Saponins From Sea Chromatography Lab Report
Materials and methods Preparation of saponins from sea cucumber:
Saponins were isolated from the sea cucumber, Holothuria thomasi (purchased from Hurghada, Red sea, Egypt and identified by Zoology department, Faculty of science, Tanta, Egypt), according to the method described by Hu et al., (2010) with some modification. Air-dried body walls (750 g) of sea cucumber were grinded into powder and extracted with eight liter with refluxing ethanol. The combined extracts were evaporated with rotary evaporator and further partitioned between water and chloroform. The water layer was extracted with n-butanol and the organic layer was evaporated with rotary evaporator to yield n-butanol extracts. The n-butanol extracts were concentrated in drying oven 60 ºC.
Identification of extracted …show more content…
The aluminum sheets paper were sprayed with sulfuric acid in ethanol (10:90 v/v) and heated for 10 min at 110 0C to visualize saponins (Kerem et al., 2005 ).
Fourier tansform infrared (FTIR):
Dried sea cucumber saponin extract and standard saponin(Fisher Scientific UK. Saponin, extra pure, SLR by Laboratory Reagents, Legacy Product Code: S/0380/48, CAS Number:8047-15-2) were powdered and analyzed as potassium bromide (KBr) pellets using FTIR (Model-JASCO FT/IR 4100 LE, made in Japan; Range: 4000-400 cm-1) Prabhu et al. (2013).
Gas Chromatography–Mass Spectrometry (GC–MS) analysis:
Acid hydrolysis of saponin was carried out according to the method described by Metwally et al. (2012) with some modification. Saponin was hydrolyzed with 2 N HCL (6 hr at 100 o C) under reflux, the residue was evaporated and the mixture was dissolved in water and extracted with CHCl3. The CHCl3 layer was evaporated to afford the
Saponins were isolated from the sea cucumber, Holothuria thomasi (purchased from Hurghada, Red sea, Egypt and identified by Zoology department, Faculty of science, Tanta, Egypt), according to the method described by Hu et al., (2010) with some modification. Air-dried body walls (750 g) of sea cucumber were grinded into powder and extracted with eight liter with refluxing ethanol. The combined extracts were evaporated with rotary evaporator and further partitioned between water and chloroform. The water layer was extracted with n-butanol and the organic layer was evaporated with rotary evaporator to yield n-butanol extracts. The n-butanol extracts were concentrated in drying oven 60 ºC.
Identification of extracted …show more content…
The aluminum sheets paper were sprayed with sulfuric acid in ethanol (10:90 v/v) and heated for 10 min at 110 0C to visualize saponins (Kerem et al., 2005 ).
Fourier tansform infrared (FTIR):
Dried sea cucumber saponin extract and standard saponin(Fisher Scientific UK. Saponin, extra pure, SLR by Laboratory Reagents, Legacy Product Code: S/0380/48, CAS Number:8047-15-2) were powdered and analyzed as potassium bromide (KBr) pellets using FTIR (Model-JASCO FT/IR 4100 LE, made in Japan; Range: 4000-400 cm-1) Prabhu et al. (2013).
Gas Chromatography–Mass Spectrometry (GC–MS) analysis:
Acid hydrolysis of saponin was carried out according to the method described by Metwally et al. (2012) with some modification. Saponin was hydrolyzed with 2 N HCL (6 hr at 100 o C) under reflux, the residue was evaporated and the mixture was dissolved in water and extracted with CHCl3. The CHCl3 layer was evaporated to afford the