GROUP : AS244 TEAM : 9

MEMBERS:

NO STUDENT ID NAME MOBILE NO.

1 2022622702 NUR SYAKIRAH BINTI KAMARUDIN 0135151352

2 2022851066 NUR ALIA ZULAIKHA BINTI SOBRI 0105471376

3 2022455406 VANESSA MICHELLE ANAK EDWARD 01140074171

4 2022868246 NUR ILYANA BINTI ISMAIL 0193592747

5 2022818914 EVANGELINA MERCY ANAK MARTIN 01114012614

LAB REPORT SUBMISSION

Practical #: 2

Title of Experiment: HOW TO USE A MICROPIPETTE

Date of Experiment: 27 MARCH 2023

Submitted to: DR NURUL AILI ZAKARIA

 (include this with every report)

Exceeds
Attribute Meet Expectations Below Expectations Marks
Expectations
Marks 5 3 1
1. Punctuality Submitted early Submitted on time Submitted late
Was the report submitted on time?
2. Organization of report Fully adhere to Mostly adhere to Not organized
Is the prescribed format used? format format

3. Originality of report Original, with creative Mostly written in own More than 50% of
Is the report original, or are most insights words, attempts to report was duplicated
parts duplicated from the manual? summarise verbatim
4. Introduction Good, concise OK. Materials included Inadequate summary
a. Did the students provide summary of but not summarised or /duplicated from
background materials? background too long. sources.
b. Is the objective well described? Clear and accurate Clear, but not accurate Not clear/ inaccurate

5. Methods Well written, clear Acceptable, some High number of

a. Was the methods rewritten in the and organised. organisation. mistakes/ simply
past tense? duplicated the
manual
b. Was the methods accurately Almost all steps were Acceptable, most steps Little attempt to
described? accurately described. were described. describe/ mostly
duplicated the
manual
6. Results
a. Is Figure in proper format? Well formatted, clear Proper format No.

b. Is a title provided? Title is in proper Title is adequate No.

format and accurate
c. Is a caption provided? Well described and Described. No caption or
informative irrelevant caption.
7. Discussion Data were well and Some attempts. mainly No attempt to
a) Analysis - are the data well creatively discussed. descriptive. discuss.
explained?
b) Interpretation - are the significance Significance well Some attempts. mainly No attempt to
of the data discussed? discussed. descriptive. discuss.

8. Conclusion Correct conclusion, Conclusion is provided, No conclusion/

Was a conclusion provided? related to the but not directly related conclusion not
objective to the objective related to objective(s)
9. Grammars, spellings Few, less than 3 Three to 10 mistakes More than 10
(per page) mistakes mistakes
10. References Used proper software Manually with mistakes No or inadequate
Is the report correctly cited? citation
SUM
TOTAL MARKS

Evaluated by :_________________________________________ Date : ________________________

INTRODUCTION

A micropipette is a lab instrument used to transfer small amounts of liquid accurately and
precisely. It has an upper and lower limit and can be classified by this limit ;

● P10 : 1.0 – 10.0 μL

● P20 : 2.0 – 20.0 μL
● P200 : 20 – 200 μL
● P1000 : 200 – 1000 μL

Exceeding these limits will put the pipette out of calibration. To attain the highest level of
precision, always use the lowest size pipette. As we use the excessively big pipette for
small volumes, accuracy drops. The tips used also vary, with the larger P1000 using the
blue tips and the P200 and other small pipettes using clear tips. Every micropipette has a
plunger that stops at two different positions when it is pressed. The first position is known
as the load volume and it is used when filling the pipette with liquid and the second
position is used for the full discharge of solutions from the tip. Press the discharge slider
on the back of the grip while holding the tip over an appropriate waste container.

OBJECTIVE

● To learn the proper use of micropipettes to measure a tiny volume of liquid

samples.

MATERIALS

● Micropipette (100 - 1000 μL)

● Analytical balance
● Beaker
● Distilled water (Stored at same temperature as micropipette - Room temperature)

METHODS

1. A micropipette that was adjustable was chosen to perform pipetting.

2. The analytical balance that was correctly calibrated and well-maintained was chosen
and was put at a vibration-free zone.

3. The room temperature, the micropipette serial number, the micropipette volume range,
the name of the person calibrating and the calibration date and time were recorded.

4. The micropipette was set at the maximum volume, which was 1000 μL.

5. A beaker was put on the balance and the balance was tared.

6. A volume of water was measured and put into the beaker and the weight measurement
was recorded right away.

7. Steps 5 to 6 were repeated to obtain the value of 10 weight measurements in total.

These steps were done quickly to avoid the water from evaporating.

