A simple high-performance liquid chromatography (HPLC) method to determine the content of betulinic acid (BA) in rat plasma collected at different times (0-8 h) after oral administration of Orthosiphon stamineus leaf extract was developed. The features of the assay include protein precipitation using acetonitrile and isocratic elution using reverse phase C-18 column with ultraviolet (UV) detection. The recovery of BA from plasma varied from 98.4 to 102.5%. The R.S.D of intra- and inter-day precision from rat plasma ranged from 4.2 to 9.8%. The maximum concentration of BA in the plasma was 1.2 +/- 0.3 microg/ml at 1 h after oral administration of the extract.
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