Aim: To separate components from black ink

Apparatus required:chromatogram paper,black ink,jar,fine capillary tube,glass rod,water,pencil,scale

Theory

 The word chromatography originated from two greek words 'chroma' meaning 'colour' and 'graphine'
meaning 'to write'. Chromatography means colour writing and it was first employed by a Russian
scientist Mikhail Tsvet. This method was first used for the separation of coloured substances in
plants.
 In the chromatographic technique, the mixture of substances is applied onto a phase called the
stationary phase. The stationary phase may be solid or liquid. A moving phase that can be a pure
solvent or a mixture of solvents, or a gas is allowed to move slowly over the stationary phase. This
moving phase is called the mobile phase. When the mobile phase is moved over the mixture on the
stationary phase, the components of the mixture gradually separates from one another.
 Depending on the basic principle involved in chromatography, it is mainly classified into two: Adsorption
chromatography and Partition Chromatography
 Adsorption chromatography is based on the differential adsorption of the components on the
adsorbent (stationary phase). This means that different compounds are adsorbed on an adsorbent at
different degrees. Partition Chromatography is based on principle that there is the continuous
differential partitioning of components of a mixture between the stationary phase and the mobile phase.
An important partition chromatography is Paper Chromatography.
 In paper chromatography, the stationary phase is a special quality paper called chromatography paper.
Mobile phase is a solvent or a mixture of solvents. A solution of the mixture is spotted on a line about 2
cm above from the bottom of the paper, called original line or base line and then suspended in a
chromatography chamber containing suitable solvent. The solvent rises up the paper by capillary action
and flows over the spot. The paper selectively retains different components according to their differing
partition in the two phases. The paper strip so developed is called Chromatogram. The spots of the
separated coloured compounds are visible at different heights from the position of initial spot on the
chromatogram. The spots of the separated colourless components may be observed either under
ultraviolet light or by the use of an appropriate spray reagent. The distance travelled by the solvent from
the original line is called solvent front. The relative adsorption of each component of the mixture is
expressed in terms of its Retardation factor (Rf) (Retention factor).

 Rf values are always less than 1. An R f value of 1 or too close to it means that the spot and the solvent
front travel close together and are therefore unreliable.
 The type of paper chromatography in which the solvent rises up is called Ascending paper
chromatography. Alternatively, the solvent may be taken on the top in a container and be allowed to
come down, in which case it is termed as Descending paper chromatography.
 Some factors that affect the Rf value of paper chromatography are the solvent
system, Composition of the mobile phase, The working temperature of the system, The
quality of the paper used, The distance through which the solvent runs, The quality and
nature of solvents used, The polarity of components, The pH of the solvent or mobile
phase, The thickness of the stationary phase and The concentration of the sample or spot.
  In the case of paper chromatography, the retention time is inversely proportional to the
temperature. If temperature increases, retention time decreases and vice versa. The
temperature increases the solubility of the substance in paper chromatography.

Procedure:

 A thin strip of filter paper is cut out and a line is drawn on it approximately 3 cm above the lower
end of the filter paper. This line is called reference line.
 A small dot of black ink is placed in the centre of the line drawn.
 Once the dot of black ink is dry, it is lowered into the chromatography jar containing water.
 The filter paper should be immersed in such a way that the dot of black ink is above the water level
in the jar.
 The set up should be left undisturbed for some time.
 As the water starts to rise up from the lower end of the filter paper, the component of black ink
which is more soluble in water rises faster and higher up on the filter paper. The level up to which
water rises is known as water front.
 Depending on the number of components present in the black ink, some coloured spots are observed
corresponding to the separated components of the black ink.
 Calculate the Rf values of the red and blue inks using the formula,

Observation
Observations can be recorded as shown.

SI Distance travelled by the component Distance travelled by the solvent

Components Rf value
No. from the original line (cm) from the original line (cm)

1. Red

2. Blue

3, Yellow

Inference

 Rf value of red colour = ……………

 Rf value of blue colour = …………..
 Rf value of yellow colour = …………..

Precautions

 Use good quality pencil for drawing the reference line so that the mark does not dissolve in the solvent in
which the chromatography is carried out.
 Always use a fine capillary tube.
 Keep the jar undisturbed and covered during the experiment.
 The spot should be small and rich in mixture.
 Allow the spot to dry before putting the strip in the jar.
 Keep the strip erect. Do not let it curl.
 Do not allow the spot to dip in the solvent.