New hyphenated technique for the extraction and determination of isoflavones in sea and freshwater algae and cyanobacteria was developed. The method consists of sonication sample pretreatment, extraction by supercritical CO(2) modified by 3% (v/v) of MeOH/H(2)O mixture (9:1, v/v) at 35 MPa and 40°C for 60 min, fast chromatography analysis by the means of Agilent 1200 Series Rapid Resolution and MS/MS determination. Agilent 1200 Series RRLC was used with Zorbax SB-CN chromatographic column (100 mm × 2.1mm, particle size 3.5 μm), 3μl injection volume, mobile phase consisting of 0.2% (v/v) acetic acid in water (solvent A) and acetonitrile (solvent B) and used with linear gradient (30% B at 0 min, from 0 min to 3 min up to 50% B, from 3 to 6 min up to 80% B and from 6 to 10 min down to 30% B). The flow-rate was 0.4 mL/min, column oven temperature 35°C. MS detector Agilent Technologies 6460 Triple quadrupole LC/MS with Agilent Jet Stream was used in a negative ESI mode under following conditions: gas temperature 350°C, gas flow 13 L/min, nebulizer gas pressure 50 psi, sheath gas temperature 400°C, sheath gas flow 12L/min, capillary voltage was 4 kV. Samples were analysed in the multiple reaction monitoring (MRM) mode. Eight isoflavone compounds were found for the first time in seven real samples of sea algae and in three control samples of freshwater algae and cyanobacteria. Usual optimisation study of extraction parameters was performed. Pressure and temperature optima for algae matrix are different from those obtained sooner for other matrices for most of the analytes, but the results of modifier optimisation study are in good accordance with those obtained sooner for spiked samples and red clover matrix. It seems that matrix has very small or no effect on the modifier selection. Two different approaches of sonication pretreatment were tested: sonication bath and the thorn instrument. In longer extraction time experiments, thorn sonication was more efficient and recovery of following supercritical fluid extraction was higher.
- MeSH
- chromatografie kapalinová metody MeSH
- isoflavony analýza izolace a purifikace MeSH
- Phaeophyceae chemie MeSH
- Rhodophyta chemie MeSH
- superkritická fluidní chromatografie přístrojové vybavení metody MeSH
- tandemová hmotnostní spektrometrie metody MeSH
- ultrazvuk MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- Názvy látek
- isoflavony MeSH
A new extraction technique based on the off-line combination of pressurized-liquid with solid-phase extraction (PLE-SPE) is described. The method was used for the extraction of bioactive phenolic acids (protocatechuic, p-hydroxybenzoic, 2,3-dihydroxybenzoic, chlorogenic, vanillic, caffeic, p-coumaric, salicylic acid), cinnamic acid and hydroxybenzaldehydes (p-hydroxybenzaldehyde, 3,4-dihydroxybenzaldehyde, vanillin) from in vitro culture of two freshwater algae (Anabaena doliolum and Spongiochloris spongiosa) and from food products of marine macroalgae Porphyra tenera (nori) and Undaria pinnatifida (wakame). For the identification and quantification of the compounds the molecular ions [M-H](-) and specific fragments were analyzed by quadrupole mass spectrometry analyzer connected on-line with a reversed-phase HPLC system. Our analysis showed that the freshwater algae and marine algal products contained submicrogram or microgram level of above-mentioned phenols per gram of lyophilized sample. In addition, the total phenol content (Folin-Ciocalteu assay) and antioxidant activity (TEAC assay, Trolox equivalent antioxidant capacity assay) of the PLE-SPE extracts were determined and discussed.
