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PubMed1989

Voltammetric determination of cyanide and thiocyanate in small biological samples.

Practical methods for the quantitative measurement of cyanide and thiocyanate content in small biological samples are presented. The basis of the procedures is electrochemical determination of CN- with a silver rotating disk electrode at 90 mV, relative to the Ag/AgCl reference electrode, or with a dropping mercury electrode in differential pulse mode at -240 mV. Thiocyanate is oxidized to CN- with permanganate under controlled conditions prior to its analysis. Endogenous cyanide and cyanide from SCN- oxidation are recovered, separately, by volatilization and trapping. Emphasis is placed on careful handling of specimens prior to analysis. Precautions include a provision for sequestering cyanide in storage, the use of an antioxidant, and an avoidance of the spontaneous sulfide formation from proteins that occurs in even mildly alkaline solutions.

Cyanides↗
PubMed1984

Thiocyanate inhibition of active chloride absorption in Aplysia intestine.

This investigation was principally undertaken to examine the mechanism of active chloride absorption across the Aplysia californica intestine by using various inhibitors of ion transport. Isolated intestine, mounted between identical oxygenated sodium-free seawater solutions, maintained stable transmural potential differences (serosa negative) and short-circuit currents for several hours at 25 degrees C. The metabolic inhibitors, 2,4-dinitrophenol and fluoride, reduced both transmural potential difference and short-circuit current; however, the electrical characteristics were predominantly dependent upon glycolytic energy. The addition of thiocyanate to the mucosal solution inhibited both electrical characteristics in parallel, and this inhibition could be titrated according to the thiocyanate concentration. The short-circuit current was carried wholly by a net active chloride transfer from mucosa to serosa as determined by flux measurements. These results suggest that active chloride absorption may be mediated by a primary active transport process.

Animals↗
PubMed1988

The measurement of relative antibody affinity by ELISA using thiocyanate elution.

A variation of the standard ELISA assay was used to determine the relative affinities of six murine monoclonal anti-dinitrophenol (DNP) antibodies for DNP-bovine serum albumin (DNP-BSA). The procedure involved exposing replicate wells containing antibody bound to antigen to increasing concentrations of the chaotropic thiocyanate ion. Resistance to thiocyanate elution was utilized as a measure of affinity and an index (affinity index) representing a 50% reduction in initial absorbance was used to rank the anti-DNP antibodies in order of increasing affinity. For comparison, the affinity constants of the six monoclonals were determined by equilibrium dialysis with 3H-epsilon-DNP-L-lysine as antigen. A significant correlation (P less than 0.02) was observed between the ranking orders obtained using the two methods, thereby demonstrating the applicability of the elution technique for the measurement of relative antibody affinity.

Animals↗
PubMed1995

The assessment of antibody affinity distribution by thiocyanate elution: a simple dose-response approach.

We describe a simple dose-response approach to assess the affinity distribution of polyclonal antibodies. The proportion of antigen-specific antibodies dissociated by increasing concentrations of the mild chaotropic agent ammonium thiocyanate (NH4SCN) was measured by enzyme immunoassay, and the distribution of tolerances to this agent was presented in a histogram form. Such 'tolerance distribution', which is analogous to that described in classical dose-response bioassays, is proposed as a representation of the actual antibody affinity distribution. To test this approach, we assessed affinity maturation patterns of anti-Plasmodium falciparum IgG antibodies in paired sera obtained from 22 malaria patients during the acute infection and convalescence. We obtained patterns of antibody affinity distributions consistent with those previously described in immunization experiments with the aid of more complex laboratory and computational approaches. Therefore, we suggest the thiocyanate elution technique as an alternative method for rapid assessment of affinity distributions of polyclonal antibodies elicited against complex antigens, readily applicable to large number of serum samples.

Animals↗
PubMed1984

A kinetic study of the reaction between human myeloperoxidase, hydroperoxides and cyanide. Inhibition by chloride and thiocyanate.

