Determination of Hexestrol in Ointment by High-pressure Liquid Chromatography

High-pressure liquid chromatography (HLC) was evaluated in determination of hexestrol in ointment. Sample ointments were mixed with ethanol and isooctane. Hexestrol in the Hexron ointment was extracted into ethanol layer. Butyl p-hydroxybenozate was used as the internal standard. HLC was performed under the following conditions: model, Shimadzu LC-2F; detector, UV visible spectrophotomer Shimadzu SPD-1; wave length, 280nm; column, Permaphase ODS (2.1φ×0.5m); mobile phase, gradient elution system, 10% to 40% MeOH in H2O at 3% min; flow rate, 1 ml/min (at room temperature). The calibration curve was traced by plotting the ratio of the peak weight against the amount of each hexestrol. Satisfactory linearity was observed in the range of 0.125 to 1μg and quantitative determination was found to be possible. Other components in the Hexron ointment did not interfere in determination of hexestrol in this method. Hexestrol in the Hexron ointment was recovered at the rate 94% on an average by ethanol extraction. The quantitative determination of hexestrol in an ointment by HLC was carried out simply and rapidly.