PROCEDURE TO FOLLOW FOR THE THIRD IN-PERSON EXPERIMENT (THIN-LAYER

CHROMATOGRAPHY)
PLEASE PRINT THESE PAGES OUT TO TAKE INTO LAB.

1) You must be properly attired as discussed in the safety rules in the syllabus.
2) You must be wearing appropriate goggles.
3) You must be wearing nitrile gloves when carrying out the experiments and handling chemicals.
4) You must have your own dishwashing gloves to use when washing and handling wet glassware
5) You will need a mobile device to take pictures where indicated. Remember to remove your nitrile
gloves when you are handling your personal items. Do not put your personal items (like cell phones and
printed lab report) in the fume hood.
6) You will need to take down data in the data sheets (print them out to bring them to lab) IN PEN that
you will find at the end of this document – YOU MUST LABEL THEM WITH YOUR NAME, ID#
AND SECTION NUMBER.
7) You will need a RULER and a PENCIL for this experiment – please make sure you have these
items.

PLEASE NOTE: ANYONE FOUND NOT ATTIRED PROPERLY AND MISSING GOGGLES OR
MASK OR DISREGARDING ANY OTHER SAFETY PROTOCOLS WILL BE ASKED TO LEAVE –
THERE CAN BE NO EXCEPTIONS FOR THIS.

As you carry out the following steps, please do so mindfully. Ask yourselves why these criteria are
important, what would you observe if you did not carry out these steps.

ONCE IN LAB:

YOUR LAB INSTRUCTOR WILL TAKE ATTENDANCE. YOU WILL THEN DO A QUICK
CHECK-IN TO SEE THAT ALL ITEMS ARE IN THE LOCKER THAT YOU WILL BE USING.
ANYTHING THAT IS MISSING YOU SHOULD GET FROM THE STOCKROOM.

YOU WILL BE RUNNING A TLC OF AN UNKNOWN AND THE REFERENCE SOLUTION (ON
THE SAME PLATE) TO FIGURE OUT THE IDENTITY OF THE UNKNOWN THAT YOU HAVE.
THIS IS ASIDE FROM THE ANALYSIS PROCESS YOU WILL USE IN POSTLAB FOR TLC.

1) Obtain a TLC plate, two spotters (microcapillary) – 1 with smaller diameter and 1 with larger diameter
(if available. If not, obtain two spotters of the same size), 2 test tubes and filter paper from the bench
where the common equipment resides. Obtain the jar with the top in your locker to use as a developing
chamber and a graduated cylinder. Your TA will distribute a test tube of a small amount of solid
containing your unknown. Make note of the number and appearance of your unknown in your data sheet.

2) Draw a line 1 cm from the bottom of the TLC plate (on the adsorbent side) parallel to the bottom with
a pencil. This is your spotting line. Make sure to this lightly so you don’t flake off the adsorbent. Place
two equidistant hash marks on this line (with about 1 cm from the edge on either side) to indicate where
you will be spotting your reference solution and unknown. (This will be like the picture of the second
TLC plate you draw for your sketch from Figure 1 from the TLC Handout). Draw a line 5 mm from the
top of the TLC plate (on the adsorbent side) with a pencil. This is the solvent front. Please see Figure 1
from the TLC Handout to get an idea of how this is going to look.

3) Obtain a pipette full of the reference solution in the hood (that contains a mixture of acetaminophen,
aspirin and caffeine) and place it in a disposable test tube. This reference solution is the same as the one
used in the postlab data provided for you. Obtain about 1 mL of methylene chloride (also named

Copyright © 2024. Govindarajoo, G. Rutgers, The State University of New Jersey. All rights reserved.
dichloromethane) – about half a pipetteful - and place it in another disposable test tube. You will also
need to measure out about 5 mL of ethyl acetate from the hood in the graduated cylinder. Have all your
items in front of you.

4) Dissolve your unknown in the test tube you received it in with less than 1 mL of methylene chloride
(around half pipetteful). Please note, not all the solid will dissolve and that is fine – it is important not to
use too much methylene chloride.

