Brožura se zaměřuje na výzkumnou činnost Ústavu analytické chemie AV ČR. Určeno odborné veřejnosti.

• Keywords
• Praha (Česko),
• MeSH
• Academies and Institutes MeSH
• Chemistry, Analytic MeSH
• Chemistry history   MeSH
• History, 20th Century MeSH
• History, 21st Century MeSH
• Science MeSH
• Research MeSH
• Check Tag
• History, 20th Century MeSH
• History, 21st Century MeSH
• Geographicals
• Czech Republic MeSH

• Conspectus
• Analytická chemie
• NML Fields
• chemie, klinická chemie
• věda a výzkum
• NML Publication type
• brožury

• About
• Ústav analytické chemie (Akademie věd ČR) Authority

• Publication type
• Meeting Abstract MeSH

Exhaled breath condensate (EBC) is an attractive, non-invasive sample for clinical diagnostics. During EBC collection, its composition is influenced by the collection temperature, a factor that is often not thoroughly monitored and controlled. In this study, we assembled a novel, simple, portable, and inexpensive device for EBC collection, able to maintain a stable temperature at any value between -7 °C and +12 °C. The temperature was controlled using a microcontroller and a thermoelectric cooler that was employed to cool the aluminum block holding the glass tube or the polypropylene syringe. The performance of the novel sampler was compared with the passively cooled RTube™ and a simple EBC sampler, in which the temperature was steadily increasing during sampling. The developed sampler was able to maintain a stable temperature within ±1 °C. To investigate the influence of different sampling temperatures (i.e., +12, -7, -80 °C) on the analyte content in EBC, inorganic ions and organic acids were analyzed by capillary electrophoresis with a capacitively coupled contactless conductivity detector. It was shown that the concentration of metabolites decreased significantly with decreasing temperature. The portability and the ability to keep a stable temperature during EBC sampling makes the developed sampler suitable for point-of-care diagnostics.

• MeSH
• Biomarkers MeSH
• Breath Tests * MeSH
• Electrophoresis, Capillary MeSH
• Point-of-Care Testing MeSH
• Temperature MeSH
• Exhalation * MeSH
• Publication type
• Journal Article MeSH

An early diagnosis of cystic fibrosis (CF) through newborn screening programmes (NBSP) is very important to initiate the treatment, genetic counseling, and specialized medical services. NBSP consists of immunoreactive trypsinogen (IRT) analysis, DNA analysis and sweat test. Sweat test is a golden standard, but many patients fall into the inconclusive diagnosis (CFSPID) due to the intermediate chloride levels (30-60 mmol/L) and unclear DNA analysis. We propose the validation of an improved method, based on non-invasive skin wipe sampling and analysis of all ions in the samples. We propose the use of ion ratios of selected ions (Cl-, K+, Na+, HCO3-) as a relevant diagnostic parameter and aim at evaluation of sensitivity, selectivity and repeatability of the method. We further propose creation of a ion ratio database of a large cohort of subjects during the NBSP as well as adult CF patients and controls to identify possible correlations between the type of mutation, severity and other symptoms with data obtained from the skin wipe samples to improve classification of CF patients.

Diagnóza cystické fibrózy (CF) novorozeneckým screeningem je důležitá, protože umožňuje včasné zahájení léčby, genetického poradenství a specializované terapie. Novorozenecký screening se skládá z analýzy imunoreaktivního trypsinogenu (IRT), genetické analýzy DNA a obsahu chloridů pomocí potního testu. Potní test je považován za tzv. zlatý standard v diagnostice CF. Výsledky potního testu v tzv. šedé zóně (Cl-: 30-60 mmol/L) a genetické analýzy ovšem někdy vedou k nejednoznačnému diagnostickému závěru (CFSPID). V tomto projektu navrhujeme validaci nové metody analýzy potu, založené na neinvazivním, rychlém a jednoduchém stěru předloktí. Ve vzorku budou analyzovány poměry vybraných iontů (Cl-, K+, Na+, HCO3-) které mají významný potenciál pro zlepšení diagnostiky CF. Je plánováno na statisticky významném souboru pacientů a zdravých kontrol vyhodnotit důležité parametry (citlivost, specificita, opakovatelnost, atd.) a vytvořit databázi iontových poměrů , určení korelace s typem genetické mutace, vážností onemocnění a dalšími symptomy, umožňujícímu zlepšení klasifikace CF pacientů.

