Spd 20a hplc

For the isolation of active compounds, both Anastatica hierochuntica and Aerva javanica plant extracts/fractions were analyzed using the preparative HPTLC and HPLC. The HPTLC was conducted with a CAMAG^® HPTLC scanner III, using hexane and ethyl acetate in the ratio of 7:3 as the solvent. The HPLC was completed on a Shimadzu SPD 20A HPLC instrument with a C18 column (4.6 mm × 250 mm; 0.5 micron), acetonitrile and water (15:85), with 0.1% phosphoric acid as the solvent at a flow rate of 1 mL/min. The UV spectra were then recorded on a PDA detector. The compounds were further characterized with IR and NMR spectroscopic techniques. The NMR spectra were recorded using a Bruker 400 MHz spectrometer for ¹H and ¹³C using deuterated chloroform (CDCl₃). The chemical shifts were expressed in δ (ppm) down-field from the tetramethylsilane as the internal standard. The IR spectra were recorded by a Bruker FT-IR spectrophotometer from 4000 to 500 cm⁻¹.