A simple and cheap all-in-one concept for at-line coupling of hollow fiber liquid-phase microextraction (HF-LPME) to commercial capillary electrophoresis (CE) is demonstrated, which enables the direct analysis of complex samples. A disposable microextraction device compatible with injection systems of Agilent CE instruments is proposed, which consists of a short segment of a porous HF attached to a tapered polypropylene holder. The holder maintains a constant position of the HF in a CE vial during extraction and simultaneously guides the injection end of a separation capillary into the HF lumen for automated CE injection and analysis. In a typical analytical procedure, the HF is impregnated with a water-immiscible solvent, its lumen is filled with 5 μL of an aqueous acceptor solution, and the microextraction device is placed in a 2 mL glass CE vial containing 550 μL of a donor solution. The vial is agitated at 750 rpm for 10 min, and the resulting acceptor solution is injected directly from the HF lumen into the commercial CE. No additional manual handling is required, except for the transfer of the CE vial to the CE autosampler. Multiple complex samples can be simultaneously pretreated in a multiple-well plate format, thus significantly reducing the total analysis time. Suitability of the analytical method is demonstrated by the direct determination of model basic drugs (nortriptyline, haloperidol, loperamide, and papaverine) in physiological solutions, urine, and dried blood spot (DBS) samples. Repeatability of the method is better than 12.8% (%RSD), extraction recoveries range between 34 and 76%, and enrichment factors are 37-84. The method is linear in a range of 2 orders of magnitude (R2 ≥ 0.9977) with limits of detection of 0.7-1.55 μg/L. The method has a high potential for the direct analysis of DBS samples since DBS elution and HF-LPME are performed simultaneously during the 10 min agitation. The manual DBS handling is thus reduced to inserting the DBS punch into the CE vial only. Moreover, the universal character of the HF-LPME might extend the applicability of the method to a wide range of analytes/matrices, and combination with other commercial detectors might improve the selectivity/sensitivity of the CE analysis.

• MeSH
• elektroforéza kapilární MeSH
• haloperidol analýza   MeSH
• lidé MeSH
• loperamid analýza   MeSH
• mikroextrakce kapalné fáze * MeSH
• nortriptylin analýza   MeSH
• papaverin analýza   MeSH
• poréznost MeSH
• povrchové vlastnosti MeSH
• tělesné tekutiny chemie   MeSH
• velikost částic MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• haloperidol MeSH
• loperamid MeSH
• nortriptylin MeSH
• papaverin MeSH

OBJECTIVE: Genetic and environmental factors interact in the development of major depressive disorder (MDD). While neurobiological correlates have only partially been elucidated, altered levels of calcitonin gene-related peptide (CGRP)-like immunoreactivity (LI) in animal models and in the cerebrospinal fluid of depressed patients were reported, suggesting that CGRP may be involved in the pathophysiology and/or be a trait marker of MDD. However, changes in CGRP brain levels resulting from interactions between genetic and environmental risk factors and the response to antidepressant treatment have not been explored. METHODS: We therefore superimposed maternal separation (MS) onto a genetic rat model (Flinders-sensitive and -resistant lines, FSL/FRL) of depression, treated these rats with antidepressants (escitalopram and nortriptyline) and measured CGRP-LI in selected brain regions. RESULTS: CGRP was elevated in the frontal cortex, hippocampus and amygdala (but not in the hypothalamus) of FSL rats. However, MS did not significantly alter levels of this peptide. Likewise, there were no significant interactions between the genetic and environmental factors. Most importantly, neither escitalopram nor nortriptyline significantly altered brain CGRP levels. CONCLUSION: Our data demonstrate that increased brain levels of CGRP are present in a well-established rat model of depression. Given that antidepressants have virtually no effect on the brain level of this peptide, our study indicates that further research is needed to evaluate the functional role of CGRP in the FSL model for depression.

