The experiment that will be performed this week deals with melting point determination and thin layer

chromatography (TLC). An unknown will be given to each student in addition to information about its possible identity. Melting point and TLC data will be used to identify the unknown. Required reading: chapter 27 pp. 221-223 chapter 28 pp. 225-237 o we will not be preparing our own TLC plates nor will we prepare our own spotters. o the section concerning preparative TLC is not necessary.

Identification of an Unknown Using Melting Point/TLC1

The melting point of a solid is a physical property that is useful in its identification. Since many compounds have the same or similar melting points, most often melting point data alone is inadequate for compound identification unless it is combined with other data that suggests the compounds identity. An experimental technique that will be employed along with melting point determination for this experiment is Thin Layer Chromatography (TLC). TLC is useful in identifying compounds when a standard of known identity is available. TLC is a low tech analysis of a compounds polarity based on different interactions (hydrophilic, hydrophobic, etc.) the molecules experience with the TLC plate and the developing solvent. These two techniques will be used in conjunction to identify an unknown. Experimental Obtain an unknown from your instructor and record the unknown number in your notebook. This compound is a pure substance and is one of the 12 compounds in the table below. Perform melting point determination on the unknown compound to the best of your ability. It is extremely important that you get quality and reliable melting point data because this will make it possible to narrow your choices to one of four compound groups. Be sure to pulverize and homogenize your compound completely for the best results. Record the compound group that your unknown belongs to in your notebook. Notice, however, how each compound in a common group has a very similar melting point. Even compounds that have melting ranges
1

Levine, S. G. J. Chem. Educ 1990, 67, 972

that vary by 2 C can be easily mistaken for one another. TLC analysis will make it possible to identify which compound in the group is your unknown. TLC analysis of each known in the compound group along with an analysis of your unknown should reveal the identity of your unknown. When performing TLC analysis there are many variables that can cause the analysis to yield poor and inconclusive results. Quite possibly the variable that is most responsible for poor TLC data is the amount of sample used for the analysis. Solutions of the compounds to be analyzed must be made using small test tubes and small amounts of compound. One or two milligrams of the substance is all that is needed for a proper TLC analysis. It is not necessary to use a balance to obtain the correct mass, simply approximate the amount needed using a spatula. A milligram is approximately 50 grains of salt. Place one to two milligrams of each known and unknown into a labeled test tube and dissolve the solids using approximately 1 mL of a volatile solvent such as acetone or dichloromethane. TLC can be performed on each compound individually or simultaneous analysis can be run also. The optimal method for TLC analysis of the unknown and the three known compounds is to spot them all on the came TLC plate. The TLC plate can be set up as shown below.

baseline

unknown

known 1

known 2

Be sure to draw a feint pencil line representing the baseline taking care to not break the silica away from glass plate. Spot the samples on the TLC plate one at a time using an open ended capillary. After placing one spot of compound onto the TLC plate, use a UV lamp to make sure that enough sample has been spotted. When the TLC plate is exposed to UV light the sample, if enough has been spotted, show as a black/purple spot. If more sample needs to be added simply spot over the original spot again until enough compound is on the plate for analysis. Repeat this procedure with the three known samples and develop the plate using the appropriate solvent. Continue developing the TLC plate until the solvent front is approximately 1 cm from the top of the plate. Remove the plate from the developing chamber and allow solvent to evaporate from the plate. Exposure of the plate to UV light will reveal the results of your analysis. Dark spots of compound will appear at various places on the TLC plate based on polarity. Provided that a sufficient analysis has been completed, the identity of your unknown will be clear. If not, additional TLC analyses may be necessary to confirm the identity of your unknown. At this point if you question the results of your analysis a mixed melting point of your unknown and the known that you believe to be your compound can be performed.

known 3

Compound Table Group I biphenyl 2,5-dimethyphenol 4-nitrobenzylchloride Group II 1-chloro-4-nitrobenzene naphthalene 4-hydroxy-3-methoxybenzaldehyde Group III acenaphthene m-toluamide methyl-4-nitrobenzoate Group IV benzoic acid trans-stilbene succinimide

mp (C) 69-72 68-71 70-73

83-84 80-82 81-83

93-95 94-96 94-96

122-123 122-123 122-124

***Dont include all of these compounds in your prelab since you will be working with an unknown compound***

Data Table for MP/TLC Exp

Unknown # Melting Point of Unknown Group Number of Unknown Rf of Unknown Rf of Compounds in Group Identity of Unknown Mixed Melting Point of Unk/Known (optional) Draw diagrams of your TLC results. _______________ _______________ _______________ _______________ _______________ _______________ _______________