8. The above steps were repeated with the micropipette set at half-maximum volume,
which was 500 μL.

9. The mean weight delivered (x̅) of the water for the value of 10 weight measurements
were calculated for volume setting 1000 μL and 500 μL respectively.
10. The mean volume delivered (Vt) at the mean temperature of water of 25.0℃ was
calculated using the formula:

Vt = (x̅) * Z

Z = Conversion factor (in μL/mg) for the mean temperature of water

In this experiment, Z = 1.0041.
Table shows the values for Z factor as a function of temperature and pressure of distilled water

11. The inaccuracy was determined by calculating the percentage error of the
micropipette at each volume respectively by using the following formula:

𝑉𝑡 − 𝑒𝑥𝑝𝑒𝑐𝑡𝑒𝑑 𝑣𝑜𝑙𝑢𝑚𝑒 𝑥 100

Percentage error = 𝑒𝑥𝑝𝑒𝑐𝑡𝑒𝑑 𝑣𝑜𝑙𝑢𝑚𝑒

The percentage error may be lower (negative percentage error) or higher (positive
percentage error) than the intended volume setting.

12. The standard deviation (SD) was first calculated using the 10 value of weight
measurement as follows to determine the precision (repeatability) of the micropipette at
each volume setting:

2
Σ (𝑥 −𝑥̅)
𝑆𝐷 = (𝑛 − 1)

Xi = each weight measurement

x̅ = mean delivered weight measurement
n = number of measurements = 10

13. The coefficient of variation (CV%) was calculated to determine the imprecision as
follows:

CV = SD x 100%

14. The experimental results for the inaccuracy (percentage error) and the imprecision
(CV) were compared with the manufacturer’s specifications from the table below whether
it is within the manufacturer’s inaccuracy and imprecision limits. The comparisons were
recorded whether the results were out or within the manufacturer’s limits.
RESULTS

SETTING 1 : FULL VOLUME

SETTING 2 : HALF VOLUME
DISCUSSION

For the first setting, the maximum volume of micropipette was 1000 μL. The total
weight measurement was 9.8878 g, which would be used to calculate the mean weight
delivered, 0.98878 g and converted to 988.78 mg. Since the mean water temperature in
this experiment was about 25℃, the value of Z for this experiment was 1.0041 according
to the table of values of Z factor as a function of temperature and pressure for distilled
water. Using the formula given, the mean volume delivered can be determined, which was
992.83 μL. This value was used to calculate the percentage error of the micropipette in
order to determine the inaccuracy, which was - 0.717 %. The 10 values of weight
measurements were used to calculate the standard deviation to determine the precision
or repeatability of the micropipette. The value of standard deviation obtained was 2.369 x
−3
10 . The imprecision obtained by calculating the coefficient of variation and the value
obtained was 0.2396 %. Based on the manufacturer’s specifications for the micropipette,
the manufacturer’s inaccuracy limits is - 0.717 % ± 0.6 %, which was out of range,
whereas the manufacturer’s imprecision limits was 0.2396 % ≤ 0.2%, which was within
the range.

Next, the second setting of this experiment was to pipette the half volume which is
500 μL. The total weight obtained was 5.035 g and the mean weight calculated from it
was 0.5035 g which had been converted into a mean volume of 505.41 μL. Same as the
first experiment, the value of Z used in this experiment was also 1.0041. The percentage
of error calculated was used to indicate the inaccuracy in this experiment. The percentage
of error obtained was 1.082 %. The manufacturer’s inaccuracy limits were dependent on
the percentage of error. Based on the date given, the manufacturer’s inaccuracy limits
obtained for the half volume setting of 500 μL was 1.082 % ± 1.0 % which is within the
range. Meanwhile, the manufacturer's imprecision limit was dependent on the coefficient
of variation (CV %). CV % obtained was 0.4917 % and the manufacturer’s imprecision
limits for the half volume setting of 500 μL were 0.4917 % ≤ 0.2 % which is out of range.

In handling the micropipette during the experiment, there were a few precautions
taken to ensure high accuracy and high imprecision of measurements taken. For
example, make sure not to exceed the volume range of the micropipette when setting the
volume to use. Exceeding the volume of range of the micropipette can damage the piston
of the micropipette. Next, make sure that the size of the tip used for the micropipette was
correct to ensure the adequate seal and also to avoid leakage of liquid during pipetting.
Apart from that, avoid placing the micropipette on the table or positioning the micropipette
upside down when there is some residue of liquid inside the tip. The residue in the tip
might flow into the interior of the micropipette which can cause contamination.
CONCLUSION

The experiment was conducted to educate students about micropipette. The

students were able to learn the proper usage of micropipette by measuring volume of the
liquid samples. When the maximum volume of the P1000 micropipette was measured,
findings that were out of range were obtained and when half of the maximum volume of
the P1000 micropipette was measured, findings of inaccuracy limit was within a range
whereas imprecision limit was out of range were obtained. The results showed that the
micropipette is not calibrated. It is crucial to ensure the micropipette is well calibrated to
obtain results that are high precision and high accuracy.
REFERENCES

Batista, E., Pinto, L., Filipe, E., & Van der Veen, A. M. H. (2007). Calibration of micropipettes: Test
methods and uncertainty analysis. Measurement, 40(3), 338-342.

de Groot, M. (2018). Calibrating a micropipette. Int. J. Metrol, 25(1), 19-25.

Pipette Calibration - Eppendorf Handling Solutions. (n.d.). Handling-Solutions.eppendorf.com.

https://handling-solutions.eppendorf.com/liquid-handling/pipetting-facts/pipette-calibration/
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