- MeSH
- antioxidancia analýza chemie MeSH
- biologie sladkých vod metody MeSH
- cinnamáty analýza chemie MeSH
- Eukaryota chemie MeSH
- extrakce na pevné fázi metody MeSH
- fenoly analýza chemie MeSH
- kyselina benzoová analýza chemie MeSH
- mořská voda chemie MeSH
- on-line systémy * MeSH
- referenční standardy MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antioxidancia MeSH
- cinnamáty MeSH
- cinnamic acid MeSH Prohlížeč
- fenoly MeSH
- kyselina benzoová MeSH
In the present paper a new extraction technique based on the combination of solid-phase/supercritical-fluid extraction (SPE/SFE) with subsequent reversed-phase HPLC is described. The SPE/SFE extractor was originally constructed from SPE-cartridge incorporated into the SFE extraction cell. Selected groups of benzoic acid derivatives (p-hydroxybenzoic, protocatechuic, gallic, vanillic and syringic acid), hydroxybenzaldehydes (4-hydroxybenzaldehyde and 3,4-dihydroxybenzaldehyde) and cinnamic acid derivatives (o-coumaric, p-coumaric, caffeic, ferulic, sinapic and chlorogenic acid) were extracted. Cyclic addition of binary extraction solvent system based on methanol:water (1:1, v/v) and methanol/ammonia aqueous solution was used for extraction at 40MPa and 80 degrees C. The p-hydroxybenzoic, protocatechuic, vanillic, syringic, caffeic and chlorogenic acid; 4-hydroxybenzaldehyde and 3,4-dihydroxybenzaldehyde were identified by HPLC-electrospray mass spectrometry in SPE/SFE extracts of acid hydrolyzates of microalga (Spongiochloris spongiosa) and cyanobacterial strains (Spirulina platensis, Anabaena doliolum, Nostoc sp., and Cylindrospermum sp.). For the identification and quantification of the compounds the quasi-molecular ions [M-H](-) and specific fragments were analysed by quadrupole mass spectrometry analyzer. Our analysis showed that the microalgae and cyanobacteria usually contained phenolic acids or aldehydes at microg levels per gram of lyophilized sample. The proposed SPE/SFE extraction method would be useful for the analysis of different plant species containing trace amount of polar fraction of phenols.
- MeSH
- aldehydy analýza chemie MeSH
- Chlorophyta chemie MeSH
- design vybavení MeSH
- extrakce na pevné fázi metody MeSH
- fenoly analýza chemie MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací MeSH
- kinetika MeSH
- methanol chemie MeSH
- reprodukovatelnost výsledků MeSH
- sinice chemie MeSH
- teplota MeSH
- tlak MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- aldehydy MeSH
- fenoly MeSH
- methanol MeSH
Complete separation of aglycones and glucosides of selected isoflavones (genistin, genistein, daidzin, daidzein, glycitin, glycitein, ononin, sissotrin, formononetin, and biochanin A) was possible in 1.5 min using an ultrahigh-pressure liquid chromatography (U-HPLC) on a different particular chemically modified stationary phases with a particle size under 2 microm. In addition, selected separation conditions for simultaneous determination of isoflavones together with a group of phenolic acids (gallic, protocatechuic, p-hydroxybenzoic, vanillic, caffeic, syringic, p-coumaric, ferulic, and sinapic acid) allowed separation of all 19 compounds in 1.9 min. Separations were conducted on a non-polar reversed phase (C(18)) and also on more polar phases with cyanopropyl or phenyl groups using a gradient elution with a mobile phase consisting of 0.3% aqueous acetic acid and methanol. Chromatographic peaks were characterised using parameters such as resolution, symmetry, selectivity, etc. Individual substances were identified and quantified using UV-vis diode array detector at wavelength 270 nm. Limits of detection (3S/N) were in the range 200-400 pg ml(-1). Proposed U-HPLC technique was used for separation of isoflavones and phenolic acids in samples of plant materials (Trifolium pratense, Glycine max, Pisum sativum and Ononis spinosa) after acid hydrolysis of the samples and modified Soxhlet extraction.
- MeSH
- genistein chemie izolace a purifikace MeSH
- Glycine max chemie MeSH
- hrách setý chemie MeSH
- hydroxybenzoáty chemie izolace a purifikace MeSH
- isoflavony chemie izolace a purifikace MeSH
- kyselina gallová analogy a deriváty chemie izolace a purifikace MeSH
- kyselina vanilová chemie izolace a purifikace MeSH
- kyseliny kávové chemie izolace a purifikace MeSH
- kyseliny kumarové chemie izolace a purifikace MeSH
- molekulární struktura MeSH
- propionáty MeSH
- rostlinné extrakty chemie izolace a purifikace MeSH
- Trifolium chemie MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- biochanin A MeSH Prohlížeč
- caffeic acid MeSH Prohlížeč
- daidzein MeSH Prohlížeč
- daidzin MeSH Prohlížeč
- ferulic acid MeSH Prohlížeč
- formononetin MeSH Prohlížeč
- genistein MeSH
- genistin MeSH Prohlížeč
- glycitein MeSH Prohlížeč
- glycitin MeSH Prohlížeč
- hydroxybenzoáty MeSH
- isoflavony MeSH
- kyselina gallová MeSH
- kyselina vanilová MeSH
- kyseliny kávové MeSH
- kyseliny kumarové MeSH
- p-coumaric acid MeSH Prohlížeč
- phenolic acid MeSH Prohlížeč
- propionáty MeSH
- protocatechuic acid MeSH Prohlížeč
- rostlinné extrakty MeSH
- sinapinic acid MeSH Prohlížeč
- syringic acid MeSH Prohlížeč
Isoflavones belong to the natural substances exhibiting a number of physiological effects in living organisms. The substances are synthesized in plant tissues as protective agents against biotic stress (i.e. bacterial infection). Isoflavones are also an important dietary constituent in human nutrition. The review discusses modern trends in the studies of isoflavones in plant materials and foodstuffs and procedures for chemical analyses of isoflavones in human body fluids and plant tissues. Highly effective extraction and purification techniques, i.e. solid-phase extraction (SFE), accelerated-solvent extraction (ASE), and Soxhlet extraction, are presented. Latest procedures in chromatographic separation of isoflavones that apply different types of stationary phases are described. Immunochemical analysis, electrochemical sensing of isoflavones, spectrometric and other analytical techniques and their applications are also mentioned. Special attention is focused on a highly selective and sensitive technique of mass spectrometry and its application for identification of isoflavones and their glucosides in plants. Studies of interactions of isoflavones with cell receptors and a number of biologically active substances such as DNA and proteins are described. The reason for the presentation of the review was not to give a full overview of the presented topics but mainly to show modern and the most recent methods in the studies of isoflavones.
