silicon and silicomolybdate

The sensitive differential pulse anodic stripping voltammetry (DPASV) proposed originally by Ishiyama et al. (2001) has been revised and improved to allow the accurate measurement of silicon on a hanging mercury drop electrode (HMDE) instead of a glassy carbon electrode. We assessed the rate of formation of the partially reduced β-silicododecamolybdate and found that metallic mercury promotes the reaction in the presence of a large concentration of Fe(3+). The scope of the method has been broadened by carrying out the measurements in the presence of a constant amount of Fe(3+). The limit of detection (LOD) of the method described in the present paper is 100 μg Sig(-1) of steel, with a relative precision ranging from 5% to 12%. It can be further enhanced to 700 ng Sig(-1) of steel provided the weight of the sample, the dilution factors, the duration of the electrolysis and the ballast of iron are adequately revised. The tolerance to several interfering species has been examined, especially regarding Al(3+), Cr(3+) and Cr VI species. The method was validated using four low-alloy ferritic steels certified by the National Institute of Standards and Technology (NIST). Its application to nickel base alloys as well as to less complicated matrixes is straightforward. It has also been successfully applied to the determination of free silicon into silicon carbide nano-powder.

Topics: Acetone; Electrochemical Techniques; Electrodes; Ferric Compounds; Hydrogen-Ion Concentration; Indicators and Reagents; Limit of Detection; Mercury; Molybdenum; Silicon; Silicon Compounds; Solutions; Steel

An indirect method for the determination of silicon in blood samples has been developed. The proposed method overcame interference from a large amount of salts and phosphate in blood samples, and enabled us to determine the silicon contents in serum and whole blood by the same operation. After blood samples were digested by microwave heating, silicon, present as silicate in the sample solution, was reacted with molybdate to form a silicomolybdate complex. The complex was then separated from unreacted molybdate by a cation-exchange resin column. The molybdate liberated from the complex was spectrophotometrically determined in place of silicon. Since the method is not affected the composition of matrices between serum and whole blood, it could achieve good precision and accuracy, and could also estimate the silicon contents in erythrocytes from those in serum and whole blood. The sensitivity of the method was almost equal to that of the conventional silicomolybdenum blue method, and the calibration curve was linear up to 50 micromol l(-1) of silicon with a detection limit of 1.1 micromol l(-1) in whole blood. The mean concentrations of silicon in five healthy subjects were 11 micromol l(-1) for serum, 28 micromol l(-1) for whole blood and 50 micromol l(-1) for erythrocytes. Thus, the obtained distribution ratio between serum and erythrocytes was in the range of 0.15-0.39, and was found to be included in a narrow range.

Topics: Calibration; Erythrocytes; Humans; Molybdenum; Reference Standards; Sensitivity and Specificity; Silicon; Silicon Compounds; Spectrophotometry; Time Factors

We show that CCCP, known as an uncoupler of photophosphorylation and an ADRY agent, inhibits FeCy photoreduction and coupled O2 evolution by isolated chloroplasts equally (I50 approximately 2 microM), but is practically without effect on the O2 evolution coupled with SiMo reduction within the 0.2-10 microM concentration range. CCCP has no effect on the nanosecond chlorophyll fluorescence in chloroplasts incubated at low light intensity, but decreases it at high light intensity. The electron transfer from reduced TMPD or duroquinol to methylviologen is resistant to CCCP. The efficiency of the CCCP inhibitory action on the FeCy photoreduction depends on the rate of electron flow, which is controlled by the light intensity. The data obtained show that CCCP is oxidized by the photosystem II donor side and is reduced by QP, competing for electrons with FeCy and the cytochrome blf complex.

