Bioanalytical methods
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Magnetic particles play an important role in current technology, and this field of technology extends to a broader progression. The term magnetic particles typically cover the paramagnetic particles and super-paramagnetic particles. Various materials like iron oxide are common, but other materials are available as well; a survey of such materials has been included in this work. They can serve for technological purposes like separation and isolation of chemical products or toxic waste, their use in the diagnosis of pathologies, drug delivery and other similar applications. In this review, biosensors, bioanalytical devices and bioassays, have been discussed. Materials for magnetic particles preparation, methods of assay, biosensors and bioassays working in stationary as well as flow-through arrangements are described here. A survey of actual literature has been provided as well.
- MeSH
- biosenzitivní techniky * MeSH
- biotest MeSH
- lékové transportní systémy MeSH
- lidé MeSH
- magnetické jevy MeSH
- magnetismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Současné možnosti diagnostiky karcinomu prostaty neumožňují odlišit agresivní (signifikantní tumory) od klinicky latentních (nesignifikantních forem). Práce se zaměřuje na stanovení hladiny klinicky využitelných tumorových markerů karcinomu prostaty: alfa-metyl CoA-racemáza (AMACR), caveolin-1, metalothionein (MT), p53, NF-?B, c-FOS, c-JUN, Ki-67, prostatický specifický antigen (PSA), ZIP1 a ZnT-1 na úrovni prostatické tkáně (reprezentované buněčnými liniemi 22Rv1 a PNT1A) a v séru pacientů s histologicky ověřeným adenokarcinomem (82 karcinomových a 51 kontrolních sér). Na úrovni mRNA bylo zjištěno snížení exprese Cav-1, NF-?B, c-FOS a c-JUN a zvýšení exprese metalothioneinu, AMACR, PSA, Ki-67, MMP-9 a zinkových transportérů ZIP1 a ZnT-1. V séru je popsáno signifikantní zvýšení metalothioneinu nad hladinu 1 µM. Hladina caveolinu-1 se významně zvyšuje u high-grade tumorů, což jej staví do pozice možného markeru agresivní formy karcinomu prostaty.
Current methods for diagnosis of prostate carcinoma do not enable us to distinguish aggressive tumours (significant tumours) from clinically latent tumours (non-significant tumours). This study aims to determine levels of potential clinically important tumour markers such as alpha-methyl CoA-racemase (AMACR), caveolin-1, metallothionein (MT), p53, NF-?B, c-FOS, c-JUN, Ki-67, prostate-specific antigen (PSA), ZIP1 and ZnT-1 in prostatic tissue represented by 22Rv1 (tumour) and PNT1A (healthy) cell lines and in blood serum of patients with histologically evaluated adenocarcinoma (82 tumour patients and 51 healthy volunteers). Based on mRNA expression, it was found that Cav-1, NF-?B, c-FOS and c-JUN were down-regulated and MT, AMACR, PSA, Ki-67, MMP-9 and zinc transporters ZIP1 and ZnT-1 were up-regulated. In serum, the level of MT was significantly enhanced above 1 µM. Caveolin-1 levels were significantly increased in high-grade tumours, which points to the possibility of using this protein as a marker for the aggressive form of prostate carcinoma.
- Klíčová slova
- nádorová onemocnění, imunodetekce, molekulárně-biologické techniky,
- MeSH
- 2D gelová elektroforéza využití MeSH
- diagnostické techniky molekulární metody MeSH
- elektrochemie metody MeSH
- financování organizované MeSH
- kaveolin 1 diagnostické užití MeSH
- lidé MeSH
- nádorové biomarkery krev MeSH
- nádorové buněčné linie chemie MeSH
- nádory prostaty genetika krev MeSH
- polymerázová řetězová reakce využití MeSH
- prostatický specifický antigen izolace a purifikace krev MeSH
- senzitivita a specificita MeSH
- stupeň závažnosti nemoci MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
sv.
