An Introduction to chromatography Chromatography in Lab.

Experiment No. 3: Chromatographic Separation of Pigments of Two

Commercial Inks

Background
A pigment is a dry, powdery substance that must be mixed with a liquid like water to
leave behind black, white or color. The pigment is not necessarily water soluble, but it
may remain suspended in the liquid (much as is the case for a “colloid"). Pigment
particles tend to bond to edges within the chosen medium, a reason why these inks
tend to last longer and resist fading over time.

Pigment inks are generally better-suited to printing on slicker surfaces, such as

transparencies and stickers. They are also a little more expensive to produce and their
colors are not near as brilliant as with dyes. If you are printing an important
image/document that you want to last for a long time, pigment inks are your best bet.
If you're looking for bright color at a lower cost, dye ink may be a better fit for you
Procedure
1. Prepare a strip of filter paper to fit inside a beaker 500 ml.
2. Take a line with a pencil at 1 cm from the lower end of the strip.
3. Pipette a spot of the ink at the center of the line and allow it to dry.
4. Prepare the solvent as following (30 ml n-butanol + 7.5 ml acetic acid glacial +
12.5 ml distiled water) into a dry and clean 500 ml beaker.
5. Hang the paper inside the beaker without allowing the lower end of the paper to
touch the solvent then cover it with foil.
6. Leave the paper with the solvent in the beaker for 15 min.
7. Allow the lower end of the paper to touch the solvent, but not the cylinder walls.
8. Leave the system in contact for one hour.
9. The developing solvent will ascend through the paper carrying the component
pigments with it which after a period of time appear as colored zones.
10. Remove the chromatogram and mark the solvent front with a pencil.
11. Leave the chromatogram to dry.
12. Determine the number of pigments of the ink.
13. calculate the Rf for each pigment as following

Rf = Distance moved by the pigment (cm) / Distance moved by the

solvent (cm)

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An Introduction to chromatography Chromatography in Lab.

PART III: SOLVENT EXTRACTION

Experiment No. 1: Extract DNA from Anything Living

Background:
Since DNA is the blueprint for life, everything living contains DNA. DNA isolation
is one of the most basic and essential techniques in the study of DNA. The extraction
of DNA from cells and its purification are of primary importance to the field of
biotechnology and forensics. Extraction and purification of DNA are the first steps in
the analysis and manipulation of DNA that allow scientists to detect genetic disorders,
produce DNA fingerprints of individuals, and even create genetically engineered
organisms that can produce beneficial products such as insulin, antibiotics, and
hormones.

DNA can be extracted from many types of cells. The first step is to lyse or break
open the cell. This can be done by grinding a piece of tissue in a blender. After the
cells have broken open, a salt solution such as NaCl and a detergent solution
containing the compound SDS (sodiumdodecyl sulfate) is added. These solutions
break down and emulsify the fat & proteins that make up a cell membrane. Finally,
ethanol is added because DNA is soluble in water. The alcohol causes DNA to
precipitate, or settle out of the solution, leaving behind all the cellular components
that aren't soluble in alcohol. The DNA can be spooled (wound) on a stirring rod and
pulled from the solution at this point.

Materials:
Strawberry, plastic bag, salt, detergent, water, alcohol, filter paper, funnel, conical
flask, test tube, glass stirring rod

Procedure:
1. First, you need to find something that contains DNA such as strawberry, kiwi,
banana, fresh spinach, chicken liver, or onion.

2. Squeeze well two strawberries in a plastic bag.

3. Prepare the lysis solution (0.5 cup of water + 2 tablespoons of detergent + one
tablespoons of NaCl) and add it to the squeezed strawberries. Wait for 5-10
minutes.

8. Filter the resultant mixture and keep the filtrate for the next step.

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9. Pour the mixture into test tubes or other small glass containers, each about 1/3
full.

11. Tilt your test tube and slowly pour rubbing alcohol (95% ethyl alcohol) into
the tube down the side until you have about the same amount of alcohol in the
tube as pea mixture.

12. Alcohol is less dense than water, so it floats on top forming two separate
layers.

13. All of the grease and the protein that we broke up in the first two steps move
to the bottom, watery layer.

14. DNA will rise into the alcohol layer from the pea layer. You can use a glass
stirring rod or a wooden stick to draw the DNA into the alcohol.

15. Slowly turning the stirring rod will spool (wrap) the DNA around the rod so it
can be removed from the liquid.

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An Introduction to chromatography Chromatography in Lab.

PART IV: ION EXCHANGE CHROMATOGRAPHY

Experiment No. 1: Determination of Cation Exchange Resin Capacity by Batch

Method

Background
Ion exchange is a reversible chemical reaction wherein an ion (an atom or molecule
that has lost or gained an electron and thus acquired an electrical charge) from
solution is exchanged for a similarly charged ion attached to an immobile solid
particle. These solid ion exchange particles are either naturally occurring inorganic
zeolites or synthetically produced organic resins. The synthetic organic resins are the
predominant type used today because their characteristics can be tailored to specific
applications.

An organic ion exchange resin is composed of high-molecular-weight polyelectrolytes

that can exchange their mobile ions for ions of similar charge from the surrounding
medium. Each resin has a distinct number of mobile ion sites that set the maximum
quantity of exchanges per unit of resin (resin capacity).

Procedure
1. Weight out 0.2g of the cation exchange resin.
2. Regenerate the resin with 20ml 1N HCl.
3. Filter the mixture and wash the resin with distilled water to get rid of acidity.
4. The resin is now ready to use.
5. Prepare 50ml 0.01N NaOH and 50ml 0.25N Na2SO4.
6. Add 50ml of 0.25N Na2SO4 solution to the resin and stir well for 20 min.
7. Take 10ml from the solution (Na2SO4) after the mixing period and titrate it
against 0.01N NaOH with ph.ph. indicator.
8. calculate the capacity of the resin (C) as following

C = a×V ⁄ W

Where a Normality of the titrating solution

V Volume of the titrating solution
W Weight of the resin

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