scholarly journals Efficient Cleavage of Conjugates of Drugs or Poisons by Immobilized β-Glucuronidase and Arylsulfatase in Columns

1999 ◽  
Vol 45 (12) ◽  
pp. 2173-2182 ◽  
Author(s):  
Stefan W Toennes ◽  
Hans H Maurer

Abstract Background: Cleavage of conjugates is an important step in toxicological analysis, especially of urine samples. The aim of this study was to combine the advantages and to reduce the disadvantages of acid hydrolysis and conventional enzymatic hydrolysis procedures. Methods: β-Glucuronidase (GRD; EC 3.2.1.31) and arylsulfatase (ARS; EC 3.1.6.1) were purified and coimmobilized on an agarose gel matrix and packed into columns. Results: In columns packed with GRD and ARS, the test conjugates 4-nitrophenyl glucuronide and 4-nitrophenyl sulfate added into urine could be completely cleaved within 25 min. Even the relatively stable morphine conjugates could be completely hydrolyzed within 60 min in authentic urine samples. Therefore, an incubation time of 1 h is recommended. Enzyme inhibition by matrix or by rather high concentrations of acetaminophen conjugates was tested and found to be up to 50%. However, a large excess of GRD and ARS was used. The immobilizate columns could be reused for at least 70 incubations and had a storage stability of at least 12 weeks. Carryover of analytes in reused columns could be avoided by rinsing with 200 mL/L methanol in acetate buffer. Thus, five drugs known to be contaminants added in very high concentrations into urine could be completely removed from the columns. A study on the applicability in systematic toxicological analysis showed that 120 different drugs and/or their metabolites could be detected in 35 different authentic urine samples. Conclusions: Use of immobilized and column-packed GRD and ARS is an efficient alternative for the cleavage of urinary conjugates in clinical toxicology.

1984 ◽  
Vol 49 (5) ◽  
pp. 1061-1078 ◽  
Author(s):  
Jiří Čeleda ◽  
Stanislav Škramovský

Based on the earlier paper introducing a concept of the apparent parachor of a solute in the solution, we have eliminated in the present work algebraically the effect which is introduced into this quantity by the additivity of the apparent molal volumes. The difference remaining from the apparent parachor after substracting the contribution corresponding to the apparent volume ( for which the present authors suggest the name metachor) was evaluated from the experimental values of the surface tension of aqueous solutions for a set of 1,1-, 1,2- and 2,1-valent electrolytes. This difference showed to be independent of concentration up to the very high values of the order of units mol dm-3 but it was directly proportional to the number of the free charges (with a proportionality factor 5 ± 1 cm3 mol-1 identical for all studied electrolytes). The metachor can be, for this reason, a suitable characteristic for detection of the association of ions and formation of complexes in the solutions of electrolytes, up to high concentrations where other methods are failing.


Water ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 1607
Author(s):  
Mariano Venturini ◽  
Ariana Rossen ◽  
Patricia Silva Paulo

To produce nuclear fuels, it is necessary to convert uranium′s ore into UO2-ceramic grade, using several quantities of kerosene, methanol, nitric acid, ammonia, and, in low level, tributyl phosphate (TBP). Thus, the effluent generated by nuclear industries is one of the most toxic since it contains high concentrations of dangerous compounds. This paper explores biological parameters on real nuclear wastewater by the Monod model in an ORP controlled predicting the specific ammonia oxidation. Thermodynamic parameters were established using the Nernst equation to monitor Oxiders/Reductors relationship to obtain a correlation of these parameters to controlling and monitoring; that would allow technical operators to have better control of the nitrification process. The real nuclear effluent is formed by a mixture of two different lines of discharges, one composed of a high load of nitrogen, around 11,000 mg/L (N-NH4+-N-NO3−) and 600 mg/L Uranium, a second one, proceeds from uranium purification, containing TBP and COD that have to be removed. Bioprocesses were operated on real wastewater samples over 120 days under controlled ORP, as described by Nernst equations, which proved to be a robust tool to operate nitrification for larger periods with a very high load of nitrogen, uranium, and COD.


Author(s):  
Mario Vincenzo Russo ◽  
Ivan Notardonato ◽  
Alberto Rosada ◽  
Giuseppe Ianiri ◽  
Pasquale Avino

This paper shows a characterization of the organic and inorganic fraction of river waters (Tiber and Marta) and ores/soil samples collected in the Northern Latium region of Italy for evaluating the anthropogenic/natural source contribution to the environmental pollution of this area. For organic compounds, organochloride volatile compounds in Tiber and Marta rivers were analyzed by two different clean-up methods (i.e., liquid–liquid extraction and static headspace) followed by gas chromatography–electron capture detector (GC-ECD) analysis. The results show very high concentrations of bromoform (up to 1.82 and 3.2 µg L−1 in Tiber and Marta rivers, respectively), due to the presence of greenhouse crops, and of chloroform and tetrachloroethene, due to the presence of handicrafts installations. For the qualitative and quantitative assessment of the inorganic fraction, it is highlighted the use of a nuclear analytical method, instrumental neutron activation analysis, which allows having more information as possible from the sample without performing any chemical-physical pretreatment. The results have evidenced high levels of mercury (mean value 88.6 µg g−1), antimony (77.7 µg g−1), strontium (12,039 µg g−1) and zinc (103 µg g−1), whereas rare earth elements show levels similar to the literature data. Particular consideration is drawn for arsenic (414 µg g−1): the levels found in this paper (ranging between 1 and 5100 µg g−1) explain the high content of such element (as arsenates) in the aquifer, a big issue in this area.


