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Determination of organophosphorus and triazine pesticides in ground- and drinking water by solid-phase extraction and gas chromatography with nitrogen-phosphorus or mass spectrometric detection.

Trace enrichment and determination of ethoprophos, fenamiphos, fenthion, isophenphos, mevinphos, monocrotophos, atrazine and simazine were performed by solid-phase extraction on XAD-2 columns and Sep-Pak C18 cartridges, subsequent elution with an organic solvent and determination by GC with nitrogen-phosphorus detection (NPD) and mass spectrometry in the selected-ion monitoring mode (MS-SIM). Ground- and drinking water volumes of 1-2.5 l at concentrations levels of 0.1-5 micrograms/l were used for application of the method. Both adsorbents provided recoveries of 75-95%. The limits of detection were 0.08-0.60 micrograms/l with NPD and 0.03-0.13 micrograms/l with MS-SIM.

Gas Chromatography-Mass Spectrometry↗

UV detection of triazine herbicides and their hydroxylated and dealkylated degradation products in well water.

The aim of this work was to develop a simple method to confirm the presence of hydroxytriazine products (hydroxyatrazine, hydroxysimazine, hydroxyethylterbutyltriazine and hydroxydiaminotriazine) in water and to apply it to well water samples. The hydroxytriazines were concentrated on a Sep-Pak C18 cartridge. Analysis was performed by HPLC using an RP8-DB column with phosphate buffer (pH 4.7)-acetonitrile (72:28, v/v) as the mobile phase and photodiode-array detection at 233 nm. Hydroxyatrazine, hydroxysimazine and hydroxyethylterbutyltriazine were detected at ppb levels in samples from two shallow wells under a very sandy soil citrus orchard taken at three different times in a 1-year period.

Chromatography, High Pressure Liquid↗

Metal ion-promoted binding of proteins to immobilized triazine dye affinity adsorbents.

Low concentrations of metal ions, particularly those of the first row transition series such as Zn2+, Co2+, Mn2+, Ni2+, Cu2+, and, to a lesser extent, the group IIA ions, Ca2+ and Mg2+, promotes binding of carboxypeptidase G2, alkaline phosphatase and yeast hexokinase to immobilized Procion Red H-8BN, Procion Yellow H-A and Cibacron Blue F3G-A respectively. The binding of ovalbumin to immobilized Cibacron Blue F3G-A and Procion Orange MX-G is selectively enhanced in the presence of AI3+. With ovalbumin and alkaline phosphatase, the effect is almost totally specific for both the metal ion and dye, whereas with carboxypeptidase G2 and hexokinase, metal ions such as Co2+, Ni2+, Mn2+, Cu2+, Ca2+ and Mg2+ also promote binding to varying degrees. Almost all other monovalent and trivalent metal ions appear to be ineffective. Metal ion-bound enzymes can subsequently be eluted with appropriate chelating agents of the amine, aminocarboxylate or substituted pyridine classes.

Alkaline Phosphatase↗

Interaction of alpha 2-HS-glycoprotein with immobilized triazine dyes.

We studied the interaction of alpha 2-HS-glycoprotein with immobilized Cibacron blue F3-GA (Blue A) and Procion red HE-3B (Red A). When whole plasma was applied on the Blue A, alpha 2-HS-glycoprotein remained unbound, together with other plasma proteins. In contrast, when this fraction was applied on the Red A, alpha 2-HS-glycoprotein was shown to bind tightly and was eluted with a linear sodium chloride gradient between 0.5 and 0.8 M. This proved to be a useful two-step technique for the purification of alpha 2-HS-glycoprotein. Further characterization revealed that the protein appeared homogeneous by immunoelectrophoresis and SDS-polyacrylamide gel electrophoresis with yields greater than 30%. A small (less than 5%) but significant fraction of alpha 2-HS-glycoprotein with a same molecular weight as the native protein was consistently found in the wash of the Red A column, and may correspond to alpha 2-HS-glycoprotein bound to a yet unidentified ligand.

Blood Proteins↗

Interaction of NADPH and triazine dyes with ferredoxin-NADP+ oxidoreductase.