5) Use your smaller diameter spotter to take up the reference solution and spot it gently on the spotting
line on one hash mark on the adsorbent side of the TLC plate. Make sure the spots are small and compact
(check under the UV lamp). Make sure to allow the spot to dry in between repeated spottings (if
necessary). Use a paper towel to absorb the remaining reference solution from your spotter. Now, tip your
test tube slightly, use the larger diameter spotter (if available, or your 2nd spotter) to take up the
dissolved unknown (avoiding the undissolved solid as much as possible) and spot it on the other hash
mark on the adsorbent side of the TLC plate.

6) Place the filter paper in the developing chamber (like in Figure 8.9 and how it is shown in the
experiment video) – if the filter paper is larger that can be added to the developing chamber, you can cut
or fold the filter paper to enable it to fit. Add ethyl acetate (eluent) to the developing chamber (make sure
that the level of the eluent is not higher than 4 mm from the bottom as indicated in Fig. 8.9 in the
textbook and the experiment video). Show both your prepared and spotted TLC plate and your
developing chamber with the eluent and filter paper inside and the cover on the chamber to your
TA. Your TA then will ask you a few questions on how you will carry out the developing of the TLC
plate that you will have to answer.

7) Remove the cover from the developing chamber, place your TLC plate in the developing chamber
(using tweezers if they are available in your locker or handling with fingers along the thin edges avoiding
touching the adsorbent side if there are no tweezers) with the adsorbent side facing up and adhering to
all the precautions you learned about in lecture and discussed in the experiment video, including placing
the cover back on the chamber. Note: NEVER pick up a TLC chamber and carry it around while a plate
is developing - same reason you don't want to jostle the chamber while a TLC plate is developing. Watch
the solvent rise to the solvent front of the TLC plate (DO NOT WALK AWAY). Remove the plate from
the developing chamber as soon as the solvent reaches the solvent front.

8) Place plate in your hood while it dries (it will take a few seconds). Label the plate with your initials
with a pencil at the top of the TLC plate. Take the plate and pencil to the bench where the UV lamp is
plugged in. Call your TA over at this time before shining the UV light on your plate. Shine the UV
lamp directly on the plate (DO NOT SHINE THE LIGHT INTO YOUR EYES!). Trace the spots you see
on your plate with a pencil while the light is shining on the plate. Turn of the UV lamp and return with
your plate to your hood.

9) Take all the necessary measurements using your ruler (distance from spotting line to solvent front,
distance from spotting line to all spots found on the TLC plate). Record them in the data sheet. Sketch the
TLC plate on the data sheet with all the measurements showing on the diagram.

10) Take a picture of your TLC plate with your initials and the traced spots showing to be uploaded on
your postlab. Dispose of your TLC plate in the solid waste. Please see below where all the other
disposable items you have for this experiment should be disposed.

11) Clean up:

Copyright © 2024. Govindarajoo, G. Rutgers, The State University of New Jersey. All rights reserved.
You will need to clean the glassware from your locker with soap and water (use your dishwashing gloves
over your nitrile gloves). Make sure the glassware is dried completely prior to placing them back to the
locker.

Locations to dispose other items:

Pipettes and TLC spotters – in Sharps container

Test tubes and any broken glass (cleaned free of chemicals) – Broken glass container
Paper towels (free of chemicals) – Regular Trash
Filter paper (after drying) is disposed in the regular trash
TLC plates would be disposed in the solid waste in the hood
Organic Solvents – Solvent/Liquid Waste
Water – Sink/Drain
Make sure your hood and bench area is wiped down and free of residue.

Data sheets for TLC on next page

Copyright © 2024. Govindarajoo, G. Rutgers, The State University of New Jersey. All rights reserved.
Name:

Student ID:

Chem 311 Section number:

TLC DATA SHEET (Make all entries in ink!)

Unknown #:

Appearance of the Unknown:

Appearance of the Reference solution:

Appearance of the Dissolved Unknown:

What is measured Measurements in cm

Distance from spotting line to solvent front
Distance from spotting line to farthest spot of
reference solution
Distance from spotting line to middle spot of
reference solution
Distance from spotting line to closes spot of
reference solution
Distance from spotting line to 1st spot of
unknown
Distance from spotting line to 2nd (IF
APPLICABLE)
Distance from spotting line to 3rd of unknown (IF
APPLICABLE)

Sketch of the TLC plate with all measurements listed: (DRAW PLATE BELOW)

Copyright © 2024. Govindarajoo, G. Rutgers, The State University of New Jersey. All rights reserved.