING

A REPORT ON CALIBRATION OF A PIPETTE

BY

PAPPOE RICHLOVE PAPPOE (SPE.41.017.084.22)

FOR EXPERIMENT CONDUCTED ON 1ST FEBRUARY,2024.

CH 2
COURSE NAME: CHEMISTRY LABORATORY PRACTICES (CH 273)

COURSE LECTURER: MR. AMI JOHANNES

3RD FEBRUARY,2024.
GROUP 14
PAPPOE RICHLOVE ADOTEY
OWIREDU DENISE OFOSU
OWUSU AFRIYIE JOSEPH
PEPRAH PERRY FOKUO
QUASHIE ANASTASIA
QUAYSON JEFFREY NYAMEKYE
SARPONG BELINDA ACHIAA

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TABLE OF CONTENTS

INTRODUCTION......................................................................................................................3
PURPOSE OF EXPERIMENT............................................................................................5
PROCEDURES........................................................................................................................ 5
EQUIPMENT/MATERIALS USED FOR THE EXPERIMENT....................................... 6
SAFETY IN LABORATORY DURING EXPERIMENT.................................................. 6
DATA COLLECTION............................................................................................................... 8
RESULTS AND DISCUSSION..................................................................................... 9
ANSWERS TO POST LAB QUESTIONS..............................................................................10
QUESTION 1 ANSWERS.......................................................................................... 10
QUESTION 2 ANSWER....................................................................................... 11
QUESTION 3 ANSWER....................................................................................... 11
QUESTION 4 ANSWER....................................................................................... 11
QUESTION 5 ANSWER....................................................................................... 11
CONCLUSION....................................................................................................................... 12
REFERENCES.......................................................................................................................12

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ABSTRACT ;

In order for a pipette to be in range with the actual value expected from an experiment, it
must be calibrated. Precision and accuracy are key factors to consider when handling a
pipette. Pipettes become inaccurate over time, resulting in incorrect measurements, which is
why calibration is necessary. In order to prevent this ,the pipette must be calibrated on a
regular basis.The calibration process involves testing the pipette against a known reference
standard and then adjusting the calibration as necessary to ensure accuracy. Note: error in
pipette measurements could arise from failure or due to the misuse by the operator. Changes
in the temperature and pressure can influence the accuracy of measurements.
Some calculations that would be carried out are Percentage error,standard deviation and
average value of pipette.

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CHAPTER 1

INTRODUCTION

Pipettes are one of the most extensively used lab tools. They are designed to measure small
amounts of liquid and/or transfer it between containers.Pipettes also play an important role in
achieving accurate and precise measurements in laboratory work. They're available in a wide
variety of types, from simple serological and Pasteur pipettes to more complex instruments
containing a body, with a plunger and piston mostly known as micropipettes.
Pipettes can either be Manual or Electronic ,but most scientists prefer using electronic
pipettes since it has some few advantages such as;
• Better precision and accuracy
• Straightforward calibration
• Multiple pipetting modes , but Manual pipettes are more budget friendly.

BRIEF STEPS ON HOW TO USE A PIPETTE

• Pre-wet the pipette tip by aspirating and dispensing the nominal volume 3 times to humidify
the air cushion.
• Always hold the pipette at an angle not exceeding 20 degrees to ensure that the hydrostatic
Pressure inside the tip stays consistent.
• When immersing the tip into a liquid, it shouldn't be more than 2-3 mm below the surface,
to
minimise the amount of liquid retained on the outside of the tip.
• Following each dispense, there may be a liquid droplet clinging to the end of the tip. To
remove it, you should perform a touch off.
• When you repeat dispensing multiple aliquots, you should discard the first and last
dispense of the series, as they have the largest associated volume errors.

Pipettes should be cleaned frequently and stored vertically on a stand.This would ensure that
the liquid residue trapped inside the pipette body drians out and it also prevents piston
misalignment.

