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Twenty-seven isolates of Phytophthora infestans collected in Hungary in 2001 were tested for mating type, response to metalaxyl, isozyme geno­type at glucose-6-phosphate isomerase (Gpi) and peptidase A (Pep) loci and nuclear DNA fingerprints with probe RG57. The ratios of the mating types A1 to A2 were 5:6 and 9:7 among isolates from potato and tomato, respectively. Seventeen isolates were sensitive to metalaxyl, 1 isolate responded intermediately and 9 isolates were resistant. No novel combi­nations of isozyme alleles were found; all isolates were Gpi 100/100, and genotypes at the Pep locus were 96/96 (63%), 83/96 (11%) and 100/100 (26%). In contrast, all of the 22 RG57 fingerprints exhibited patterns that have not been reported in Hungary before. On the basis of combined traits, 22 multilocus genotypes, unnoted elsewhere in Europe, were con­struct­ed among the 27 isolates analysed. These results indicate that varia­bi­lity in the Hungarian P. infestans populations is likely due to local events (asexual and sexual interactions) rather than migration from other countries.

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JPC - Journal of Planar Chromatography - Modern TLC
Authors: Mieczysław Sajewicz, Łukasz Wojtal, Michał Hajnos, Monika Waksmundzka-Hajnos, and Teresa Kowalska

In a previous paper we discussed the possibility of fractionating the essential oils of different sage species by low-temperature preparative layer chromatography (PLC), followed by preparative isolation of the contents of each fraction and further analysis by GC-MS. In that way we attempted to emphasize the practical usefulness of lowtemperature planar chromatography for investigation of volatile compounds. In this study, we explore a possibility of fractionating essential oils contained in the different sage species by low-temperature analytical TLC followed by direct mass spectrometric analysis of the separated fractions. This objective can be achieved by TLC-MS with on-line transfer of the eluted fractions. The densitograms obtained from five different sage species (i.e., S. lavandulifolia, S. staminea, S. hians, S. triloba , and S. nemorosa ) are compared. Each densitogram is accompanied by mass spectra recorded for each peak. Videoscans of the chromatograms are also presented. In this way multiple fingerprints of the analyzed plant material, each comprising a densitogram and a selection of mass spectra, were obtained. Advanced chemometric treatment of these multiple fingerprints can be used to reveal statistically significant differences between the plant species. Analytical and chemotaxonomic advantages and further aspects for this kind of approach are discussed.

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Summary  

We have developed cleanroom compatible techniques for processing bone samples for characterization of their uranium and plutonium content. The bone samples are dried and ashed in quartz crucibles placed inside cleanroom compatible thermal ashing furnaces. The bone ash is dissolved in ultra-pure acids prepared by sub-boiling distillation. The uranium and plutonium in the samples are isolated and purified by ion-exchange chromatography and measured by thermal ionization mass spectrometry. The technique is capable of detecting 74 picograms of 238U and 8 femtograms of 239Pu in 100 mg bone ash samples. If the ash contains larger amounts of uranium and plutonium, the technique can be used to isotopically fingerprint the material to identify potential origins.

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Sixteen pumice samples produced by the youngest eruption sequences of Mt. Pelato (Island of Lipari, Italy) were analyzed with instrumental neutron activation analysis (INAA) for their major and trace element contents, in particular Al, Ba, Ca, Ce, Co, Cr, Cs, Dy, Eu, Fe, Hf, K, La, Lu, Mn, Na, Nd, Rb, Sb, Sm, Ta, Tb, Th, Ti, U, V, Yb, Zn, and Zr. A pumice from the archaeological excavation in Tel Megadim, Israel, could be correlated to this volcanic source, using its chemical fingerprint. This result, together with the background information about the well dated eruption cycles of this volcano, lead to the assumption that trade connections existed between cultures in Palestine and the Tyrrhenian region during the Persian Period (approx. between the 6th and 3rd century B.C.), in spite of the long distance of over 2000 km.

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Coffee, due to its common consumption, is one of the main sources of polyphenols in human diet. Coffee species and coffee-related products differ in composition and content of main components, such as chlorogenic acid and caffeine. Chemical and biological fingerprints of various Coffea arabica L. extracts were obtained in order to check and compare their antibacterial and antioxidant properties. The antibacterial activity of green and roasted coffee seeds and pomace was evaluated against Bacillus subtilis using thin-layer chromatography (TLC)-direct bioautography. TLC-2,2-diphenyl-1-picrylhydrazyl (DPPH) test was used to determine antioxidant properties of the afore-mentioned extracts. Furthermore, different solvents and several extraction methods such as simple maceration, maceration under stirring, and ultrasonic accelerated extraction were tested. The most efficient method of extraction of caffeine and chlorogenic acid was chosen based on quantitative TLC analysis. Additionally, these two main components of coffee were quantitatively determined in commercial products of green coffee.

