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    Report - #1 - Micropipetting Exercise

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    Edvair Filho
    The student performed a micropipetting exercise in their BIOL 10016 Lab. They used a micropipette to measure different volumes of a dye solution and water, which were added to cuvettes. The absorbance of each solution was measured using a UV-visible spectrophotometer. The student created a calibration curve of absorbance versus concentration and calculated relevant statistics. While their results did not produce a precise calibration curve, the student identified sources of error and discussed how to improve technique for absorbance analysis.

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    Report - #1 - Micropipetting Exercise

    Uploaded by

    Edvair Filho
    46 views5 pages
    The student performed a micropipetting exercise in their BIOL 10016 Lab. They used a micropipette to measure different volumes of a dye solution and water, which were added to cuvettes. The absorbance of each solution was measured using a UV-visible spectrophotometer. The student created a calibration curve of absorbance versus concentration and calculated relevant statistics. While their results did not produce a precise calibration curve, the student identified sources of error and discussed how to improve technique for absorbance analysis.

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    Report_#1_Micropipetting Exercise

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    Report_#1_Micropipetting Exercise

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    The student performed a micropipetting exercise in their BIOL 10016 Lab. They used a micropipette to measure different volumes of a dye solution and water, which were added to cuvettes. The absorbance of each solution was measured using a UV-visible spectrophotometer. The student created a calibration curve of absorbance versus concentration and calculated relevant statistics. While their results did not produce a precise calibration curve, the student identified sources of error and discussed how to improve technique for absorbance analysis.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    46 views5 pages

    Report - #1 - Micropipetting Exercise

    Uploaded by

    Edvair Filho
    The student performed a micropipetting exercise in their BIOL 10016 Lab. They used a micropipette to measure different volumes of a dye solution and water, which were added to cuvettes. The absorbance of each solution was measured using a UV-visible spectrophotometer. The student created a calibration curve of absorbance versus concentration and calculated relevant statistics. While their results did not produce a precise calibration curve, the student identified sources of error and discussed how to improve technique for absorbance analysis.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    of 5

    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.

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