Background: Brassino-steroids as hormonal plants are of the most important components that have shown the biological effects. These compounds may be act as inducer of growth in the plant and their concentrations is increased under some stress conditions. Such clinical studies show that these hormones have also anabolic activities similar to the human anabolic hormones hence, may be administrated as supplements in athletes. Their increasing consumption as supplement suggests introducing a precise and accurate analytical method for determination of brassino-steroids in pharmaceutical dosage forms and also in some biological fluids.

Objective: In the current study an HPLC method for detect and determination of brassino-steroids in capsules was developed and optimized in aspects of analytical conditions.

Methods: In order to analysis of the sample a reversed phase HPLC system including a C₁₈ column, and a mobile phase including water: Acetonitrile in an acidic media were used. Detection was carried out at 210 nm by an UV detector.  

Results: In this study, we attempted to optimize some analytical aspects e.g: solvent, stationary phase and other assay parameters to obtain best condition to assay. Also the validation parameters such as LOD, LOQ, accuracy, precision and linearity of the method were also studied and were satisfactory.

Conclusion: Obtained data indicated that HPLC is a suitable analytical method for assay of brassino-steroids in pharmaceutical dosage forms.