The article contains information on the properties of fentanyl derivatives and a method for their simple orienta-tion identification. Thin-layer chromatography (TLC) was deliberately chosen to perform the experiment because it is the most suitable method for rapid analysis of the col-lected sample under the conditions of the field chemical laboratory operated by the Army of the Czech Republic. On the basis of the experimental work performed, it was found that the most suitable system for the identification of the fentanyl derivatives of interest by TLC is a system where the stationary phase consists of "alumina 60 F254 neutral" and the mobile phase is comprised of ethyl ace-tate and hexane in a ratio of 7:3. The findings are fully usable in practice and will be included in the standard operating procedures for the determination of these types of substances in the Field Transport Chemical Laboratory PPCHL-AL2/ch, most likely in the form of acertified methodology.
Pancreatic ductal adenocarcinoma (PDAC) is one of the most common causes of cancer-related deaths worldwide, accounting for 90% of primary pancreatic tumors with an average 5-year survival rate of less than 10%. PDAC exhibits aggressive biology, which, together with late detection, results in most PDAC patients presenting with unresectable, locally advanced, or metastatic disease. In-depth lipid profiling and screening of potential biomarkers currently appear to be a promising approach for early detection of PDAC or other cancers. Here, we isolated and characterized complex glycosphingolipids (GSL) from normal and tumor pancreatic tissues of patients with PDAC using a combination of TLC, chemical staining, carbohydrate-recognized ligand-binding assay, and LC/ESI-MS2. The major neutral GSL identified were GSL with the terminal blood groups A, B, H, Lea, Leb, Lex, Ley, P1, and PX2 determinants together with globo- (Gb3 and Gb4) and neolacto-series GSL (nLc4 and nLc6). We also revealed that the neutral GSL profiles and their relative amounts differ between normal and tumor tissues. Additionally, the normal and tumor pancreatic tissues differ in type 1/2 core chains. Sulfatides and GM3 gangliosides were the predominant acidic GSL along with the minor sialyl-nLc4/nLc6 and sialyl-Lea/Lex. The comprehensive analysis of GSL in human PDAC tissues extends the GSL coverage and provides an important platform for further studies of GSL alterations; therefore, it could contribute to the development of new biomarkers and therapeutic approaches.
- MeSH
- chromatografie kapalinová MeSH
- chromatografie na tenké vrstvě MeSH
- duktální karcinom pankreatu diagnóza patofyziologie MeSH
- gangliosidy chemie MeSH
- glykosfingolipidy * analýza chemie MeSH
- lidé MeSH
- nádorové biomarkery metabolismus MeSH
- nádory slinivky břišní * diagnóza patofyziologie MeSH
- sulfoglykosfingolipidy chemie MeSH
- tandemová hmotnostní spektrometrie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Passiflora species, mangosteen, and cherimoya peels are a source of bioactive phenolic compounds. Nevertheless, a significant fraction of polyphenols, called non-extractable polyphenols (NEPs), are retained in the extraction residue after a conventional extraction. Thus, alkaline, acid, and enzymatic-assisted extractions to recover high contents of antioxidant NEPs from the extraction residue of fruit peels, were compared in this work. A high-performance thin-layer chromatography method with UV/Vis detection was developed in order to obtain the phenolic profile for the extracts. The most intense bands were further analyzed by direct analysis in real-time-high-resolution mass spectrometry to tentatively identified NEPs in fruit peel extracts. Total phenolic and proanthocyanidin contents and antioxidant capacity of the extracts were measured to carry out a multivariate statistical analysis. Alkaline hydrolysis was the most efficient treatment to recover NEPs from fruit peels as well as a promising treatment to obtain antioxidant extracts along with EAE. Cherimoya peel extracts were the richest in antioxidant NEPs. This work highlights that many NEPs remain on the extraction residue of fruit peels after conventional extraction and are not usually taken into account.
