EXPERIMENT 19:

Gas Chromatography
Jennifer Driskell February 26, 2009 Chem. 113, Section 101 TA: Todd Day Group Members: Arina Earnest Michael Schiavone Jenna Doud

Introduction:
One thing many people do not know or understand is how or more importantly what is depleting the environment in which we are living. There are many problems that affect the environment from industrial smog to the depletion of the ozone but what most people fail to understand is what exactly is causing this destruction. Using gas chromatography one can answer these problems with pretty good accuracy. The process of gas chromatography is a technique that separates substances by passing a gaseous mobile phase through a porous stationary phase (1). Gas chromatography is very useful and can be applied in many situations such as: determining the drugs present in a persons blood, breath, saliva, and urine samples, determining trace hydrocarbons and other pollutants in the air, and determining the pheromone sex attractants in insects (1). This idea of gas chromatography was first discovered in the 1906, when Mikhail Tswett, the Russian botanist, published his findings describing the separation of plant pigments using column liquid chromatography (2). Then in the 1930s, English chemists Martin and Synge developed a form of chromatography called partition chromatography. This method is more commonly referred to as paper chromatography and was originally used by Martin and Synge to analyze the structure of proteins in wool (1). Gas chromatography was not thought of until 1952 when Martin and James discovered the use of chromatography using a gaseous mobile phase (2). This discovery led to the widespread use and popularity of chromatography. Restating the idea of gas chromatography, it is the separation of a gaseous mobile phase through a stationary phase to separate the gas into its simple components. This can be done through gas-solid chromatography (GSC) or gas-liquid chromatography (GLC) (2). A small volume of sample is passed into the stationary phase by a carrier gas at a constant rate. The carrier gas is an inert gas such as nitrogen or helium which flows through the column carrying the sample with it (3). In our lab the carrier gas was natural gas out of the gas jets at the lab stations. The choice for which stationary phase to use most often is based upon the polarity of the substance being passed through the stationary phase, using the like

dissolves like principle (1). Once the separated components exit the column they are quantitatively detected by chosen detectors. In our lab the detector was the Beilstein detector that changed the color of the flame produced by the natural gas. Many commercial GCs use a couple detectors at once including, thermal conductivity (TCD), flame ionization (FID), and electron capture (ECD), which feed information to a recorder (1). Chromatography is important because it allows for the separation of complex compounds into their individual components so they can be identified and evaluated. This method is widely used for the reason that it is fast and can be done usually within a few minutes to a few seconds. Chromatography is used by many professions for many different purposes. Such as environmentalist can use chromatography to test soil and water contamination or air pollution, forensics can use it to test for the presence of drug in urine or blood samples, other areas that use chromatography are clinical, pharmaceutical, and biochemical industries (4). The environment is something that needs to be protected especially the ozone layer since it protects us from harmful ultraviolet radiation. Chemists have found the detrimental effects that halocarbons, such as CFCs, have done to our environment so they have created chemicals such as HFCs and HCFCs to replace CFCs (1). In this lab the substances being separated are Freon 22 and Freon 123 to determine the differences between the two. The goal for this experiment is to examine the differences between Freon 22 and Freon 123 as well as the differences between the homemade gas chromatograph and the commercial gas chromatograph.

Procedure: (The full procedure can be found under Experiment 19 in the PSU Chemtrek
2008-2009)
This lab used both a homemade gas chromatograph and a commercial chromatograph. However before being able to use the homemade gas chromatograph it was first created in Section A- The Construction of a Gas Chromatograph. This section goes through the construction of the column, the detector, and the overall construction of the apparatus. Section B- Measurement of the Retention Time of Air and Gas Flow Rate, was to measure how

long it took for a blast of air to make its way through the column of tide along with the carrier gas (natural gas) to briefly change the strength of the flame. The retention time allows data gathered from other homemade gas chromatographs to be compared. Section C- Measurement of the Retention Times of Halocarbons, was to measure how long it took for Freon 123 and Freon 22 to make it through the column. Once they made it through the column to the detector the flame became green, then a brighter more intense green, and then the color disappeared. These three results were used calculate the retention time, and the band width of the halocarbon peak, W b. Section D- GC Separation of Halocarbon Mixtures, sent a mixture of Freon 22 and Freon 123 into the column. When the gas mixture made its way through the column to the detector it produced two sets of times for the appearance of the green flame, an intense green color and the disappearance of the green color because it contained both Freon 22 and Freon 123. This data was then compared to the data obtained from Section C, when the gases were detected separately. Section E- Quantitative Analysis Using a Commercial GC, uses an actual commercial gas chromatograph to measure the retention times and area under the peaks for Freon 22 and Freon 123. Using the calibration data provided by the GC Technician Michael Bogdan, two separate graphs can be constructed providing an equation for a line of best fit that was used to determine the concentration of Freon 22 and Freon 123 in the unknown samples.

