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Fish-Kill Self Directed Lab

Samuel Delgado Gomez

Tyler Woicicki

Lab Section 1

October 15, 2016


The following experiment will be executed to distinguish if the simulated water sample contains

toxic levels of Cu2+ and Fe3+ ions. By comparing other water samples with known concentrations and

ions, the student will be able to distinguish whether or not the sample contains toxic levels of the

copper and iron ions, and be able to identify other ions found in the simulated water sample. The known

water samples will be tested using a spectrophotometer that will examine the absorption spectrum of

each of the solutions. With the data from each of the solutions, the concentration will be determined

based upon their absorption spectra. The samples will also be burned to better analyze their emission

spectra, and the data will be compared in the same way.

Certain salts or high ion concentrations in most environments are usually not very good unless

the environment needs a large supply or mining will take place. Otherwise, a high concentration in

certain salts can cause a lot of damage to an already established ecosystem. Such was the case in Clark

Fork of the Columbia River during a major fish kill. The water had very high levels of metal ions, and that

increased the salinity of the water. This in turn made the river inhabitable to some of the marine life.

This lab is to use skills learned in previous labs to analyze the simulated water sample and point out the

ions present along with the concentrations of iron and copper. The concentrations and the ions present

will reveal what killed the fish.

The experiment has two major objectives; to find the presence of Group 1A and IIA metal ions,

and to determine the concentration of the iron and copper ions. With the data collected, the cause of

death for the fish will be easily determined.

This experiment will demonstrate knowledge and competence in both interpreting data

collected, and the use of lab equipment. It will also show that the student understands what they should

be looking for and how to get there. This experiment also serves as a small demonstration of how

electrons can jump energy levels and drop back down. This is how the emission spectrum is produced.

The absorption spectrum is what light is not being emitted but absorbed.

The most plausible outcomes for the experiment will be most of the IA and IIA salts will be

present in the solution and the Cu concentration will have been too high in the simulated water sample.

1. List of reagents

a. 0.5 M solution of Nacl, LiCl, KCl, Ca(Cl)2, Ba(Cl)2, and Sr(Cl)2

b. Fe/Cu SCN- solution containing 400 ppm Cu2+ and Fe3+

c. 0.1M Iron (III) Nitrate and Copper (II) Nitrate solutions

d. Simulated/Unknown lake sample

e. Distilled water

2. Making the Serial Dilutions

a. Acquire a small beaker and rinse out with distilled water.

b. Dry beaker with paper towel.

c. Fill beaker to roughly 30 ml. with the 400 ppm [CuSCN]+ or [FeSCN]2+

d. Acquire necessary Milliliters of solution (Use table A to find the milliliters of solution

needed to dissolve in the distilled water.)

e. Slowly add solution to the distilled water in a glass vial.

i. Again, refer to table A to know how much water to add vs. how much solution

f. Cu should have four dilutions as well as iron for a grand total of eight dilutions to put in

the cuvettes.

g. The following solutions should already be prepared

i. 0.5M Nacl, LiCl, KCl, Ca(Cl)2, Ba(Cl)2, and Sr(Cl)2 solutions

ii. Fe/Cu in SCN- solution (400 ppm)

iii. 0.1M Iron (III) Nitrate and Copper(II) Nitrate solutions (already mixed with SCN-


iv. And the simulated water sample

3. Burning solutions (Emissions)

a. Pre checks

1. Have a lab partner at a station ready to record data.

2. Turn on spectrometer

b. Carefully take beaker with solution and pour roughly 5 ml onto watch glass, or enough

so that a Nichrome wire may be partially submerged.

c. Set up spectroscopy instrument, If spectrometer is already set up, skip to step d

i. Press main menu -> press F5 Spectroscopy -> and the press F1 emission.

ii. Press Set Up to establish Rest of steps in red chem book on page 425

d. Take nichrome wire in right hand

e. Place left hand on spectrometer and hit enter when ready to scan

f. If the recording of the spectra looks good (it has peaks in the data, and the data is set to

the right parameters) then save on the station and move onto next sampled

g. To best avoid cross contamination, follow these steps after each trial.

i. Dump out watch glass contents into waste container

ii. Rinse out with distilled water and dry off with a paper towel.

iii. Pour some distilled water into watch glass

iv. Heat up nichrome wire to an orange glow and submerge into watch glass with

distilled water

v. *after a couple of trials (about 10) use the spatter method to clean out the

Bunsen Burner

vi. Empty watch glass

vii. Rinse off watch glass with distilled water and wipe off using a paper towel.

viii. Rinse off stir pill with distilled water and dry with a paper towel 2 times after

each solution
ix. Take new solution and repeat steps a-e

1. Solutions to Burn ; Na, Li, K, Ca, Ba, Sr, Iron (III) Nitrate, Copper (II)

Nitrate, and the unknown or simulated water sample.

