Solvent n-hexane, ethyl acetate and acetone will use to elute the column in isolation of chemical constituents of figeroots chloroform partition. The solvent system of different ratios of hexane, hexane/ethyl acetate, hexane/acetone, and acetone will use. Silica gel 60 (mesh 230-400 ASTM) will use to pack CC. In order to achieve good separation, mass ratio of silica gel to compound 20:1and 50:1 will use.
3.10.2Column Packing
Before starting to pack a column, a small piece of cotton is gently will insert into the centre hole of column with the aid of a long stick. A small amount of n-hexane will add into the column. Silica gel weighted is mixed with n-hexane to form a mobile slurry solution and then slowly transferred into column. The side of the column will gently tape to prevent the formation of air bubbles in the column. Additional solvent will use to add the remaining silica gel into the column. Once all silica gel will add into column will gently tapped again to make the silica gel layer is flat. One precaution when packing a column is that never let the solvent to
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The sample will prepare by using the KBr pelleting method. The sample will mix with the KBr pellet in the mortar and pestle to grind until become fine powder form. The mixture will then press in a special die at 10,000 lb in-2 to yield a transparent disc. Before running the test of the sample, the background spectra will collected under identical conditions is subtracted from the sample spectra automatically. The spectrometer analysed the compound in the KBr disc form and a spectrum was displayed after the testing of the sample was completed. Based on the peaks resolved in the spectra, the possible IR functional groups which relevant to the chemical constituent isolated were identified by table of characteristic IR absorption (refer Appendix