Spec20 Serial Dilution Lab

By Annette Jordan,2014-01-20 03:51
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Spec20 Serial Dilution Lab

    Serial Dilution & Spectronic-20 Activity Name: _____________________

     Date: ___________ Hour: _____

    Dilution Background:

    Acids and bases usually come in concentrated form, as “concentrates,” but they

    are rarely used in this form. In order to obtain a certain concentration of acid or base,

    dilutions are made. A dilution is prepared by adding a specific amount of a concentrate

    to water to obtain a new volume and concentration. One common unit of concentration

    is molarity (M), which is equal to moles of solute per liters of solution. In order to

    calculate dilutions of solutions the equation:

    MV = MV 1122

where M is concentration in molarity (mol solute/L solution) and V is volume (L), is used.

    Spectroscopy Introduction:

    Spectroscopy is one method of determining the concentration of an unknown

    solution. By measuring the respective absorbance values for solutions with known

    concentrations, a calibration curve can be constructed. The absorbance of an

    unknown can be used to determine its concentration through use of this calibration

    curve. In this lab we will use a Spectronic-20 to measure the concentration of solutions

    in order to construct a calibration curve for the diluted solutions prepared. Finally, an

    unknown solution will be analyzed to determine its concentration using the calibration

    curve created.

    Data Analysis:

Part 1:

    Unknown Letter: _____ Matching Cuvette/Dilution #: _____

Part 2:

Wavelength used: ________ nm

    Cuvette/Dilution # Absorbance











     unknown solution

    Procedure: Part 1: Dilutions and Eye Analysis

    ? Students will pair up into groups of four and obtain a test tube rack containing 9

    cuvettes, a test tube rack, a pipet, a pipet bulb, a beaker, and one cuvette filled with

    10.00 mL of simulated nitric acid.

CAUTION: This is not actual nitric acid so it will not burn you; however, it is not

    food coloring, but a laundry dye so it WILL stain you and your clothes. Please be

    careful when transferring and transporting solutions.

? To each of the 9 empty cuvettes students should add 3.00 mL of water from a

    beaker using a pipet and bulb.

? We will be diluting the original acid solution by a factor of 2 into each of the

    remaining 9 cuvettes. The group should begin by transferring 3.00 mL of the

    contents of cuvette 1 into cuvette 2 using the pipet and bulb.

? Once the solution in cuvette 2 has been mixed thoroughly, as shown by the teacher,

    the group should transfer 5.00 mL of the solution in cuvette 2 to cuvette 3 using the

    same technique.

? This procedure is repeated until all of the dilutions are complete.

? Each group will then obtain an unknown solution of simulated nitric acid.

? The letter of the unknown should be recorded and the group should decide which

    cuvette (1-10) the unknown matches most closely and record its number.

Part 2: Spectronic-20 Analysis

    ? Each group will use the Spectronic-20 instrument to measure the absorbance of the

    solutions from part one. The Spectronic-20 should be set at a wavelength of _____

    nm for these measurements.

? First, students should measure a blank solution (just water) and set the absorbance

    measurement at zero to calibrate the instrument.

? Each cuvette should be measured and their absorbance values should be recorded.

    Later these values will be used to prepare a calibration plot of absorbance vs.

    dilution number.

? The absorbance should be measured and recorded for the unknown solution used in

    Part 1. Make sure you have recorded the unknown letter.

? The dilution number and concentration of the unknown will be determined based on

    the previously constructed calibration curve.

    Post-Lab Questions: Name: _____________________

     Date: ___________ Hour: _____

    1. What is the name of the point at which you measure the volume of a solution in a

    pipet or graduated cylinder?


2. If the initial concentration of your acid in cuvette 1 was 15 M, what was the nd cuvette? What was the concentration in concentration of the solution in the 2ththe 5 cuvette?

3. Why do you need to calibrate the Spectronic-20 with a blank first?

4. If I forgot to wipe the cuvette with a Kim Wipe and there were fingerprints of the

    glass, how could this affect the value of the absorbance read by the

    spectrophotometer? Would it increase or decrease the absorbance value?


5. On the graph paper provided, plot the absorbance vs. the cuvette/dilution number

    for each of the solutions measured. Start with the most dilute solution near the y-

    axis as shown in class. From this plot, determine the cuvette/dilution number of

    the unknown solution and label the point on your graph.

    Unknown Letter: ______ Matching Cuvette/Dilution # from graph: _____

    Explain how you found this.

    6. Calculate the concentration of the unknown cuvette assuming that the original

    acid solution had a concentration of 15 M. (Show your work for full credit)


    7. What volume of the original acid solution (15 M) would you need to use to make

    a 20.00 mL solution with a concentration of 4 M? (Show your work for full credit)


    8. Explain the procedure you would use to prepare the solution in question 7 using

    the techniques learned in lab.

Name: _____________________

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