Accuracy and Precision in Micropipettor Measurement

Balmores, Cabildo, Guerra, Sebastian (3MB4); Boctuanon, Chua, Diolata, Gordola

(3MB5); Canete, Taniedo, Torres, C. (3MB6)

Abstract
Micropipette is an instrument used for transferring microvolumes of liquid
solutions. The experiment aimed to determine the following: to distinguish
accuracy from precision of measured volumes of water using micropipettes on an
analytical balance, to operate micropipettes correctly, and to compute the
percent error. After collecting the data of each group, average weight of water
samples and percent error was computed. Experimental results of the average
weights turned out to be accurate and precise since the values obtained were
close to the theoretical values. Percent error was achieved by obtaining values
ranging from 0.31% to 6.93%.
Keywords: micropipette, precision, accuracy, percent error

I. Introduction
Pipettes are used to measure and transfer accurate but small volumes of liquid. It
can transfer 10 milliliters of liquids or less (JoVE Science Education Database, 2017).
The two types of pipettes are serological pipettes and mechanical pipettes or
micropipettes. Serological pipettes are long, narrow, cylindrical, glass rod with
gradations used to measure the amount of liquid, it requires the use of an aspirator bulb
to draw in and dispense liquid while mechanical pipette or micropipette operates by
piston-driven air displacement which creates a vacuum that draws in or dispenses the
liquid and it also uses plastic, color coded, disposable tips that hold the liquid.
Mechanical pipettors have different sizes that are also color coded namely, P20,
P200, and P1000. These sizes are used to measure and transfer different volumes of
liquids. P20 uses white disposable tips and can hold volumes from 0.5 to 20μL, P200
uses yellow disposable tips and are used for 20-200μL, and P1000 which uses blue
disposable tips are used for volumes from 200-1000μL. The volume that the
micropipette draws in is regulated by the volume window which can be adjusted by the
volume control knob.
 The objective of this exercise is to distinguish accuracy from precision using the
micropipettes and to compute the percent (%) error.

II. Materials and Method

100 of distilled water was dispensed on a beaker using an analytical balance. An
aluminium foil both was also folded. With P20 micropipette, 50 μL of the water from the
beaker was withdrawn then deposited on to the both. The both was then weighed with
analytical balance. The reading was then recorded as part of the results. To cancel the
previous weight, the tare was pressed. This process of pipetting and weighing 50 μL
was done three times to test for the micropipette’s accuracy. For the 100 μL and 500
μL, the procedures mentioned above were repeated.
For the precision test, the same processed used for the accuracy test was done.
However, three members did the experiment; one member executed one trial, and so
on. The data gathered were then tabulated and standard deviation was calculated.
Moreover, the percent error and coefficient variations were computed using the
following formulas:
𝑎𝑐𝑐𝑒𝑝𝑡𝑒𝑑 𝑣𝑎𝑙𝑢𝑒 – 𝑒𝑥𝑝𝑒𝑟𝑖𝑚𝑒𝑛𝑡𝑎𝑙 𝑣𝑎𝑙𝑢𝑒
% 𝑒𝑟𝑟𝑜𝑟 = 𝑥 100
𝑎𝑐𝑐𝑒𝑝𝑡𝑒𝑑 𝑣𝑎𝑙𝑢𝑒

𝑠𝑡𝑎𝑛𝑑𝑎𝑟𝑑 𝑑𝑒𝑣𝑖𝑎𝑡𝑖𝑜𝑛
𝐶𝑉 = 𝑥 100
𝑚𝑒𝑎𝑛

The accepted value was computed by taking the density of water at 17°C.
According to Weast (1984), the density is 0.99877 g/mL. Then using the process of
dimensional analysis, the density was converted to the respective masses of the
volumes pipetted. The following accepted values are as follows: 0.499 g for 50 μL,
0.0999 g for 100 μL, and 0.4994 g for 500μL.

