Irgasan DP 300 Research Articles

Identification of methyl triclosan and halogenated analogues in male common carp ( Cyprinus carpio) from Las Vegas Bay and semipermeable membrane devices from Las Vegas Wash, Nevada

Methyl triclosan and four halogenated analogues have been identified in extracts of individual whole-body male carp ( Cyprinus carpio) tissue that were collected from Las Vegas Bay, Nevada, and Semipermeable Membrane Devices (SPMD) that were deployed in Las Vegas Wash, Nevada. Methyl triclosan is believed to be the microbially methylated product of the antibacterial agent triclosan (2, 4, 4'-trichloro-4-hydroxydiphenyl ether, Chemical Abstract Service Registry Number 3380-34-5, Irgasan DP300). The presence of methyl triclosan and four halogenated analogues was confirmed in SPMD extracts by comparing low- and high-resolution mass spectral data and Kovats retention indices of methyl triclosan with commercially obtained triclosan that was derivatized to the methyl ether with ethereal diazomethane. The four halogenated analogues of methyl triclosan detected in both whole-body tissue and SPMD extracts were tentatively identified by high resolution mass spectrometry. Methyl triclosan was detected in all 29 male common carp from Las Vegas Bay with a mean concentration of 596 μg kg − 1 wet weight (ww) which is more than an order of magnitude higher than previously reported concentrations in the literature. The halogenated analogs were detected less frequently (21%–76%) and at much lower concentrations (< 51 μg kg − 1 ww). None of these compounds were detected in common carp from a Lake Mead reference site in Overton Arm, Nevada.

Formation of antibiotic, biodegradable polymers by processing with Irgasan DP300R (triclosan) and its inclusion compound with β‐cyclodextrin

AbstractThe inclusion compound (IC) between the FDA‐approved antibacterial Irgasan DP300 (Trichlosan), and β‐cyclodextrin (CD) has been formed. When the Irgasan–β‐CD–IC is embedded in biodegradeable/bioabsorbable films of poly(ϵ‐caprolactone) (PCL) at low levels (a few wt %), they are rendered resistant to the growth of E. coli bacteria. When these same PCL films embedded with Irgasan–β‐CD–IC are used as the adhesive for laminating cotton fabrics, we observe the resulting cotton laminates to also be resistant to the growth of E. coli bacteria. These results hold promise for the fabrication of bacteria‐resistant polymer films and fibers, as well as antibacterial fabrics, by means of simple melt processing with Irgasan–β‐CD–IC. © 2001 John Wiley &amp; Sons, Inc. J Appl Polym Sci 82: 300–309, 2001

Inhibition of light emission from the bioluminescent bacterium Vibrio fischeri after exposure to triclosan and related hygiene care products.

The affect of the anti-microbial agent triclosan (alternative names Microban and Irgasan DP300) on the light emission by the bioluminescent bacterium Vibrio fischeri was determined. Triclosan at concentrations greater than 0.2% (w/v) caused cell lysis and immediate (< 5 s) loss of light emission. Exposure to triclosan at lower concentrations caused a decrease in light output over time. The rate of the decrease in light output followed a cuboid relationship, of which the initial rate (first 60 s) of light loss was proportional to the concentration of triclosan. The effect on light output by two commercially available hygiene products containing triclosan also caused a similar response in light loss.

Effect of 2,4,4′-trichloro-2′-hydroxydiphenyl ether on cytochrome P450 enzymes in the rat liver

