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1)To identify semi-quantitatively Ciprofloxacin hydrochloride in a

tablet dosage form using Thin layer chromatography(TLC)

Apparatus required:
1)UV cabinet(with shorter/longer wavelength), 2)Silica gelG coated plate, 3)Micro pipette/Micro syringe, 4) Beakers, 5)Clock glass or aluminum foil.

Chemicals required:
1)Dichloromethane, 2)Methanol, 3)Strong ammonia solution, 4)Acetonitrile.

Theory:
The technique thin-layer chromatography(TLC) is open bed technique substances are separated by the differential migration that occurs when a solvent flow along a thin layer of fine powder spread on a glass or plastic plate(TLC). In this technique a solute undergoes distribution between two phases, a stationary phase acting through adsorption and a mobile phase in the form of a liquid. The adsorbent is a relatively thin, uniform layer of dry finely powdered material applied to a glass, plastic or metal sheet or plate. Glass plates are most commonly used. Glass plates are most commonly used. Separation may also be achieved on the basis of partition or a combination of partition and adsorption, depending on the particular type of support, its preparation and its use with different solvent. If the coating material is silica or alumina activated by heating to 110C for to 1hr. and the solvent system is a mixture of organic solvent, the principal mechanism is adsorption chromatography. If solvent system contains organic solvents and water, then the principal mechanism becomes partition chromatography. In the test for identification, The principal spot in the chromatogram obtained with the test solution is visually compared to the corresponding spot in the chromatogram obtained with the reference solution in respect of the colour , the size and the Rf of the spots. The spots are detected either by UV irradiation or other chemical techniques.

Procedure:

Mobile phase: A mixture of 40 volumes of dichloromethane, 40 volumes of methanol, 20 volumes of strong ammonia solution and l0 volumes of acetonitrile. Test solution: Shake a quantity of the powdered tablets containing about 0. 15 g of ciprofloxacin with 15 ml of water for 20 minutes, dilute to 100.0 ml with water, mix, centrifuge and use the clear supernatant liquid Reference solution: A 0.15 per cent w/v solution of ciprofloxacin hydrochloride RS in water. Dry a coated plated at 110C for to 1 hour. Pour the mobile phase into the tank to form a layer of solvent 5 to l0 mm deep. Apply 5l of solutions being examined in the form of circular spots about 2 to 6 mm in diameter, or in the form of 1cm-bands on a line parallel with, and 20 mm from, one end of the plate, and not nearer than 20 mm to the sides; the spots should be 15 mm apart. If necessary the solutions may be applied in portions, drying between applications. Mark the sides of the plate l2 cm from the starting line. Allow the solvent to evaporate .Place the plate in an atmosphere of ammonia for about 15 minutes and finally in the unsaturated tank, ensuring that it is as nearly vertical as possible and that the spots or bands are above the level of the mobile phase. Close the tank partially with its lid or clock glass or aluminum foil. and allow to stand at room temperature, until the mobile phase has ascended to the marked line. Remove the plate and dry it. Examine in ultravioiet light at 254 nm and 365nm. The principal band in the chromatogram obtained with the test solution corresponds to that in the chromatogram obtained with the reference solution. Wt. of 5 tablets of Ciprofloxacin hydrochloride= Avg. Wt. = Wt. of Sp .= 100ml(in water) Wt.of Ref.=

Observation:
The chromatogram obtained with the test solution corresponds/does not correspond to that in the chromatogram obtained with the reference solution

i)Distance sample moves from origin = ii)Distance reference moves from origin= iii)Distance solvent moves from origin=

Calculation:
Rf (Sample) = Distance sample moves from origin Distance solvent moves from origin Rf (Sample) = Distance sample moves from origin Distance solvent moves from origin =