INTRODUCTION There are several ways for organic chemists to determine the identity of unknown compound.

One of the ways is through the thin layer chromatography (TLC) technique. Qualitative analysis of small amounts of material and rapid separation plays the major role in this technique. Chromatography is a separation technique which involves a compound to be resolve between two phases, mobile phase and stationary phase. Depending on the compounds interaction with the stationary phase, it will either being adsorbed by by the stationary phase or reacting chemically with it. Various types of chromatography is available such as liquid-liquid chromatography and liquid-solid chromatography

OBJECTIVES Objectives of the experiment are to identify and study: 1. 2. 3. 4. The technique of thin layer chromatography (TLC). The principle of liquid-solid chromatography (adsorption chromatography). The effect of intermolecular forces in TLC. The unknowns by Thin Layer Chromatography and Melting Point Combination.

PROCEDURE An unknown sample was obtained and its melting was measured and determined its group.

1 mL of dichloromethane was added to the remainder of the unknown and the mixture was swirled until dissolved.

3 silica gel TLC plate was obtained and a horizontal line of 0.5 cm was mark at bottom plate.

A spot of unknown solution was put on the plate alongside with the compound from the determined group.

About 5 mL of developing solvent was put in the developing chamber. 4.5 ml of Hexane and 0.5 ml of Ethanol

1 TLC plates was placed in the developing chamber to allow the solvent to rise to within 1 cm of the top of the plate

The plate was removed and the solvent front was marked.

Ultraviolet (UV) ray was used to visualize the components and the outline spots was marked with a pencil.

The distance traveled by each components was measured and retardation factor (Rf) values was calculated. Every components on each two plate was compared to find the Steps 4 to 9 was repeated by using the unknown solution together with the suspected compound.

All the results were recorded on the data sheet.

TABLE OF REAGENTS AND CHEMICALS No 1 2 3 Reagents Chemicals

RESULTS

(I)

(II)

(III)

DISCUSSION

During chromatography, a mobile phase (eluent) distributes the compounds present in a mixture over a stationary phase (adsorbent). Thin Layer Chromatography (TLC) is one type of chromatography, where: - the mobile phase is a solvent or mixture of solvents (eluent) - the stationary phase is a solid adsorbent (e.g., silica or alumina) coated on a stationary support such as glass or plastic (TLC plate). The chromatogram is then developed by placing the TLC plate into a TLC chamber (a beaker containing the mobile phase, i.e., the eluent). The eluent will travel up the adsorbent by capillary action to the top of the TLC plate (known as the solvent front), carrying the sample with it. This process is generally referred to as "running your TLC plate". Depending on the relative polarity of the sample and the eluent, the sample either adsorbs to the solid phase or is eluted by the mobile phase (eluent). If the eluent is very polar relative to your compound, it will dissolve your sample and the sample will move with the mobile phase. Overall, the eluent and your sample will compete for a space (an active site) on the adsorbent (stationary phase) coated on the TLC plate. The more polar compound will win this competition and adhere (adsorb) to the adsorbent, while the rest of the mixture will move. In this experiment, firstly we find the melting point of the unknown sample to determine the group it belong. Since its melting point range within 95.8-99.6, we found out that it is from group 3. In group 3, there are three compounds which are acenaphthene, benzil and methyl 4-nitrobenzoate and for simplicity, we name them A, B and C respectively. After the spots position was determined with the aid of UV ray, we observed that compound A traveled higher than other while compound C traveled lower than the rest. This leave only compound B that traveled about the same level as the unknown sample. The reason behind this is because of the polarity of the compound. The more polar the compound, the more it will adhere to the adsorbent and the smaller the distance it will travel from the baseline. Likewise, the less polar the compound, the higher the distance it travels. The third plate was used to confirm that the unknown sample is compound B which is methyl-4-nitro. One of the spot on the plate is the mixture of unknown sample and compound C. After the spots were marked, we observed that the spot of mixture of unknown sample and compound C produce a single spot and was at the same level with the other spots. Hence, it can be concluded that the unknown has been purifed, and that the unknown is possibly the same compound as the authentic compound which is methyl-4nitro.

There are some errors that we make that might affected the outcome of the results. Firstly is about the solvent front. For the first plate, we forgot to mark the solvent front after removing it from the developing chamber. Besides, the solution evaporated quickly so that we cannot see the solvent front anymore. As the result, we just approximate where was the solvent front when marking it. This might affect the calculation for retardation factor (Rf). To overcome this error, we need to be ready to mark the solvent front whenever the plate is being taken out from the developing chamber. The second error is about the quantity of eluent used. Our lab instructor have already told us to use 4.5 mL of hexane and 0.5 ml of ethanol. However we made a mistake here that we put few drops more of ethanol. Excessive drop of ethanol may prompt the spots to move higher than the solvent front. Thanks to Allah, the spots did not and we are able to finish our lab smoothly. Therefore, we should listen carefully to the instructor and use the apparatus appropiately so that we can prevent this type of error in the future. Some precautions that we need to take into account are like we must use pencil and draw lightly when marking on the plate. Usage of other marking tools might change the result of this experiment. Besides, we should always keep the developing chamber closed since the eluent might react with air if it is being exposed too frequently. Other than that, we must use the UV ray apparatus with extra careful since any mistake can cause bad condition to others. Many steps can be taken to improve this experiment. First, the lab assistant need to be more alert of excessive use of material, as we would not want anything goes to waste. Secondly, we need to have better melting point apparatus that can help us to determine the melting point more efficiently. CONCLUSION Alhamdulillah, we have successfully finish the experiment and completed the report individually. For conclusion, we are able to fulfill all the objectives of this experiment such as to learn the technique of thin layer chromatography (TLC), to study the principle of liquidsolid chromatography (adsorption chromatography) , to learn the effect of intermolecular forces in TLC and to identify the unknown sample by thin layer chromatography and melting point combination. The unknown sample is actually methyl-4-nitro with the Rf value of 0.4571.