Alendronic acid is widely used for the treatment of osteoporosis.
According to the Japanese Pharmacopeia (JP; 17^th edition*), quantification of alendronate sodium injection should be carried out with a column filled with styrene divinyl benzene copolymer gel and meets following requirements. The method also mentions to pretreat the alendronic acid by adding 9-fluorenylmethyl chloroformate to let the amino groups to form 9-fluorenylmethyloxycarbony (FMOC)-alendronic acid. The RSpak DS-413 confirmed the requirements were met.

System suitability requirements:
   Theoretical plate number of FMOC–alendronic acid: ≥ 1,500
   Symmetry: ≤ 1.5
   Relative standard deviation (RSD) of six analyses: ≤ 1.0%

*The version at the time of the application acquisition.


Sample : 50 μL
prepared according to JP
1. FMOC-Alendronic acid

Column       : Shodex RSpak DS-613 (6.0 mm I.D. x 150 mm)
Eluent       : 50 mM Trisodium citrate + 50 mM K2HPO4 aq.(adjusted to pH 8.0 with H3PO4)
               /CH3CN/CH3OH=75/20/5
Flow rate    : 0.9 mL/min
Detector     : UV (265 nm)
Column temp. : 40 °C