We examined the effect of 2,4,4′-trichloro-2′-hydroxydiphenyl ether (Irgasan DP300) on microsomel cytochrome P450 (P450) enzymes in rat liver. Rats were treated intraperitoneally with Irgasan DP300 daily for 4 days, at doses of 0.2, 0.4 and 0.8 mmol/kg. Among the P450 dependent monooxygenase activities, 7-benzyloxyresorufin O-debenzylase (BROD) and 7-pentoxyresorufin O-depentylase (PROD) in rats, which are associated with CYP2B1, were remarkably induced by all doses of Irgasan DP300. The relative induction to each control activity were from 5.6- to 22.3-fold and 4.9- to 20.2-fold, respectively. Furthermore, immunoblotting showed that CYP2B1/2 protein level in rat liver microsomes was increased from 10.8- to 34.4-fold by Irgasan DP300. In addition, 7-ethoxycoumarin O-deethylase (ECOD) and p-nitrophenol hydroxylase (PNPH) activities were significantly increased by Irgasan DP300 at all doses (from 1.4- to 4.9-fold). Although the activities of other P450-dependent monooxygenases, namely aminopyrine N-demethylase (APND), aniline p-hydroxylase (ANPH), erythromycin N-demethylase (EMND), lauric acid w-hydroxylase (LAOH) and testosterone 6β-hydroxylase (TS6BH) were increased at high doses (≥0.4 mmol/kg) of Irgasan DP300, the relative level was lower than those of the CYP2B1-dependent monooxygenases such as BROD and PROD. However, 7-ethoxyresorufin O-deethylase (EROD), 7-methoxyresorufin O-demethylase (MROD), testosterone 2a-hydroxylase (TS2AH) and testosterone 7a-hydroxylase (TS7AH) activities were not affected by any doses of Irgsan DP300. Immunoblotting showed that CYP3A2/1 and CYP4A1 protein levels were significantly induced from 1.3- to 2.2-fold by Irgasan DP300 (≥0.4 mmol/kg), whereas those of CYPlA1/2, CYP2C11/6 and CYP2E1 were not affected by any doses of Irgasan DP300. These results suggest that Irgasan DP300 induces the P450 isoforms of CYP2B subfamily in the rat liver, and that the induced P450 isozymes closely relates to the toxicity of Irgasan DP300 or its chlorinated derivatives.

Interaction of 2,4,4′-trichloro-2′-hydroxydiphenyl ether with microsomal cytochrome p450-dependent monooxygenases in rat liver

We studied the effects of 2,4,4′-trichloro-2′-hydroxydiphenyl ether (Irgasan DP300) on the kinetics of the cytochrome P450 (P450)-dependent monooxygenases in rat liver microsomes. The activities of 7-ethoxyresorufin 0-deethylase (EROD) and 7-pentoxyresorufin O-depentylase (PROD) in rat liver microsomes exposed to 3-methylcholanthrene (MC) and phenobarbital (PB) respectively, were substantially inhibited by Irgasan DP300. The inhibition profile of EROD was competitive, whereas that of PROD was noncompetitive; the Ki values from Hanes plots were 0.24 and 1.48 μM for EROD and PROD, respectively. Phenacetin O-deethylase (PCOD) and 4-nitrophenol hydroxylase (4NPH) activities in rats exposed to PB were also inhibited by Irgasan DP300, at Ki values lower than those for other microsomes. Irgasan DP300 slightly inhibited testosterone 6β-hydroxylase (TS6BH) activities in some microsomes. No effect of Irgasan DP300 on lauric acid ω-hydroxylase (LAOH) activity was evident in any microsomal preparations. These results indicated that Irgasan DP300 inhibits MC- and PB-inducible P450-dependent monoxygenase in vitro competitively or noncompetitively, and that the P450 enzymes of the CYP1A or CYP2B subfamily may contribute to Irgasan DP300 toxicity.

Mutagenicity of Polychlorophenoxyphenols (Predioxins) and Their Photo-degradation Products in Aqueous Solution.

Polychlorinated phenoxyphenols (PCPPs), including predioxins and isopredioxins, and their photo-degradation products in aqueous solution were tested for mutagenicity on Salmonella typhimurium TA98 and TA100 strains. Mutagenic effects of Irgasan DP 300 [5-chloro-2-(2, 4-dichlorophenoxy) phenol] as a typical predioxin and PCPPs (chlorophenol dimers) could not be observed on both TA98 and TA100 strains with and without S9 mix, because of thier great toxicity to tester strains. No mutagenic effects were also observed for 2-alkylated PCPPs (chloro-2-alkylphenol dimers) on both tester strains. However, some 4-alkylated PCPPs (chloro-4-alkylphenol dimers) were mutagenic on TA100 strain without S9 mix, and great mutagenicity was detected for 4-methylated PCPPs. PCPPs in aqueous solution were found to be converted to compounds with mutagenicity to TA100 strain without S9 mix, when the solution was subjected to photo-irradiation in the presence of chlorine for 5 h. In addition, a mutagenic effect of 4-methylated PCPPs solution on TA100 strain without S9 mix was shown to be remained about 50% of the original activity, even after treatment with chlorine alone and photo-irradiation in the presence of chlorine for 5 h. However, no mutagenic effects of Irgasan DP300 in aqueous solution were observed after any treatments.