III. Results and Discussion

The precision and accuracy of micropipette were tested by measuring water as a
sample. The tests for accuracy and precision were done by weighing 50 µL, 100 µL,
and 500 µL of water. For the accuracy test, the first member of each group weighed the
aforementioned masses of water three times. The weight of water at the mentioned
volumes was averaged and standard deviation was computed. Standard deviation is a
measure of dispersion of the data set from its mean. These values were tabulated as
follows.
Average Average Average
Group Standard Standard Standard
Weight of Weight of Weight of
Number Deviation Deviation Deviation
50 µL 100 µL 500 µL
1 0.0525 g 0.0050 0.1014 g 0.0080 0.4982 g 0.0040
2 0.0518 g 0.0079 0.0932 g 0.0057 0.4913 g 0.0010
3 0.0485 g 0.0003 0.1035 g 0.0018 0.5004 g 0.0007
4 0.0521 g 0.0101 0.1095 g 0.0021 0.4979 g 0.0093
5 0.0499 g 0.0007 0.0983 g 0.0021 0.5015 g 0.0009
6 0.0492 g 0.0016 0.1015 g 0.0026 0.4943 g 0.0032
7 0.0499 g 0.0007 0.0983 g 0.0021 0.5015 g 0.0009
8 0.0498 g 0.0002 0.1001 g 0.0008 0.4981 g 0.0021
9 0.0485 g 0.0008 0.0979 g 0.0002 0.5021 g 0.0053
Table 1. Average Weights and Standard Deviations of the Accuracy of Water Samples
The table shows the summary of the average weight and standard deviation of
the measured volumes of water for the accuracy test. For the 50 µL, Group 8 had the
lowest SD value at 0.0002, while Group 4 had the highest value at 0.0101. The mean
closest to the true value was 0.0499 g, which is a result from groups 5 and 7. For the
100 µL, Group 9 had the lowest SD value, meanwhile Group 1 has the highest value at
0.0080. Only Group 8 had a mean that was closes to the true value, which is 0.1001 g.
Lastly, for the 500 µL,the lowest value of SD is from Group 3 at 0.0007 while the
highest is at 0.0093 from Group 4. Only Group 3 had the mean that was closest to the
true value which is 0.5004 g.
Accuracy refers to the nearness of experimental value to the theoretical value.
Experimental results of the average weights of 50 µL,100 µL, and 500 µL all turned out
to be accurate as the experimental value obtained by all groups are close to the
theoretical value, 0.0500 g, 0.1000 g, and 0.5000 g, respectively.
For the precision test, the remaining members of each group weighed the
masses of a specified volume of water. Each member had measure the volume
assigned to them three times and recorded. The results are as follows.
Group Average Standard Average Standard Average Standard
 Number Weight of Deviation Weight of Deviation Weight of Deviation
50 µL 100 µL 500 µL
1 0.0473 g 0.0108 0.1173 g 0.0069 0.5053 g 0.0042
2 0.0534 g 0.0109 0.1107 g 0.0107 0.4961 g 0.0025
3 0.0476 g 0.0006 0.0987 g 0.0004 0.4980 g 0.0018
4 0.0523 g 0.0040 0.0995 g 0.0720 0.4983 g 0.0065
5 0.0501 g 0.0002 0.1002 g 0.0004 0.5035 g 0.0003
6 0.0486 g 0.0020 0.0959 g 0.0015 0.4898 g 0.0022
7 0.0486 g 0.0006 0.0971 g 0.0001 0.5061 g 0.0009
8 0.0478 g 0.0033 0.1006 g 0.0004 0.5009 g 0.0046
9 0.0482 g 0.0045 0.0970 g 0.0002 0.5006 g 0.0009
Table 2. Average Weights and Standard Deviations of the Precision of Water Samples