Topics: Carbonyl Cyanide m-Chlorophenyl Hydrazone; Chloroplasts; Cytochrome b Group; Dibromothymoquinone; Diuron; Electron Transport; Fabaceae; Ferricyanides; Hydroxyquinolines; Light-Harvesting Protein Complexes; Molybdenum; Oxidation-Reduction; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Plants, Medicinal; Silicon; Silicon Compounds; Tetramethylphenylenediamine; Trinitrobenzenes

Oxygen evolution by photosystem II membranes was inhibited by Cu(II) when 2,6-dichlorobenzoquinone or ferricyanide, but not silicomolybdate, was used as electron acceptor. This indicated that Cu(II) affected the reducing side of the photosystem II. The inhibition curves of Cu(II), o-phenanthroline and 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), were compared; the inhibitory patterns of Cu(II) and o-phenanthroline were very similar and different in turn from that of DCMU. Cu(II) did not eliminate or modify the electron paramagnetic resonance signal at g = 8.1 ascribed to the non-heme iron of the photosystem II reaction center, indicating that the inhibition by Cu(II) was not the result of the replacement of the iron by Cu(II). Controlled trypsin digestion of thylakoid membranes inhibited oxygen evolution using 2,6-dichlorobenzoquinone, but had no effect when using ferricyanide or silicomolybdate. Using ferricyanide, oxygen evolution of trypsin-treated thylakoids was insensitive to DCMU but became even more sensitive to Cu(II) and o-phenanthroline than nontreated thylakoids; however, trypsinized thylakoids were insensitive to inhibitors in the presence of silicomolybdate. We conclude that Cu(II) impaired the photosystem II electron transfer before the QB niche, most probably at the pheophytin-QA-Fe domain.

Topics: Benzoquinones; Binding Sites; Copper; Ferricyanides; Iron; Light-Harvesting Protein Complexes; Molybdenum; Oxidation-Reduction; Pheophytins; Photosynthetic Reaction Center Complex Proteins; Photosystem II Protein Complex; Plants; Quinones; Silicon; Silicon Compounds; Trypsin

Two possible 3-(3,4-dichlorophenyl)-1,1-dimethylurea-insensitive sites were found in PS II of spinach chloroplasts, depending on the pH of the assay medium used. The low site (pH 6) can be inhibited by certain quinolines, such as 8-hydroxyquinoline at concentrations less than 50 microM. The high pH site (pH 8) can be inhibited by disodium cyanamide, folic acid, or 5,6-benzoquinoline at concentrations from 50 microM to 5 mM. With the exception of orthophenanthroline, which stimulates the high pH site but does not show much inhibition at low pH, all other inhibitors gave opposite effects at the pH values used, i.e., they stimulated at low pH or inhibited at high pH, or vice versa. Several mechanisms for the observed effects are discussed.

Topics: Ammonium Chloride; Chloroplasts; Diuron; Hydrogen-Ion Concentration; Kinetics; Molybdenum; Oxidation-Reduction; Photosynthesis; Plants; Silicon; Silicon Compounds

Topics: Chloroplasts; Electron Spin Resonance Spectroscopy; Electron Transport; Kinetics; Light; Molybdenum; Photosynthesis; Plants; Silicon; Silicon Compounds; Spectrophotometry

Three sites of silicomolybdate reduction in the electron transport chain of isolated tobacco chloroplasts are described. The relative participation of these sites is greatly influenced by the particular reaction conditions. One site (the only site when the reaction medium contains high concentrations of bovine serum albumin (greater than 5 mg/ml) is associated with Photosystem I, since it supports phosphorylation with a P/e2 value close to 1 and the reaction is totally sensitive to both plastocyanin inhibitors and 3-(3,4-dichlorophenyl)-1,1-dimethylurea. Two other sites of silicomolybdate reduction are associated with Photosystem II. One site is 3-(3,4-dichlorophenyl)-1,1-dimethylurea insensitive and supports phosphorylation when the reaction mixture contains dimethyl sulfoxide and glycerol (protective agents). The P/e2 value routinely observed is about 0.2. Bovine serum albumin (1-2 mg/ml) can also act as a protective agent, but the efficiency of Photosystem II phosphorylation observed is lower. Silicomolybdate reduction supports virtually no phosphorylation, regardless of the reduction pathway, when the reaction mixture contains no protective agents. This is due to irreversible uncoupling by silicomolybdate itself. The silicomolybdate uncoupling is potentiated by high salt concentrations even if the presence of protective agents. Exposure of chloroplasts to silicomolybdate in the absence of protective agents rapidly inactivates both photosystems.

