Gas Chromatography In a previous lab you attempted to purify a mixture of ethyl acetate and n-butyl acetate using fractional

distillation. You collected three fractions; each successive fraction should theoretically be richer in the higher boiling component. You will analyze the three fractions with gas chromatography and refractive index – each method allows you to determine the ratio of ethyl acetate to n-butyl acetate. Based on the ratios you will determine how successful your distillation was. Chromatography Overview Chromatography is the general name for a variety of techniques used in the physical separation of mixtures. Chromatography separates compounds based on their differing affinities for a medium known as the stationary phase. The stationary phase is a solid material that organic compounds will adhere to in to varying degrees depending mostly on their polarity and molar mass. Chromatography techniques also have a mobile phase that pushes the mixture through the stationary phase. Substances with a low affinity for the stationary phase will move through the medium quickly, those with a high affinity for the stationary phase will move slowly. Common chromatography techniques include Gas Chromatography (GC), Column Chromatography, High Performance Liquid Chromatography (HPLC), and Thin Layer Chromatography (TLC). The Gas Chromatography Machine The basic parts of a GC (GC is used to refer to both the technique and the apparatus) are the injection port, the column and the detector. The separation depends on the type of stationary phase inside the column, the rate of flow of the mobile phase and temperature of the column and the length of the column. GC can only be used with compounds that can be vaporized and do not decompose in the gas phase.

gas tank

injection port recorder

detector column

The Column GCs contain a long metal column that contains the stationary phase. The column can be wide bore (1/4-1/8” diameter) or capillary. The stationary phase in GC is a wax or thick oil that coats the inside the capillary column or the solid packing inside a wide bore column. GC columns are 3 ft or longer, and are therefore coiled so they will fit inside the machine. The longer the column, the better the separation. The column is kept a specific temperature during each experiment. The temperature can be adjusted based on how volatile the components are. The Carrier Gas The mobile phase in GC is an inert gas, usually helium or nitrogen. The gas carries the sample through the column. The flow rate of the gas must be optimized for different types of samples. If the flow rate is too fast it may lead to poor separation. The Microsyringe Typically only 1 to 5 microliters of sample are injected into the GC. The sample is measured in a microsyringe. The needle and plunger of the microsyringe are very thin pieces of metal that are prone to bending; once bent this $50+ piece of equipment becomes useless. Move the plunger slowly and carefully – do not let the plunger come out of the syringe. To measure a sample, submerge the needle in your sample and pull back the plunger until you have the

appropriate amount. If you get mostly air, start again. Once you have measured, lift the needle from the sample and pull the plunger back a bit more so that there is no liquid in the needle. The Injection Port The sample is introduced and vaporized at that injection port then gets carried by the inert gas to the column to be separated. To inject a sample, hold the syringe horizontally with two hands and carefully push the needle all the way into the port. When you are ready, carefully (but not too slowly!) push the plunger all the way in, then remove the needle from the port. Start the detector as soon as you have injected. You want the entire sample to be vaporized and enter the column at the same time. Releasing the sample slowly can lead to broadening of the peaks. Retention Time The essence of a chromatographic separation is the partitioning of a component between the mobile and stationary phases. If the sample likes to stick to the stationary phase, it will take a longer time to reach the end of the column. The time it takes for a compound to go from the injection port to the detector is known as the retention time. The retention time in GC is analogous to the Rf in TLC. A compound will have a characteristic retention time under a set of GC conditions (flow rate, column temperature, etc.) therefore compounds can be identified by their retention time. The Detector and the Recorder The detector works by the fact that sample gases have a different thermal conductivity than the carrier gas. The detector will lose heat a different rate when substances other than the carrier gas pass by. This information is converted into an electric signal that is sent to the recorder. Peaks are plotted versus time and appear on the recorder as each component of the sample passes by the detector. Quantitative Analysis Gas chromatography offers an opportunity to determine the relative amounts of each mixture component while keeping sample size down to microgram amounts. Other techniques allow quantization, but most require much more material for analysis. The output of the GC is a graphed series of peaks. The relative areas of the peaks are proportional to the relative amounts of the compounds in the mixture. The integrating recorder digitizes the voltage from the GC, calculates the relative areas of each peak, and prints the results. The relative areas appear as percentages on the chart paper. Ideally, only two peaks will appear for your fractions, one for ethyl acetate and one for n-butyl acetate. Since the detector response increases for compounds with larger molecular weights and/or increasing polarity, a correction factor must be applied. The correction factor for n-butyl acetate is 0.74 and for ethyl acetate is 0.89. Notice that the higher molar mass butyl acetate has a lower correction factor than ethyl acetate. After application of the correction factors, the resulting percentages must be renormalized to 100% Example: The recorder claims that a mixture is 50% ethyl acetate and 50% n-butyl acetate. Application of the correction factors gives (50)(0.74) = 37.0% for n-butyl, and (50)(0.89) = 44.5% for ethyl. Clearly a mixture of only ethyl and n-butyl acetates can't be 44.5% one and 37.0% the other. What's the other 18.5% ?! We normalize the values to add to 100% by dividing each percentage by the sum of the percentages: 37.0/(44.5 + 37.0) = 45.4% n-butyl acetate and 44.5/(44.5 + 37.0) = 54.6% ethyl acetate