RP-HPLC ANALYSIS OF HYDROPHOBIC ALENDRONATE AMIDATED DERIVATIVES

Abstract

The reversed phase high performance liquid chromatography (RP-HPLC) was evaluated for its application in the analysis of the quantiation of the alendronate derivatives. The anti-phase Agilent CLC-ODS C18 column was used at the temperature of 50°C. The RP-HPLC trace was monitored with a differential refractive index detector for the lack of ultraviolet (UV) absorbing groups in the derivative molecule. The mobile phase was an aqueous solution mixture of MeOH and H2O at pH 7.2, containing small amounts of dodecoulde tributyl bromide and disodium ethylente diamine tetraacetic acid. The chromatographic conditions, such as the concentration of ion-pairing agent, pH, and methanol content in mobile phase, were studied. This RP-HPLC method provides high separation and specificity to derivative analysis. In quantitative analysis, the method showed satisfactory precision (less than 0.62%) and recovery (higher than 98.3%), good linearity (r2 = 0.9994) with the linear range of 0.5 ∼ 250 μg·mL−1. The limit of quantification (0.5 μg·mL−1) and limit of detection (0.2 μg·mL−1) were also achieved.