The table shows the summary of the average weight and standard deviation of
the measured volumes of water for the precision test. For 50 µL, the highest SD value is
0.0109 of Group 2. Meanwhile the lowest value is 0.0002 from Group 5.For 100µL, this
is 0.0720 from Group 4 and 0.0001 from Group 7, respectively. Lastly for 500 µL, we
can observe that 0.0065 from Group 4 have the highest, while 0.0003 from Group 5 was
the lowest.
Precision, on the other hand, is the closeness of the experimental values to each
other. This shows variability of measurements when measured repeatedly with the
same instrument and is independent of accuracy. It could be established that the
experimental results of the average weight of 50 µL, 100 µL and 500 µL are precise as
all the experimental values are close to one another.
The table below presents the percent error and coefficient variations of each
groups for the precision test.
Group 50 μL 100 μL 150 μL
Number % Error CV % Error CV % Error CV
1 5.2839% 22.83% 1.5245% 7.89% 0.2377% 0.80%
2 6.9311% 15.25% 6.6856% 6.12% 1.6194% 0.20%
3 2.8809% 0.62% 3.6270% 1.74% 0.2028% 0.14%
4 4.3279% 19.39% 9.6344% 1.92% 0.2978% 1.87%
5 0.0775% 1.40% 1.5793% 2.14% 0.4231% 0.18%
6 1.4792% 3.25% 1.6246% 2.56% 1.0186% 0.65%
7 0.0775% 1.40% 1.5793% 2.14% 0.4231% 0.18%
8 0.2777% 0.40% 0.2229% 0.78% 0.2577% 0.42%
9 2.8809% 1.65% 1.9798% 0.20% 0.5433% 1.06%
Table 3. Percent Error and Coefficient Variation of the Accuracy of Water Samples
 For the percent error, all groups were within acceptable parameters of error,
which is less than 10%. The coefficient variations are also low, meaning there is lesser
dispersion around the mean.
The table on the next page shows the summary of each group’s percent error
and coefficient variation for the precision test.
Group 50 μL 100 μL 150 μL
Number % Error CV % Error CV % Error CV
1 5.2839% 22.83% 17.4440% 5.88% 1.1841% 0.83%
2 6.9311% 20.41% 10.8359% 9.67% 0.6582% 0.50%
3 4.6831% 1.26% 1.1788% 0.41% 0.2777% 0.36%
4 4.7284% 7.65% 0.3779% 72.36% 0.2177% 1.30%
5 0.3230% 0.40% 0.3230% 0.40% 0.8236% 0.06%
6 2.6807% 4.12% 3.9823% 1.56% 1.9198% 0.45%
7 2.6807% 1.23% 2.7808% 0.10% 1.3442% 0.18%
8 4.2827% 6.90% 0.7235% 0.40% 0.3030% 0.92%
9 3.4817% 9.34% 2.8809% 0.21% 0.2429% 0.18%
Table 4. Percent Error and Coefficient Variation of the Precision of Water Samples
The same could be said for the precision. The percent errors for the precision
test were still within the acceptable range, hence the precision of the micropipette is
high. However, for the coefficient variation, group 4 had a very high value for 100uL.
This signifies that their data are heavily dispersed around the mean. The remaining
groups, on the other hand, had low coefficient variations.
The graphs below showed multiple differences in their accuracy and precision
which were represented by bar graphs In the determination of error between accuracy
and precision the 50ul, 100ul and 500ul showed different bar graphs such as low
accuracy and high precision, high accuracy high precision and high accuracy and low
precision. The accuracy and precision were measured using a 200ul and 1000ul
micropipettor.
 0.07 0.07