• MeSH
• Chlorides MeSH
• Cystic Fibrosis diagnosis   MeSH
• Genetic Testing MeSH
• Ions MeSH
• Clinical Chemistry Tests MeSH
• Infant, Newborn MeSH
• Neonatal Screening MeSH
• Sweat MeSH
• Reproducibility of Results MeSH
• Check Tag
• Infant, Newborn MeSH

• Conspectus
• Pediatrie
• NML Fields
• perinatologie a neonatologie
• genetika, lékařská genetika
• pneumologie a ftizeologie
• NML Publication type
• závěrečné zprávy o řešení grantu AZV MZ ČR

Bile acids are a group of steroid compounds essential for lipid digestion. However, when bile acids are refluxed into the stomach and the esophagus, during the so called duodenogastroesophageal reflux, they can have a detrimental effect on the esophageal epithelium and cause pathological changes of esophageal tissue, e.g., Barrett's esophagus (BE). The levels of bile acids in saliva could therefore serve as possible biomarkers for the diagnostics of BE. In this work, we focused on optimization of sample collection and preparation by solid-phase extraction and subsequent quantification of 11 bile acids (unconjugated, glycine-conjugated) in saliva from healthy volunteers and BE patients by ultra-high-performance liquid chromatography coupled to triple-quadrupole tandem mass spectrometry. Moreover, high resolution MS (Orbitrap-MS) was utilized for identification of new bile acids in saliva. Methods for saliva collection including simple spitting and the Salivette® saliva collection system were compared; the latter was found to be unsuitable due to excessive retention of bile acids in the cotton swab. Methanol with 0.1% formic acid were selected for protein precipitation and bile acid extraction prior to SPE. Separation was performed in gradient elution of methanol and 0.1% formic acid in less than 10 min. Saliva from BE patients contained higher levels of almost all bile acids, and the tested groups could be distinguished by principal component analysis. In untargeted analysis by high resolution MS, taurine-conjugated bile acids and glycine-conjugated dihydroxy-bile acid sulfate were identified in saliva from healthy volunteers. We propose that analysis of salivary bile acids including taurine conjugates could be applicable in diagnostics of BE, following a larger clinical study.

• MeSH
• Barrett Esophagus * metabolism   MeSH
• Chromatography, Liquid MeSH
• Formates MeSH
• Glycine analysis   MeSH
• Mass Spectrometry MeSH
• Humans MeSH
• Methanol analysis   MeSH
• Saliva chemistry   MeSH
• Taurine analysis   MeSH
• Bile Acids and Salts analysis   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

Gastroesophageal reflux disease (GERD) is a highly prevalent disease, affecting 10-20% individuals worldwide. It can lead to esophageal and extraesophageal (EER) symptomatology. GERD and EER are two unique, but related disease entities, with different risk factors, symptoms, pathophysiology and response to therapy. Diagnostic methods to confirm or reject GERD/EER do not achieve high sensitivity and specificity. In routine clinical practice, there is a need to distinguish patients with symptoms that are due to reflux (either increased reflux or hypersensitivity to a normal amount of reflux) from patients with symptoms suggestive of reflux but not due to reflux (ie, FH). Better tools for GERD/EER diagnosis are needed, particularly, if they have low cost, are non-invasive and can be performed in a primary care setting. The development and assessment of non-invasive methods based on monitoring exhaled breath condensate and saliva is proposed to improve the diagnosis of GERD/EER.

Refluxní choroba jícnu (GERD) je rozšířené onemocnění postihující 10-20% světové populace. Může vést k jícnové a mimojícnové (EER) symptomatologii. GERD a EER jsou dvě propojené, ale specifické nemoci s rozdílnými rizikovými faktory, symptomy, patofyziologií a odpovědí na terapii. Diagnostické metody pro potvrzení nebo vyvrácení GERD / EER nedosahují vysokou senzitivitu a specificitu. V klinické praxi je třeba rozlišit pacienty s příznaky v souvislosti s refluxem (zvýšený počet refluxních epizod, nebo hypersenzitivita k normálnímu množství refluxních epizod) od pacientů s příznaky podobnými refluxním, ale ne v důsledku refluxu (funkční poruchy). K detekci GERD je zapotřebí lepších diagnostických metod zejména v případě, že mají nízké náklady, jsou neinvazivní a mohou být prováděny v rámci primární péče. Současný návrh se zaměřuje na vývoj a hodnocení neinvazivních metod založených na analýze kondenzátu vydechovaného vzduchu a slin s cílem zlepšit diagnostiku GERD / EER.

• MeSH
• Breath Tests methods   MeSH
• Gastroesophageal Reflux diagnosis   MeSH
• Humans MeSH
• Pilot Projects MeSH
• Reproducibility of Results MeSH
• Saliva MeSH
• Check Tag
• Humans MeSH
• Publication type
• Evaluation Study MeSH

• Conspectus
• Patologie. Klinická medicína
• NML Fields
• gastroenterologie
• NML Publication type
• závěrečné zprávy o řešení grantu AZV MZ ČR

This article describes the history, development, and current state of two analytical methods to which Czech scientists have made significant contributions. It describes the first steps of these methods in Czechoslovakia during the 1970s and 1980s, the development of the methods through innovation, and the establishment of cooperation with foreign scientists. Today, Czech scientists from the Department of Analytical Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University, and Institute of Analytical Chemistry of the Czech Academy of Sciences in Brno are among the major contributors to these fields of analytical chemistry. The milestones in the development and the prominent scientists mentioned in the individual chapters give a chance for a bright future for these methods, not only in the Czech Republic.

