E Extract 1 g of the sampleสารสกัดจ

E Extract 1 g of the sampleสารสกัดจาก 1 กรัมของกลุ่มตัวอย่าง
A Reflux 1 g of the sample กรดไหลย้อน 1 กรัมของกลุ่มตัวอย่าง
A. Reflux 1 g of the sample , in powder , with 30 ml of ethanol for 15 minutes, and filter. To 2 ml of the filtrate, add a few drops of amnanium malybdate TS : a bright yellow precipitate is produced.
B. To 2 ml of the filtrate obtained from test A, add a greenish brown colour is produced.
C. To 2 ml of the filtrate obtained from test A, add a few drops of ninhydrin TS and warm on a water-bath for a few minutes: a reddish purple colour is produced.
D. Moisten 1 g of the sample, in powder, with 0.5 ml of strong ammonia solution, add 5 ml of chloroform, occasionally shaking for 10 minutes, add filter. Evaporate the filtrate until dryness and dissolve the residue in1 ml of ethanol, and add a few drops of modified Dragendorff TS 2: an orange precipitate is produced.
E. Extract 1g of the sample, in powder, with 10 ml of chloroform, shank occasionally for 20 minutes, and filter. Evaporate 2 ml of the filtrate to dryness, dissolve the residue in 2 ml of acetic anhydride, and add slowly 1ml of sulfuric acid to make two layers: a brown colour forms at the zone of contact and the upper layer is green.
F. Carry out the test as described in the “Thin-layer Chromatography” (Appendix 3.1),using silica gel 60 F.254 as the coating substance and chloroform as the mobile phase but allowing the solvent front to ascend 15 cm above the line of application. Apply to the plate, 5 ul of the test solution prepared by moistening 1 g of the sample, in powder, with 1 ml of strong ammonia solution for awhile, exracing with 10 ml of chloroform by occasionally shaking for 15 minutes, filtering, and concentrating the filtrate to 1 ml. After removal of the plate, allow it to dry in air, and examine under ultraviolet light (254 nm, 366 nm), locating the spots (Table 1); see also Fig. 3. Spray the plate with anisaldehyde TS and heat at 105 for 5 minutes. Several spots of different colours are observed (Table 1); see also Fig
. 3.
Repeat the same procedure on other two plates but use a mixture of 20 volumes of chloroform and 1 volume of methanol as the mobile phase and allow the solvent front to ascend 12 cm above the line of application. After removal of the plates, allow them to dry in air, and spray one plate with modified Dragendorff TS2: two orange spots are observed. Spray another plate with indoplatinate TS: one dark blue spot and one brown spot are observed (Table 1); see also Fig. 4.