- MeSH
- chemické techniky analytické přístrojové vybavení metody MeSH
- isoflavony analýza chemie izolace a purifikace MeSH
- lidé MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- isoflavony MeSH
A rapid-resolution HPLC/UV-VIS DAD separation method (which takes <1 min) for the determination and identification of genistin, genistein, daidzein, daidzin, glycitin, glycitein, ononin, formononetin, sissotrin and biochanin A in fmol quantities in submicroliter sample volumes was optimized. A linear gradient elution (0 min 22% B, 1.0 min 80% B, 1.4 min 100% B, 1.8 min 22% B) using a mobile phase containing 0.2 % (v/v) acetic acid (solvent A) and methanol (solvent B) was applied on a Zorbax SB C18 column (1.8 microm particle size) at 80 degrees C. The method was verified using samples of bits of soy and methanolic extracts from Trifolium pratense, Iresine herbstii and Ononis spinosa plants. Pseudobaptigenin glucoside, irilone, prunetin, texasin, tlatlancuayin and other isoflavones, in addition to aglycones of isoflavones and their beta-glucosides and malonyl and acetyl derivatives, were identified by UV-VIS DAD and electrospray mass spectrometric (ESI-MS) detection in the extracts.
- MeSH
- časové faktory MeSH
- Glycine max chemie MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací MeSH
- isoflavony analýza chemie MeSH
- methanol chemie MeSH
- molekulární struktura MeSH
- rostlinné extrakty chemie MeSH
- rostlinné přípravky chemie MeSH
- spektrofotometrie MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- Názvy látek
- isoflavony MeSH
- methanol MeSH
- rostlinné extrakty MeSH
- rostlinné přípravky MeSH
The effects of demineralized water (DEMI H(2)O) and 0.5 M ammonium acetate (0.5 M AAc) on losses of L-glutamic acid and L-arginine in the course of shaking and filtration at low temperature (6 degrees C) were tested. The concentration of L-glutamic acid decreased by 6.3% in DEMI H(2)O and by 4.9% in 0.5 M AAc, whereas the L-arginine concentration decreased by 6.0% (DEMI H(2)O) and 10.7% (0.5 M AAc). We found a significantly (P < 0.05) higher degradation of L-arginine in 0.5 M AAc compared with that of DEMI H(2)O.
Influence of saccharose in the presence or absence of polyethylene glycol (PEG), methyl jasmonate, and an inactivated bacterial culture of Agrobacterium tumefaciens in cultivation medium on morphology of Hypericum perforatum L. and production of hypericin and hyperforin was studied under in vitro conditions. Production of hypericin and hyperforin was influenced by the presence of different concentrations of saccharose (10-30 g L(-1)) in cultivation medium. Addition of PEG (1.25-5 g L(-1)) in the presence of saccharose (10-30 g L(-1)) increased production of hypericin and hyperforin in the H. perforatum in vitro culture. Synthesis of hypericin and hyperforin was unchanged or reduced for most of the experimental plants at higher contents of PEG (10 and 15 g L(-1)). Concentrations of hypericin and hyperforin in the H. perforatum were on the order 100 and 103 microg g(-1) of dry plant material, respectively. Production of hypericin and hyperforin was stimulated either in the presence of a chemical elicitor (methyl jasmonate) or an inactivated bacterial culture of A. tumefaciens. Morphological changes induced by the abovementioned substances were observed and described in detail. The obtained results will be applied in experimental botany and in the technology of H. perforatum cultivation for pharmaceutical applications.
