Experiment 4

Color Test and Microscopy: Bring Invisible Clues to Light

Jessica Nicholson and Madi Kreitz

10/3/2017

Mike Miller
Introduction:

Our experiment focuses on the ways that are used to attempt to separate an unknown compound

to discover that is contained within. The color test section includes three major tests: Marquis

Field Test, Cobalt Thiocyanate Reagent Test, and the Ferric Chloride Test. Color tests are

utilized to define powders or drugs by the colors whey appear when being presented with

different wavelengths. The marquis field test is where we use a marquis reagent to react with the

powder use of the carbonium ion which reacts with the aromatic structure of our compound. The

cobalt thiocyanate test is a color test which uses the compound in its title to bring out colors in

the substance and is used mainly for cocaine. The ferric chloride test is used to test for the

presence of salicylates (aspirin). The microscopy section contains two major tests:

microcrystalline test and hair/fiber examination with a microscope. The microcrystalline test is a

type of screening tool that is used to again identify powders. A small amount of the sample is

viewed in a microscope and it is compared and contrasted with others. The hair/fiber

examination is used to identify the unique aspects of the cuticle, medulla, and the cortex n a

piece of hair or a string of fiber. We place the sample under a microscope and sketch the image,

highlighting the important aspects. The infrared spectroscopy only consists of one test. This test

is used to analyze powders and fibers. It is used to identify the functional groups in an organic

molecule.

Experimental Procedure:

A. Color Tests

Perform a standard test using diphenhydramine as the sample for the Marquis Field Test to

confirm that it is good to use. Result should be positive.

 1. Start by taking a small amount of diphenhydramine and place it in the pouch and close it.

2. Next, break the ampoules in order from left to right, observing color changed after each

break.

Marquis Field Test and Cobalt Thiocyanate Reagent Test

1. Add a small amount of diphenhydramine to the Marquis Field Test Kit

2. Tap to the bottom of the pouch and replace top

3. Break reagent ampoule. Mix and record color changes. Observations should be noted.

4. Repeat the above steps with the unknown sample. Observations should be noted.

5. Repeat steps 1-3 with diphenhydramine and the unknown sample using cobalt

thiocyanate reagent. Observations should be recorded.

Ferric Chloride Test

1. Add a small amount (~ 2 mL) of a 1% aqueous ferric chloride solution into a small test

tube.

2. Add a small spatula tip of the unknown sample into the test tube containing aqueous

ferric chloride solution and mix with a glass rod.

3. Observe and record color of the solution before and after the sample is added. Does the

unknown contain a phenol group?

B. Microscopy Tests

Microcrystalline tests

1. Obtain a clean microscope slide

 2. Add a few solid crystals to the slide and then add a small drop of mineral oil onto the

crystal.

3. Place the cover over the slide carefully.

4. Analyze the shape, size, color, etc. of the crystal under the microscope.

5. Adjust microscope until each crystal appears clearly and then record their appearance

from each standard compound.

6. Determine identity of the sample.

Hair/Fiber Examination under the Microscope

1. Repeat steps 1-4 of the previous test, using hair and not crystals

2. Determine the identity of the unknown sample by comparing it to the standard hair

sample

C. Infrared Spectroscopy

1. Collect IR spectrum for each of the standard samples (powders and hair/fibers) and the

unknown sample of hair/fiber and powder.

2. Make a layover spectrum for each set of all the samples along with the unknown so each

standard sample is compared to an unknown.

3. Once the unknown powder is identified, label each peak on the spectra, and explain it.

Chemical Hazards:

 Gloves, goggles, and lab coats must be worn at all times in lab.

 Avoid breathing in any chemical vapors that are used in this lab or any contact with

the skin or the eyes should also be avoided. Some are toxic and can cause skin and

respiratory tract irritation.

  Ethanol is flammable and contact with eyes and skin should be avoided.

 All liquid and solid waste should be disposed of in the designated waste containers.

