Determination of Related Substances in Estradiol Valerate Tablet by Use of HPLC Method

Background: Estradiol valerate tablet is used for ovarian dysfunction, menopause, menopausal syndrome, breast and prostate cancer.

Introduction: HPLC method coupled with photodiode-array detection has been used to develop a simple and sensitive reversed-phase method for quantitative determination of all possible related impurities in Estradiol valerate tablet.

Methods: Zorbax eclipse XDB C8(150mmx4.6mm,5μm) column was used. The mobile phase was gradient prepared from 30:10(v/v) acetonitrile-methanol (component A) and water (component B); the gradient program (time (min)% B) was 0.01/40, 40.0/40, 40.1/15, 50.0/15, 50.1/40, 55.0/40, 55.1/40, 60.0/40. Eluting compounds were monitored at UV detection (220nm) and fluorescence detection (Ex:280nm, Em 310nm).

Result: All standard curves obtained exhibited good linear regression (r0.9996) within the tested range. All the average recovery rates were in the range of 99.2%-100.8% and RSD were less than 2.0%. The method was successfully validated according to ICH guidelines acceptance criteria for robustness, selectivity, linearity, accuracy and precision.

Conclusion: Estradiol valerate was subjected to oxidative, acid, base, thermal and photolytic stress, and analysis was conducted to determine the amounts of related impurities.