The reaction between myeloperoxidase (donor:hydrogen-peroxide oxidoreductase, EC 1.11.1.7), hydrogen peroxide and ethyl hydroperoxide was investigated using the stopped-flow technique. Like other peroxidases, myeloperoxidase forms two sequential peroxide compounds. The pH-dependence of the apparent second-order rate constant of compound I formation shows that there is an acid/base group on the enzyme with a pKa of 4.30 +/- 0.15, which - when protonated - prevents the reaction of the enzyme with peroxides. The rate constants for the formation of compound I by hydrogen peroxide and ethyl hydroperoxide are (2.3 +/- 0.1) X 10(7) M-1 X s-1 and (2.8 +/- 0.3) X 10(5) M-1 X s-1, respectively. The binding of cyanide to myeloperoxidase (k1 = (1.30 +/- 0.05) X 10(6) M-1 X s-1) is also regulated by an acid/base group with a pKa of 4.00 +/- 0.05 as is the case with hydrogen peroxide; also, only the protonated uncharged form of cyanide reacts with the enzyme. From their effects on the binding of cyanide to the enzyme it is concluded that chloride and thiocyanate bind to myeloperoxidase only when the acid/base group is protonated. The pH-dependence of the dissociation constant of the myeloperoxidase-chloride complex obtained from the spectral changes induced by chloride is the same as observed in the inhibition by chloride of the binding of cyanide. It is concluded that hydrogen peroxide, cyanide, chloride and thiocyanate bind at the same site on the enzyme.

Chlorides↗
PubMed1994

1H- and 15N-NMR study of the binding of thiocyanate to chemically modified horseradish peroxidase and involvement of salt bridge.

The chemical modification of native horseradish peroxidase (HRP) has been carried out by esterification of the heme propionic group. 15N- and 1H-NMR studies on binding of thiocyanate ion to chemically modified HRP have been utilized to demonstrate the existence of salt bridge between the heme propionic acid and distal amino acid group. The catalyzed oxidation of thiocyanate by the native HRP, and the chemically modified HRP has also been studied at different pH, and the significance of the salt bridge discussed.

Binding Sites↗
PubMed1985

Expired air carbon monoxide and saliva thiocyanate: relationships to self-reports of marijuana and cigarette smoking.

This study examined the relationship between self-reports of marijuana and cigarette smoking, and the physiological measures of expired air carbon monoxide (CO) and saliva thiocyanate (SCN) in a sample of 1,130 seventh, ninth, tenth, eleventh, and twelfth graders. Subjects who reported marijuana smoking were likely to also report cigarette smoking. The correlation between self-reported marijuana smoking and SCN was negligible. The correlation between CO and self-reported marijuana smoking was statistically significant, but when the variance due to cigarette smoking was removed, this relationship also became negligible. The existence of a sizeable number of marijuana smokers in this sample of adolescents did not alter the correlation between CO and self-reports of cigarette smoking. However, in adult samples, where marijuana and cigarette smoking are less highly correlated, marijuana smoking could affect the relationship between CO and self-reported cigarette smoking. Carbon monoxide predicted self-reported cigarette smoking better than did saliva thiocyanate. There was an interaction between grade and the CO/cigarette smoking correlation. The correlations were generally higher in upper grades.

Adolescent↗
PubMed2006

Some properties of a sequencing batch reactor for treatment of wastewater containing thiocyanate compounds.

Thiocyanate (SCN) compounds in photo-processing wastewater (PPWW) could be treated by an SBR system without any release of thiocyanate to the atmosphere during the aeration step. An SCN loading greater than 84 g m(-3)d(-1) showed negative effects on the growth of bio-sludge and removal efficiencies of the system. The acclimatization period of the system was increased with an increase in SCN concentration or loading. The COD, BOD(5), TKN, and SCN removal efficiencies were 96.0 +/- 1.6%, 72 +/- 2%, 49 +/- 5%, and 82 +/- 3%, respectively, under an SCN loading of up to 84 g m(-3)d(-1). The removal efficiency of the system was repressed by SCN due to the repressed growth rate of nitrification bacteria. However, the removal efficiency could be increased with an increase in HRT or a decrease in SCN loading. Also, increases in HRT or decreases in SCN loading led to increased sludge age or solid retention time (SRT) and decreased the sludge volume index (SVI) value. The SRT and SVI of the system with synthetic wastewater containing 840 mg l(-1) SCN under an HRT of 3 days (SCN loading of 280 g m(-3)d(-1)) were 3.9 +/- 0.7 days and 65 +/- 4 ml g(-1), respectively, while they were 11.2+/-0.8 days and 55 +/- 6 ml g(-1), respectively under an HRT of 10 days (SCN loading of 84 g m(-3)d(-1)).