Acute toxicity, percutaneous absorption and effects on hepatic mixed function oxidase activities of 2,4,4'-trichloro-2'-hydroxydiphenyl ether (Irgasan DP300) and its chlorinated derivatives.

Acute toxicity of 2,4,4'-trichloro-2'-hydroxydiphenyl ether (Irgasan DP300) (I) and its three chlorinated derivatives, 2',3,4,4'-tetrachloro-2-hydroxydiphenyl ether (II), 2',4,4',5-tetrachloro-2-hydroxydiphenyl ether (III) and 2',3,4,4',5-pentachloro-2-hydroxydiphenyl ether (IV), in mice were examined by intraperitoneal injection. The LD50 values of Irgasan DP300, II, III and IV were 1,090, 710, 650 and 430 mg/kg, respectively. The percutaneous absorptions of these tritiated compounds were also examined by the application on the backs of mice. The radioactivities in most tissues reached to the maximal levels at 12 h or 18 h after dosing, which corresponded to 11-76% of the maximal levels given by the oral administration (Kanetoshi et al. 1988a). These results show the high percutaneous absorbability of Irgasan DP300 and its chlorinated derivatives. The intraperitoneal administrations of III and IV to rats induced hepatic microsomal aminopyrine N-demethylase and aniline 4-hydroxylase activities similarly to phenobarbital. These chlorinated derivatives also increased cytochrome P-450 content, and the activities of aminopyrine N-demethylase and N-methylaniline N-demethylase in hepatic microsomes from mice. The extents of the increases were similar to those by phenobarbital and 3-methylcholanthrene.

Disposition and excretion of Irgasan DP300 and its chlorinated derivatives in mice.

The distributions of radioactivity were examined by whole body autoradiography and liquid scintillation spectrophotometry in male ddY mice following oral administration of tritium-labelled Irgasan® DP300 (2,4,4′-trichloro-2′-hydroxydiphenyl ether) (I) and its three chlorinated derivatives; 2′,3,4,4′-tetrachloro-2-hydroxydiphenyl ether (II), 2′,4,4′,5-tetrachloro-2-hydroxydiphenyl ether (III) and 2′,3,4,4′,5-pentachloro-2-hydroxydiphenyl ether (IV). The autoradiograms at 6 or 24 hr showed that the radioactivity distributed in the gall, liver, lung, heart, and kidneys. Among these tissues the radioactivity was most concentrated in the gall, suggesting the enterohepatic circulation of these compounds. A much higher level of radioactivity in each tissue was observed in mice receiving [3H]-III than the other compounds tested. Most of the radioactivity disappeared from each tissue in 24 hr due to [3H]-Irgasan DP300, [3H]-II or [3H]-IV, but in 96 hr it was due to [3H]-III.

Formation of polychlorinated dibenzo- p-dioxin from 2,4,4′-trichloro-2′-hydroxydiphenyl ether (irgasan® DP300) and its chlorinated derivatives by exposure to sunlight