- MeSH
- acetáty farmakologie MeSH
- Agrobacterium tumefaciens metabolismus MeSH
- anthraceny MeSH
- cyklopentany farmakologie MeSH
- floroglucinol analogy a deriváty metabolismus MeSH
- můstkové bicyklické sloučeniny metabolismus MeSH
- oxylipiny MeSH
- perylen analogy a deriváty metabolismus MeSH
- polyethylenglykoly farmakologie MeSH
- sacharosa farmakologie MeSH
- semena rostlinná růst a vývoj metabolismus MeSH
- terpeny metabolismus MeSH
- třezalka růst a vývoj metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- acetáty MeSH
- anthraceny MeSH
- cyklopentany MeSH
- floroglucinol MeSH
- hyperforin MeSH Prohlížeč
- hypericin MeSH Prohlížeč
- methyl jasmonate MeSH Prohlížeč
- můstkové bicyklické sloučeniny MeSH
- oxylipiny MeSH
- perylen MeSH
- polyethylenglykoly MeSH
- sacharosa MeSH
- terpeny MeSH
Sanguinarine is an alkaloid with known antibiotic and anti-inflammatory activity and its pharmacokinetics have been studied in the rat after a single oral dose (10 mg kg(-1) body weight). Alkaloid determination in the plasma and liver was carried out by high-performance liquid chromatography-electrospray ionization mass spectrometry (HPLC/ESI-MS). The pharmacokinetic parameters (t(max), c(max), AUC(0-->t) and AUC(0-->infinity)) were determined for sanguinarine and dihydrosanguinarine, the major components detected in plasma. The first step in sanguinarine metabolism in the rat was the reduction of the iminium bond resulting in formation of the less toxic dihydrosanguinarine. Both compounds were completely eliminated from the plasma and liver after 24 h and not detected in urine. After a single oral dose of (3)H-sanguinarine, more than 42% of the ingested radioactivity was present in gastrointestinal tract. Benz[c]acridine, up to date the only sanguinarine metabolite referred to in the literature, was not detected in the plasma, liver or urine.
- MeSH
- akridiny chemie MeSH
- alkaloidy aplikace a dávkování krev chemie farmakokinetika MeSH
- antiinfekční látky aplikace a dávkování krev chemie farmakokinetika MeSH
- aplikace orální MeSH
- benzofenantridiny aplikace a dávkování krev chemie farmakokinetika MeSH
- časové faktory MeSH
- hmotnostní spektrometrie MeSH
- isochinoliny aplikace a dávkování krev chemie farmakokinetika MeSH
- krysa rodu Rattus MeSH
- potkani Wistar MeSH
- tkáňová distribuce MeSH
- tritium MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- akridiny MeSH
- alkaloidy MeSH
- antiinfekční látky MeSH
- benz(c)acridine MeSH Prohlížeč
- benzofenantridiny MeSH
- dihydrosanguinarine MeSH Prohlížeč
- isochinoliny MeSH
- sanguinarine MeSH Prohlížeč
- tritium MeSH
St. John's Wort (Hypericum perforatum L.) is commonly accepted as a medicinal plant. The data on the physiological activities of the individual substances that are produced in different organs of H. perforatum are well known at present. The highest attention is focused on the characterization and phytochemical properties of hypericin and hyperforin. These organic compounds are used as antidepressant, anticarcinogenic (photodynamic), antimicrobial and virostatic agents. The review paper surveys the present knowledge of chemical and analytical methods for their identification and quantification, physiological activity, and pharmacological and biomedical applications of hypericin and hyperforin.
- MeSH
- anthraceny MeSH
- antidepresiva terapeutické užití MeSH
- depresivní poruchy farmakoterapie MeSH
- floroglucinol analogy a deriváty izolace a purifikace farmakologie terapeutické užití MeSH
- fytoterapie MeSH
- léčivé rostliny chemie MeSH
- lidé MeSH
- můstkové bicyklické sloučeniny izolace a purifikace farmakologie terapeutické užití MeSH
- perylen analogy a deriváty izolace a purifikace farmakologie terapeutické užití MeSH
- rostlinné extrakty izolace a purifikace farmakologie terapeutické užití MeSH
- terpeny izolace a purifikace farmakologie terapeutické užití MeSH
- třezalka chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- anthraceny MeSH
- antidepresiva MeSH
- floroglucinol MeSH
- hyperforin MeSH Prohlížeč
- hypericin MeSH Prohlížeč
- můstkové bicyklické sloučeniny MeSH
- perylen MeSH
- rostlinné extrakty MeSH
- terpeny MeSH
