Paper chromatography experiment


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Paper chromatography experiment

  1. 1. Clarinda clare linusDiploma In Medical Sciences – January 2011
  2. 2.  At the end of this laboratory activity, the students should be able to: Be introduced to the principles and terminology of chromatography and demonstrate separation of the dyes in colored pens with paper chromatography.
  3. 3.  Paper chromatography is an analytical chemistry technique for separating and identifying color mixtures. In paper chromatography, substances are distributed between stationary phase and a mobile phase. Stationary phase is usually a piece of filter paper and mobile phase is the colors that travels up the stationary phase. Components of the samples will separate readily according to how strongly they absorb on the stationary phase vs. how readily they dissolve in the mobile phase.
  4. 4.  6 beakers of jars 6 covers of lids Distilled water Isopropanol Graduated cylinder 6 strips of filter paper Different colors of water color pens Pencil Ruler Scissors tape
  5. 5.  Prepare 15 ml of isopropanol solutions in labeled beakers0%, 5%, 10%, 20%, 50% and 100% Cut 6 strips of filter paper- Draw a line 1cm above the bottom edge of the strip with the pencil- Label each strip with its corresponding solution- Place a spot from each pen on your starting line
  6. 6.  Place each strip in each beaker- Cover the beakers- Let the strips absorb the solutions about 2 cm from the top of the strips- Remove strips and let them dry Observe the colors separated in low concentrations of isopropanol and high concentrations of propanol. Record data.
  7. 7. Distilled water 5% alcohol
  8. 8. 10% alcohol 20% alcohol
  9. 9. 50% alchohol 100% alcohol
  10. 10.  1st – black 2nd – red 3rd – green 4th – blue 5th – purple 6th - pink
  11. 11.  According to the conducted experiment, the results are as follows: Distilled water : Colors does not rise up to stationary phase, dissolve Alcoholic solution: Colors rise up to stationary phase and spread its colors1. Black = purple and yellow color2. Red = bright red and white3. Green = blue and yellow4. Blue = bright blue5. Purple = bright purple and pink6. Pink = Bright pink and white
  12. 12.  When a colored sample is placed on a filter paper, the colors separate from the sample by placing one of the paper in a low-leveled solvent. The solvent diffuses up to the filter paper, dissolving the various molecules in the sample according to polarities of the molecules and the solvent. If the sample contains more than one color, it means that it have more than one kind of molecule.
  13. 13.  Because of the different chemical structures of each kind of molecules, the chances are very high that each molecule will have at least a slightly different polarity, giving each molecule a different solubility in the solvent. The unequeal solubilities cause the various color molecules to leave solution at different places as the solvent continues to move up the paper.
  14. 14.  Hence, the more soluble is a molecule, the higher it will migrate up the paper.
  15. 15.  Chromatography has many uses. It is commonly used in laboratories to isolate new compounds, analyze subtle differences between different environmental samples, and even in the sequencing of DNA.
  16. 16.  To perform chromatography, one need two things; a matrix and a color indicator. A matrix is simply the materials to which a sample is applied, the material is often porous, action as filter paper that affects the rate of flow of a sample. Special beads with different pore sizes are commonly used to separate proteins in biochemistry labs.
  17. 17.  The matrix may interact with a sample, and depending on the chemical properties of both, and is commonly used to analyze mixtures of sugars.
  18. 18.  The color indicator may be the compound itself. Some organic compounds are bright yellow or orange, which makes them easy on identifying on a chromatogram. Others need to react with another chemicals to become colored. Dyes specific for particular chemical substances properties can be used to identify samples. For examples, rhodamine dye can be applied to visualize fats and oils.