Background: In this investigation, we developed a new, rapid, novel HPLC technique for simultaneous
routine analysis of amitriptyline and clomipramine. For this method, desipramine was used as
an internal standard.
Experiment: Amitriptyline, clomipramine and desipramine (IS) were separated in a X-Terra RP-18 column
(250mm×4.60mm ID×5µm) and DAD detector set at 210 nm. The composition of the mobile phase
was acetonitrile:water 50:50 (v/v) at a flow rate of 1.0 mL min-1
. Isocratic separation was performed in
less than 6 min. Additionally, pKa
values of studied compounds were evaluated by HPLC method in order
to find the optimum pH value of separation. pKa
values were determined at 45 and 50% (v/v) acetonitrilewater
mixtures in order to find the effect of the mobile phase composition on the ionization constants.
program was used to calculate pKa
values of drugs. The proposed method also has some advantages
such as a number of cost and time saving benefits and can be routinely used for the analysis of
Result: The results show that developed method is suitable for the simultaneous analysis of these compounds
in tablets without any interference from excipients.