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CALIBRATION
Calibration is the pillar and fundamental bedrock when it comes to scientific research
instrumentation. Calibration ensures that accurate and precise measurements and results are
obtained during experiments. With calibration,you simply adjust instruments to align with
known references. It enhances credibility of experimental results by minimising errors and
enhancing the repeatability and accuracy of experimental outcomes.

CALIBRATION OF A PIPETTE.
Calibration of pipettes is an important task for researchers in many fields, including
molecular,biology and chemistry. Calibration of pipettes is essential to ensure the accuracy
;the ability of a measuring instrument to give responses to a true value, and precision;
expresses the degree of reproducibility, or agreement between repeated
measurement of volume measurements. Errors in pipette measurement could arise from
device failure or due to the misuse by the operator. That's why calibration is needed.

PURPOSE OF EXPERIMENT.
The purpose of this experiment is to calibrate a pipette and to use this pipette to
determine the density of an unknown liquid of which the volume of the pipette is initially
known.
Density=Mass/Volume is the equation used to help us find the volume of the pipette. Under
constant temperature and atmospheric pressure,the density of the distilled water used in the
experiment is constant.
The method used for this experiment is GRAVIMETRIC METHOD.
The calculated volume of the water is compared with the theoretical volume to determine the
accuracy and precision of the pipette.

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CHAPTER 2

PROCEDURES

NB. The necessary instruments needed were cleaned before the experiment began to avoid
dirt from interfering with the experiment
1. Determined the appropriate pipette volume for the test.
2. Recorded the temperature of the water.
3. Placed a beaker filled with distilled water on the analytic balance, waited for vapour
equilibrium, and closed the door.
4. Zeroed the weight with a plastic medication cup on the balance pan.
5. Attached a tip to the pipette, set the test volume, and pre-rinsed the tip by aspirating and
dispensing the specified volume three times.
6. Pressed the "frenzy" button on the second stop to clear any remaining liquid.
7. Aspirated the liquid by holding the pipette vertically, immersing the tip 1-4mm in the
liquid, and slowly releasing the rush button.
8. Waited a second, withdrew the tip, wiped away droplets, and positioned the tip against the
inner wall of the plastic cup at a 10-40° angle.
9. Pressed the push-button smoothly to the first stop, waited a second, changed the site, and
pressed the button to the second stop.
10. While keeping the rush-button pressed, removed the pipette, drawing the tip along the
inner surface. Recorded the value on the data sheet.
11. Repeated the process four more times.
12. Calculated the pipette volume using the water mass and corresponding sample density.
13. Utilised the table for various calculations, including standard deviation, percentage error,
etc.

EQUIPMENT/MATERIALS USED FOR THE EXPERIMENT.

1• PIPETTES 5• BEAKER 9.PLASTIC M. CUP

2• DISTILLED WATER. 6• NAPKIN
3• PIPETTE BULB 7.THERMOMETER

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4• ANALYTICAL BALANCE 8.ANALYTICAL BALANCE

SAFETY IN LABORATORY DURING EXPERIMENT

1. Be in your PPEs during experiment ino order to be protected during any accidents
2. No mouth pipetting ,a pipette bulb should be used instead to avoid swallowing the liquid
being pipetted.
3. Always make sure your working surroundings in the lab are clean to avoid contaminating
4. The balances are delicate instruments and should be treated with care
5.Handle glass apparatus with care to avoid breakage.