Exposure to sunlight in the solid state of 2,4,4′-trichloro-2′-hydroxydiphenyl ether (Irgasan® DP300) ( I) produced dichlorodibenzo- p-dioxin(s) (di- CDD) and a trace amount of trichlorodibenzo- p-dioxin (tri- CDD) together with three chlorinated derivatives of Irgasan DP300, 2′,3,4,4′-tetrachloro-2-hydroxydiphenyl ether ( II), 2′,4,4′,5-tetrachloro-2-hydroxydiphenyl ether ( III) and 2′,3,4,4′,5-pentachloro-2-hydroxydiphenyl ether ( IV). These chlorinated derivatives gave various polychlorinated dibenzo- p-dioxins ( PCDDs) upon exposure to sunlight; i.e., II gave 1,2,8-tri- CDD and a tetrachlorodibenzo- p-dioxin (tetra- CDD); III gave di- CDD(s), 2,3,7-tri- CDD and a tetra- CDD and IV gave three pentachlorodibenzo- p-dioxins (penta- CDDs) with two tri- CDDs and four tetra- CDDs. Upon exposure to sunlight of commercial textile products containing Irgasan DP300, 0.02–0.03% of the agent was converted into di- CDD(s). These results suggest that Irgasan DP300 is one of the precursors of various PCDDs in the environment.

Formation of polychlorinated dibenzo- p-dioxins upon combustion of commercial textile products containing 2,4,4′-trichloro-2′-hydroxydiphenyl ether (Irgasan® DP3000

Commercial textile products containing 2,4,4′-trichloro-2′-hydroxydiphenyl ether (Irgasan DP3000 as an antimicrobial agent gave dichlorodibenzo- p-dioxin(s) (di-CDD) upon combustion. The extent of conversion of Irgasan DP300 into di-CDD(s) reached 19–43% at 600°C. Upon bleaching the textile products with sodium hypochlorite, Irgasan DP300 was chlorinated to 2′-3,4,4′-tetrachloro-2-hydroxydiphenyl ether, 2′,4,4′,5-tetrachloro-2-hydroxydiphenyl ether and 2′-3,4,4′-5-pentachloro-2-hydroxydiphenyl etherl. These chlorinated derivatives were converted into trichlorodibenzo- p-dioxins and tetrachlorodibenzo- p-dioxin upon combustion, which are more toxic than di-CDD(s). These results suggest that the bleaching and incineration of textile products containing Irgasan DP300 result in environmental pollution by PCDDs.

Chlorination of Irgasan DP300 and formation of dioxins from its chlorinated derivatives.

Irgasan DP300 (2,4,4'-trichloro-2'-hydroxydiphenyl ether) (I), an antimicrobial agent for use with fabrics, was easily chlorinated with sodium hypochlorite to give 2',3,4,4'-tetrachloro-2-hydroxydiphenyl ether (II), 2',4,4',5-tetrachloro-2-hydroxydiphenyl ether (III) and 2',3,4,4',5-pentachloro-2-hydroxydiphenyl ether (IV). Irgasan DP300 and its chlorinated derivatives were readily converted into polychlorinated dibenzo-p-dioxins (PCDDs) by heating: Irgasan DP300 was converted into dichlorodibenzo-p-dioxin(s) (di-CDD, 42%); II into two trichlorodibenzo-p-dioxins (tri-CDDs, 22%) and three tetrachlorodibenzo-p-dioxins (tetra-CDDs, 46%); III into two tri-CDDs (44%), more than two tetra-CDDs (25%) and pentachlorodibenzo-p-dioxin(s) (penta-CDD, 1%); and IV into two tetra-CDDs (16%), trace amounts of penta-CDD(s) and four hexachlorodibenzo-p-dioxins (hexa-CDDs, 40%). Although UV irradiation of Irgasan DP300, II and III gave PCDDs, the amounts of PCDDs formed were much smaller than those obtained by heating. Moreover, PCDD was not detected in the UV irradiation of IV. The identified products suggested that disproportionation of chlorine atom(s) occurred in the photolysis.

Studies on the chemicals used in household necessities. I. Analysis of antimicrobial agents in fabrics.