Bioreactors↗
PubMed2006

Simultaneous removal of phenol, ammonium and thiocyanate from coke wastewater by aerobic biodegradation.

A laboratory-scale activated sludge plant composed of a 20 L volume aerobic reactor followed by a 12 L volume settling tank and operating at 35 degrees C was used to study the biodegradation of coke wastewater. The concentrations of ammonium nitrogen (NH(4)(+) -N), phenols, chemical oxygen demand (COD) and thiocyanate (SCN(-)) in the wastewater ranged between 504 and 2,340, 110 and 350, 807 and 3,275 and 185 and 370 mg/L, respectively. The study was undertaken with and without the addition of bicarbonate. The addition of this inorganic carbon source was necessary to favour nitrification, as the alkalinity of the wastewater was very low. Maximum removal efficiencies of 75%, 98% and 90% were obtained for COD, phenols and thyocianates, respectively, without the addition of bicarbonate. The concentration of ammonia increased in the effluent due to both the formation of NH(4)(+) as a result of SCN(-) biodegradation and to organic nitrogen oxidation. A maximum nitrification efficiency of 71% was achieved when bicarbonate was added, the removals of COD and phenols being almost similar to those obtained in the absence of nitrification. Batch experiments were performed to study the influence of pH and alkalinity on the biodegradation of phenols and thiocyanate.

Aerobiosis↗
PubMed2006

Spectrophotometric determination of some anti-tussive and anti-spasmodic drugs through ion-pair complex formation with thiocyanate and cobalt(II) or molybdenum(V).

Two rapid, simple and sensitive extractive specrophotometric methods has been developed for the determination of anti-tussive drugs, e.g., dextromethorphan hydrobromide (DEX) and pipazethate hydrochloride (PiCl) and anti-spasmodic drugs, e.g., drotaverine hydrochloride (DvCl) and trimebutine maleate (TM) in bulk and in their pharmaceutical formulations. The proposed methods depend upon the reaction of cobalt(II)-thiocyanate (method A) and molybdenum(V)-thiocyanate ions (method B) with the cited drugs to form stable ion-pair complexes which extractable with an n-butnol-dichloromethane solvent mixture (3.5:6.5) and methylene chloride for methods A and B, respectively. The blue and orange red color complexes are determined either colorimetrically at lambdamax 625 nm (using method A) and 467 or 470 nm for (DEX and PiCl) or (DvCl and TM), respectively (using method B). The concentration range is 20-400 and 2.5-50 microg mL-1 for methods A and B, respectively. The proposed method was successfully applied for the determination of the studied drugs in pure and in pharmaceutical formulations applying the standard additions technique and the results obtained in good agreement well with those obtained by the official method.

1-Butanol↗
PubMed2000

Determination of thiosulfate, thiocyanate and polythionates in a mixture by ion-pair chromatography with ultraviolet absorbance detection.

A sensitive ion chromatographic method has been developed for the determination of mixtures of thiosulfate, thiocyanate and polythionates (tri-, tetra-, penta- and hexathionate). The proposed method is based on the separation of the sulfur anions on an octadecylsilica (ODS) column with an acetonitrile-water mobile phase containing tetrapropylammonium salt (TPA) as an ion-pairing reagent and the ultraviolet absorption detection of the sulfur anions. When an acetonitrile-water (20:80, v/v) solution (pH 5.0) containing 6 mM TPA was used as a mobile phase at flow-rate of 0.6 ml min(-1), the sulfur anions were resolved within 22 min. The detection limits defined at S/N=3 and 230 nm were very low for all anions, except trithionate: 30 nM for thiosulfate, 60 nM for thiocyanate, 20 nM for tetrathionate, 15 nM for pentathionate and 18 nM for hexathionate. The proposed method gave recoveries ranging from 95.0 to 105.0% when applied to the determination of polythionates added to hot spring waters.

