Relative oral bioavailability of glycidol from glycidyl fatty acid esters in rats.

In order to quantify the relative bioavailability of glycidol from glycidyl fatty acid esters in vivo, glycidyl palmitoyl ester and glycidol were orally applied to rats in equimolar doses. The time courses of the amounts of glycidol binding to hemoglobin as well as the excretion of 2,3-dihydroxypropyl mercapturic acids were determined. The results indicate that glycidol is released from the glycidyl ester by hydrolysis and rapidly distributed in the organism. In relation to glycidol, there was only a small timely delay in the binding to hemoglobin for the glycidol moiety released from the ester which may be certainly attributed to enzymatic hydrolysis. In both cases, however, an analogous plateau was observed representing similar amounts of hemoglobin binding. With regard to the urinary excretion of mercapturic acids, also similar amounts of dihydroxypropyl mercapturic acids could be detected. In an ADME test using a virtual double tag (³H, ¹4C) of glycidyl palmitoyl ester, a diverging isotope distribution was detected. The kinetics of the ¹4C-activity reflected the kinetics of free glycidol released after hydrolysis of the palmitoyl ester. In view of this experimental data obtained in rats, it is at present justified for the purpose of risk assessment to assume complete hydrolysis of the glycidyl ester in the gastrointestinal tract. Therefore, assessment of human exposure to glycidyl fatty acid ester should be regarded as an exposure to the same molar quantity of glycidol.

Relative oral bioavailability of 3-MCPD from 3-MCPD fatty acid esters in rats.

In order to quantify the relative oral bioavailability of 3-chloropropane-1,2-diol (3-MCPD) from 3-MCPD fatty acid diesters in vivo, 1,2-dipalmitoyl-3-chloropropane-1,2-diol (3-MCPD diester) and 3-MCPD were orally applied to rats in equimolar doses. In both cases, the time courses of 3-MCPD concentrations were measured in blood, various organs, tissues and intestinal luminal contents. The results show that 3-MCPD is released by enzymatic hydrolysis from the 3-MCPD diester in the gastrointestinal tract and distributed to blood, organs and tissues. Based on the measurements in blood, the areas under the curve (AUC) for 3-MCPD were calculated. By comparing both AUC, the relative amount of 3-MCPD bioavailable from the 3-MCPD diester was calculated to be 86 % on average of the amount bioavailable following administration of 3-MCPD. In view of limited experimental data, it is justified for the purpose of risk assessment to assume complete hydrolysis of the diesters in the gastro-intestinal tract. Therefore, assessment of the extent of exposure to 3-MCPD released from its fatty acid esters should be performed in the same way as exposure to the same molar quantity of 3-MCPD.

Unusual polar metabolites in the groundwater of a contaminated waste site indicate a new pathway of mononitrotoluene transformation.

At a mononitrotoluene-contaminated waste disposal site, the groundwater was screened for polar transformation products of mononitrotoluenes, by means of HPLC-MS, HPLC-NMR and further off-line NMR and MS techniques. Besides expected metabolites such as aminotoluenes (ATs) and nitrobenzoic acids (NBAs), three unknowns (di- and tetrahydro-derivatives of (2-oxo-quinolin-3-yl) acetic acid) could be identified which, in the context of explosives and related compounds, are new metabolites. Evidence could be provided by microcosm experiments with 2-nitrotoluene (2-NT) that these metabolites are microbial transformation products of 2-NT under anaerobic conditions. The NMR and MS data are presented and the possible pathway for the formation of these metabolites after addition of 2-NT to fumarate is discussed.

Development and validation of an analytical method for determination of 3-chloropropane-1,2-diol in rat blood and urine by gas chromatography-mass spectrometry in negative chemical ionization mode.

We have developed a highly selective and sensitive method using gas chromatography-mass spectrometry with negative chemical ionization for measuring 3-chloropropane-1,2-diol (3-MCPD) in rat blood and urine. Samples were adsorbed on silica gel, extracted with ethyl acetate, and derivatized by chemical derivatization with heptafluorobutyric acid anhydride. For quantification, matrix-based calibration curves and 3-MCPD-d (5), as an isotope-labeled internal standard, were used. The relative recoveries of 3-MCPD were between 80 and 110% in most cases and the relative standard deviations were typically less than 10%, with some exceptions. The limit of quantification of the method was found to be about 2 ng/mL. In conclusion, a valuable, robust, and sensitive method for detection of 3-MCPD is now available for biokinetics studies.