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CHAPTER 3

DATA COLLECTION

THE GATHERED DATA FOR THE EXPERIMENT

TRIAL 1 TRIAL 2 TRIAL 3 UNKNOWN

VOLUME

Mass of empty 51.93 49.63 50.34 52.35

container/g

Mass of empty 61.91 59.63 60.29 60.49

container and
water/g

Mass of water/g 9.98 10.00 9.950 8.141

Temperature of 24 24 24 24
water/℃

Volume of 10.01 10.03 9.977 10.01

water/mL

True volume of 10.01 10.03 9.977

pipet

Average volume 10.01

of water/mL

(Table 1-1)

CALCULATIONS
At constant temperature of 24℃ for all trials ; density of water is 0.9973 g/mL
Let Volume of first,second and third trial be V1,V2 and V3 respectively
𝑚𝑎𝑠𝑠
Density= 𝑣𝑜𝑙𝑢𝑚𝑒

𝑚𝑎𝑠𝑠 9.980𝑔
For trial 1;V1= 𝑑𝑒𝑛𝑠𝑖𝑡𝑦 = 0.9973𝑔/𝑚𝑙 =10.01mL

𝑚𝑎𝑠𝑠 10.00𝑔
For trial 2; V2= 𝑑𝑒𝑛𝑠𝑖𝑡𝑦 = 0.9973𝑔/𝑚𝐿
= 9. 977𝑚𝐿

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 𝑚𝑎𝑠𝑠 9.950𝑔
For trial 3; V3 = 𝑑𝑒𝑛𝑠𝑖𝑡𝑦 = 0.9973𝑔/𝑚𝐿
= 9. 977𝑚𝐿

𝑉1+𝑉2+𝑉3
Average Volume of water = 3

(10.01+10.03+9.977)𝑚𝐿
Average volume of water = 3
= 10.01mL

Hence ;
Volume of pipette =volume of the unknown liquid=Average volume of water=10.01mL
Mass of unknown liquid=8.141g
Using;volume of the unknown liquid=10.01 mL
𝑚𝑎𝑠𝑠 𝑜𝑓 𝑢𝑛𝑘𝑛𝑜𝑤𝑛 𝑙𝑖𝑞𝑢𝑖𝑑 8.141𝑔
Density of unknown liquid = 𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑢𝑛𝑘𝑛𝑜𝑤𝑛 𝑙𝑖𝑞𝑢𝑖𝑑 = 10.01𝑚𝐿
= 0. 8133𝑔/𝑚𝐿

RESULTS AND DISCUSSION

The main aim of the experiment was to calibrate a pipette and use it to determine the density
of an unknown liquid. The exercise was carried out repeatedly (three times) to ensure a more
precise work was done.
At a constant temperature of 24℃ the volume of water recorded was 10.01 mL,10.03 mL
and 9.977 mL for Trial 1 ,Trial 2 and Trial 3 respectively.There was a slight change of in the
various trial tests signifying preciseness.
After calculations,the average volume obtained was 10.01 mL. This value clarifies that
accuracy and precision was achieved in this experiment conducted.
Mass of the unknown liquid was 8.141g which yielded a density of 0.8133g/mL.
In all,the experiment done had negligible value difference.

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CHAPTER 4

ANSWERS TO POST LAB QUESTIONS

QUESTION 1 ANSWERS

a. For a set to be precise,there has to be a degree of agreement between the repeated

trials’ measurement and for a set to be accurate, the measurement has to be close to
the true value. SET A meets the conditions for a set to be precise and accurate since it
contains three same values of the true value (325mg) after the analysis was
completed.Therefore, SET A is precise and accurate.

b. Dealing with precise and not accurate,that would be SET C. Its SET C because in
terms of agreement between the repeated trials’ measurements obtained,it satisfied
this condition. Since Sammy obtained the same mass of aspirin(400mg). But with
accuracy,SET C is not,because it's nowhere near the true value (325mg) needed.

c. SET B was not precise but accuracy was obtained,the values did not satisfy the term
precision since different masses were obtained,that is 300mg,325mg and 350 mg.BUt
in terms of accuracy ,it is ;since all the masses obtained were close to the true value
needed.

d. SET D is neither precise nor accurate since there was no degree of agreement between
the repeated measurements obtained and also there's no degree of agreement between
the measured value and the true value. 250mg,280mg and 350mg are far away from
the true value and they are also precise.

