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Chem275L | 34


EXPERIMENT 9
Gas Chromatography of Diesel Fractions
(Groups of 4 students)
Background Reading:
Christian, G. D., Analytical Chemistry; 6
th
ed.; Wiley & Sons: New York, 2003; pp 555-562, 568-570
& p. 574-588
Introduction:
The purpose of this experiment is to learn how to use a GC instrument to separate and detect the
volatile components of a Diesel fraction and how the GC instrument can be used to identify and
quantify a compound using the method of internal standard.
In GC, the liquid sample is injected into a separation column as a sharp plug. The sample is
vaporized to a degree and at a rate dependent on its boiling point. Boiling points are often used as a
measure of the relative volatilities of compounds in a mixture. However, volatility alone does not
determine the distribution between the stationary and mobile phase. Specific interactions between the
injected compound and the stationary phase play an important role as well. Since, the components of
the analyzed diesel fraction in this experiment are alkanes their separation on the GC column will be
based on the boiling point alone.
Quantization is possible in GC methods by analyzing the peak area. A larger peak area indicates a
larger amount of analyte present in the sample. Spiking samples with internal standards helps to
compensate for the imprecision inherent to GC methods (e.g. variable injection volumes). In this
method, the analyte sample is spiked with a fixed amount of an internal standard, whose retention time
is similar to (but resolved from) that of the sample.
In this experiment, first an analyte sample (X) of known concentration spiked with a fixed amount
of internal standard (S) is analyzed by GC to determine the response factor of the analyte to the
instrument. Second, an analyte of unknown concentration spiked with the same amount of internal
standard is analyzed to determine the concentration of the unknown.
Diesel fuel contains a mixture of aliphatic hydrocarbons with chain lengths ranging from 7 carbons up
to 30 carbons, with the majority in the C
10
C
20
range. For this experiment will be used a fraction of diesel
containing only ten hydrocarbons. The gas chromatographic retention time, t
R
, of an analyte depends
primarily on its volatility and its degree of interaction with the stationary phase. Based on the volatility
of the analyte, the retention time of a compound will increase with its boiling point. The strength of
interaction between analyte and the stationary phase is determined by the polarities of both the analyte
and the stationary phase. Thus, the retention of a polar analyte increases with the polarity of the
column. This principle can be used to identify an unknown analyte by determining its retention on
several stationary phases of different polarities.
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Chem275L | 35

Kovat's retention index or simply Kovat's index, I, is a method of quantifying the
relationship between the adjusted retention time of compounds in GC and their structures in such a
way as to help identify the components of a mixture. It's applicability is restricted to organic
compounds. The method takes advantage of the well-established linear relationship between the values
of log(t
r
') and the number of carbon atoms in a molecule. The time analytes spend in the stationary
phase (t
R
) increments by a constant factor per additional carbon atom or, more specifically, per
methylene group, CH
2
. This can be described to the constant change in the analyte volatility and
strength of interaction with the stationary phase per additional CH
2
. Thus, the change in the log t
R
per
carbon atom in a molecule is a constant for a particular GC system.
Note: If no any programming temperature is used, the series of n-alkane homologues elute very
precisely in logarithmic shape. Under programmed conditions - by flow or temperature programming -
the homologue series elutes nearly in a linear shape but NOT precisely linear enough.
By definition, Kovts index for a linear alkane equals 100 times the number of carbons atoms (I =
100* number of C atoms). Hence, I for n-octane is always around 800, for nonanene ~900 etc. Some
differences from these values will be register in function of the nature of the column used to elute these
compounds. A compound eluted between octane and nonane has a retention index between 800 and
900. A retention index value may be calculated for a peak of an unknown compound (C
unknown
) by
comparing its retention characteristics to those of the two closest eluting components analyzed under
identical conditions, using different mathematical equations based on the type of GC analysis. For
temperature programmed chromatography, as the one you will perform in this experiment, the Kovats
index is given by the equation:
)
1
( 1
)
1
( ) (
) (
1 1 1
) ( 100 100