Assessment of pulmonary function and serum substance levels in newborn and juvenile rats.

In the context of pharmaceutical development today, studies for pediatric drug approval are requested more and more often by the regulatory authorities. The developing lung represents a potential target in juvenile toxicity studies. Due to physiological differences in prenatal and postnatal development between humans and standard animal models, experimental methods have to be modified to assess pulmonary function, and basic data on respiratory parameters need to be provided. Daily nose-only inhalation exposure from postnatal days 4 to 21 using a model substance (verapamil HCl) and plethysmographic measurements between postnatal days 2 and 50 were performed noninvasively in conscious juvenile Wistar (WU) rats. The methods proved to be feasible and did not interfere with normal growth and development of the animals. Both techniques therefore permit new insights to support human neonatal risk assessment and therefore these animal models are suitable for regulatory studies.

Use of biocidal products (insect sprays and electro-vaporizer) in indoor areas--exposure scenarios and exposure modeling.

Five commercially available insect sprays were applied in a model room. Spraying was performed in accordance with the manufacturers' instructions and in an overdosed manner in order to simulate worst-case conditions or an unforeseeable misuse. In addition, we examined electro-vaporizers. The Respicon aerosol monitoring system was applied to determine inhalation exposure. During normal spraying (10 seconds) and during the following 2-3 minutes, exposure concentrations ranged from 70 to 590 microg/m3 for the pyrethroids tetramethrin, d-phenothrin, cyfluthrin, bioallethrin, and the pyrethrins. Calculated inhalable doses were 2-16 microg. A concentration of approximately 850 microg chlorpyrifos/m(3) (inhalable dose: approximately 20 microg) was determined when the "Contra insect fly spray" was applied. Highest exposure concentrations (1100-2100 microg/m3) were measured for piperonyl butoxide (PBO), corresponding to an inhalation intake of 30-60microg. When simulating worst-case conditions, exposure concentrations of 200-3400microg/m3 and inhalable doses of 10-210microg were determined for the various active substances. Highest concentrations (4800-8000 microg/m3) were measured for PBO (inhalable: 290-480 microg). By applying the electro-vaporizer "Nexa Lotte" plug-in mosquito killer concentrations for d-allethrin were in the range of 5-12microg/m3 and 0.5-2 microg/m3 for PBO while with the "Paral" plug-in mosquito killer concentrations of 0.4-5microg/m3 for pyrethrins and 1-7 microg/m3 for PBO were measured. Potential dermal exposures were determined using exposure pads. Between 80 and 1000microg active substance (tetramethrin, phenothrin, cyfluthrin, bioallethrin, pyrethrins, chlorpyrifos) were deposited on the clothing of the total body surface area of the spray user. Highest levels (up to 3000 microg) were determined for PBO. Worst-case uses of the sprays led to 5-9 times higher concentrations. Also a 2-hour stay nearby an operating electro-vaporizer led to a contamination of the clothing (total amounts on the whole body were 450 microg d-allethrin and 50 microg PBO for "Nexa Lotte" plug-in mosquito killer and 80 microg pyrethrins and 190 microg PBO for "Paral" plug-in mosquito killer). Human biomonitoring data revealed urine concentrations of the metabolite (E)-trans-chrysanthemum dicarboxylic acid ((E)-trans-CDCA) between 1.7 microg/l and 7.1 microg/l after 5 minutes of exposure to the different sprays. Also the use of electro-vaporizers led to (E)-trans-CDCA concentrations in the urine in the range of 1.0 microg/l to 6.2 microg/l (1-3 hours exposure period). The exposure data presented can be used for performing human risk assessment when these biocidal products were applied indoors. The airborne concentrations of the non-volatile active chemical compounds could be predicted from first principles using a deterministic exposure model (SprayExpo).

Identification of highly polar nitroaromatic compounds in leachate and ground water samples from a TNT-contaminated waste site by LC-MS, LC-NMR, and off-line NMR and MS investigations.

