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Group 4

SALMI SEPRIANTI (1310411054)

1. How to make the unseen spots from non-colored substance become visible?
And explain the steps concisely!
a. Using fluorescent, put the Chromatogram under UV light, mark the visible
spots of the substances and circle the spots by using a pencil.
b. Reacting the substances with chemicals to produce a colored product. A good
example is
thechromatogram produced from a
of amino acids. Chromatogram can
be dried and sprayedwith ninhydrin solution. Ninhydrin reacts with amino acid
s to produce colored compounds,generally brown or purple.
2. Why is in practice, normal phase ion pair partition chromatography is rather
Because of stripping of the stationary phase and necessity of a precoloumn
loaded with the aqueous phase.
3. What is the obstacles that often find in anion chromatography separation ?
In anion chromatography separation , column separators used only one to
determine anions. Application of this method is limited in a small number of
anions alone . Limitations that, when the number of common anions (common
inorganic anions ) that commonly arise in natural water samples to multiply, then
the system is not suitable anymore.


1. What is the principle of liquid chromatography?
Answer : Partition of analyte between liquid mobile phase and immobilized
liquid stationary phase; partition, adsorption, ion exchange, or size exclusion.
2. Mention and explain about the application of size exclusion chromatography!
Answer : Size exclusion chromatography commonly used for biochemical
application such to assay protein tertiary structure and SEC can be used as a
measure of both the size and the polydispersity of a synthesised polymer, that
is, the ability to be able to find the distribution of the sizes of polymer
3. Mention the advantages and disadvantages of column chromatography!
Answer :
Advantages are Less expensive procedure than other methods of separation
(HPLC, GC, etc.); Do not require the help of expensive machinery like high
pressure solvent pumps used in HPLC; Widely used in industry because it is
possible to scale it appropriately and it can be automated; Is used to separate
and purify the substance
Disadvantages are Difficult to separate two very similar compounds (often
with proteins); Need to prepare the column with the appropriate resin; This
method is very time consuming; Limited selection of stationary phase

Group 4

RAHMI ANNISA (1310411106)

1. In the high performance liquid chromatography, if it can not be used

resistance time? And also mentioned application if high performance liquid

chromatography in the environment?
In high performance liquid chromatography resistance time should be used,
because resistance is time itself is the time required for the compound
through the column to the detector. Time is measured from the time in which
the sample is injected until the samples showed a maximum peak height.
High performance liquid chromatography applications in an environment that
can be used to analyze substances contained in acid rain, such as H2SO4 and
2. Why Ion pair liquid-liquid or partition chromatography has largely been

replaced today by bonded phase chromatography and give the principle of

reversed phase ion partition chromatography
Because of ease of manipulation of bonded phases. Principle is In reserved
phase ion pair partition chromatography a chemically bonded silica support
such as ODS-silica is used alongwith an aquous phase containing the required
acidic or basic counter ions. Thus the nature or concentration of the counter
ions can be easily changed. This also overcomes the problems of reproducibly
coating the support with aquous phase and the additional problems of
bleeding ion pairs are now partitioned into the stationary phase and the free
ions eluted by the mobile phase.
3. Explain type of chromatography based on shape of stationary phase

1. Planar
Here the stationary phase is located on a flat plate or porous paper, the
mobile phase moves through the stationary phase due to capillary action
or by gravity. In the column chromatography, the stationary phase is
located within the capillary tube and the mobile phase moves through the
stationary phase due to pressure or by gravity.
2. Column
Included into the gas chromatograph column chromatography, the analyte
is injected at the injection point and then carried by the mobile phase to
the column that is located inside an oven whose temperature is
Separation occurred within a column, analyte or component that will
evaporate more quickly achieve faster exit detector, while the component
that has a higher vapor point will come out more slowly from the column.
1. How to detect the presence of the bands as in HPLC?
This using UV-Vis Spectroscopy

Group 4
2. Silica gel is the one of most popular adsorbents used. Why?
Because its high sample capacity.
3. What is other varieties of LC that have stationary liquid would be an
immiscible liquid with mobile phase?
Partition chromatography.

FAJAR NURRAHMAN (1010412050)

1. What is the difference between thin-layer chromatography paper chromatography ?
Differences thin layer chromatography with chromatography paper lies in the use phase
of the silence , in which the stationary phase thin layer chromatography has a higher
resolution for use adsorbent separate samples / compounds based on polarity , while the
paper chromatography using filter paper as the phase of the silence , where the separation
is based on the style kapilaritasnya,
In the thin layer chromatography stationary phase used the silica gel or alumina where
the silica gel so that it is highly polar polar footage bersifar be adsorbed on this adsorbent
while excerpts nonpolar will continue to flow with the mobile phase.
2. Explain what affects the strength of the ion exchange chromatography ion exchange ?
Answer :
The strength of the cation exchange is affected by various factors , the greater the load
the stronger exchange capacity . total number of exchanged cations will be proportional
to the total hydrogen ions are released resin .
3. What is the retention time in HPLC ?
Answer :
Answer : The retention time is the time required by the compound to move through the
column towards the detector . Different compounds have different retention times .