Results:
Table 1. Measuring the Gas Flow Rate: This information was gathered during Section B of the lab. It shows the number of trials and the amount of time it took for the air that was injected into the carrier gas to make its way through the column and cause the flame to dip. The time was measured in seconds using the second hand on the wall clock located in room 105C in Whitmore. Data taken from Jennifer Driskells lab notebook (5).

Trial Number 1 2 3

Time (seconds) 3 3 3

Gas Flow Rate (cm/sec) 9.33

Table 2. Injection of Sample Gases: This information was gathered during Section C of the experiment. It shows the comparison of Freon 22 and Freon 123 with regards to the first appearance, the max intensity, and the disappearance of green in the flame. The time was measured in seconds using the second hand on the wall clock located in room 105C in Whitmore. The data was taken from Jennifer Driskells lab notebook (5). Sample Freon 22 (.1 mL) Freon 22 (.1 mL) Freon 22 (.1 mL) Freon 123 (.2 mL) Freon 123 (.2 mL) Freon 123 (.2 mL) Trial Number 1 2 3 1 2 3 Appearance (sec) 3 3 3 7 8 6 Max Intensity (tR) (sec) 4 3.5 3.5 9 10 8 Disappearance (sec) 5 5 5 16 17 15

Table 3. Averages for Sample Gases: This table is to show the averages for each of the two sample gases found by averaging each of the columns from Table 3 that pertain to each Freon 22 or Freon 123. The band width of the halocarbon peak is found by subtracting the disappearance of the green color from the appearance of the green color. This data is taken from Jennifer Driskells lab notebook (5) .

Sample Freon 22 (.1 mL) Freon 123 (.2 mL)

Appearance (sec) 3 7

Max Intensity (tR) (sec) 3.7 9

Disappearance (sec) 5 16

Band Width (Wb) (sec) 2 9

Table 4. Molecular Make-up: This table is to show the molecular weight, the boiling point, and the dipole moment for each Freon 22 and Freon 123. This information helps to determine why one of the gases is quicker than the other. (6) Sample Freon 22 (.1 mL) Freon 123 (.2 mL) Boiling Point (oC) -40.7 29.5 Molecular Weight (g/mol) 86.469 152.93 Dipole Moment 1.42 1.36

Table 5. Neighbors Data: This table was the data received from Arinna Earnest and Mike Schiavones experiment. It gives the retention time and the gas flow rate for their homemade gas chromatography. The data was timed in seconds using a stopwatch (7,8). Sample Freon 22 (.1 mL) Freon 123 (.2 mL) Max Intensity (tR) (sec) 7 18.9 Gas Flow Rate (u) 6.99 6.99

Table 6. Homemade Gas Chromatography with Mixtures: This table shows data obtained from the three trials of injecting the mixture of .1 mL of Freon 22 and .2 mL of Freon 123 into the carrier gas. Since Freon 22 and Freon 123 were injected together they produced two green flames back to back one another (5).

Sample

1st Appearance (sec) 5 3 3

1st Max Intensity (sec) 6 4 4

1st Disappearance (sec) 10 6 8

2nd Appearance (sec) 11 8 9

2nd Max Intensity (sec) 13 10 10

2nd Disappearance (sec) 18 15 17

Mixture gas trial 1 Mixture gas trial 2 Mixture gas trial 3

Table 7. Averages for Mixture using the GC: This table shows the average of each of the categories for the mixture of .1 mL of Freon 22 and .2 mL of Freon 123 (5). Sample 1st Appearance (sec) 3.7 1st Max Intensity (sec) 4.7 1st Disappearance (sec) 8 2nd Appearance (sec) 9.3 2nd Max Intensity (sec) 11 2nd Disappearance (sec) 16.7

Mixture gas Freon 22 and Freon 123

Table 8. Comparison of Mixture vs. Separate: This table shows the comparison between the retention times and the band width of Freon 22 and Freon 123 both separate and combined in the mixture of the two (5). Sample Freon 22 (separate) Freon 123 (separate) Freon 22 (mixture) Freon 123 (mixture) Retention Time (sec) 3.7 9 4.7 11 Band Width (Wb) (sec) 2 9 4.3 7.4