The ability of NADPH to compete for binding with other ligands of known affinity has been used to provide values for the Kd of NADPH with ferredoxin-NADP+ oxidoreductase (EC 1.18.1.2) (FNR). When the competing ligand is procion red, which binds with a red-shift in spectrum, or Woodwards reagent K(N-ethyl-5-phenylisoxazolium 3'-sulfonate), which covalently modifies an active site carboxyl residue, the calculated Kd for the NADPH-FNR complex is greater than 8 or 0.08 mM, respectively. Because of the feeble (or non-existent) ability of NADPH to dislodge procion red, we propose that this dye and NADPH are not binding at the same site. Procion red must, however, bind additionally at the active site (presumably without spectral perturbation) as it is a competitive inhibitor of NADPH in ferricyanide reduction assays and more crucially proves to be a novel substrate itself, being reduced to a leuco form which can be reoxidised by oxygen. Although a Kd for the NADPH-FNR complex of 0.08 mM is reasonable, we point out the difficulty of interpreting this value and question its physiological significance.

Ferredoxin-NADP Reductase↗

Induction of SOS responses in Escherichia coli by Panfuran-S, 3-di(hydroxymethyl)amino-6-(5-nitro-2-furylethenyl)-1,2,4-triazine.

The inducibility of SOS responses by Panfuran-S, which has been used as an antimicrobial medicine in Japan, was studied in Escherichia coli cells having different DNA-repair capacities for UV lesions. Panfuran-S induced mutations at high frequencies in uvrA and the wild-type strains, and significant killing effects of Panfuran-S were detected in DNA-repair-deficient strains, uvrA and recA. The effective prophage induction was detected in two kinds of lambda-lysogenized cells treated with Panfuran-S. The expression of the umuC+ gene was apparently induced in uvrA and the wild-type strains, but not induced in lexA and recA strains. In particular, high inducibility of the gene expression was detected in uvrA strain as compared with the wild-type strain. From these results, we conclude that Panfuran-S is a DNA-damaging agent and may induce the error-prone SOS responses.

Anti-Bacterial Agents↗

The induction of bladder stones by terephthalic acid, dimethyl terephthalate, and melamine (2,4,6-triamino-s-triazine) and its relevance to risk assessment.

Terephthalic acid (TPA), dimethyl terephthalate (DMT), and melamine (MA) induced calculi and transitional cell hyperplasia in urinary bladders of rats. A high incidence of calculi was induced in weanling rats, but the incidence was much lower in adult rats ingesting the same dietary concentration of the chemical. The dose-response curves for the induction of urolithiasis in weanling rats were extremely steep, consistent with the fact that the formation calculi can occur in urine that is supersaturated, but not in urine that is undersaturated with respect to the stone components. In the cases of TPA and DMT, stones were composed primarily of calcium terephthalate (CaTPA). By determining the solubility of CaTPA, the concentration of TPA that would be required to achieve urinary saturation was calculated, and a conservative estimate of the amount of TPA or DMT that would have to be absorbed in order to induce calculi was derived. TPA and MA induced bladder tumors in rats in chronic feeding studies. However, it is likely that these tumors were secondary to the development of calculi. TPA and MA are apparently nongenotoxic, and they do not appear to be metabolized. Increased cell replication in the urothelium of the bladder caused by chronic physical injury was probably a major factor in the mechanism of induction of bladder tumors by bladder stones. Bladder neoplasms occurred primarily in the high dose groups, and they were usually, although not invariably, associated with stones. The possibility that stones were passed or were lost during processing of tissues for histopathologic examination could explain the absence of calculi from some of the neoplastic bladders. The formation of bladder calculi is an example of a threshold effect. Although there is strong evidence linking bladder stones with the induction of tumors, the existence of thresholds in chemical carcinogenesis continues to be controversial. A decision by the U.S. Environmental Protection Agency concerning the levels of MA allowed to occur in the food chain indicates that data regarding thresholds, even in the case of urolithiasis, are not being utilized in the risk assessment process.

Animals↗

Triazines.

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Animals↗

A triazine dye, cibacron blue 3G-A induces Staphylococcus aureus to form giant clusters.

Cibacron blue 3G-A (CB), one of the triazinyl dyes commonly used as a ligand for affinity chromatography, induced staphylococci to form giant clusters without affecting cell viability. It was demonstrated that the cluster formation was not a mechanical aggregation of cells by CB, but a consequence of its biological effect on staphylococcal growth. Clusters induced by CB did not form a regular arrangement, and were different from clusters induced by SDS or polyanethole sulfonate.

Cell Division↗