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QUESTION 2 ANSWER

This experiment would likely result in an overestimation of the volume of the pipette. Since
the student is unaware of the residual liquid in the pipette, the readings would be higher than
the actual volume of liquid dispensed. This error could be caused by a number of factors,
such as liquid clinging to the sides of the pipette or not being fully expelled after use. To
avoid this error, it is important to ensure that the pipette is cleaned and dried properly after
each use, and that any residual liquid is removed before taking a measurement.

QUESTION 3 ANSWER

The volume becomes inaccurate.This is because the value for the true volume of the pipette
will increase if it is to be determined with density.This would tend to make the volume of the
pipette lower than the actual value.This is because the mass of the reference standard will be
higher than it should be,which would affect the calculation of the volume.

QUESTION 4 ANSWER

Precision and accuracy can be extended by using a properly calibrated pipette.Also,attention

needs to be paid to the information on how to use the equipment properly and also increasing
the sample length.Avoid parallax error when reading meniscus of pipette and also make sure
analytical balance is place on a levelled surface when in use.

QUESTION 5 ANSWER

2
Σ(𝑥−𝑥 )
Standard deviation(S) = 𝑛−1

where;Average volume of water (𝑥 ) = 10. 01𝑚𝐿 𝑎𝑛𝑑 𝑥1 ,𝑥2 and 𝑥3 are the respective

volumes for Trial 1,Trial 2 and Trial 3.

2 2
(𝑥1 − 𝑥) = (10. 01 − 10. 01) = 0𝑚𝐿
2 2 −4 2 2
(𝑥2 - 𝑥 ) = (10. 03 − 10. 01) = 4 × 10 𝑚 𝐿
2 2 −3 2 2
(𝑥3 − 𝑥) = (9. 977 − 10. 01) = 1. 089 × 10 𝑚 𝐿
2 −4 −3 2
∑(𝑥1 − 𝑥 ) = (0 + 4 × 10 + 1. 089 × 10 )(𝑚𝐿)
−3 2 2
=1. 489 × 10 𝑚 𝐿

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 −3 2 2
1.489×10 𝑚 𝐿
S= 3−1

S=0.02729mL

𝑀𝑒𝑎𝑠𝑢𝑟𝑒𝑑 𝑣𝑎𝑙𝑢𝑒 −𝑅𝑒𝑎𝑙 𝑣𝑎𝑙𝑢𝑒

Percentage error(σ) = 𝑅𝑒𝑎𝑙 𝑣𝑎𝑙𝑢𝑒
× 100%

where;Measured value =Average volume of water

(10.01−10.00)𝑚𝐿
σ= 10.00𝑚𝐿
× 100%

σ =0.1000%

𝑠
Relative standard deviation in ppt (RSD) = × 1000 𝑝𝑝𝑡
𝑋

0.02729𝑚𝐿
= 10.01𝑚𝐿
× 1000 𝑝𝑝𝑡
=2.726ppt

CONCLUSION
To ensure precise and accurate measurements in the laboratory settings,pipette calibration
is crucial.Regular calibration not only minimises errors in experiments but also upholds result
validity,reliability and compliance with quality assurance standards.This process involves
comparing the actual volume delivered by the pipette to the expected volume,making
necessary adjustments to achieve precision with acceptable limits. A successful pipette
calibration program should incorporate consistent schedules.

REFERENCES
Elsa Batista,Calibration of micropipette’,www.researchgate.net
Clinical Biochem.Res.Lab.(April 14, 2007)
https://osha.europa.eu/en/tools-and-publications/publications/factsheets/80
https://www.integra-biosciences.com/en/manual-pipettes/evolve#downloads
Ami Johannes,Lab manual.(November 2021)
Zeman GH,Mathewson NS.”Necessity of pre rinsing disposable polypropylene pipette tips,”CLIN
Chem 1974;20(4)497-8

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