Table 9. Neighbors Date for Mixture Gas: This table shows Arinna Earnest and Mike Schiavones data for the retention times and band widths of Freon 22 and Freon 123 in the mixture gas composed of .1 mL of Freon 22 and .2 mL of Freon 123 (7,8). Sample Freon 22 (mixture) Freon 123 (mixture) Retention Time (sec) 5.8 18.1 Band Width (Wb) (sec) 3.4 12.2

Table 10. Resolution Comparison: This table shows both the resolution of our homemade GC and our neighbors homemade GC that was obtained from the mixture of .1 mL Freon 22 and .2 mL Freon 123 (5,7,8,and 9). Sample Mixture (our data) Mixture (neighbors data) Resolution 1.08 1.60

Table 11. Calibration Table for Peak 1-Freon 22: This data was provided by the GC Technician Mike Michael Bogdan who showed the students how to correctly use the Commercial Gas Chromatograph. It was used to figure out the concentration of the unknown sample of Freon 22 (10). Concentration (ppm) 125 250 500 Area (mv-sec) 4431 8951 17671

Graph 1. The Graph of the Freon 22 Peak from the Commercial GC:

Peak #1 - Freon 22
20000 18000 A r 16000 e 14000 a 12000 ( m 10000 v 8000 . s 6000 e 4000 c 2000 0 0 100 200 300 400 500 600 Concentration (ppm) Series1 Linear (Series1)

y = 35.246x + 71

Table 12. Calibration Table for Peak 2- Freon 123: This data was provided by the GC Technician Michael Bogdan who showed the students how to correctly use the Commercial Gas Chromatograph. The data was used to figure out the concentration of the unknown sample of Freon 123 (10). Concentration (ppm) 125 250 500 Area (mv-sec) 9891 18558 33933

Graph 2. The Graph of Freon 123 Peak from the Commercial GC:

Peak #2 - Freon 123

40000 A 35000 r e 30000 a 25000 ( y = 63.739x + 2203.5

m 20000 v 15000 . s 10000 e c 5000

) 0 0 100 200 300 400 500 600 Concentration (ppm)

Series1 Linear (Series1)

Table 13. Concentration of Freons in Unkown: This shows the concentration of Freon 22 and Freon 123 that were in the unknown sample put through the commerical gas chromatograph. This information can be obtain by using the equation gather from the line of best fit from each of the graphs that pertain to the respective Freon (5). Sample Freon 22 Freon 123 Concentration (ppm) 349.1 391.5

Equation 1. for Gas Flow Rate in Table 1: U=

= 9.33 cm/sec

Equation 2. for Resolution in Table 10: Resolution=

= 1.08

Equation 3. For Figuring Out Concentration of Unknown Freon Samples in Table 13: Freon 22: y = 35.246x + 71 12376=35.246x + 71 12305=35.246x x= 349.1 ppm Freon 123: y = 63.739x + 2203.5 27158=63.739x +2203.5 24954.5=63.739x x= 391.5 ppm

Discussion:
The beginning, or the majority of the results from this lab experiment resulted from the homemade gas chromatograph created in section A of the lab procedure. The first thing done to the homemade gas chromatograph was the calculation of the gas flow rate. This rate was needed later in the experiment for comparing our data to our neighbors data. The gas flow rate was calculated by dividing the length of the gas chromatograph column in centimeters by the average retention time for air in seconds. The length of our column was 28cms and the average retention time for air was 3seconds which gave us a gas flow rate of 9.33 cm/sec. The gas flow rate determines how fast the carrier gas and thus the sample gas will flow through the column. If this rate was too fast it would be hard to determine when the first appearance of green was, from the maximum intensity, from the disappearance of green in the flame. It also does not allow for any of the sample to separate inside the column because it is being pushed through so quickly. The optimum speed for the gas flow rate was somewhere under 10 seconds. This was to give enough time so a change could be noticed. Once the gas flow rate was calculated it was time to experiment with the two samples of Freon, Freon 22 and Freon 123. Freon 22 had a first appearance of 3 seconds, a max intensity of 3.7 seconds, and a halocarbon bandwidth of 2. The boiling point was -40.7 degrees Celsius, the