+ =
+
n
C n
n
C
R C R
R R
n n n
t t
t t
C C C I
unknown
C
(Eq. 1)
Where: t
R
refers to the adjusted retention times,
C
unknown
is the number of carbon atoms of the alkane of interest (unknown)
C
n-1
is the number of carbons in the alkane eluting before the component of interest,
C
n+1
is the number of carbon atoms in the alkane eluting after alkane of interest.
) (
1 ) ( )
1
( +
< <
n n
C
C R R R
t t t
unknown
C
(Eq. 2)
The Kovts index is not absolute. It depends on flow rate and column stationary phase. When
quoting retention indexes, the stationary phase used in the column must be quoted. Use of GC retention
indexes, particularly in combination with mass spectrometry can provide substantial information about
trace amounts of unknowns. Comparison of Kovts indexes (I) of an unknown with those of standards
can provide an unambiguous identification, or at least, limit the number of possible structures to a few
compounds.
Example of calculation:
A temperature programmed chromatogram contains three peaks: the first peak (t
R
=1.10 min) is for
C
10
, the second peak (t
R
=1.53 min) is for C
12
and the third peak (t
R
=2.10 min) is for C
14
. The
unretained peak is at 0.76 min. The Kovats index, I, for C
10
cannot be calculated, since there is not a
peak before C
10
, and the Kovats index, I, for C
14
cannot be calculated, since there is not a peak after
C
14
. The Kovats index, I, for C
12
will be:
1172 )
10 . 1 10 . 2
10 . 1 53 . 1
)( 10 14 ( 100 ) 10 ( 100 =
(

+ = I

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Chem275L | 36

Procedure:
Sample preparation:
You will be using three samples in this experiment but no preparation needed from your part::
Sample A: is a standard fraction type of diesel fuel (UST Modified DRO, cat. No. 48166, Sigma-
Aldrich) containing 10 hydrocarbons (in the range C
10
-C
28
), listed in the Table 1.
Sample B: is a mixture in hexane of an analyte compound (X) and an internal standard (S), both of
them with the concentration of 1000 g/mL.
Sample C: is a mixture in hexane of the analyte compound (X) of unknown concentration and an
internal standard (S) of concentration 1000 g /mL.
Instrument Preparation & Sample Injections:
Follow the instructions for the use of the Hewlett-Packard 5890A Gas Chromatograph
instrument presented in your laboratory manual to analyze the samples you received. Make two
injections for the sample A and only one injection for each of the samples B and C.

Place the following information in the Observations section of your report.
a. Name and Model of Instrument: _________________________
b. Column type: _____________________________________________
c. Carrier Gas (mobile phase): _____________________________
d. Column dimensions: length (L) =________ & interior diameter (d) =__________
e. Volume Injected: _______________
f. Injection port temperature: __________
g. Detector Temperature: ___________________
h. Flow rate in mL/min:_______________________
i. Unretained (void) time (t
M
):
min) / (
) ( ) ( 500 . 0
2
2
mL rate flow
cm d cm L
t
M
=
=____________________, min
j. Parameter for the Temperature Program:
(1) Oven initial temperature: ______________
(2) Initial hold time: __________
(3) Ramp rate : __________
(4) Final oven temperature: ______________
(5) Final hold time: __________________
Treatment of Data: (place this section in the Results section of the Report)
A. Use thesamples A chromatograms to do the followings:
a) Mark clearly the identity of the components (i.e. write down the name of each component for
the corresponding peak on the chromatogram) in each chromatogram obtained from the sample
A injection and determine the retention times for each compound present in the mixture. Attach
your marked Chromatograms for Sample A and GC reports to the final report.
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b) Record the retention times (t
R
) of the identified components in the column 4 of Table 1.
Calculate the adjusted retention times (t
R
) and logt
R
and fill out column 6 and 7 of Table1.
c) Determine the Kovts indexes, I, using the equation (Eq.1) and fill out column 8 of Table 1.
Calculate the Kovats indexes values by multiplying the number of carbons atoms with 100 and
compared them to the one obtained using Eq. 1. [Fill out Column 9 in Table 1].
d) Construct a plot of logt
R
versus number of carbons of the alkane homologue series you
identified in diesel fuel. Insert your graph in the results section of the report.
e) Construct a plot of the Kovats index (I) versus number of carbons of the alkane homologue
series you identified in diesel fuel. Insert your graph in the results section of the report.
f) Compare your graphs form (i) and (ii) and comment on them.

