Comparison of types of compound in different methods of extraction from Caesalpinia sappan (Leguminosae) extracts

Caesalpinia sappan, is a medicinal plant from Leguminosae family. It is called as Sappan wood and traditionally used during postpartum period in Malaysia and other Asia countries. C. sappan is widely used for medicinal purpose due to its various immunosuppressive activities present in the various us types of secondary metabolites of C. sappan. 300. g of Caesalpinia sappan was exiotralcoted using two methods; which are Soxhlet extraction with 250ml ethanol for 72 h (100 g) and maceration method of extraction of 200 g of sample using methanol for 3 days with 1:20 ratio. The residue of Soxhlet extraction (4.18 g) and maceration method of extraction (6.28 g) were partitioned with hexane, dichloromethane, ethyl acetate and methanol giving 8 fractions in total. The fractions were then subjected to thin layer chromatography with different types of mobile phase. For the hexane fraction, hexane-ethyl acetate 9: 1 and 7: 3 were used as the mobile phase, while for dichloromethane fraction, the same solvent were used as the mobile phase but with different ratio; 6: 4, 4: 6 and 3: 7. The mobile phases used for the ethyl acetate fraction were chloroform-methanol 5: 5, 9: 1 and 6: 4. All the TLC plates were sprayed with four types of spray reagents namely sulphuric acid, vanillin, ammonia and dragendorff. Further separation using column chromatography was used for separation of dichloromethane fraction (6:3) of maceration method of extraction and eluted with hexane-ethyl acetate (9:1, 8:2, 7:3, 6:4, 4:6, 2:8). Both of the extraction methods showed the presence of 5 types of phytoconstituents identified as alkaloid, flavonoid, phenols, terpenes and amino acid. Out of the selected methods of extraction, the maceration method of extraction showed the most visible spot of compounds when subjected to the thin layer chromatography. Further separation using column chromatography and High Performance Liquid Chromatograp (HPLC) indicated the presence of the major compound of C. sappan, which are probably flavonoids after comparing .with the color from the TLC plates. Numerous types of compound observed in maceration method of extraction, indicating that the secondary metabolites of this plant are more methanol than ethanol (Soxhlet extraction)