• MeSH
• History, 20th Century MeSH
• History, 21st Century MeSH
• Electrophoresis, Capillary * history   methods   MeSH
• Humans MeSH
• Flow Injection Analysis MeSH
• Check Tag
• History, 20th Century MeSH
• History, 21st Century MeSH
• Humans MeSH
• Publication type
• Historical Article MeSH
• Geographicals
• Czech Republic MeSH

A simple and fast method for the analysis of lactate from a single drop of blood was developed. The finger-prick whole blood sample (10 µL) was diluted (1:20) with a 7% (w/v) solution of [tris(hydroxymethyl)methylamino] propanesulfonic acid and applied to a blood plasma separation device. The device accommodates a membrane sandwich composed of an asymmetric polysulfone membrane and a supporting textile membrane that allows the collection of blood plasma into a narrow glass capillary in less than 20 s. Separated and simultaneously diluted blood plasma was directly injected into a capillary electrophoresis instrument with a contactless conductivity detector (CE-C4D) and analyzed in less than one minute. A separation electrolyte consisted of 10 mmol/L l-histidine, 15 mmol/L dl-glutamic acid, and 30 µmol/L cetyltrimethylammonium bromide. The whole procedure starting from the finger-prick sampling until the CE-C4D analysis was finished, took less than 5 min and was suitable for monitoring lactate increase in blood plasma during incremental cycling exercise. The observed lactate increase during the experiments measured by the developed CE-C4D method correlated well with the results from a hand-held lactate analyzer (R = 0.9882). The advantage of the developed CE method is the speed, significant savings per analysis, and the possibility to analyze other compounds from blood plasma.

• MeSH
• Exercise * MeSH
• Electric Conductivity * MeSH
• Electrophoresis, Capillary methods   MeSH
• Lactic Acid blood   MeSH
• Humans MeSH
• Limit of Detection MeSH
• Athletes psychology   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

A fast method for the determination of tropane alkaloids, using a portable CE instrument with a capacitively coupled contactless conductivity detector (CE-C4D) was developed and validated for determination of atropine and scopolamine in seeds from Solanaceae family plants. Separation was obtained within 5 min, using an optimized background electrolyte consisting of 0.5 M acetic acid with 0.25% (w/v) β-CD. The limit of detection and quantification was 0.5 µg/mL and 1.5 µg/mL, respectively, for both atropine and scopolamine. The developed method was validated with the following parameters-precision (CV): 1.07-2.08%, accuracy of the assay (recovery, RE): 101.0-102.7% and matrix effect (ME): 92.99-94.23%. Moreover, the optimized CE-C4D method was applied to the analysis of plant extracts and pharmaceuticals, proving its applicability and accuracy.

• MeSH
• Solanaceous Alkaloids analysis   MeSH
• Atropine analysis   MeSH
• Electrophoresis, Capillary MeSH
• Limit of Detection MeSH
• Scopolamine analysis   MeSH
• Solanaceae chemistry   MeSH
• Publication type
• Journal Article MeSH

Analýza biologických markerů v kondenzátu vydechovaného vzduchu (KVV) je považována za bezpečnou neinvazivní techniku k hodnocení zánětu v dýchacích cestách u pacientů všech věkových kategorií. Zánět v dýchacích cestách je charakteristickým znakem řady chronických plicních onemocnění (chronické obstrukční plicní nemoci, bronchiálního astma, cystické fibrózy, bronchopulmonální dysplázie a primární ciliární dyskineze). Tato metoda by mohla být užitečná pro rozlišení přítomnosti zánětu, k monitorování efektu léčby a k predikci exacerbace onemocnění. Cílem této práce bylo podat souhrnný přehled poznatků z vybraných studií, které se věnovaly detekci cytokinů v kondenzátu vydechovaného vzduchu (KVV) u pacientů s bronchiálním astmatem (AB) a idiopatickou plicní fibrózou (IPF). Popisujeme klinický význam vybraných cytokinů, metodologii odběru KVV, analytické metody a možné důvody, proč tato metoda nebyla dosud zavedena do klinické praxe.

• MeSH
• Biomarkers analysis   MeSH
• Asthma * diagnosis   MeSH
• Cytokines * therapeutic use   MeSH
• Pulmonary Elimination MeSH
• Idiopathic Pulmonary Fibrosis * diagnosis   MeSH
• Humans MeSH
• Exhalation MeSH
• Check Tag
• Humans MeSH