0.06 0.06

0.05 0.05
Accuracy (g)

0.04 0.04

0.03 0.03

0.02 0.02

0.01 0.01

0 0
1 2 3 4 5 6 7 8 9
Group Number

Figure 1. Accuracy-Precision Graph for 50 μL

For the 50ul, Group 1 obtained an accuracy of 0.0525g and a 0.0191g for
precision range indicating a low accuracy and low precision, group 2 got 0.0478g for
accuracy and 0.0187g for precision also indicating a low accuracy and low precision.
Group 3 obtained 0.0476g for accuracy and 0.003 for precision which a high accuracy
and precision while group 4 gathered an accuracy of 0.0520g and precision of 0.015g
with a low accuracy and low precision. Groups 5,6,7 & 8 got similar high accuracy and
good precision, obtaining a 0.0439g,0.04865g,0.04856g and 0.0534g for accuracy
respectively and 0.004g, 0.0031g, 0.0030g and 0.0172g for precision

0.12 0.12

0.1 0.1

0.08 0.08
Accuracy (g)

0.06 0.06

0.04 0.04

0.02 0.02

0 0
1 2 3 4 5 6 7 8 9
Group Number
 Figure 2. Accuracy-Precision Graph for 100 μL
For 100ul, Group 1 obtained a 0.1014g for accuracy and 000137g for precision
indicating a low accuracy and low precision while group 2 obtained a high accuracy and
low precision. Groups 3,4,5 obtained 0.0987g, 0.1098g and 0.999g for accuracy and
0.0238g, 0.0134g, and 0.0321g for precision. Groups 3,4,5 got low accuracy and high
precision. Groups 4,7,8,9 obtained both high in accuracy and precision getting 0.0238g,
0.4999g, 0.5061g, 0.4791g and 0,4891g for accuracy and 0.0055g, 0.0031g, 0.0038g,
0.0053g for precision.

0.5 0.5

0.4 0.4
Accuracy (g)

0.3 0.3

0.2 0.2

0.1 0.1

0 0
1 2 3 4 5 6 7 8 9
Group Number

Figure 3. Accuracy-Precision Graph for 500 μL

For 500ul, Group 1 obtained a low accuracy and low precision on the group,
having a 0.04982g for accuracy and 0.00094g for precision, while group 2 garnered a
high accuracy and low precision with a 0.500g accuracy and 0.00071g for precision.
Groups 3,4,7,and 8 obtained a low accuracy and high precision with 0.498g, 0.5015g,
0.5061g, 0.4791g respectively for accuracy and 0.0055, 0.0043g, 0.0053g, 0.0081g for
precision. Groups 4 and 9 got 0.04978g and 0.04999g for accuracy and 05056g and
0.5043g for precision.

IV. Conclusion
Micropipettes are used to accurately transfer small volumes of liquids. The
experiment aims to produce accurate and precise quantitative data using micropipettes.
The results showed that accuracy and precision was achieved by obtaining percent
error values ranging from 0.31% to 6.93%. The highest coefficient variation was
72.46%, indicating there is high dispersion of data from the mean.. In conclusion,
micropipettes are accurate and precise for measuring smaller volumes.

V. References
JoVE Science Education Database. (2017). Introduction to serological pipettes and
pipettors [Video file]. Retrieved October 30, 2017, from
https://www.jove.com/video/5034
Weast, R. C. (1984). Appendix A. In Handbook of chemistry and physics (64th ed.).
Boca Raton, FL: CRC Press.

VI. Guide Questions

1. How precise are the microliter pipettes? The standard pipettes?
The microliter pipettes used by groups 5 and 3 for 50 µL resulted with low standard
deviation. For 100 µL, groups 3, 5, and 8 resulted with low standard deviation. And
lastly, for 500 µL groups 3, 5, and 6 resulted with low standard deviation. The lower the
standard deviation among the measured volumes the more precise the micropipette is.
The researchers were not able to perform the experiment with the standard pipettes. It’s
possible for a group of values to be precise without being accurate, or to be accurate
without being precise.

2. Does it make a difference if one transfers 1mL using a 10mL pipette as opposed to
pipetting 1mL with a 1mL pipette?
Pipettes with different calibration should only be used for its specific volume range. For
10mL graduated pipettes measured with 1mL, the results might be not as accurate as a
1mL volumetric pipette with 1mL of any measured liquid in it. Graduated pipets are not
as accurate as volumetric pipets, since each graduation line is not individually calibrated
and any imperfection in the internal diameter will have a greater effect on the volume
delivered. On the other hand, volumetric pipettes are used to measure a single volume
accurately, up to 4 significant figures.
3. Compare the absorbance (or % transmission) of the dilute solutions prepared with the
pipettes and the micropipette. Do they agree?
The researchers were not able to get the absorbance of the volumes measured to
compare with a pipette and a standard pipette.