4. Absorption

a. Pre checks

i. Have a lab partner at a station ready to record data.

ii. Turn on spectrophotometer

b. Carefully take a sample of either the Fe or Cu dilution and clean off with the kimwipe

c. To set up the Spectrophotometer, if set up, skip to step d

i. Press on/ off

ii. Press Main Menu-> F5 Spectroscopy -> F2 absorbance

iii. Press Setup to establish Rest of steps in red chem book on page 423

d. *This lab requires two different cuvettes to be scanned prior to the serial Dilutions and

unkown sample

i. Insert a cuvette containing a black mixture and press ZERO

ii. Wait until the ZERO-ing process is complete

iii. Remove the black cuvette and insert a clear cuvette (containing distilled water)

1. Wipe off clear cuvette before putting it inside the sample compartment.

iv. Make sure cuvette is inserted with the clear side (two opposite sides are a little

cloudy, and the other two are clear) facing the light source.


vi. Wait until reference is complete

Once complete, the serial dilutions along with the unknown water sample can be

run through
e. Dilutions to check absorption spectra; 100, 200, 300, 400 ppm Cu and Fe dilutions along

with the unknown/simulated water sample.

5. Comparing Results

a. Take Results and organize them into a graph using excel

b. Lambda max is determined after data has been graphed

c. Look at which wavelength peaks most closely resemble the ones in the unknown


d. Plot dilutions on a graph (one graph for copper and another for iron)

e. Find the line of best fit

f. Determine the concentration of copper and iron in the unknown sample by using the

wavelengths for copper and iron as x (in their respective equations for line of best fit)

6. Table A

a. Red and black color code separates the dilutions

400ppm Distilled 300ppm Distilled 200ppm Distilled 100ppm Distilled Water

Cu 10Ml 0 Ml 7.5Ml 2.5Ml 5ml 5ml 2.5ml 7.5Ml

Fe 10 ML 0Ml 7.5Ml 2.5 Ml 5ml 5ml 2.5ml 7.5Ml

7. Formulas

a. (M1) (V1) = (M2) (V2)

b. Regression Equation (y) = mx + b (line of best fit)

i. *will not be done by hand but on excel

ii. Slope(b) = (NXY - (X)(Y)) / (NX2 - (X)2)

iii. Intercept(a) = (Y - b(X)) / N

iv. Where, x and y are the variables. (points on the graph)

1. m = The slope of the regression line

2. b = The intercept point of the regression line and the y axis.

3. N = Number of values or elements

4. X = x axis value

5. Y = y axis value

6. XY = Sum of the product of first and Second values

7. X = Sum of x values

8. Y = Sum of y values

9. X2 = Sum of square x values

8. Equipment

o Looks like a giant old calculator
o Has an open ended silver pointed object that can record emission or absorption spectra
Tells you what color it is and shines a light
Bunsen burner
o Looks like an open ended ratchet with a tube connected to it.
o Tube supplies gas to the open end where the fire
Makes fire
Watch glass
o Looks like an oversized monocle, but only the glass piece
o Will be used as a container for a solution in which the nichrome wire will be able to be
submerged to acquire the solution.
Mini bowl with liquid
Nichrome wire
o Looks like solder wire
o Dont poke yourself or others with it, it will be used acquire solution to burn
Wire so you dont burn your fingers or have to spatter
o Listed above on reagents
Distilled water
o Use what is necessary
o Look like tiny glass rectangular prisms
o Containers for the serial Dilutions
Glass vials
o Look like vials that would go on a centrifuge
o This is where the dilutions will be mixed

Experiments in General Chemistry book
Previous lab data