MOHAWK COLLEGE OF APPLIED ARTS AND TECHNOLOGY

CHEMICAL ,ENVIRONMENTAL & BIOTECHNOLOGY DEPARTMENT

Lab Report
ROOM NO: FE E309

Class : BIOL 10016 Lab.

EXPERIMENT NO : #1

TITLE : Micropipetting Exercise

Submitted by : Edvair Moreira (000837578)

Partners : (1) Claudia Leonce (000842066)

(2) Fatima Alhubaishi (000817060)
(3) Rachel Joseph (000795867)

Instructor : Farag Soliman

Date lab performed : MAY 27, 2021

Date of submission : JUN 03, 2021

FENNELL CAMPUS
HAMILTON, ONTARIO
Purpose

The purpose of this experiment is to learn how to use a micropipette properly to achieve good
reproducibility with acceptable accuracy.

Materials/Method

The method send by professor Soliman was followed without any change.
The materials used to perform this exercise were:

 Cuvettes
 Distilled water
 Micropipette
 Parafilm
 Solution of dye
 UV-visible Spectrophotometer
 Volumetric dispenser

Quantitative observation

Volume of dye Volume of water Total volume (mL) Mass of dye

(0.5 μg/μL) needed in cuvette in cuvette (μg)
Stock Sol's (μL) (mL)
0 4 4 0
2 4 4.002 1
5 4 4.005 2.5
10 4 4.01 5
Table 1. Mass of dye in cuvette.

μg of dye A620 Reading of A620 Reading of Mean value of Standard

sample A sample B duplicate samples Deviation
0 0.000 0.000 0.00000 0.00000
1 0.028 0.027 0.02750 0.00071
2.5 0.064 0.059 0.06150 0.00354
5 0.100 0.115 0.10750 0.01061
Table 2. Absorbance results.

The dye solution used was blue and throughout the experiment, it was possible to observe that
the more concentrated the dye sample, the darker the solution and the higher the measured absorbance
value.
Sample calculations

Considering the mass of the dye equal to 5 μg:

0.100+ 0.115
Mean value( x́)= =0.10750
2

N
Standard deviation( s)=
√ 1
N −1 ∑
i=1
( xi − x́)2

2
s=
√1
∑ (x −0.10750)2
2−1 i=1 i

s= √(0.100−0.10750)2 +(0.115−0.10750)2

s=0.01061

Graph

Graph 1. Absorbance vs mass.

Discussion
 The A UV-VIS spectrophotometer is an instrument able to measure the intensity of a light beam

that through a solution. Since the dye solutions contains molecules that are capable to absorb a specific

wavelength of light energy, was possible to measure its intensity and observes that, the more

concentrated the solution, the more light was absorbed. According to Beer’s-Law, exist a linear

relationship between the amount of light a solution can absorb and its concentration, therefore a graph of

absorbance versus concentration was plotted making it possible to use the straight-line equation to

calculate the concentration of an unknown solution from its absorbance value. This graph is known as

the calibration curve (Skoog et al., 2017).

The calibration curve plotted from the data measured showed a straight-line equation with a

correlation coefficient (R2) equal to 0.9926. As closer this value to one, as precise is our measures, and

we can consider a good precision when this value is among 1.000 - 0.9998 (Skoog et al., 2013).

Therefore, it was noticed that the measures were not precise enough to obtain a reliable calibration

curve. Some errors can justify this result, such as personal errors (parallax error; do not invert the

volumetric flask 17 times to mix solution properly; do not rinse the cuvette 3 times with the working

standard to remove residual water), technique errors (do not read the absorbance using the same side of

the cuvette, do not avoid fingerprints on the cuvette glass), instrument errors (do not keep calibration

and cleaning up to date; do not use UPS to avoid electrical current variation), or a combination of these.

It is important to highlight that random errors ever must be avoided because they can compromise the

experiment, but small systematic errors can be fixed with some calculation adjustments (Skoog et al.,

2017).

To improve the personal technique when performing an absorbance analysis, it is important ever

review the recommendations of sample preparation and handling of instruments listed above, besides

practicing whenever possible. It is also important to remember that each kind of analyte absorbs light

energy at a specific wavelength, which must be considered when setting the UV-VIS spectrophotometer

before reading samples (Skoog et al., 2017).

 Conclusion

Since there are several compounds that have chromophore groups, the UV-VIS spectrophotometer

can be applied in numberless study fields, from quality control to research and development (Lambert et

al., 2010). The experiment performed in this report for instance could be applied in a paint industry into

a quality control laboratory to ensure that all manufactured products have the same shade of color.

During the performance of this experiment was possible to observe the importance of each procedure

step, from the preparation of dilutions to UV-VIS reading, once small errors can make a great impact on

the calibration curve.

References

Lambert, J. B., Gronert, S., Shurvell, H. F., Lightner, D., & Cooks, R. G. (2010). Organic Structural

Spectroscopy (2nd ed.). Pearson.

Skoog, D. A., Holler, J. F., & Crouch, S. R. (2017). Principles of Instrumental Analysis (7th ed.).

Cengage Learning.

Skoog, D. A., West, D. M., & Holler, F. J. (2013). Fundamentals of Analytical Chemistry (9th ed.).

Cengage Learning.