TITLE: Gas Chromatography OBJECTIVES OF THE EXPERIMENT:

1) To conclude the relation between tR and the number of atomic carbon in homolog series. 2) To explain the Van Deemter equation. 3) To describe the effect of flow rate on the efficiency of column. INTRODUCTION: Gas chromatography specifically gas-liquid chromatography - involves a sample being vaporized and injected onto the head of the chromatographic column. The sample is transported through the column by the flow of inert, gaseous mobile phase. The column itself contains a liquid stationary phase which is adsorbed onto the surface of an inert solid. In gas chromatography (GC), the stationary phase is a high-boiling liquid and the mobile phase is an inert gas. In the organic chemistry teaching labs at CU Boulder, GC is used as an analytical tool to find out how many components are in a mixture. It can also be used to separate small amounts of material. In gas chromatography, the moving phase ( "mobile phase") is a carrier gas, usually an inert gas such as helium or an unreactive gas such as nitrogen. The stationary phase is a microscopic layer of liquid or polymer on an inert solid support, inside a piece of glass or metal tubing called a column (a homage to the fractionating column used in distillation). The instrument used to perform gas chromatography is called a gas chromatograph (or "aerograph", "gas separator"). The gaseous compounds being analyzed interact with the walls of the column, which is coated with different stationary phases. This causes each compound to elute at a different time, known as the retention time of the compound.

The comparison of retention times is what gives GC its analytical usefulness. Gas chromatography is in principle similar to column chromatography (as well as other forms of chromatography, such as HPLC, TLC), but has several notable differences. Firstly, the process of separating the compounds in a mixture is carried out between a liquid stationary phase and a gas moving phase, whereas in column chromatography the stationary phase is a solid and the moving phase is a liquid. Secondly, the column through which the gas phase passes is located in an oven where the temperature of the gas can be controlled, whereas column chromatography (typically) has no such temperature control. Thirdly, the concentration of a compound in the gas phase is solely a function of the vapor pressure of the gas. Gas chromatography is also similar to fractional distillation, since both processes separate the components of a mixture primarily based on boiling point (or vapor pressure) differences. However, fractional distillation is typically used to separate components of a mixture on a large scale, whereas GC can be used on a much smaller scale (microscale). Gas

chromatography is also sometimes known as vapor-phase chromatography (VPC), or gas-liquid partition chromatography (GLPC). The carrier gas is an inert gas, helium. The flow rate of the gas influences how fast a compound will travel through the column; the faster the flow rate, the lower the retention time. Generally, the flow rate is held constant throughout a run. (The GCs at CU Boulder are set at a flow rate of 55 mL/min). The gas chromatography can be divided into column, detector, gas (flow rate) and temperature. The detector can be divided into FID, ECD, TCD, FTD that functioning based on the type of sample. The detector will combined all the compound in the peak form. Gas as mobile phase, for example helium and nitrogen. Column known as capillary which are different based on the polarity of sample used. The longer size of column, more better separation will

occur. It also based on the polarity of sample to known the time taken for separation. Other than that, the column can be divided into polar, semi polar and non polar. In the column has stationary phase, the sample will go through it and this phase will separated it. RESULTS: Part I Determination of tR for the homolog series Homolog Series Methanol Ethanol Butanol Number of Carbon 1 2 4 Mr (g/mol) 32.04 46.07 74.12 Boiling Point (C) 64.7 78.1 117.7 log tR -0.0969 -0.0980 0.0326

Mixture of alcohol Mixture Butanol + Methanol Butanol + Ethanol 0.802 1.002 -0.0958 0.001 6 120.19 tR1 (min) 0.803 tR2 (min) 1.035 log tR1 -0.096 log tR2 0.015 No. of C 5 Mr (g/mol) 106.16

Part II The effect of flow rate on efficiency column Flow Rate ( mL/min ) 20 30 40 50 80 tR (min) 1.278 1.078 0.918 0.815 0.650 w (min) 0.294 0.183 0.471 0.179 0.130 n 302.33 555.21 60.78 331.69 400.00 HETP (cm) 9.9 5.4 49.4 9.0 7.5

Effect of temperature on retention time of 2-propanol Temperature ( C ) 90 120 tR (min) 0.807 0.765

Part III Quantitative procedure in GC analysis A. Standard Sample Sample Octane Decane Napthalene Hexadecane Weight (mg) 6.6 3.3 3.1 5.0 tR (min) 8.323 11.43 15.688 21.282 Peak Area 42232 20997 27143 31051 Concentration (M) 12.5346 6.2321 8.0562 9.21620

B. Unknown Sample Sample 1 2 3 4 tR (min) 8.318 11.428 15.695 21.275 Peak Area 13261 17575 30400 18560 Concentration (M) 4.5452 6.024 10.42 6.3618

CALCULATION: Part II 1) =

2) n = 16 ( tR/w )2 3) HETP = ( L/n ) i. When flow rate = 20 mL/min 1) = w = 0.294 min 2) n = 16 3) HETP =
2

= 302.33

= 9.9 cm

ii. When flow rate = 30 mL/min 1) = w = 0.183 min 2) n = 16 3) HETP =

2

= 555.21

= 5.4 cm

iii. When flow rate = 40 mL/min 1) = w = 0.471 min 2) n = 16 3) HETP =

2

= 60.78

= 49.4 cm

iv. When flow rate = 50 mL/min 1) = w = 0.179 min 2) n = 16 3) HETP =

2

= 331.69

= 9.0 cm

v. When flow rate = 80 mL/min 1) = w = 0.130 min 2) n = 16 3) HETP = Part III A. Standard Sample Peak Ratio from Standard = 1) For naphthalene: Peak ratio = 2) For octane: Peak ratio = 3) For decane: Peak ratio = = 0.77 = 1.56 = 1.00
2

