give a herbal extract one hundred milligrams of this extract was dissolved in 50 ml 100 methanol the solution was sonicated for 30 minutes and filtered through a 0 45 u03bcm membrane it was subjected to hplc and eluted with 75 methanol six compounds were detected and separated by hplc chromatographic conditions for the analytical hplc were mobile phase a h 2 o ch 3 cn acoh 800 ml 200 ml 10 ml b h 2 o ch 3 cn meoh acoh 100 ml 450 ml 450 ml 1 ml flow rate 0 8 ml min detection wavelength 515 nm column temperature 40 u00b0 c gradient linear from 10 b to 100 b run time 60 minutes for each compound the fractions containing it were pooled the solvent in the pooled fractions was removed under vacuum to give a residue each residue was partitioned between ethyl acetate and water the aqueous fraction was discarded ethyl acetate was then removed under vacuum to give r 5 8 dihydroxy 2 1 hydroxy 4 methylpent 3 enyl naphthalene 1 4 dione u201c sk u201d r 1 5 8 dihydroxy 1 4 dioxo 1 4 dihydronaphthalen 2 yl 4 methylpent 3 enyl acetate u201c ask u201d 5 8 dihydroxy 2 4 methylpent 3 enyl naphthalene 1 4 dione u201c dsk u201d r 1 5 8 dihydroxy 1 4 dioxo 1 4 dihydronaphthalen 2 yl 4 methylpent 3 enyl isobutyrate u201c isk u201d r 1 5 8 dihydroxy 1 4 dioxo 1 4 dihydronaphthalen 2 yl 4 methylpent 3 enyl u03b2 u03b2 dimethylacrylate u201c bsk u201d and 1 5 8 dihydroxy 1 4 dioxo 1 4 dihydronaphthalen 2 yl 4 methylpent 3 enyl 2.