1960 ◽  
Vol 38 (1) ◽  
pp. 25-32 ◽  
Author(s):  
Ivan T. Beck ◽  
E. Pinter ◽  
R. D. McKenna ◽  
H. Griff

Acute hemorrhagic pancreatitis in humans is thought to be perpetuated by the autolytic processes catalyzed by trypsin and lipase. This study is an integral part of our search for trypsin and lipase inhibitors to be used in the treatment of this disease.Benzethonium chloride was found to inhibit tryptic activity in vitro. The proteolytic activity of rabbit's serum was inhibited, and the inhibition was most pronounced 6 to 12 hours after the subcutaneous injection of the compound. Fibrinolysin was also inhibited in vitro but benzethonium chloride had no inhibitory action on chymotrypsin, pepsin, or lipase.Serum proteins in vitro were precipitated only with very high concentrations of the compound. No significant protein changes were observed in sera of rabbits after the subcutaneous injection of the compound.


2007 ◽  
Vol 13 (2) ◽  
pp. 213-218
Author(s):  
Thomas W. Stief

Serine protease inactivators (serpins) are important regulators in biochemistry. Often it is necessary to block the serpin action, that is, to stabilize the sample. The guanidine group of arginine is the ligand for the active center pocket of many serine proteases. Arginine or guanidine inhibits serine proteases, and arginine belongs to the reactive P1-P1' center of many serpins. The plasmatic antithrombin, antiplasmin, or anti-C1-esterase activity was determined: A total of 20 µL of pooled normal plasma or 7% human albumin was added to 100 µL of 0—2.67 M arginine, pH 8.6, 10 µL of 26 mIU/mL thrombin in 7% human albumin, and 30 µL of 1.7 mM CHG-Ala-Arg-pNA (37°C). ΔA at 405 nm was determined, by using a microtiter plate reader. Thrombin was substituted by plasmin or C1-esterase, and the chromogenic peptide substrates <Glu-Phe-Lys-pNA or MeOC-Lys(eCBO)-Gly-Arg-pNA, respectively, were used. The IC50 of arginine against plasmatic antithrombin activity is 580 mM; the IC 25 is 440 mM. The IC25 of arginine against plasmatic α 2-antiplasmin or C1-inactivator is 1650 mM. The amidolytic activity of thrombin, plasmin, and C1-esterase is inhibited similarly by arginine: the IC50 for arginine against the amidolytic activity of these proteases is about 400 mM. Arginine at very high concentrations inhibits serpins. This is important, if stabilization of a biological fluid is a prerequisite for valid activities of serine proteases. In addition, these high concentrations of arginine might be a new gentle principle to inhibit pathogens that need serpins for their pathophysiology.


Blood ◽  
1975 ◽  
Vol 45 (1) ◽  
pp. 71-82 ◽  
Author(s):  
E Gimpert ◽  
M Jakob ◽  
WH Hitzig

Abstract Some characteristics of vitamin B12 binding and transport in the serum of an infant with congenital hereditary transcobalamin II (TC II) deficiency were studied using the following parameters and methods: vitamin B12 level and binding capacity; electrophoretic mobility in polyacrylamide gel electrophoresis; various immunodiffusion and absorption experiments, using a specific anti-TC II antiserum and the patient's serum as antigen. The results of these studies point to a deficient synthesis of TC II. Parenteral administration of high doses of vitamin B12 was followed by rapid and complete clinical remission and the appearance of vitamin B12 binder in the alpha 2 region which is similar to “fetal binder.” Thus, very high concentrations of vitamin B12, either carrier free or bound to this alpha 2 binder, were able to correct the disturbed physiology of TC II deficiency, presumably by normalization of DNA-thymine synthesis.


This work analyzes the state of snowpack in Nizhny Novgorod on the basis of certain chemical performances and integral biological toxicity. Snow samples were obtained in February 2018 along major highways of Nizhny Novgorod. A snow-covered area in Dubrava forestry was selected as reference. The studies demonstrated that the snowpack was characterized by very high concentrations of chlorides and sulfides: in sampling points of the Lower City, the content of chlorides and sulfates varied in the ranges of 24.67–62.36 mg/l and 30.16–62.09 mg/l, respectively, and in sampling points of the Upper City, this variability was 416.82–988.45 mg/l and 280.11–879.22 mg/l, respectively. The content of lead in snowpack in both the Lower City and the Upper City was approximately the same (0.0053 and 0.0048 mg/l). The minimum content of pollutants in snow samples from reference site was characterized by toxicity (10%, V = 6.0%) which was estimated as allowable (toxicity class 1). Snowpack water from the Lower City was characterized generally by medium toxicity (class 2), and sampled in the Upper City – by acute toxicity (59%, V = 26.5%), with regard to the reference (class 3).


2021 ◽  
Vol 26 (51) ◽  
pp. 27-32
Author(s):  
Biljana Sikirić ◽  
Vesna Mrvić ◽  
Olivera Stajković-Srbinović ◽  
Vladan Ugrenović ◽  
Darko Jaramaz ◽  
...  

During the regular control of soil fertility in the Rasina District, it was established that the plots of land were distributed across Vertisol, Eutric Cambisol and Fluvisol types of soils, and to a lesser extent on Pseudogley and Ranker. The tested samples had different textures - sandy loam and loam, clay-sandy loam and clay loam, and clay. Plots of land that were of very acidic and acidic reactions were predominant, with medium amounts of humus, very low amounts of available phosphorus, and high amounts of available potassium. High or very high cation absorption capacity was found in about half of the examined fields; a deficient content of exchangeable Ca was recorded in 22% of plots and that of exchangeable Mg in 16% of plots, while an unfavorable Ca/Mg ratio was measured in 44% of plots. The overall sensitivity to acidification was mainly moderate (50.6% of plots) and strong (20.2% of plots). Very high concentrations of mobile Al, which could be toxic to plants, were found in 5 field plots.


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