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
The impact of manganese excess using naturally contaminated soil (Mn-soil, pseudototal Mn 6494 vs 675 μg g(-1) DW in control soil) in the shoots of four crops was studied. Mn content decreased in the order Brassica napus > Hordeum vulgare > Zea mays > Triticum aestivum. Growth was strongly depressed just in Brassica (containing 13696 μg Mn g(-1) DW). Some essential metals (Zn, Fe) increased in Mn-cultured Brassica and Zea, while macronutrients (K, Ca, Mg) decreased in almost all species. Toxic metals (Ni and Cd) were rather elevated in Mn-soil. Microscopy of ROS, NO, lipid peroxidation, and thiols revealed stimulation in all Mn-cultured crops, but changes were less visible in Triticum, a species with low shoot Mn (2363 μg g(-1) DW). Antioxidative enzyme activities were typically enhanced in Mn-cultured plants. Soluble phenols increased in Brassica only while proteins rather decreased in response to Mn excess. Inorganic anions (chloride, sulfate, and phosphate) were less accumulated in almost all Mn-cultured crops, while the nitrate level rather increased. Organic anions (malate, citrate, oxalate, acetate, and formate) decreased or remained unaffected in response to Mn-soil culture in Brassica, Hordeum, and Triticum but not in Zea. However, the role of organic acids in Mn uptake in these species is not assumed. Because control and Mn-soil differed in pH (6.5 and 3.7), we further studied its impact on Mn uptake in solution culture (using Mn concentration ∼5 mM deducted from water-soluble fraction of Mn-soil). Shoot Mn contents in Mn-treated plants were similar to those observed in soil culture (high in Brassica and low in Triticum) and pH had negligible impact. Fluorescence indicator of "general ROS" revealed no extensive or pH-dependent impact either in control or Mn-cultured roots. Observed toxicity of Mn excess to common crops urges for selection of cultivars with higher tolerance.
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
A new method for the electrophoretic separation of nine phenolic acids (derivatives of benzoic and cinnamic acids) with contactless conductometric detection is presented. Based on theoretical calculations, in which the mobility of the electrolyte co- and counterions and mobility of analytes are taken into consideration, the electrolyte composition and detection mode was selected. This approach was found to be especially valuable for optimization of the electrolyte composition for the separation of analytes having medium mobility. Indirect conductometric detection mode was superior to the direct mode as predicted theoretically. The best performance was achieved with 150 mM 2-amino-2-methylpropanol electrolyte at pH 11.6. The separation was carried out in a counter-electroosmotic mode and completed in less than 6 min. The LODs achieved were about 2.3-3.3 microM and could be further improved to 0.12-0.17 microM by using a sample stacking procedure. The method compares well to the UV-Vis detection.
