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CHROMATOGRAPHY QUESTION

1. You inject your sample into the GC and then wait several minutes, but you see no
peak. What should you do?

2. How would you use gas chromatography to detect compounds with melting point
greater than 250 dgrees C?

3. Why would gas chromatography be unsuitable for certain vitamins or


substances?
4. In gas chromatography, what would the effect of raising the column temperature
or increasing the carrier gas flow rate on the ability to resolve two closely spaced
peaks?
Temperature programming separates species with wide range of polarities or
vapor pressure, once the temperature programming is used, sample with broad
boiling range can be separated well.
The high flow rate could reduce molecular diffusion
5. Why are the plates in column chromatography called 'theoretical plates'?
6. What is the Significance of N, the Number of Theoretical Plates for a Given
Column?
The N is does not really exist, they are a figment of the imagination that helps
us understand the processes at work in the column. They are also serves as a
way of measuring column efficiency, either by stating the no of theoretical
plates in a column, N (the more plates the better), or by stating the plate
height, the height equivalent to a theoretical plate (the smaller the better).

7. In GC why is it important to use temperature programming when using a splitless


injection?
8. What is the effect of physical adsorption on polar analyte species in Gas
Chromatography?
A real problem in both LC and GC has been the physical adsorption of polar
or polarizable analyte species such as alcohols or aromatic hydrocarbons, on the silicate
surfaces of column packings or capillary walls. Adsorption results in distorted peaks, which
are broadened and are tailed. Adsorption is the consequence of residual silanol groups that
are present on the surface of silicates. The silanol (SiOH) groups on the support surface or
walls of capillary columns have a strong affinity for polar organic molecules and tend to
retain them by adsorption. Adsorption on support materials can be deactivated
by silanization with dimethylchlorosilane(DMCS).
9. What is the function of sample splitter in Gas Chromatography?

10. Shown below are two chromatograms of the same set of analytes obtained using
the same GC system.

a. Which separation above was conducted using isothermal conditions and


which was conducted using programmed temperature GC?
Upper temperature programming
Below isothermal conditions
b. Explain why the two different temperature conditions during the analysis give
widely different retention times, especially for the late-eluting compounds.
Gas chromatographs are usually capable of performing what is known as temperature
programming gas chromatography (TPGC). The temperature of the column is changed
according to a preset temperature isotherm. TPGC is a very important procedure which is used
for the attainment of excellent looking chromatograms in the least time possible. For example,
assume a chromatogram obtained using isothermal GC at 80 oC, as shown below:

Temperature isotherms of practically any shape can be constructed to affect suitable


separations. If it is required to separate some overlapping peaks, it is always wise to

reduce the temperature rather than flow rate because flow rate is troublesome to
adjust. It is also observed that broad peaks obtained in isothermal separations are
changed into sharp looking peaks in TPGC which is also an additional advantage of
the technique.