Results:

A. Color Tests

Marquis Field Test and Cobalt Thiocyanate Reagent Test

Diphenhydramine and Unknown Compound (Marquis Field Test)

Yellow Orange/Red Brown

Unknown Compound (Cobalt Thiocyanate Reagent Test)

Blue Blue

No Change

Ferric Chloride Test

Yellow Yellow

No Change

Table 1

Record your observations and results for both tests. (Marquis Field Test and Cobalt Thiocyanate

Reagent Test)
In the marquis field test, the unknown solution started yellow, switched to orange/red, and

ultimately ended at a rusty brown color as seen in table 1. These results revealed that the

unknown contained a carbonyl group in its structure. In the Cobalt Thiocyanate Reagent Test, the

solution started blue and had no color change what so ever, this proved that the unknown was not

cocaine.

Record your observations and results. Does your unknown contain a phenol group? (Ferric

Chloride Solution)

The solution in this test was yellow when the unknown was added with the chemical, there

proceeded to be no change in color therefore showing that the unknown contains no phenol

group.

B. Microscopy Tests

Microcrystalline tests

Unknown Diphenhydramine

Ibuprofen Acetaminophen
Table 2

Record your observations and results. Determine the identity of your unknown sample. Which of

the standard sample is your unknown?

As seen in table 2 (above), our unknown resembles the crystals of diphenhydramine. Both

compounds have crystals that are slightly round in shape but still have an edge to them, their

crystals also have very similar sizes throughout.

Hair/Fiber Examination under the Microscope

Unknown Dyed Hair

Black Hair Brown Hair

Table 3

Record your observations and results. Determine the identity of your unknown sample by

comparing to standard hair samples. Which of the standard sample is your unknown?
As seen in table 3 (directly above) the two hair samples that resemble each other closely are the

unknown and the natural black hair. These two samples both have a very thick medulla that is a

dark black color and is surrounded by a slightly lighter, shaded outer edge of the hair.

C. Infrared Spectroscopy

Unknown compound

Key: Table 4

Red – Acetaminophen (peak 1 – N-H; peak 2 – C=C)

Black - Unknown

Green – Diphenhydramine

(Peaks: O-H  2500 – 3300; -C-H  1350-1480)

Identify the unknown powder, label each peak on the spectra and explain how you determine the

identity of your known.

The unknown powder can by identified by this test to be diphenhydramine. They have almost

matching peaks other than being a very slight distance of each other on the y axis. Their peaks all

happen at the same time. They have two main peaks that indicate an O- H bond as well as a -C-H

bond.

Unknown Hair/Fiber

Key: Table 5

Cyan – Natural Black

Red - Natural Blonde

Green - Natural Red

Black – Unknown

Conclusion:

In this experiment, both unknowns that were present were successfully identified by a

variety of different tests. To begin, let’s look at the unknown compound. The first tests looked at

for our unknown compound would be the various color tests. Here our results are seen in table 1.

The Marquis Field Test gave us a positive result which gave us an insight on which compounds

that our unknown could possibly be. The other two tests gave us negative results which also

helped us to root out some of the compounds that our compound would not be able to be. Next,

we looked at the crystals that could be our unknown based on the previous tests under the

microscope. In table 3 you can see that the only compound that really resembled our unknown

was diphenhydramine, they had crystals that were round at points but also had an edge to them,

they also had similarly sized crystals. Lastly, we used IR spectroscopy to compare the same

compounds as compared on the microscopes and the results are seen in table 4. In this section we

see that again, diphenhydramine and our unknown are the most similar compounds. They have

major peaks at the same points to show a -C-H bond as well as a O-H bond somewhere within

these two molecules. So, through these tests, it has been successfully determined that the

unknown compound is diphenhydramine. Now, there was also an unknown hair/fiber that had to

be analyzed. The hair/fiber only had two tests performed on it. The microscope comparison as

well as IR spectroscopy. In the microscope comparison (ass seen in table 3) three hairs that

looked similar to our unknown were viewed. Due to things such as the medulla, pigmentation,

and texture the unknown hair and the natural black hair appeared to be the closest two hairs to

one another. In the IR spectroscopy, the same hairs were looked at and compared to one another
but no data can be taken from this section of the results (table 5) because hair is all mainly made

of the same compounds so the hairs are very close in composition and do not show any help for

finding the unknown. So, the unknown hair/fiber is able to be, possibly, tied to the natural black

hair, but jo real result was reached due to the incapability of the IR test to help out.