Electron paramagnetic resonance (EPR) spectroscopy represents an established tool to study properties of microenvironments, e.g. to investigate the structure and dynamics of biological and artificial membranes. In this study, the partitioning of the spin probe 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) in ex vivo human abdominal and breast skin, ex vivo porcine abdominal and ear skin as well as normal and inflammatory in vitro skin equivalents was investigated by EPR spectroscopy. Furthermore, the stratum corneum (SC) lipid composition (as determined by high-performance thin-layer chromatography), SC lipid chain order (probed by infrared spectroscopy) and the SC thickness (investigated by histology) were determined in the skin models. X-band EPR measurements have shown that TEMPO partitions in the lipophilic and hydrophilic microenvironment in varying ratios in different ex vivo and in vitro skin models. Ex vivo human abdominal skin exhibited the highest amount of TEMPO in the lipophilic microenvironment. In contrast, the lowest amount of TEMPO in the lipophilic microenvironment was determined in ex vivo human breast skin and the inflammatory in vitro skin equivalents. Individual EPR spectra of epidermis including SC and dermis indicated that the lipophilic microenvironment of TEMPO mainly corresponds to the most lipophilic part of the epidermis, the SC. The amount of TEMPO in the lipophilic microenvironment was independent of the SC lipid composition and the SC lipid chain order but correlated with the SC thickness. In conclusion, EPR spectroscopy could be a novel technique to determine differences in the SC thickness, thus suitably complementing existing methods.
- MeSH
- břicho MeSH
- buněčné mikroprostředí MeSH
- chromatografie na tenké vrstvě MeSH
- cyklické N-oxidy chemie MeSH
- dospělí MeSH
- elektronová paramagnetická rezonance MeSH
- epidermis chemie MeSH
- kůže chemie cytologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- lipidy chemie MeSH
- mladý dospělý MeSH
- prasata MeSH
- prsy MeSH
- senioři MeSH
- spektrofotometrie infračervená MeSH
- spinové značení MeSH
- tloušťka kožní řasy MeSH
- zevní ucho MeSH
- zvířata MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The lipid composition of sperm membranes is crucial for fertilization and differs among species. As the evolution of internal fertilization modes in fishes is not understood, a comparative study of the sperm lipid composition in freshwater representatives of externally and internally fertilizing fishes is needed for a better understanding of taxa-specific relationships between the lipid composition of the sperm membrane and the sperm physiology. The lipidomes of spermatozoa from stingray, a representative of cartilaginous fishes possessing internal fertilization, and sterlet, a representative of chondrostean fishes with external fertilization, have been studied by means of nuclear magnetic resonance (NMR), matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), electrospray MS, gas chromatography-(GC) MS, and thin-layer chromatography (TLC). NMR experiments revealed higher cholesterol content and the presence of phosphatidylserine in stingray compared to sterlet sperm. Unknown MS signals could be assigned to different glycosphingolipids in sterlet (neutral glycosphingolipid Gal-Cer(d18:1/16:0)) and stingray (acidic glycosphingolipid sulpho-Gal-Cer(d18:1/16:0)). Free fatty acids in sterlet sperm indicate internal energy storage. GC-MS experiments indicated a significant amount of adrenic acid, but only a low amount of docosahexaenoic acid in stingray sperm. In a nutshell, this study provides novel data on sperm lipid composition for freshwater stingray and sterlet possessing different modes of fertilization.