- MeSH
- chemické techniky analytické MeSH
- Publikační typ
- periodika MeSH
- Konspekt
- Chemie. Mineralogické vědy
- NLK Obory
- chemie, klinická chemie
- chemie, klinická chemie
sv.
- MeSH
- elektroforéza MeSH
- klinická chemie metody MeSH
- Publikační typ
- periodika MeSH
- Konspekt
- Chemie. Mineralogické vědy
- NLK Obory
- chemie, klinická chemie
- chemie, klinická chemie
The development of bioanalytical methods has become more and more challenging over the past years due to very demanding requirements in terms of method reliability, sensitivity, speed of analysis and sample throughput. LC-MS/MS has established itself as a method of choice for routine analysis of biological materials. A development of such method consists of several steps including sample preparation and clean-up step, efficient chromatographic separation, sensitive and selective detection of analytes in complex matrices, a choice of convenient data processing and calibration approach and finally method validation. Each of these steps has its own constraints and challenges, which are discussed in detail in this review. Novel and modern approaches in sample preparation, chromatography and detection are especially emphasized. Attention is paid to proper calibration approach and matrix effects that can seriously affect method accuracy and precision.
- MeSH
- časové faktory MeSH
- chromatografie kapalinová ekonomika přístrojové vybavení metody MeSH
- hmotnostní spektrometrie ekonomika přístrojové vybavení metody MeSH
- lidé MeSH
- validační studie jako téma MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Cefazolin je širokospektré antibiotikum používané zejména jako profylaktické zajištění pacientů při chirurgických výkonech. U specifických skupin pacientů se setkáváme s variabilitou farmakokinetiky oproti populačnímu průměru. K optimalizaci dávkovacích režimů je třeba využívat spolehlivé bioanalytické metody schopné rychlého stanovení vzorku. V současnosti nejvyužívanější metodou pro měření plazmatických koncentrací je kapalinová chromatografie s UV nebo MS detekcí. K preanalytické přípravě vzorku lze využít proteinovou precipitaci a extrakci na pevné fázi. Při potřebě stanovení volné frakce lze využít rovnovážnou dialýzu či ultrafiltraci. Nežádoucí faktory, ovlivňující správnost stanovení, mohou být matricové efekty nebo nízká stabilita vzorku.
Cefazolin is a broad-spectrum antibiotic used primarily as a prophylactic care for patients during surgery. In specific groups of patients, we encounter variability in pharmacokinetics from the population average. To optimize dosing regimens, reliable bioanalytical methods capable of rapid sample determination should be used. Currently, the most widely used method for measuring plasma concentrations is liquid chromatography with UV or MS detection. Protein precipitation and solid phase extraction can be used for pre-analytical sample preparation. Equilibrium dialysis or ultrafiltration can be used if the free fraction needs to be determined. Undesirable factors affecting the accuracy of the assay may be matrix effects or low sample stability.
- MeSH
- cefalosporiny antagonisté a inhibitory farmakokinetika krev MeSH
- cefazolin * analýza farmakokinetika krev MeSH
- imunoanalýza metody MeSH
- lidé MeSH
- monitorování léčiv MeSH
- odběr biologického vzorku metody MeSH
- vazba proteinů MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
Cell-based bioassays are very sensitive and allow integrative effect screening of the whole environmental sample, which is usually composed of a mixture of agonists and antagonists. Measured toxicity is usually expressed as a bioanalytical equivalent concentration. So far, it is not possible to distinguish which part of this value is caused by the agonists and which by the antagonists. In this article, we present a simple method to analyze the dose-response curve of a mixture and to determine an agonistic bioanalytical equivalent concentration: a concentration of a reference chemical that would elicit the same effect as do only agonists in an unknown mixture. The method has been validated using several artificially prepared mixtures of agonists and competitive antagonists measured in a recombinant yeast assay. No difference was observed between the calculated equivalent concentrations and the used concentrations of the agonist in the mixture.