Leachate and ground water samples from a trinitrotoluene-contaminated waste disposal site near a former ammunitions plant in Stadtallendorf, Germany, were analyzed by liquid chromatography (LC)-mass spectrometry (MS) and LC-NMR hyphenated techniques to comprehensively characterize the range of highly polar nitroaromatic compounds. Wherever unknown components could not be identified by comparison with a multistandard, the spectroscopic data obtained on-line were used to make initial structure proposals, which were later confirmed by comparison with authentic reference materials. In those cases where reference materials were not commercially available, unknown compounds were isolated by HPLC cuts and their structures were elucidated by off-line NMR and MS investigations. A variety of previously unknown compounds, including nitrophenols, nitrobenzyl alcohols, methylnitrobenzoic acids, and hydroxynitrobenzoic acids, could be identified. The NMR and MS data are presented here. The main polar compounds were additionally quantified.

Cross validation of multiple methods for measuring pyrethroid and pyrethrum insecticide metabolites in human urine.

The objective of our study was to compare three vastly different analytical methods for measuring urinary metabolites of pyrethroid and pyrethrum insecticides to determine whether they could produce comparable data and to determine if similar analytical characteristics of the methods could be obtained by a secondary laboratory. This study was conducted as a part of a series of validation studies undertaken by the German Research Foundation's Committee on the Standardization of Analytical Methods for Occupational and Environmental Medicine. We compared methods using different sample preparation methods (liquid-liquid extraction and solid-phase extraction with and without chemical derivatization) and different analytical detection methods (gas chromatography-mass spectrometry (single quadrupole), gas chromatography-high resolution mass spectrometry (magnetic sector) in both electron impact ionization and negative chemical ionization modes, and high-performance liquid chromatography-tandem mass spectrometry (triple quadrupole) with electrospray ionization). Our cross validation proved that similar analytical characteristics could be obtained with any combination of sample preparation/analytical detection method and that all methods produced comparable analytical results on unknown urine samples.

Aircraft disinsection: exposure assessment and evaluation of a new pre-embarkation method.

A new "pre-embarkation" method for aircraft disinsection was investigated using two different 2% d-phenothrin containing aerosols. Five experiments in aircrafts of the type Airbus 310 (4x) and Boeing 747-400 (1x) were performed. In the absence of passengers and crew the d-phenothrin aerosol was sprayed under the seat rows and in a second step at the height of approximately 1.60 m by moving from one end of the cabin to the other. Concentration levels of d-phenothrin were determined at different time periods after application of the aerosol spray. In a B 747-400 with the air conditioning system operating the concentrations ranged between 853 and 1753 microg/m3 during and till 5 min after the beginning of spraying at different locations in the cabin. Within 5-20min after the end of the spraying concentrations of 36-205 microg/m3 and 20-40 min thereafter only ca. 1 microg d-phenothrin/m3 were detectable (average values in relation to each period of measurement). On cabin interior surfaces the median values for mainly horizontal areas ranged from 100 to 1160 ng d-phenothrin/cm2. d-Phenothrin concentrations in the air were sufficient to kill flying insects like house flies and mosquitoes within 20 min. Horizontal surfaces were 100% effective against insects up to 24 h after spraying. Doses inhaled by sprayers determined by personal measurements were calculated to be 30-235 microg d-phenothrin per 100 g spray applied (30% in the respirable fraction for Arrow Aircraft Disinsectant; 10% for Aircraft Disinsectant Denka). If passengers will board, e.g., 20 min after the end of the disinsection operation, inhalation exposure is estimated to be practically negligible. Also possible dermal exposure from residues in seats and headrests is very low for passengers during the flight. Therefore any health effects for passengers and crew members are very unlikely.

Inhalational and dermal exposures during spray application of biocides.