Anions↗
PubMed2001

Analysis of thiocyanate in biological fluids by capillary zone electrophoresis.

A new sensitive and simple method has been developed for the determination of thiocyanate in human serum, urine and saliva. The determinations were performed in a fused-silica capillary [64.5 cm (56 cm effective length) x 75 microm] using 0.1 M beta-alanine-HCl (pH 3.50) as a background electrolyte, separation voltage 18 kV (negative polarity), temperature of capillary 25 degrees C and direct detection at 200 nm. Serum samples were 10-times diluted with deionised water and deproteinised with acetonitrile in the ratio 1:2. Urine and saliva samples need only 20-fold dilution with deionised water. The proposed method was successfully applied to the determination of thiocyanate in various human serum, saliva and urine samples.

Electrophoresis, Capillary↗
PubMed2000

Thiocyanate mediated antifungal and antibacterial property of goat milk lactoperoxidase.

Goat milk lactoperoxidase was purified using CM-Sephadex-C-50 ion exchange chromatography and Sephadex G-100 gel filtration. The purified protein was found to have high antifungal and antibacterial activity in a thiocyanate-H2O2 medium. The results are relevant enough to suggest the exploitation of LP-thiocyanate-H2O2 system as an effective agent against many of the disease causing organisms in plants and animals.

Animals↗
PubMed2002

Determination of urinary thiocyanate and nitrate using fast ion-interaction chromatography.

A simple and rapid chromatographic method for determination of nitrite, nitrate and thiocyanate is reported, and applied to the analysis of non-, medium and heavy smokers' urine samples. Ion-interaction liquid chromatography was carried out on a short 30 mm x 4.6 mm C18 column (3 microm particle size) with a mobile phase of 10 mM tetrabutylammonium phosphate in 20% MeOH. The chromatography was performed at an elevated temperature of 45 degrees C, at a flow-rate of 1 ml/min. Detection was by direct UV absorption at 230 nm. Sample preparation involved centrifugation and dilution, followed by sample clean-up on a C18 solid-phase extraction cartridge. The developed method proved both precise (% RSD <2%) and sensitive (standard detection limits <0.1 mg/l), and yielded total run times of under 10 min when applied to urine analysis of smokers and non-smokers, with thiocyanate eluting in under 5 min.

Chromatography, Liquid↗
PubMed1999

Narrowbore high performance liquid chromatography of berberine and palmatine in crude drugs and pharmaceuticals with ion-pair extraction using cobalt thiocyanate reagent.

For the simultaneous determination of berberine and palmatine from Phellodendri Cortex, Coptidis Rhizoma and pharmaceuticals, a new narrowbore HPLC method was developed with a simple and selective sample clean-up using cobalt thiocyanate reagent. Samples were sonicated with 2% hydrochloric acid for 30 min and protoberberine-type alkaloids in the resulting mixture were extracted with cobalt thiocyanate reagent and dichloroethane. The aliquot of dichloroethane layer was evaporated and the residue was dissolved for HPLC analysis. The recoveries of berberine and palmatine from Phellodendri Cortex, and Coptidis Rhizoma were better than 90%. Calibration curves for berberine and palmatine were linear over the concentration range of 0.1-50 microg/ml. Limits of quantitation for berberine and palmatine were 0.5 ng. This sample preparation process can be used for the identification of protoberberine-type alkaloids from crude drugs and oriental pharmaceutical preparations.

Alkaloids↗
PubMed2000

Indirect atomic absorption spectrometric determination of pindolol, propranolol and levamisole hydrochlorides based on formation of ion associates with manganese thiocyanate and potassium ferricyanide.

