Implementation of separation by paper chromatography method is dividedinto three stages, namely the stage of spoting, stage of development, and theidentification or the appearance of stains.At this stage of spoting, first prepared acertain size of filter paper. Following thatis a line beginning at a distance of 2-3 cmwith one end of the paper, and then spottedthe solution samples using a micropipetteor capillary tube at the beginning of theline was then dried. In practice this time,kind of a solution containing variousamino acids will be tested by paper chromatography.In the identification phase or appearance of stains, if the stain wascolored can be directly examined anddetermined its value Rf. The value of Rf stated degrees retention of a component inthe stationary phase. The distance traveled by each compound from baseline relativeto the distance the solvent/ eluent isdefined as Rf can be formulated asfollows.
outlineeluent the fromce Dis
outlin samplethe fromce Dis
Rf value of a compound on paper chromatography system depends on manyvariables, including solvent system,temperature, duration of elution, and paper type. Because it is influenced by manyvariables, the Rf of a compound that isknown to be used as a standard or benchmark to determine Rf other compounds. For qualitative analysis,certain compounds that are known to beused in conjunction with the compound to be identified. Two different compounds ina given solvent system can have the sameRf value. Therefore, the analysis of theresults obtained by paper chromatographytechniques, must be justified by other methods.Compounds were analyzed by paper chromatography techniques can bedetermined location on the paper invarious ways. If the tested compoundsabsorb UV light or fluoresce with UVrays, stains or spots can be detected byirradiation with UV light in a dark place.Stains or spots can also be specifiedlocation on paper using dye compounds,for example with ninhydrin (Tika, 2010).
MATERIALS AND METHODS
Experiment of identification of amino acids by chromatographictechniques was conducted at theLaboratory of Organic ChemistryDepartment of Chemistry, University of Education Singaraja Ganesha on March19, 2013.The equipments used were beaker glass 100 mL (5 units), beaker glass 25 mL(1 unit), separation funnel 500 mL (1 unit),ring (1 unit), stative and clamp (1 pair),ruler 30 cm (1 unit), spatula (1 unit), glassroad (1 unit), drop pippete (3 units),capillary pipe (3 units), chromatographyroom (1 unit), pinset (1 unit), scissor (1unit), beaker glass 250 mL (2 units).The materials used were n- buthanol solution (100 mL), distilled water (500 mL), acetic acid glacial (25 mL),glicine solution (25 mL), sample solution 4(25 mL), alcohol 50 mL Phenol solution50mLMethodsIn the preparation of elutionsolution, 100 mL of n-butanol was addedwith 100 ml of distilled water and 24 mLof glacial acetic acid. The third solutionwas placed in a separating funnel andshaken. The two layers were formed andthen separated.Process of chromatography usingeluent phenol in the filter paper was prepared with the size of the container adapted to chromatography place. In thesection about 1.5 cm from the bottom edgeof the paper was marked with a pencil.Filter paper with a size of 15 cm x 25 cmspotted with a solution of tryptophan,leucine, tyrosine, methionine, glycine, asample solution 4 using capillary pipette.Spot distance between each other is 1.5cm. Things to note, that each droplet must