Methods for the qualitative analysis and determination of antimicrobial agents in fabrics are described. At present, antimicrobial agents mainly used are 2, 4, 4'-trichloro-2'-hydroxydiphenyl ether (Irgasan DP300), 3-(trimethoxysilyl) propyldimethyloctadecyl ammonium chloride (DC5700) and thiabendazole (TBZ). The procedure of qualitative analysis is as follows : (1) DC-5700 in fabrics is checked by the adsorption of bromophenol blue (BPB) on the strips packed in column. (2) Irgasan DP300 eluted with ether from fabrics is detected by Gibbs reagent on silica gel thin layer chromatography (TLC). (3) TBZ is eluted with methanol from the fabrics after the ether-elution and detected by UV 254 nm after spraying sulfuric acid on silica gel TLC. DC-5700 is determined from the amount of 1-octadecene, formed in Hofmann degradation, measured by gas-liquid chromatography (GLC). The determination of Irgasan DP300 in fabrics is carried out by high performance liquid chromatography (HPLC) on the acidic fraction of extract with methanol : acetic acid (9 : 1) under reflux and TBZ is determined by GLC on the alkaline fraction of the abovementioned extract. By using the qualitative and quantitative system, Irgasan DP300, DC-5700 and TBZ were detected in 12, 4 and 2 samples, respectively, of 23 commercial fabrics treated with antimicrobial agents.

The acute toxicity of penta-, hexa-, and heptachlorohydroxydiphenyl ethers in mice.

The acute intraperitoneal LD50 values of various hydroxychlorodiphenyl ethers (HO-ClX-DPEs; X = 5-7) in mice have been determined. The acute toxicities observed were on the order of, or slightly less than, that observed previously for 2-hydroxy-2',4,4'-trichlorodiphenyl ether (2-HO-Cl3-DPE; Irgasan DP-300; Triclosan), a commonly used bactericide. However, the acute toxicities determined for these compounds were substantially less than have been observed for HO-Cl9-DPEs and pentachlorophenol. The HO-ClX-DPEs had a marked hypothermic effect, similar to that produced by 2-HO-Cl3-DPE. Symptomatology following exposure to the HO-ClX-DPEs (X = 5-7) suggested a nonspecific depressant effect on the central nervous system.

The acute toxicity of nonachloropredioxin and 3‐ and 4‐hydroxynonachlorodiphenyl ether in mice

The acute intraperitoneal LD50 values of hydroxynonachlorodiphenyl ethers (HO-Cl9-DPEs) in mice have been determined. The acute toxicity of each of these compounds is compared with that of pentachlorophenol (PCP) and 2-hydroxy-2',4,4'-trichlorodiphenyl ether (2-HO-Cl3-DPE; Irgasan DP-300; Triclosan), a commonly used bactericide. The order of acute toxicity observed was: 2-HO-Cl9-DPE greater than technical PCP approximately equal to pure PCP greater than 3-HO-Cl9-DPE greater than 4-HO-Cl9-DPE greater than 2-HO-Cl3-DPE. Symptomatology following exposure to the HO-Cl9-DPEs was similar to that observed for PCP, a known uncoupler of oxidative phosphorylation. 2-HO-Cl3-DPE, however, produced clinical signs suggestive of a suppressive effect on the central nervous system. Data on time response following a lethal dose of each of these compounds was also obtained.

L-Arabinose-ornithine-Irgasan medium for differentiating Serratia species

A semisolid medium (designated Serratia differentiation medium) containing L-arabinose, ornithine, and selective inhibitor was used to differentiate three clinically encountered Serratia species. The inhibitor, Irgasan DP-300, was incorporated to eliminate false-positive reactions from most remaining Enterobacteriaceae. The suspected Serratia colony was inoculated as a stab into the medium. Serratia marcescens was indicated by a change in color from olive to purple following 18 h of incubation, whereas S. rubidaea (not listed in Bergey's Manual of Determinative Bacteriology) was indicated by a change to bright yellow. S. liquefaciens (described in Bergey's Manual of Determinative Bacteriology [8th ed., 1974] as Enterobacter liquefaciens) produced a small purple band at the top of the medium and a yellow or yellow-green butt. Absence of growth and color change following incubation indicates that the suspected colony is a non-Serratia. Thirty-six Serratia strains and 97 other Enterobacteriaceae and Pseudomonadaceae strains were tested. Two strains of the non-Serratia Enterobacteriaceae (one each of Citrobacter freundii and Proteus morganii) and two strains of Pseudomonas aeruginosa produced a color change in the medium. All of the Serratia strains tested were correctly identified using this medium, while 96% of the other species tested were inhibited.