A new simple, accurate, precise and sensitive indirect method for the determination of pindolol HCl (1), propranolol HCl (2) and levamisole HCl (3) using atomic absorption spectrometry has been developed. The method is based on precipitation of the ion associates formed from the reaction of (1), (2) or (3) with manganese thiocyanate and/or potassium ferricyanide. The solubility of the solid complexes at the optimum conditions of pH and ionic strength values have been studied. Saturated solutions of each ion-associate were prepared under the optimum conditions and the metal ion content in the supernatant was determined. The method has been used for the determination of 1.14-17.07, 1.18-17.75 and 1.08-16.24 microg/ml of (1), (2) and (3), respectively using manganese thiocyanate and 1.71-25.60, 1.77-26.62 and 1.62-24.36 microg/ml of (1), (2) and (3), respectively using potassium ferricyanide. The method developed applied for analysis of bulk drugs and some of their pharmaceutical preparations.

Adjuvants, Immunologic↗
PubMed2000

Spectral and kinetic studies on the formation of eosinophil peroxidase compound I and its reaction with halides and thiocyanate.

Compound I of peroxidases takes part in both the peroxidation and the halogenation reaction. This study for the first time presents transient kinetic measurements of the formation of compound I of human eosinophil peroxidase (EPO) and its reaction with halides and thiocyanate, using the sequential-mixing stopped-flow technique. Addition of 1 equiv of hydrogen peroxide to native EPO leads to complete formation of compound I. At pH 7 and 15 degrees C, the apparent second-order rate constant is (4.3 +/- 0.4) x 10(7) M(-1) s(-1). The rate for compound I formation by hypochlorous acid is (5.6 +/- 0.7) x 10(7) M(-1) s(-1). EPO compound I is unstable and decays to a stable intermediate with a compound II-like spectrum. At pH 7, the two-electron reduction of compound I to the native enzyme by thiocyanate has a second-order rate constant of (1.0 +/- 0. 5) x 10(8) M(-1) s(-1). Iodide [(9.3 +/- 0.7) x 10(7) M(-1) s(-1)] is shown to be a better electron donor than bromide [(1.9 +/- 0.1) x 10(7) M(-1) s(-1)], whereas chloride oxidation by EPO compound I is extremely slow [(3.1 +/- 0.3) x 10(3) M(-1) s(-1)]. The pH dependence studies suggest that a protonated form of compound I is more competent in oxidizing the anions. The results are discussed in comparison with those of the homologous peroxidases myeloperoxidase and lactoperoxidase and with respect to the role of EPO in host defense and tissue injury.

Eosinophil Peroxidase↗
PubMed1982

Anation of laccase type 2 copper by azide and thiocyanate ions.

A mechanistic study of the anation of type 2 Cu(II) in fully oxidized laccase by azide and thiocyanate ions (X-) is reported. The rate data support a mechanism involving rapid formation of an outer-sphere complex (laccase . X-) followed by rate-limiting dissociative interchange to give the inner-sphere complex (laccase-X-) product: (formula; see text) Rate parameters for the laccase-azide reaction are k'2 = 1.25 X 10(-1) s-1 and k'-2 = 1.50 X 10(-2) s-1; those for the laccase-thiocyanate reaction are k'2 = 2.0 X 10(-2) s-1 and k'-2 = 1.1 X 10(-2) s-1 (25 degrees C, pH 6.1 phosphate buffer, I = 0.5 M). Although the rate law for anation of type 2 Cu(II) by N3- in 2-(N-morpholino)ethanesulfonic acid (Mes) or acetate medium (kobsd = k3 + k4[N3-]) differs from that observed in phosphate buffer, the data may still be accounted for in terms of the above mechanism, with Kos[N3-] much less than 1 [in Mes, k3 = 2.2 X 10(-2) s-1 and k4 = 3.3 X 10(-1) M-1 s-1; in acetate, k3 = 2.1 X 10(-2) s-1 and k4 = 3.4 X 10(-1) M-1 s-1 (25 degrees C, pH 6.0, I = 0.25 M)]. The equilibrium and spectroscopic characteristics of the laccase type 2 Cu(II)-N3- complex are compared with those of low molecular weight copper(II)-azide species, and the factors responsible for the very low substitutional reactivity of the type 2 cupric ion are discussed.

Anions↗