QUANTITATION OF AN ALCOHOL BY GAS CHROMATOGRAPHY

PURPOSE Utilize gas chromatography to separate butanol from methanol and analyze for the volume percent composition of butanol in an unknown sample. TECHNIQUES Gas chromatography Preparation and use of a standardized curve THEORY - GAS CHROMATOGRAPHY Gas chromatography (GC) is an analytical technique that allows volatile sample mixtures to be separated and detected as they pass through a coiled column of packing material. An inert carrier gas (often helium) serves as the mobile phase to push the now gaseous substances through the porous column packing (stationary phase). Substances that spend more time on the stationary phase will take longer to travel through the column. They will have a longer retention time than substances that move quickly through the column. It is often seen that lower boiling point components travel more rapidly while substances with higher boiling points will stay on the column longer. A detector can measure how much of a component exits the column at any given time. By comparing the "area counts" of each component, it is often possible to quantitate the amount of each substance. BACKGROUND STANDARDIZED CURVE The detectors used with gas chromatographs do not always respond equally to chemicals of different structures. A series of samples of known concentrations can be carefully prepared and analyzed by GC. By plotting the detector response to these known quantities of a particular chemical, a standardized curve can be created. This curve can then be used to predict the concentration of a substance based on how the detector responds to a sample of unknown concentration. PROCEDURE 1) To cut down on the time it takes to analyze samples, it is best if you work in a group of three students. While two of your group starts to prepare your standards, one of you should approach the instructor and ask to have an unknown alcohol mixture injected. You and your teammates should record the unknown code. The printed chromatogram will be given to your group when it is ready. 2) Use the burets preloaded with methanol or butanol to quantitatively prepare the following series of standards. Each solution should be mixed in a small labeled flask and stoppered shut. You should be able to easily calculate the "volume percent of butanol in each standard." (continued on other side)
S '08 v2 M. Hauser (Survival Manual 7e) Gas Chromatography of Alcohols

Identity Standard 1 Standard 2 Standard 3 2 5 8

Volume Butanol (mL) 8 5 2

Volume Methanol (mL)

Volume Percent Butanol

GAS CHROMATOGRAPHY 3) Your instructor will make a random draw of groups to determine the order that your set of standard samples will be injected. Take the standards to the instructor at the gas chromatograph when your turn is announced. He will instruct you on the use of the GC and your samples will be analyzed. You should record the gas chromatograph parameters in your lab notebook. 4) When your chromatograms are returned to you, you should be able to logically determine the identity of each of the two major peaks. CALCULATIONS 5) From the data for each standard, identify the peaks corresponding to butanol and methanol. Record the area counts associated with each of these two peaks. Divide the butanol area count by the methanol area count to get the BuOH/ MeOH ratio for each standard. Put your data in a table similar to this one: Standard 1 2 3 6) Each of you should use graph paper to prepare a graph. The vertical axis should indicate the BuOH/MeOH ratio for each standard, while the horizontal axis should show volume % butanol. Plot the three standards and draw the best-fit straight line. 7) Analyze the chromatogram for your unknown sample to get the BuOH/MeoH ratio as before. Use your newly generated standardized curve to predict the volume percent of butanol in your unknown. The samples may be discarded in the "nonhalogenated waste." RESULTS In your notebook Conclusion, list the typical retention time of each alcohol. Report the volume percent of butanol from your unknown sample (dont forget the unknown code). State what physical property you could use to tell which of your major peaks was butanol and which was methanol. Tape your Standardized Curve into your notebook. A member Vol % of Std. BuOH area counts MeOH area counts BuOH/MeOH ratio

S '08 v2 M. Hauser (Survival Manual 7e) Gas Chromatography of Alcohols

of your group should staple and submit the chromatograms to the instructor; write each group member's name on top of this packet. QUESTIONS 1) Which one of the following gases do you think would be unsuitable for use as a GC carrier gas- nitrogen, argon, or oxygen? EXPLAIN your answer. 2) If the GC oven temperature was accidentally set too low, how would this impact retention times? 3) Suppose a student left the cap off of her prepared standard sample and the sample was not analyzed immediately. How might this affect the GC results? 4) Suppose a student accidentally splashed a small amount of water into his sample and then injected this contaminated sample onto a GC operating with a Flame Ionization Detector. Would this incident impact the gas chromatogram printout? If so, how would the chromatogram be changed? 5) When you did the GC area calculations in this lab, you made a broad assumption that each peak we observed was not superimposed on top of another peak. This would make a peak show a higher area count than it actually had. [Suppose another chemical in our unknown sample had the same retention time as butanol but it was hidden from sight. Would this make the % butanol number we report higher or lower than it actually was?] (You need rewrite only the portion in brackets in your notebook before providing an answer.)

GC column

S '08 v2 M. Hauser (Survival Manual 7e) Gas Chromatography of Alcohols