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A procedure is demonstrated, through a simulation study, for the determination of the origin of unknown spent nuclear fuel, an important and challenging task in nuclear forensics. The procedure is an isotopic fingerprinting method relying on the fission product content of the unknown. The ‘unknown’ nuclear material is represented by the spent nuclear fuel of known origin in order to demonstrate the method and verify its predictive capabilities. The method is based on the comparison of the fission product compositions of the ‘unknown’ material and simulated known spent fuels from a range of commercial nuclear power stations using the multivariate statistical technique of factor analysis. Then, the provenance of the ‘unknown’ spent fuel is the commercial fuel with which it exhibits the highest similarity with respect to the fission product content.

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Quantitative evaluation of phytochemical diversity in Adhatoda vasica Nees populations from five different ecogeographical regions was performed using a highly sensitive, robust and economic high performance thin layer chromatography (HPTLC) procedure. The method was validated for specificity, precision and accuracy as per the guidelines affirmed by International Conference on Harmonization of technical requirements for registration of pharmaceuticals for human use (ICH). The findings support the existence of distinct natural chemotypes within the species. Two out of the five regions had about three times higher mean vasicine content than the remaining three regions. The comparative phytochemical fingerprint profiles were quite similar except for the concentration variation seen for different alkaloids. As the quinazoline alkaloids are the biologically active compounds in A. vasica, the samples that accumulate high levels of alkaloids seem to be promising for further propagation.

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Two-dimensional separations were performed on polar bonded stationary phase of type cyanopropyl-silica and diol-silica by use of non-aqueous eluents (polar modifier dissolved in n-heptane) as the first direction eluents and aqueous eluents (organic modifier — MeOH dissolved in water) as the second direction eluents. The chromatographic process was performed in micro scale using 5 × 5 cm plates, small volumes of eluents, and 10 μL of plant extracts to obtain satisfying separation. Plates developed in horizontal chambers were dried and observed in UV light (254 nm and 366 nm) photographed by digital camera and derivatized by 1,1-diphenyl-2-picrylhydrazyl (DPPH) to detect antioxidants (free radical scavengers) or derivatized by Naturstoff reagent to detect phenolic compounds (characteristic luminescence of some phenolic compounds). The above experiments give the possibility to construct fingerprints for investigated Mentha species and varieties and preparations containing the herb. It can be used in quality control of the plant material and its antioxidative activity.

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Acta Biologica Hungarica
Authors: Andrea Valasek, Írisz Éva Kiss, István Fodor, Márk Kovács, Péter Urbán, Éva Jámbor, Csaba Fekete, and Ildikó Kerepesi

Saccharomonospora azurea SZMC 14600 is a member of the family Pseudonocardiaceae exclusively used for industrial scale production of primycin a large 36-membered non-polyene macrolide lactone antibiotic belonging to the polyketide class of natural products. Even though maximum antibiotic yield has been achieved by empirically optimized two-step fermentation process, little is known about the molecular components and mechanisms underlying the efficient antibiotic production. In order to identify differentially expressed proteins (DEPs) between the pre- and main-fermentation stages of primycin, comparative 2D-PAGE experiments were performed. In total, 98 DEP spots were reproducibly detected, out of which four spots were excised from gels, and identified through MALDI-TOF/TOF mass spectrometry. Peptide mass fingerprint analysis revealed peptide matches to HicB antitoxin for the HicAB toxin-antitoxin system (EHK86651), to a nucleoside diphosphate kinase regulator ((Ndk; EHK81899) and two other proteins with unknown function (EHK88946 and EHK86777).

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The oleo-gum-resin known as ‘guggul’, obtained from the plant Commiphora mukul , Hook (family Burseraceae), is widely used, as a component of formulations containing other Ayurvedic drugs, for treatment of hypercholesterolemia, rheumatism, and arthritis. Chemically, guggul contains two active keto steroids, guggulsterones E and Z , which are reported to be responsible for its antihyperlipidemic activity. These cis and trans isomers have been isolated, purified by TLC and column chromatography, and characterized by determination of melting point and by acquisition of UV, IR, and NMR spectra. After confirmation of their identity, they were used as biomarkers in HPTLC fingerprinting analysis of five commercial Ayurvedic formulations, four tablets and a capsule, all of which contain guggul as a constituent. The analytical method was fully validated. Extrapolation was used to calculate probable amounts of guggul added to the formulations on the basis of E and Z isomer content.

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