Optimised method for determination of ticlopidine in serum by high performance liquid chromatography

In this study we verified whether high performance liquid chromatography with UV detection (HPLC-UV) could be optimised and hence become a useful method for measuring ticlopidine concentrations in patient blood. The extraction step was improved by adding isoamylic alcohol in the extraction mixture, resulting in a better recovery. Moreover, the extraction efficiency was consistently ameliorated by evaporation to dryness of the organic extract; we could therefore utilise a lower (213 nm) wavelength, corresponding to the maximum of absorption, achieving a better sensitivity. With these technical improvements, the limit of quantitation is 0.02 microgram/ml, and the limit of detection is 0.01 microgram/ml, hence comparable to levels obtained with gas-liquid chromatography with nitrogen detection (GC-NPD). In addition, HPLC method, if compared with GC-NPD, is simpler and can be easily used for routine determinations of a lot of serum samples.