- MeSH
- chromatografie na tenké vrstvě MeSH
- druhová specificita MeSH
- fertilizace fyziologie MeSH
- glykosfingolipidy chemie MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací MeSH
- kyseliny dokosahexaenové chemie MeSH
- lipidomika MeSH
- lipidy chemie MeSH
- magnetická rezonanční spektroskopie MeSH
- plynová chromatografie s hmotnostně spektrometrickou detekcí MeSH
- ryby fyziologie MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- spermie chemie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Background/Aim: Radiolabelling of monoclonal antibodies (mAbs) could be beneficial in cancer diagnosis and therapy, however it may cause structural changes and consequently deteriorate their immunoreactivity. Materials and Methods: The therapeutic mAb ramucirumab (RAM) was technetium-99m labelled using either a direct or an indirect method with the use of two bifunctional chelating agents (HYNIC, DTPA). The radiochemical purity was assessed using instant thin-layer chromatography (ITLC) and high-performance liquid chromatography (HPLC) technique. The affinity of radiolabelled RAM was tested on human cancer cell lines. Results: The radiolabelling provided the following stable compounds: [99mTc]RAM, [99mTc]HYNIC-RAM and [99mTc]DTPA-RAM. Their radiochemical purity was over 95%. All prepared radiopharmaceuticals showed moderate affinity to the targeted receptor, in vitro. However, their affinity was one order lower compared to that of the natural mAb. Moreover, directly and DTPA-radiolabelled RAM demonstrated less favourable binding kinetics. Conclusion: Radiolabelling negatively affected the affinity of RAM to its targeted receptor.
- MeSH
- chelátory chemie MeSH
- chromatografie na tenké vrstvě MeSH
- inhibiční koncentrace 50 MeSH
- inhibitory angiogeneze chemie farmakologie MeSH
- lidé MeSH
- monoklonální protilátky chemie farmakologie MeSH
- nádorové buněčné linie MeSH
- nádory prostaty patologie MeSH
- poločas MeSH
- radiofarmaka farmakologie MeSH
- receptor 2 pro vaskulární endoteliální růstový faktor metabolismus MeSH
- technecium MeSH
- tkáňová distribuce MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
This study reports the development and validation of a new, simple, and accurate high-performance thin-layer chromatography (HPTLC)-densitomeric method for the determination of nandrolone decanoate in a commercially available injection formulation. Chromatographic analysis was performed on glass CN modified silica gel 60F254 plates developed using n-hexane-ethyl acetate in volume ratio 42.5:7.5 as the mobile phase. Densitometric scanning was carried out at the wavelength of 245 nm. This chromatographic system gave compact spot and a symmetrical peak of nandrolone decanoate with retardation factor (RF) value at 0.57 (±0.02). The linearity of this method with the high correlation coefficient of calibration plot ranges from 0.780 to 12.500 μg/spot. The developed method is characterized by good precision (coefficient of variation CV < 2%) and high accuracy close to 100.3% (R = 99.0%). Values of limits of detection and quantification equal to 0.231 and 0.700 μg/spot, respectively, confirm the sensitivity of the developed method. The analysis of the pharmaceutical formulation of nandrolone decanoate indicates drug content of 50.5 mg/mL and 101.0% in relation to the label claim. This is in good agreement with the recommendation of the International Council for Harmonisation (ICH) guidelines as well as the pharmacopoeial requirements. The low CV value (<1%) of nandrolone decanoate content in the tested injection formulation confirms the suitability of the proposed HPTLC-densitometric method for routine control of this compound in examined pharmaceuticals.
- MeSH
- chromatografie na tenké vrstvě * metody normy MeSH
- denzitometrie * metody normy MeSH
- léčivé přípravky analýza MeSH
- nandrolon dekanoát analýza MeSH
- příprava léků MeSH
- reprodukovatelnost výsledků MeSH
- senzitivita a specificita MeSH
- vysokoúčinná kapalinová chromatografie * metody normy MeSH
- Publikační typ
- časopisecké články MeSH
In this work we present a simple and cost-effective approach for the determination of selenium species in algae and yeast biomass, based on a combination of thin-layer chromatography (TLC) with diode laser thermal vaporization inductively coupled plasma mass spectrometry (DLTV ICP MS). Extraction of freeze-dried biomass was performed in 4M methanesulphonic acid and the selenium species were vaporized from cellulose TLC plates employing a continuous-wave infrared diode laser with power up to 4 W using a simple laboratory-built apparatus. Selenomethionine and selenocysteine were quantified with limits of detection 3 μg L-1 in a Se-enriched microalgae Chlorella vulgaris and yeast certified reference material SELM-1. Results delivered by TLC-DLTV ICP MS were consistent with those obtained by a routine coupling of high-performance liquid chromatography (HPLC) to ICP MS. In addition, the TLC approach is capable of analyzing extract containing even undiluted crude hydrolysates that could damage HPLC columns.