Topics: Binding Sites; Chloroplasts; Darkness; Dibromothymoquinone; Dimethyl Sulfoxide; Diuron; Electron Transport; Ferricyanides; Glycerol; Hydrogen-Ion Concentration; Mercury; Molybdenum; Nicotiana; Photophosphorylation; Plants, Toxic; Serum Albumin, Bovine; Silicon; Silicon Compounds; Sodium Chloride; Temperature

1. The rate of electron transport from H2O to silicomolybdate in the presence of 3-(3-4-dichlorophenyl)-1,1-dimethylurea (diuron) (which involves the oxygen-evolving enzyme, the photochemistry of photosystem 2 and the primary electron acceptor of photosystem 2) is controlled by internal pH. This is based on the shift of the pH profile of the rate of electron transport upon addition of uncouplers, or by using EDTA-treated chloroplasts. Both stimulation and inhibition of electron transport by addition of uncouplers (depending on external pH) could be observed. These effects are obtained in the diuron-insensitive photoreductions of either silicomolybdate or ferricyanide. These experiments provide strong evidence that a proton translocating site exists in the sequence of the electron transport H2O leads to Q (the primary acceptor of photosystem 2). 2. The photoreduction of silicomolybdate in the presence of diuron causes the formation of delta pH. The value of delta pH depends on the external pH and its maximal value was shown to be 2.4. The calculated internal pH at different external pH values was found to be rather constant, namely between 5.1 -- 5.2. 3. Electron transport from H2O to silicomolybdate (in the presence of diuron) does not support ATP formation. It is suggested that this is due to the fact that the delta pH formed is below the "threshold" delta pH required for the synthesis of ATP. By adding an additional source of energy in the form of a dark diffusion potential created in the presence of K+ and valinomycin, significant amounts of ATP are formed in this system.

Topics: Adenosine Triphosphate; Chloroplasts; Diuron; Electron Transport; Gramicidin; Hydrogen-Ion Concentration; Kinetics; Molybdenum; Oxygen; Photophosphorylation; Plants; Silicon; Silicon Compounds

1. The electron transport in isolated chloroplasts with silicomolybdate as electron acceptor has been reinvestigated. The silicomolybdate reduction has been directly measured as deltaA750 or indirectly as O2 evolution (in the presence or absence of ferricyanide). 2. Silicomolybdate-dependent O2 evolution is inhibited to a similar extent by 3-(3,4-dichlorophenyl) 1, 1-dimethylurea (DCMU) or dibromothymoquinone (DBMIB), indicating the existence of two different sites of silicomolybdate reduction: one before the DCMU block (i.e. at Photosystem II) and one after the DBMIB block (i.e. at Photosystem I). 3. Silicomolybdate-dependent O2 evolution is coupled to ATP synthesis with an ATP/2e- ratio of 1.0 to 1.1. The presence of ferricyanide inhibits this ATP synthesis (ATP/2e- ratio then is about 0.3). 4. Silicomolybdate-dependent O2 evolution is also coupled to ATP-synthesis in the presence of DCMU with an ATP/2e- ratio of 0.6-0.8 characteristic of Site II; in this case the electron transport itself is not affected by uncouplers or energy-transfer inhibitors. 5. The data are interpreted as a further demonstration that the water-splitting reaction is responsible for the conservation of energy at Photosystem II.

Topics: Adenosine Triphosphate; Ammonium Chloride; Chloroplasts; Dibromothymoquinone; Diuron; Electron Transport; Ferricyanides; Hypotonic Solutions; Molybdenum; Oxygen; Phlorhizin; Plants; Silicon; Silicon Compounds; Tetramethylphenylenediamine; Water