molecular weight was about 86 g/mol and it has a dipole moment of 1.42 (5,6). Freon 123 had a first appearance of 7 seconds, a maximum intensity of 9 seconds, and a halocarbon bandwidth of 9. The boiling point was 29.5 degrees Celsius, the molecular weight was about 153 g/mol, and it has a dipole moment of 1.36. This data presented a huge inside to why these two samples of Freon behave the way they do. There is a correlation between the boiling points and the retention time for these gases. Freon 22 has a boiling point of -40.7 degrees Celsius and Freon 123 has a boiling point of 29.5 degrees Celsius which explains why the retention times were 3.7 seconds and 9 seconds for Freon 22 and Freon 123 respectively. The lower the boiling point the quicker the substance moves. Thus Freon 22 had a much lower boiling point so it moved through the column much quicker than Freon 123. The molecular polarity and weight as play a role in determining the retention time. Freon 22 has a lower molecular weight and a bigger dipole moment, this means that it takes less energy to make the reaction happen, thus it will occur quicker. Freon 123s molecular weight and dipole moment were greater than that of Freon 22 resulting in more energy needed to cause the reaction, thus it occurred slower. To compare retention times to another groups data would be useless unless the gas flow rates are also considered. Our neighbors gas flow rate, calculated the same way as our just using different numbers, was 6.99 cm/sec. Their retention times for Freon 22 and Freon 123 were 7 seconds and 18.9 seconds respectively (7,8). The faster the gas flow rate the quicker or smaller the retention times should be because it takes less time for the sample to make its way through the column. When our neighbors data was compared to our own, the numbers made sense. Our flow rate was faster than theirs so our retention times were smaller than theirs. Their flow rate is slower so it takes more time for the Freon samples to make their way through the column so the retention times would be longer or higher. Their numbers also matched our

pattern of Freon 22 being quicker than Freon 123. The previous numbers were all calculated with separate samples of Freon 22 and Freon 123, however during the lab data was also calculated with a mixture of Freon 22 and Freon 123. Table 8 shows the comparison between the retention times and halocarbon bandwidth for the Freon samples separated and for the mixture of the two Freon samples. When the mixture of Freon gases was injected into the column the detector was able to detect and exhibit both of the Freon samples separately. The retention times for the Freons in the mixture were different than the retention times of the Freons separated from each other. Also noticed in Table 8, the width of the halocarbon bandwidth increases as the retention time increases. Since the retention time is longer, the longer time the sample has for its molecules to interact with the tide and separate further increasing the distance for which they exit the column thus increasing the width of the peak. When dealing with a mixture, an important parameter to examine would be the resolution obtained between the two components. The resolution is the difference between retention times divided by the average peak widths. A sample calculation is shown in Equation 2 and the data obtained in this lab is shown in Table 10. The resolution obtained between Freon 22 and Freon 123 in the experiment was 1.08. The resolution states how much time was between the disappearance of Freon 22 and the appearance of Freon 123. A resolution of 1 says that there is no dead space between the peaks, that they occur one right after the other. A resolution of greater than one means there was dead time between the peaks, and a resolution less than one means that the peaks over lap and the flame does not go back to blue at all in between the two samples. Our data compared with our neighbors data is shown in Table 10. The numbers make sense, our gas flow rate is faster and should result in quicker numbers, whereas theirs in slower

and should result in slower numbers. This was what occurred. The next part of the experiment dealt with the commercial gas chromatograph. The homemade GCs injection was done using a syringe and it was depressed by the individual. The injection spot was marked by a dot on the latex tubing. The column was made of glass and was packed with tide, the length of the glass was 8mm. The detector was 40cm of copper wire coiled and stuck into the right angled glass tubing. The process for using this was that once the gas was injected and made its way through the glass column of tide, it would react with the Cu 0 and make the flame green. One of the partners timed how long it took to turn green, the time it took for make intensity and then the time for the color to disappear. The commercial GCs injection was also done using a syringe and was depressed by the individual. The injection spot was a copper pipe on the front of the GC. The column was a 2meter, polar graphite tube surrounded by stainless steel. The detector was an FID, Flame Ionization Detector that was 200oC and -690 voltages. The process for using the commercial GC was that you first made sure the handle was in the load position, then using the syringe collect 2mL head space of the sample. Then increase the syringe to 10mL (collecting air to dilute it), then depress the syringe to 3mL and then take in another 10mL of air to dilute it once more. Inject the sample into the injection spot, switch the handle to inject and press the spacebar on the keyboard. The computer then calculated the data and printed off the results. Components used in the commercial GC but not the homemade GC included, a GC machine, a computer, nitrogen gas, graphite column, a printer, an oven, and high temperature. Possible sources of error for the homemade instrument included, not packing the tide column well, not using exact injection spot every time, error in measuring the length of the instrument, the coiling of the copper wire, incorrect timing of intervals, incorrect amount of

