Development of an HPLC method for the monitoring of tricyclic antidepressants in biofluids

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 C₈ analytical column (250x4 mm, 5 μm) was...

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Published in:Journal of separation science 2007-10, Vol.30 (15), p.2391-2400
Main Authors: Samanidou, Victoria F, Nika, Maria K, Papadoyannis, Ioannis N
Format: Article
Language:English
Subjects:
Amitriptyline - analysis
Amitriptyline - pharmacokinetics
Analysis
Antidepressive Agents, Tricyclic - analysis
Antidepressive Agents, Tricyclic - pharmacokinetics
Biological and medical sciences
Biological fluids
Chemistry Techniques, Analytical - methods
Chromatography, High Pressure Liquid - methods
Clomipramine - analysis
Clomipramine - pharmacokinetics
Doxepin - analysis
Doxepin - pharmacokinetics
Drug Monitoring - methods
General pharmacology
HPLC
Humans
Imipramine - analysis
Imipramine - pharmacokinetics
Medical sciences
Medicinal chemistry
Models, Chemical
Monitoring
Pharmacology. Drug treatments
Reproducibility of Results
Sensitivity and Specificity
Spectrophotometry - methods
Time Factors
Tricyclic antidepressants
Urinalysis - methods
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description 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 C₈ analytical column (250x4 mm, 5 μm) was used for the separation, with a mobile phase consisting of 0.05 M CH₃COONH₄ and CH₃CN (45:55 v/v) delivered at 1.5 mL/min isocratically. Quantification was performed at 238 nm, with bromazepam (1.5 ng/μL) 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
doi_str_mv 10.1002/jssc.200700142
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Sep. Science</addtitle><description>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 C₈ analytical column (250x4 mm, 5 μm) was used for the separation, with a mobile phase consisting of 0.05 M CH₃COONH₄ and CH₃CN (45:55 v/v) delivered at 1.5 mL/min isocratically. Quantification was performed at 238 nm, with bromazepam (1.5 ng/μL) 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 (&gt; 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. 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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 &lt;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.</abstract><cop>Weinheim</cop><pub>Wiley-VCH Verlag</pub><pmid>17625794</pmid><doi>10.1002/jssc.200700142</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record>
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subjects Amitriptyline - analysis
Amitriptyline - pharmacokinetics
Analysis
Antidepressive Agents, Tricyclic - analysis
Antidepressive Agents, Tricyclic - pharmacokinetics
Biological and medical sciences
Biological fluids
Chemistry Techniques, Analytical - methods
Chromatography, High Pressure Liquid - methods
Clomipramine - analysis
Clomipramine - pharmacokinetics
Doxepin - analysis
Doxepin - pharmacokinetics
Drug Monitoring - methods
General pharmacology
HPLC
Humans
Imipramine - analysis
Imipramine - pharmacokinetics
Medical sciences
Medicinal chemistry
Models, Chemical
Monitoring
Pharmacology. Drug treatments
Reproducibility of Results
Sensitivity and Specificity
Spectrophotometry - methods
Time Factors
Tricyclic antidepressants
Urinalysis - methods
title Development of an HPLC method for the monitoring of tricyclic antidepressants in biofluids
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