- MeSH
- chemické techniky analytické ekonomika metody MeSH
- Chlorella vulgaris chemie MeSH
- chromatografie na tenké vrstvě * MeSH
- hmotnostní spektrometrie * přístrojové vybavení MeSH
- lasery polovodičové MeSH
- Saccharomyces cerevisiae chemie MeSH
- selenocystein analýza MeSH
- selenomethionin analýza MeSH
- spektrální analýza MeSH
- volatilizace MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Publikační typ
- časopisecké články MeSH
The rise of the mosquitoes-transmitted diseases, like dengue, zika and chikungunya in Brazil in the last years has increased concerns on protection against mosquitoes bites. However, the prohibitive prices of the commercially available repellents for the majority of the Brazilian population has provoked a search for cheaper solutions, like the use of the homemade ethanolic extract of Indian clove (Syzygium aromaticum L.) as repellent, which has been reported as quite efficient by the local press. In order to verify this, we performed here the quantification of the main components of this extract through high-performance thin-layer chromatography (HPTLC)-densitometry and evaluated its efficiency as a repellent and its acetylcholinesterase (AChE) inhibition capacity. Our results have proved HPTLC-densitometry as an efficient and appropriate method for this quantification and confirmed the repellency activity, as well as its capacity of AChE inhibition.
- MeSH
- acetylcholinesterasa metabolismus MeSH
- Aedes účinky léků MeSH
- cholinesterasové inhibitory * analýza chemie farmakologie MeSH
- chování zvířat účinky léků MeSH
- chromatografie na tenké vrstvě metody MeSH
- denzitometrie MeSH
- dospělí MeSH
- eugenol analýza chemie farmakologie MeSH
- lidé MeSH
- limita detekce MeSH
- lineární modely MeSH
- mladý dospělý MeSH
- repelenty proti hmyzu * analýza chemie farmakologie MeSH
- reprodukovatelnost výsledků MeSH
- rostlinné extrakty * analýza chemie farmakologie MeSH
- Syzygium chemie MeSH
- zvířata MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
In this study, 14 commercial products (dietary supplements) containing alga Chlorella vulgaris and cyanobacteria Spirulina platensis, originated from China and Japan, were analysed. UV-vis spectrophotometric method was applied for rapid determination of chlorophylls, carotenoids and pheophytins; as degradation products of chlorophylls. High Performance Thin-Layer Chromatography (HPTLC) was used for effective separation of these compounds, and also Atomic Absorption Spectrometry for determination of heavy metals as indicator of environmental pollution. Based on the results obtained from UV-vis spectrophotometric determination of photosynthetic pigments (chlorophylls and carotenoids), it was confirmed that Chlorella vulgaris contains more of all these pigments compared to the cyanobacteria Spirulina platensis. The fastest mobility compound identified in Chlorella vulgaris and Spirulina platensis using HPTLC method was β-carotene. Spectral analysis and standard calibration curve method were used for identification and quantification of separated substances on Thin-Layer Chromatographic plate. Quantification of copper (Cu2+, at 324.7 nm) and zinc (Zn2+, at 213.9nm) was performed using Flame Atomic Absorption Spectrometry with air-acetylene flame atomization. Quantification of cadmium (Cd2+, at 228.8 nm), nickel (Ni2+, at 232.0nm) and lead (Pb2+, at 283.3nm) by Electrothermal Graphite Furnace Atomic Absorption Spectrometry; and quantification of mercury (Hg2+, at 254nm) by Cold Vapour Atomic Absorption Spectrometry.
- MeSH
- Chlorella vulgaris chemie MeSH
- chlorofyl chemie MeSH
- chromatografie na tenké vrstvě metody MeSH
- karotenoidy chemie MeSH
- potravní doplňky analýza MeSH
- spektrofotometrie atomová metody MeSH
- Spirulina chemie MeSH
- těžké kovy chemie MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Čína MeSH
- Japonsko MeSH
