Validation of the doxycycline hydrochloride determination by high-performance liquid chromatography technique

Abstract

A sensitive and reproducible chromatographic method was developed and validated for the quantification of doxycycline hydrochloride in veterinary product – Doxycyclinum 50%. The analysis was performed on HPLC (Shimadzu) apparatus with diode array detector. The separation was provided with C₁₈ column (4.6mm×100 mm, 3 μm particle size) at mobile phase flow rate 1ml/min and detection at 254 nm. The method was tested in the range 50-150% nominal DOXY concentration resulting good linearity with correlation coefficient r=0.9999. The estimated LOD and LOQ elaborated method amounted to 0.46 and 1.55 mg/L. The recovery values were from 99.70 to 100.9% with RSD 0.78%. It was confirmed that the reagents used for testing of samples and solvents do not interfere with the active substance. Moreover, the significant influence of temperature and flow rate on the retention time was ascertained. The carried out research demonstrates that the method is validated and the obtained results are accurate and reproducible. The elaborated method can be suitable to routine analysis in quality control laboratory.