• Klíčová slova
• Flinders-sensitive line, calcitonin gene-related peptide, escitalopram, maternal deprivation, nortriptyline,
• MeSH
• amygdala účinky léků   metabolismus   MeSH
• antidepresiva farmakologie   MeSH
• čelní lalok účinky léků   metabolismus   MeSH
• citalopram farmakologie   MeSH
• deprese * farmakoterapie   etiologie   metabolismus   MeSH
• hipokampus účinky léků   metabolismus   MeSH
• interakce genů a prostředí * MeSH
• krysa rodu Rattus MeSH
• maternální deprivace * MeSH
• modely nemocí na zvířatech MeSH
• mozek * účinky léků   metabolismus   MeSH
• nortriptylin farmakologie   MeSH
• peptid spojený s genem pro kalcitonin * účinky léků   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antidepresiva MeSH
• citalopram MeSH
• nortriptylin MeSH
• peptid spojený s genem pro kalcitonin * MeSH

Aqueous and organic phases in microelectromembrane extraction (μ-EME) were formed as adjacent plugs of free immiscible solutions in narrow-bore polymeric tubing, and each single phase was recovered and analyzed after μ-EME. A three-phase μ-EME setup was employed for investigation of time-dependent distribution of model basic drugs among aqueous and organic phases. Exact concentrations of nortriptyline and papaverine in donor solution, acceptor solution, and free liquid membrane (FLM) were determined by capillary electrophoresis with ultraviolet detection (CE-UV). At typical μ-EME conditions (acceptor, 1 μL of 25 mM HCl; FLM, 1 μL of 4-nitrocumene; donor, 1 μL of basic drugs in 10 mM HCl; and extraction potential, 250 V), experimentally determined distribution of the drugs confirmed the kinetic model for electrically mediated transfer of charged analytes. Rapid depletion of the drugs from donor solution (0-180 s) and rapid saturation of FLM with the drugs (15-60 s) were followed by gradual transfer of the drugs across FLM and gradual liberation into acceptor solution (30-240 s). Exhaustive transfer of the drugs from donor to acceptor solution was obtained in 15 min. A good correlation between the analytes' distribution and μ-EME electric currents was observed; the currents increased during drugs' transfer across FLM, were concentration dependent, and demonstrated transfer of the drugs across FLM in their ionized forms. Proper understanding of the fundamental principles of μ-EME transfer enabled further fine-tuning of the μ-EME process. Transfer of the drugs across FLM was controlled by optimizing the composition and pH of acceptor solution, and quantitative fractionation of nortriptyline into aqueous acceptor (96%) and of papaverine into organic FLM (95%) was achieved based on their different pKa values. μ-EME fractionation of the two drugs was compatible with raw human urine and excellent repeatability (RSD ≤ 3.9%), linearity (r2 ≥ 0.9989), and limits of detection (≤ 0.15 μg/mL) were achieved for μ-EME-CE-UV of the drugs in standard solutions and urine samples.

• MeSH
• elektroforéza kapilární metody   MeSH
• lidé MeSH
• limita detekce MeSH
• mikroextrakce kapalné fáze metody   MeSH
• nortriptylin moč   MeSH
• papaverin moč   MeSH
• reprodukovatelnost výsledků MeSH
• roztoky chemie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• nortriptylin MeSH
• papaverin MeSH
• roztoky MeSH

A simple sample injection procedure compatible with commercial capillary electrophoresis (CE) instrumentation was developed, which enables handling sample volumes as little as 250nL for analytical applications where sample volume availability is of concern. Single-use micro-sampling inserts were prepared by thermal modification of polypropylene micropipette tips and the inserts were accommodated in standard CE vials in CE autosampler carousel. To ensure direct contact of separation capillary injection end with sample solution and to avoid possible damage to the capillary, a soft compression spring was placed at the bottom of the vial underneath the micro-sampling insert. Injections from sub-μL samples were carried out in conventional as well as in short-end injection mode, were compatible with standard i.d./o.d. (25-100μm/365μm) fused silica capillaries and with various background electrolyte solutions and detection modes. Excellent repeatability of replicate injections from 250nL to 3μL was achieved based on RSD values of quantitative analytical measures (peak heights ≤2.4% and peak areas ≤3.7%) for CE-UV-vis, CE-ESI-MS and CE-contactless conductivity detection of model basic drugs. The achieved RSD values were comparable with those for replicate injections of the drugs from standard CE vials. The reported concept of injections from micro-sampling inserts was further demonstrated useful in evaluation of micro-electromembrane extraction (μ-EME) of model basic drugs. Sub-μL volumes of operational solutions resulted in reduced lengths of μ-EME phases and improved extraction recoveries (66-91%) were achieved.

