SEPARATION OF Ni2+ and Co2+ IONS IN A MIXTURE BY PAPER
CHROMATOGRAPHY
Apparatus Required:
Gas jar,
glass rod,
filter paper strip (What man No. 1 filter paper),
jar cover,
Fine capillary tube.
Chemicals Required:
Sample solution containing cobalt (II) and nickel (II) ions,
Acetone,
Concentrated aqueous ammonia,
Rubeanic acid spray reagent.
Principle:
Chromatography is a term used to describe several laboratory techniques that separate mixtures on
the basis of their differing affinities for a stationary phase. It is one of the most common chemical
and biochemical separation techniques. In a variety of chromatographic methods, chemists use
liquid solvents or mobile phase to move compounds over a stationary phase (a solid, gel, or
polymer). Paper chromatography (PC) is a technique similar to the widely used thin-layer
chromatography (TLC). In TLC, silica gel – a material commonly used as a stationary phase – is
coated onto a glass or plastic sheet. In PC, the mixture is spotted onto a sheet of paper. Both of
these techniques can be used to help to identify an unknown in solution.
Paper chromatography works because of the capillary action of aqueous solution or organic
solvent, in the paper. Capillary action is defined as the movement of liquid within the spaces of a
porous material due to the forces of adhesion, cohesion, and surface tension. The liquid is able to
move up the filter paper because its attraction to itself is stronger than the force of gravity.
Compounds that are not soluble in the solvent or are very attracted to the stationary phase stay in
place. Compounds that are very soluble and less attracted to the stationary phase travel up the
stationary phase with the solvent. Some compounds are somewhat soluble and somewhat attracted
to the stationary phase. They travel up the paper, but at a slower rate.
The chromatography process comprises of a stationary phase, a mobile phase and analyte (i.e.
chemical to be separated). The analytes are generally placed on the stationary phase, with which
they have some attractive interactions (absorption to the surface). A mobile phase is then
introduced, for which they also have an attraction. As the mobile phase moves across the stationary
phase, it carries the analytes along with it. Those with greater affinity for the mobile phase will
move farther than those having affinity for the stationary phase. For a system, retention factor Rf
is the characteristic measurement for an analyte, in terms of which the a given analyte can be
identified.
Retention factor is calculated by measuring the distance travelled by spot and the distance travelled
by the solvent, and putting in the following equation:
𝐷𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑓𝑟𝑜𝑚 𝑜𝑟𝑖𝑔𝑖𝑛 𝑡𝑜 𝑐𝑒𝑛𝑡𝑒𝑟 𝑜𝑓 𝑠𝑝𝑜𝑡 (𝐷𝑠𝑝𝑜𝑡 )
𝑅𝑓 =
𝐷𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑓𝑟𝑜𝑚 𝑜𝑟𝑖𝑔𝑖𝑛 𝑡𝑜 𝑠𝑜𝑙𝑣𝑒𝑛𝑡 𝑓𝑟𝑜𝑛𝑡 (𝐷𝑠𝑜𝑙𝑣𝑒𝑛𝑡 )
Procedure:
1. Take a What-man filter paper strip (20 x 2 cm) and draw a line with pencil above 3 cm
from one end. Draw another line lengthwise from the centre of the paper.
2. With the help of fine capillary tube, put a drop of the mixture of red and blue inks at the
point P. Let it dry in air. Put another drop on the same spot and dry again. Repeat 2-3 times,
so that the spot is rich in mixture.
3. Suspend the filter paper vertically in a gas jar containing the solvent (eluent) with the help
of a glass rod in such a way that the pencil line (and the spot) remains about 2 cm .above
the solvent level (50% alcohol + distilled water).
4. Cover the jar and keep it undisturbed. Notice the rising solvent along with the mixture of
red and blue inks. After the solvent has risen about 15 cm you will notice two different
spots of blue and red colours on the filter paper.
5. Take the filter paper out of the jar and mark the distance that the solvent has risen on the
paper with a pencil. This is called the solvent front.
6. Dry the paper. Put pencil marks in the centre of the blue and red spots.
7. Measure the distance of the two spots from the original line and the distance of the solvent
from the original line.
8. Calculate the Rf values of the blue and red inks by using the formula.
9. After elution and drying, place the paper in a large, dry, covered beaker containing a
smaller beaker of concentrated aqueous ammonia. After about two minutes, remove the
paper and spray it on both sides with rubeanic acid reagent. Allow it to dry. Nickel becomes
visible as blue purple band while cobalt becomes visible as yellow orange band. Evaluate
Rf values of the two ions.
Observation and Calculation Table:
Metal Ion Color of Distance travelled by Distance travelled by Rf value
Spot Spot solvent
Ni2+ Blue purple
Co2+ Yellow
orange
Results:
Chromatography was successfully used in identifying the components of the unknown solutions,
which were found to be Ni2+ and Co2+. These ions were identified using their respective retention
factor and color of spot they formed.