= 400

= 7.5 cm

4) For hexadecane: Peak ratio = B. Unknown Sample B a) Peak Ratio from Unknown Sample B = = 1.14

b) Concentration of Unknown Sample: = Concentration of Sample (standard)

1) By referring tR of standard for naphthalene: a) Peak ratio = b) Concentration = = 1.00 8.0562 = 8.0562 M

2) By referring tR of standard for octane: a) Peak ratio = b) Concentration = = 0.44 12.5346 = 44.44 M

3) By referring tR of standard for decane: a) Peak ratio = b) Concentration = = 0.58 6.2321 = 8.27 M

4) By referring tR of standard for hexadecane: a) Peak ratio = b) Concentration = = 0.61 9.2162 = 17.22 M

DISCUSSION: In the experiment, retention time plays big role in analyzing the sample in the gas chromatography. For the first part of the experiment, retention time are differentiated according to the number of carbon, molecular weight and the boiling point of the samples. Graphs plotted from these readings that obtained from the chromatography shown that log tR is proportionally increase when number of carbon is increased. It goes the same with molecular weight and boiling point, log tR increase when the molecular weight and boiling point of the compound increased. This data concluded to the relationship that when the number of carbon in samples tested increased, their molecular weight also increased, and so with their boiling point. Therefore, the retention time, time for a maximum of symmetrical peak to occur is also increased. In Part II, the effect of flow rate on column efficiency were determined by relating the retention time with the HETP (height equivalent to a plate). The graph of flow rates versus HETP plotted is inversely proportional which meant that the efficiency of column is greater when slower flow rate is used. Other than that, retention time is lower when the temperature of the same compound tested is increased. In Part III, quantitative procedure in gas

chromatography analysis was carried out. In this part, the concentration of the unknown samples are determined by using an internal standard as reference. The peak ratio from the standard solution prepared are used to calculate the unknown samples.
The process of gas chromatography is carried out in a specially designed instrument. A very small amount of liquid mixture is injected into the instrument and is volatilized in a hot injection chamber. Then, it is swept by a stream of inert carrier gas through a heated column which contains the stationary, high-boiling liquid. As the mixture travels through this column, its components go back and forth at different rates between the gas phase and dissolution in the high-boiling liquid, and thus separate into pure components. Just before each compound exits the instrument, it passes through a detector. When the

detector sees a compound, it sends an electronic message to the recorder, which responds by printing a peak on a piece of paper. The relationship between tR. and the number of atomic carbon in homolog series is supposed to be more increase the number of atomic carbon, more increase the tR. The retention time is corrected by subtracting the value of actual retention time with the retention time of air. Unfortunately, the peak for air is not visible when ionization flame detector is used. As can be expected, the retention timed decreases with the increase of temperature. When the temperature used is 90C, the retention time is 0.807 min while for 120C, the retention time is 0.765 min. Even though the high temperature is needed to speed the analysis, lack of efficiency would cause the peaks to close together. The effect of flow rate on the efficiency of column can be determine by the Van deemter equation which is :

The form taken by f2(k') was considered by Van Deemter to be,

where H is a column plate height (also known as height equivalent to one theoretical plate, H.E.T.P.), u is average velocity of a carrier gas, and B and C are constants that depend on column dimensions, carrier gas type, and, to a smaller degree, on other factors. Explicit equations are derived for the B and C terms in the Van Deemter equation for liquid chromatography, involving diffusion coefficients in the mobile zone (eluent outside the particles), the stagnant mobile phase (eluent inside the particles) and the stationary phase. Longitudinal diffusion rates (B term) are determined by the arrested elution method for simple aromatic solutes in methanol and methanol-water mixtures with ODS-Hypersil as packing material. When

compared to diffusion rate for the same solutes in eluent, measured by the open-tube method, the rates of diffusion in the stationary ODS phase are about half those in methanol-water mixtures. C values are obtained by measuring the band-dispersion when the same solutes are eluted from 50-m and 540-m ODS-bonded silica gels at high reduced velocities. After allowing for dispersion due to processes in the mobile zone, the C values are obtained and found to be of the magnitude predicted by the non-equilibrium theory of Giddings, taking into account the actual rates of diffusion of the solutes in the stationary zone.

PRECAUTION STEPS: 1. Shake all the solution adequately to ensure dissolution and degas for 5 min before injection into chromatograph. 2. Make sure that the column is not clogged, it maybe cause by precipitation of proteins in the column caused by removal of stabilizing agents during fractionation. 3. Make sure rinse and clean the syringe before and after each sample to prevent any contaminant and others solution that will affect the process. 4. Avoid formation of bubbles during the experiment because it will affect the reading in the column while we taken the measurement. CONCLUSION: 1) The concentration of unknown samples in this experiment were determined by using gas chromatography quantitative determination. 2) The retention time increased when the number of carbon is increased. 3) The Van Deemter equation can be used to explain the HETP and the effect of flow rate on the efficiency column. 4) The faster the flow rate, the less the columns efficiency. The HETP decreased when the flow rate increased.

REFERENCES: 1.