• Klíčová slova
• Capillary electrophoresis, Contactless conductivity detection, Mass spectrometry, Micro-extractions, Micro-sampling, UV–vis detection,
• MeSH
• elektroforéza kapilární přístrojové vybavení   metody   MeSH
• elektrolyty chemie   MeSH
• hmotnostní spektrometrie MeSH
• nortriptylin analýza   izolace a purifikace   MeSH
• papaverin analýza   MeSH
• roztoky chemie   MeSH
• spektrofotometrie ultrafialová MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• elektrolyty MeSH
• nortriptylin MeSH
• papaverin MeSH
• roztoky MeSH

An isocratic reversed-phase high-performance liquid chromatographic method with on-line solid-phase extraction for the simultaneous determination of amitriptyline and nortriptyline in serum has been developed. A 250-microliters serum sample is injected directly onto a commercially available CN cartridge and, after a washing step, the retained solutes are backflushed onto a bonded-phase CN column using a column-switching technique and a mobile phase composed of acetonitrile (26%) and 0.05 M phosphate buffer with diethylamine. Serum is diluted with 0.1 M sodium lauryl sulphate and centrifuged before the injection. Detection at 210 nm ensures sufficient sensitivity. The recovery is almost quantitative and the relative standard deviation ranges from 2.8 to 8.0% for concentrations of 200-40 ng/ml. Being rapid and simple, the method is convenient for routine use.

• MeSH
• amitriptylin krev   MeSH
• lidé MeSH
• nortriptylin krev   MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• amitriptylin MeSH
• nortriptylin MeSH

• MeSH
• amitriptylin škodlivé účinky   farmakologie   MeSH
• antidepresiva tricyklická škodlivé účinky   farmakologie   MeSH
• hypotenze chemicky indukované   MeSH
• inbrední kmeny potkanů MeSH
• kardiovaskulární systém účinky léků   MeSH
• krysa rodu Rattus MeSH
• nortriptylin škodlivé účinky   farmakologie   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• amitriptylin MeSH
• antidepresiva tricyklická MeSH
• nortriptylin MeSH

• MeSH
• amitriptylin krev   terapeutické užití   MeSH
• depresivní poruchy krev   diagnóza   farmakoterapie   MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• nortriptylin krev   terapeutické užití   MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• Názvy látek
• amitriptylin MeSH
• nortriptylin MeSH

• MeSH
• amitriptylin terapeutické užití   MeSH
• depresivní poruchy farmakoterapie   MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• nortriptylin terapeutické užití   MeSH
• thioridazin terapeutické užití   MeSH
• tranylcypromin terapeutické užití   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• srovnávací studie MeSH
• Názvy látek
• amitriptylin MeSH
• nortriptylin MeSH
• thioridazin MeSH
• tranylcypromin MeSH

• MeSH
• amitriptylin farmakologie   MeSH
• antidepresiva tricyklická farmakologie   MeSH
• deprese farmakoterapie   MeSH
• dopamin metabolismus   MeSH
• krysa rodu Rattus MeSH
• levodopa farmakologie   terapeutické užití   MeSH
• lidé MeSH
• mozek metabolismus   MeSH
• noradrenalin metabolismus   MeSH
• nortriptylin farmakologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• amitriptylin MeSH
• antidepresiva tricyklická MeSH
• dopamin MeSH
• levodopa MeSH
• noradrenalin MeSH
• nortriptylin MeSH

• MeSH
• aminokyseliny metabolismus   MeSH
• fosfolipasy metabolismus   MeSH
• králíci MeSH
• lysofosfatidylcholiny farmakologie   MeSH
• mozek enzymologie   MeSH
• nortriptylin farmakologie   MeSH
• tetrahydrokanabinol farmakologie   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• aminokyseliny MeSH
• fosfolipasy MeSH
• lysofosfatidylcholiny MeSH
• nortriptylin MeSH
• tetrahydrokanabinol MeSH