title: Development of an HPLC method for the monitoring of tricyclic antidepressants in biofluids.
abstract: A simple, rapid and sensitive HPLC method was developed and validated for the determination of four tricyclic antidepressants (TCAs): amitriptyline, doxepin, clomipramine (CLO) and imipramine, in pharmaceutical formulations and biological fluids. A Kromasil C(8 )analytical column (250 x 4 mm, 5 microm) was used for the separation, with a mobile phase consisting of 0.05 M CH(3)COONH(4) and CH(3)CN (45:55 v/v) delivered at 1.5 mL/min isocratically. Quantification was performed at 238 nm, with bromazepam (1.5 ng/microL) as the internal standard. The determination of TCAs in blood plasma was performed after protein precipitation. Urine analysis was performed by means of SPE using Lichrolut RP-18 Merck cartridges providing high absolute recoveries (> 94%). Direct analysis of urine was also performed after two-fold dilution. The developed method was fully validated in terms of selectivity, linearity, accuracy, precision, stability and sensitivity. Repeatability (n = 5) and between-day precision (n = 5) revealed RSD <13%. Recoveries from biological samples ranged from 91.0 to 114.0%. The absolute detection limit of the method was calculated as 0.1-0.6 ng in blood plasma and 0.2-0.5 ng in extracted urine or 0.4-0.7 in diluted urine. The method was applied to real samples of plasma from a patient under CLO treatment.
authors.affiliation: Laboratory of Analytical Chemistry, Department of Chemistry, Aristotle University of Thessaloniki, Thessaloniki, Greece. samanidu@chem.auth.gr
authors.forename: Victoria F
authors.initials: VF
authors.lastname: Samanidou
authors.forename: Maria K
authors.initials: MK
authors.lastname: Nika
authors.forename: Ioannis N
authors.initials: IN
authors.lastname: Papadoyannis
mesh_terms: D000639:Amitriptyline / Q000032:analysis / Q000493:pharmacokinetics; D000929:Antidepressive Agents, Tricyclic / Q000032:analysis* / Q000493:pharmacokinetics*; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002997:Clomipramine / Q000032:analysis / Q000493:pharmacokinetics; D004316:Doxepin / Q000032:analysis / Q000493:pharmacokinetics; D016903:Drug Monitoring / Q000379:methods*; D006801:Humans; D007099:Imipramine / Q000032:analysis / Q000493:pharmacokinetics; D008956:Models, Chemical; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013053:Spectrophotometry / Q000379:methods; D013997:Time Factors; D016482:Urinalysis / Q000379:methods*
chemical_list: D000929:Antidepressive Agents, Tricyclic; D004316:Doxepin; D000639:Amitriptyline; D002997:Clomipramine; D007099:Imipramine