- MeSH
- biotest metody MeSH
- endokrinní disruptory * MeSH
- estradiol farmakologie MeSH
- estriol farmakologie MeSH
- fulvestrant farmakologie MeSH
- látky znečišťující životní prostředí toxicita MeSH
- lékové interakce MeSH
- receptory pro estrogeny antagonisté a inhibitory MeSH
- Saccharomyces cerevisiae účinky léků MeSH
- teoretické modely MeSH
- Publikační typ
- časopisecké články MeSH
Prostate cancer has the highest malignancy rate diagnosed in men worldwide. Albeit, the gold standard serum prostate-specific antigen (PSA) assays reduced the mortality rate of the disease, the number of false positive diagnoses steeply increased. Therefore, there is an urgent need for complementary biomarkers to enhance the specificity and selectivity of current diagnostic methods. Information about PSA glycosylation can help to fulfill this gap as alterations of its carbohydrate moieties due to cancerous transformation may represent additional markers to distinguish malignant from benign tumors. However, development of suitable methods and instrumentations to investigate the N-glycosylation profile of PSA represents a challenge. In this paper, we critically review the current bioanalytical trends and strategies in the field of PSA glycobiomarker research focusing on separation based characterization methods.
Miniaturization continues to be one of the leading trends in analytical chemistry and one that brings advantages that can be particularly beneficial in biochemical research. Use of a miniaturized scale enables efficient analysis in a short time and requires very small amounts of samples, solvents, and reagents. This can result in a remarkable decrease in costs of enzyme kinetics studies, especially when expensive or rare enzymes and/or substrates are involved. Free zone electrophoresis is without a doubt the most common microscale separation technique for capillary and on-chip enzyme assays. Progress and applications in this field are reviewed frequently whereas other modes of separation, although successfully applied, receive only marginal interest in such publications. This review summarizes applications of less common modes of separation in capillary or chip formats, namely micellar electrokinetic chromatography, liquid chromatography, gel electrophoresis, isoelectric focusing, and isotachophoresis. Because these techniques are based on separation mechanisms different from those of free zone electrophoresis, they can be, and have been, successfully used in cases where zone electrophoresis fails. Advantages and drawbacks of these alternative separation techniques are discussed, as also are the difficulties encountered most often and solutions proposed by different research groups.
- MeSH
- chromatografie micelární elektrokinetická kapilární metody MeSH
- elektroforéza kapilární metody MeSH
- enzymatické testy metody MeSH
- isoelektrická fokusace metody MeSH
- izotachoforéza metody MeSH
- kapilární elektrochromatografie metody MeSH
- kinetika MeSH
- lidé MeSH
- miniaturizace metody MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
This study compares alternative approaches for analyzing phytocannabinoids in different plant materials. Three chromatographic analytical methods (ultra-high-performance liquid chromatography with tandem mass spectrometric detection and gas chromatography with mass spectrometric and flame ionization detection) were evaluated regarding selectivity, sensitivity, analytical accuracy, and precision. The performance of the methods was compared and all three methods were demonstrated to be appropriate tools for analyzing phytocannabinoids in cannabis. Gas chromatography coupled with mass spectrometric detection showed slightly better accuracy in determining phytocannabinoid acids, which are often difficult to quantify owing to their limited stability. Aspects of sample preparation, such as material homogenization and extraction, were also considered. A single ultrasonic-assisted ethanolic extraction of dried and powdered plant samples of cannabis was shown to be exhaustive for extracting the samples prior to analysis.
- MeSH
- Cannabis chemie MeSH
- kanabinoidy analýza MeSH
- laboratoře organizace a řízení MeSH
- limita detekce MeSH
- plamínková ionizace metody MeSH
- plynová chromatografie s hmotnostně spektrometrickou detekcí metody MeSH
- reprodukovatelnost výsledků MeSH
- tandemová hmotnostní spektrometrie metody MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