Table 1:
1 2 3 4 6 7 8 9

Compound
*

MF

b.p.
(
0
C)
**
Retention Time,
t
R
(minutes)
Adjusted, t
R

(minutes)
(t
R
= t
R
t
M
)
logt
R
(ave)

Kovt
Index
(from
Eq. 1)
Kovt
Index
(from
I = 100 x
# of C
Injection 1 2 Ave. 1 2 Ave.
Unretained
peak (t
M
)
----------- --------- ------ ---------- ---------
Hexane
(solv.)
C
614
68 ------- ---------- ---------
Decane C
10
H
22
174
Docosane C
22
H
46
369
Dodecane C
12
H
26
216
Eicosane C
20
H
42
343
Hexacosane C
26
H
54
412
Hexadecane C
16
H
34
287
Octadecane C
18
H
38
317
Octacosane C
28
H
58
432
Tetracosane C
24
H
50
391
Tetradecane C
14
H
30
254
*The compounds are listed in alphabetical order.
**B.P. collected from: http://en.wikipedia.org/wiki/Nonadecane

B. Use the sample B chromatogram and do the followings:
a) Label the compound X and the compound S on the chromatogram of sample B. Hint:
Compound S and X are compounds from the homologue series identified in the diesel fraction
of sample A and #C atoms in X are higher than #C atoms in S.
b) Calculate the values of logt
R
for each component on the chromatogram.
1. Retention time of the unknown compound (X):___________________
2. Retention time of the internal standard (S):____________________________
3. Logarithm of adjusted retention time (logt
R
) of X:________________________
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4. Logarithm of adjusted retention time (logt
R
) of S:___________________________

c) Use the retention time of each component and one of the sample A chromatogram and determine
the Kovats index for X and S:
1. Kovats index of the unknown compound (X):___________________
2. Kovats index of the internal standard (S):________________________
3. Use one of the two graphs linear equations (the one which is giving a better correlation
factor), you have prepared using data from sample A chromatograms and determine the
number of carbons and the identity of the unknown compound (X) and the internal standard
(S) in the sample B:
4. The name of the unknown compound X identified is: ____________________________
5. The name of the internal standard (S) is: _____________________________________

d) Determine the response factor (F) of the unknown compound (X) relative to the internal
standard (S):
1. Peak area of the unknown compound (A
X
) =________________.

2. Peak area of the internal standard (A
S
) =__________________.

3. Area peak ratio (A
X
/ A
S
) =______________________________________.

4. Concentration ratio (C
S
/ C
X
): =___________________________________.

5. Response factor:
X S
S X
C A
C A
F =
=_____________________________.
C. Use the data from the sample C chromatogram and the response factor (F) calculated above, to
determine the concentration of the unknown compound (X) in the sample C:
1. Peak area of the unknown compound (A
X
) =________________
2. Peak area of the internal standard (A
S
) =__________________
3. Area peak ratio (A
X
/ A
S
) =______________________________________
4. Concentration (C
X
) =_______________________, g/mL; _________, in ppm.

D. Attach your marked Chromatograms and GC reports to the final report.

Postlaboratory Questions:
1. Calculate the resolution of the column used in this experiment for the two first peaks and for the
last two peaks in the sample A chromatogram (use one of the two recorded). The formula needed is:
(5 pts)
2 / ) (
2 1
1 2
) ( b b
R R
av b
R
w w
t t
W
t
R
+

=
A
=

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Chem275L | 39

2. Give two major advantages in the use of capillary columns compared to packed columns in the
analysis of complex mixtures and explain these advantages. (4 pts)
3. What are the most important properties of a compound used in the GC analysis? (1 pt)
References:
Marques, F., McElfresh, J .S.,Millar, J .G, J. Braz. Chem. Soc., Vol.11,No.6,592-599, 2000.
http://en.wikipedia.org/wiki/Kovats_retention_index
http://www.interchromforum.com/html/body_ql_err_gc.html