R.A. van Iterson Drenthe College Emmen Holland for www.standardbase.

com

Theoretical Consideration: Qualitative Analysis Kovats index

The first thing to do when you have a sample separated into

components is to identify the peaks. The Kovats retention index is a
mathematical aid to help identify each compound.

CONTENT
1. Qualitative Analysis
2. Retention Time
3. Retention Index

1. Qualitative Analysis
Chromatography is widely used for recognizing the presence or absence
of components in mixtures that contain a limited number of possible
species whose identities are known. Numerous spectrometric methods for
identifying these compounds are available. For example, often the
compounds are passed into a mass spectrometer.

2. Retention Time
The simplest way to identify a compound that gives a chromatographic
peak is to compare its retention time with that of a sample of the
suspected compound. The most reliable way to do this is by co-
chromatography in which the compound is added to the unknown. If this
compound is a component of the unknown, the area of its peak will
increase with respect to the others in the chromatogram. This identification
is only tentative when it is carried out with a single type of column, but it is
nearly conclusive when carried out with two or more different kinds of
columns. It is unlikely that two compounds with the same retention time on
one stationary phase will have the same retention times on two different
stationary phases.

3. Retention Index
Methods based on relative retention are inaccurate for reporting
chromatographic data which is to be used for interlaboratory substance
identification. In gas chromatography, the retention index system is used
for this purpose. One common definition is the Kovats index, a
logarithmic scale on which the adjusted retention time of a peak is
compared with those of unbranched alkanes.

Kovats (1958) devised a system of indexing chromatographic retention

properties of a stationary phase with respect to the retention
characteristics of unbranched alkanes. These are used as reference
materials since they are non-polar, chemically inert and soluble in most
common stationary phases.
R.A. van Iterson Drenthe College Emmen Holland for www.standardbase.com

The retention behavior is expressed on a uniform scale determined by a

series of closely related standard substances, for example n-alkanes. The
Kovats index, I, for the unknown is calculated from the formula:

log tr' (unknown) − log t r' (n)

I = 100[n + ( N − n) ]
log tr' ( N ) − log tr' (n)
where
n is the number of carbonatoms in the smaller alkane
N is the number of carbonatoms in the larger alkane
t r' is in all cases the adjusted retentiontime (measured time minus the
time of the undelayed methane or small compound.

This definition makes the Kovats index for a linear alkane equal to 100
times the number of carbon atoms. For octane I = 800, and for nonane I =
900. A graph of log of retention time against carbon number, Z, for a
homologous series will be linear since there is an almost uniform increase
in boiling point between members of a given series. A graph of the log of
corrected retention time against the retention index, for several alkanes
with retention times similar to the compound, is constructed. Log t'R of the
compound is then noted on the graph and the retention index determined.
See figure 1 below.

The retention index of an unknown, measured on several different

columns, is useful for identifying the unknown by comparing it with
tabulated retention indexes. For a homologous series of compounds
(those with similar structures, but differing by the number of CH2 groups in
a chain, such as CH2CO(CH2)nCO2CH2CH3) log t'R is usually a linear
function of the number of carbon atoms. A further useful indication of
retention characteristics may be obtained by analysing a sample with a
dual column GC fitted with a non-polar and polar column.

It is important to note that while a chromatogram may not lead to positive

identification of the species in a sample, it often provides sure evidence of
the absence of species. Thus, failure of a sample to produce a peak at the
same retention time as a standard, obtained under identical conditions, is
strong evidence that the compound in question is absent (or present at a
concentration below the detection limit of the procedure).

Its more common to make a calibration line for at least four n-alkanes. The
unknown must lie in between these retention times. The number of C-
atoms (x100) is plotted on the x-axis and the net-retention time of the
compounds on the y-axis. So when for n-nonane a retention of 234 s. is
measured and for methane (unretained partical) 12 s. is measured, then
R.A. van Iterson Drenthe College Emmen Holland for www.standardbase.com

log t’R= log(234-12). When an unknown compound is measured, it is

necessary to calculate log t’R also and read the x-value from the line as
shown below.

Figure 1. Determination of retention index for X, Z1... Z4 are

retention index values of alkanes

Remarks
1 For better accuracy it is advised to use at least 4 n-alkanes and
calculate the Kovats index graphically.
2 The Kovats method of identification is also applicable on fatty-
acids (methylesters) to identify cheese or olive-oil
3 When you find an index of 1040 this will not state the compound
has 10,4 C-atoms. It merely tells you it has this retention index
in the Kovats-scale. As an example : 3,3-dimethyloctane will
have a different index from 2,7-dimethyloctane. although the
number of carbon atoms and molecular-mass are equal.
4 Remember the Kovats index is not absolute. It depends on
flowrate and column stationary phase.