Simple analysis of four bisphosphonates simultaneously by reverse phase liquid chromatography using n-amylamine as volatile ion-pairing agent

Volatile organic amine was used as the mobile phase addictive during the separation of four bisphosphonates (alendronate, pamidronate, zoledronic acid and etidronate). An isocratic liquid chromatography method with evaporative light-scattering detection (ELSD) was developed for these bisphosphonates which are not retained on non-polar column and lack chromophore for detection. The analytes have sufficiently separated from each other on a Phenomenex C18 column. The effects of mobile phase composition and instrumental parameters of ELSD were studied. This newly developed method enables direct measurement for analysis of bisphosphonates without the need of derivatization. This developed method provides high separation and specificity to bisphosphonate analysis. In quantitative analysis, the method showed satisfactory precision (less than 2.8%) and accuracy (higher than 94.4%), good linearity (r=0.9991-0.9997) and sufficient sensitivity (15-18 microg/ml). It can be easily and conveniently adopted for the routine quality control analysis.