Method development and validation of levonorgesterel and ethinyl estradiol in bulk and its pharmaceuticals dosage forms

Abstract

HPLC is regarded as one of the most sophisticated and reliable analytical techniques currently

available. The quantitative determination of Levonorgestrel and Ethinyl Estradiol was performed using

an optimized HPLC method. The mobile phase comprised Methanol and Phosphate buffer (pH 3.0) in

a ratio of 70:30 % v/v. Chromatographic separation was achieved on an Inertsil C18 column (4.6 ×

150 mm, 5 μm) or an equivalent stationary phase chemically bonded to porous silica particles.

Detection was carried out using a UV detector at 265 nm, with a constant flow rate of 0.7 mL/min.

The method demonstrated excellent linearity over the concentration ranges of 100–500 μg/mL for

Levonorgestrel and 1–5 μg/mL for Ethinyl Estradiol, with correlation coefficients (R²) greater than

0.999. The %RSD values for precision studies were below 2%, indicating satisfactory repeatability

and accuracy. The percentage recoveries of Levonorgestrel and Ethinyl Estradiol were found to be

within 98–102%, and both the limits of detection (LOD) and quantitation (LOQ) were within

acceptable limits.

All validation parameters complied with ICH and USP guidelines, confirming the method’s suitability

for its intended purpose. The proposed HPLC method is simple, accurate, precise, and linear, and can

be effectively employed for the routine quality control analysis of Levonorgestrel and Ethinyl

Estradiol in pharmaceutical formulations.