sample gas. Possible sources of error using the commercial GC included, error in preparing diluted sample in syringe, timing of switching handle and pressing the spacebar, and incorrect calibration of machine. For the homemade GC, the identity of the compound was determined by reading the label on the flask. It could also be determined in the mixture because the data was already collected for the samples separately. The speed, timed in seconds, was shorter for the Freon 22 than for Freon 123. For the commercial GC, we identified the concentration of the compound injected by graphing Area vs. Concentration of the two Freons using the data on the printout (10). Using the trend line on the two graphs, plug the area obtained on the data for the mixture into the equation and solve for X, or the concentration (ppm). To know which equation to plug which area into one must know which area refers to Freon 22 and Freon 123. Using the information from the homemade GC, one would know that Freon 22 was quicker than Freon 123 thus it would come out of the column the quickest. This means that the first peak, closest to the top, represents Freon 22 and the second peak, or middle peak, corresponds to Freon 123. The retention times for Freon 22 and Freon 123 from the commercial GC are 1.07 minutes and 2.52 minutes respectively. This equals 64.2 seconds for Freon 22 and 151.2 seconds for Freon 123. These results for the retention times are extremely large compared to the retention times of the homemade GC. For the homemade GC, the retention times of the gases separately equals 3.7 seconds for Freon 22 and 9 seconds for Freon 123. Some reasons for the differences include a longer column in the commercial GC, which means that the sample has a longer distance to travel to get to the detector thus a longer retention time. Another reason could possibly be that there were different amounts of sample gas used, 10mL in the commercial and 3mL in the homemade. The homemade GC also did not have very accurate timing as compared

to the commercial so the timing could be off but not nearly that far off. After the area vs. concentration for Freon 123 for the commercial GC was plotted in Graph 2, and the trend line was created it could be used to determine the concentration of Freon 123 in the unknown sample. The concentration of Freon 123 was 391.5 ppm, a sample calculation can be found in Equation 3. After the area vs. concentration for Freon 22 for the commercial GC was plotted in Graph 1, and the trend line was created, it could be used to determine the concentration of Freon 22 in the unknown sample. The concentration of Freon 22 was 349.1 ppm, a sample calculation can be found in Equation 3. Peak intensities are not necessarily the same for different compounds of the same concentration. Different compounds means, that there could be different molecular weights, which means they would have different intermolecular forces. Therefore the compounds would move through the gas chromatograph at different speeds and lengths of time creating different peak intensities.

Conclusion:
As showed through the data obtained in this experiment, Freon 22 did in fact take less time to travel through the column in the commercial gas chromatograph and a shorter amount of time to turn the flame green in the homemade gas chromatograph. This supports the hypothesis stated at the beginning of the experiment. Many things about these two compounds contributed to this result, such as their molecular weight, boiling point, dipole moments, and London dispersion forces. This experiment also proved that although the homemade GC was much simpler and inexpensive than the commercial GC, it still did a great job separating the components that made up the halocarbons. The commercial GC although it provides a more accurate and efficient way to analyze the substances, it also costs a lot more than the homemade GC. Choosing between the two gas chromatographs really depends on how specific of results one wants to obtain and how much money one has to spend.

References:
(1) Thomas, Stephen; PSU Chemtrek, Small-Scale Experiments for General Chemistry; Hayden-McNeil Publishing, Plymouth, 2009, 15ed, pg. 19-12 to 19-21. (2) Miller, James M. Chromatography: Concepts and Contrasts; Wiley-Interscience Publication, New York, 1988, pgs, 1-4. (3) Richard S. Juvet, Jr., Gas chromatography, in AccessScience@McGraw-Hill, McGraw-Hill Companies, 2008. 12 February 2009. <Http://www.accessscience.com>. (4) Scott, Raymond P. W., The Importance of Chromatography as an Analytical Technique, Journal of Chromatographic Science. 2009. Library 4 Science. 22 February 2009. <http://www.chromatography-online.org/quant/Historical-Introduction/Importance.html>. (5) Driskell, Jennifer, Chemistry 113 Laboratory Notebook, pg. 9-14. (6) Lide, David R., CRC Handbook of Chemistry and Physics, 88 th Ed. CRC Press, Taylor & Francis Group, New York, 2008, Pg. 3-98 and 3-160. (7) Earnest, Arina, Chemistry 113 Laboratory Notebook, pg. 13-14. (8) Schiavone, Michael, Chemistry 113 Laboratory Notebook, pg. 13-17. (9) Doud, Jenna, Chemistry 113 Laboratory Notebook, pg. 7-10. (10) Printouts from the lab given to us before and after we analyzed our unknown sample