Thin layer chromatography (TLC) is a chromatographic technique used to separate the components of a mixture

using a thin stationary phase supported by an inert backing. It may be performed on the analytical scale as a means
of monitoring the progress of a reaction, or on the preparative scale to purify small amounts of a compound. TLC is
an analytical tool widely used because of its simplicity, relative low cost, high sensitivity, and speed of separation.TLC
functions on the same principle as all chromatography: a compound will have different affinities for the mobile and
stationary phases, and this affects the speed at which it migrates. The goal of TLC is to obtain well defined, well
separated spots.

Retention Factor
After a separation is complete, individual compounds appear as spots separated vertically. Each spot has a retention
factor (Rf) which is equal to the distance migrated over the total distance covered by the solvent. The Rf formula is

Rf=distance traveled by sampledistance traveled by solvent

The Rf value can be used to identify compounds due to their uniqueness to each compound. When comparing two

Rf value is less polar because it does

not stick to the stationary phase as long as the polar compound, which would have a lower Rf value.
different compounds under the same conditions, the compound with the larger

Rf values and reproducibility can be affected by a number of different factors such as layer thickness, moisture on
the TLC plate, vessel saturation, temperature, depth of mobile phase, nature of the TLC plate, sample size, and
solvent parameters. These effects normally cause an increase in Rf values. However, in the case of layer thickness,
the Rf value would decrease because the mobile phase moves slower up the plate.
If it is desired to express positions relative to the position of another substance, x, the

Rx (relative retention value)

can be calculated:

Rx=distance of compound from origindistance of compound x from origin

Rx can be greater than 1.

Codeine (methyl morphine) is a strong mono acidic base and laevorotatory. It effloresces slowly in dry air and is
affected by light. The chemical name of codeine phosphate is 7,8-didehydro-4,5alpha-epoxy-3-methoxy-17methylmorphinan-6alpha-olphosphate (1:1) (salt) hemihydrate and has the empirical formula of
C 18 H 21 NO 3H 3 PO 41/2H 2 O. [1] Codeine phosphate contains not less than 98.5 percent and not more than 101.0
percent of C 18 H 21 NO 3 , H 3 PO 4 calculated on the dried basis. [2] Codeine molecular weight is 406.4. Each soluble
tablet contains 30 mg (0.074 mmol) or 60 mg (0.15 mmol) of codeine phosphate. These tablets also contain lactose
and
sucrose.
Soluble
tablets
of
codeine
phosphate
are
freely
soluble
in
water. [1]

High performance liquid chromatography is basically a highly improved form of

column chromatography. Instead of a solvent being allowed to drip through a
column under gravity, it is forced through under high pressures of up to 400
atmospheres. That makes it much faster.
It also allows you to use a very much smaller particle size for the column
packing material which gives a much greater surface area for interactions
between the stationary phase and the molecules flowing past it. This allows a
much better separation of the components of the mixture.
The other major improvement over column chromatography concerns the
detection methods which can be used. These methods are highly automated
and extremely sensitive.