Data on inhalational and potential dermal exposures during spray application of liquid biocidal products were generated. On the one hand, model experiments with different spraying devices using fluorescent tracers were carried out to investigate the influence of parameters relevant to the exposure (e.g. spraying equipment, nozzle size, direction of application). On the other hand, measurements were performed at selected workplaces (during disinfection operations in food and feed areas; pest control operations for private, public and veterinary hygiene; wood protection and antifouling applications) after application of biocidal products such as Empire 20, Responsar SC, Omexan-forte, Actellic, Perma-forte; Fendona SC, Pyrethrum mist; CBM 8, Aldekol Des 03, TAD CID, Basileum, Basilit. The measurements taken in the model rooms demonstrated dependence of the inhalation exposure on the type of spraying device used, in the following order: "spraying with low pressure" < "airless spraying" < "fogging" indicating that the particle diameter of the released spray droplets is the most important parameter. In addition inhalation exposure was lowest when the spraying direction was downward. Also for the potential dermal exposure, the spraying direction was of particular importance: overhead spraying caused the highest contamination of body surfaces. The data of inhalational and potential dermal exposures gained through workplace measurements showed considerable variation. During spraying procedures with low-pressure equipments, dose rates of active substances inhaled by the operators ranged from 7 to 230 microg active substance (a.s.)/h. An increase in inhaled dose rates (6-33 mg a.s./h) was observed after use of high application volumes/time unit during wood protection applications indoors. Spraying in the veterinary sector using medium-pressure sprayers led to inhaled dose rates between 2 and 24mga.s./h. The highest inhaled dose rates were measured during fogging (114 mg a.s./h) and after-high-pressure applications in the antifouling sector (110-300 mg a.s./h). The potential dermal exposure of spray operators was lowest (dose rates from 0.2 to 7 mg a.s./h) in the areas of food and feed disinfection and private and public hygiene during spraying with low-pressure devices. During fogging, wood protection and antifouling applications, high-potential dermal exposures of the operators were determined. Dermal dose rates varied between 100 and 34,000 mg a.s./h.

Pyrethroids used indoor-ambient monitoring of pyrethroids following a pest control operation.

House dust and airborne particles (PM) were sampled before (T1) and 1 day (T2), 4-6 months (T3) as well as 10-12 months (T4) after a pest control operation (PCO). Cyfluthrin was applied in 11, cypermethrin in 1, deltamethrin in three and permethrin in four interiors. The pyrethroid concentrations in house dust and PM were measured by GC/MS with a detection limit for all pyrethroids of 0.5 mg/kg house dust and of 1 ng/m3 PM for deltamethrin and permethrin and 3 ng/m3 PM for cyfluthrin and cypermethrin. A general background concentration of permethrin (95th percentile: 5.9 mg/kg) and cyfluthrin (95th percentile: 34.9 mg/kg) in house dust was found. In general, an appropriately performed PCO lead to an increase of pyrethroids in house dust as well as in PM, in some cases up to 1 year after application. One day after the application the cyfluthrin concentration increased significantly from 0.25 (T1) to 33.8 mg/kg house dust (T2) and up to 4.9 ng/m3 in PM. The permethrin concentration increased significantly from 4.3 to 70 mg/kg in house dust and up to 18.1 ng/m3 in PM, deltamethrin increased to 54.5 mg/kg and 20.8 ng/m3 and cypermethrin to 14 mg/kg and 45.7 ng/m3. Thereafter a continuous decrease could be observed during the time course of 1 year. After 1 year the permethrin concentration in house dust was still 1/5 of the T2 concentration, whereas for cypermethrin and cyfluthrin only 1/14 and 1/23 of the T2 concentration were found. Deltamethrin was not detected at all after T2. Moreover, the data of this study showed significant, positive correlations between pyrethroids in house dust and in airborne particles especially one day after PCO.

In-flight spraying in aircrafts: determination of the exposure scenario.

Exposure measurements were carried out in parked aircrafts during and after application of a biocide aerosol spray (simulated in-flight spraying). The aerosol product SRA spray (Standard Reference Aerosol Spray) was used for spraying. Concentrations of the pyrethrins--the active ingredients--in the air of the passenger cabin (airborne particles, measured during spray application and 40 minutes afterwards) varied from 11 to 65 microg/m3; those of the synergist piperonyl butoxide were 200-485 microg/m3. The concentrations on surfaces of the cabin furniture differed widely. Low concentrations were determined on surfaces in vertical positions (median values: pyrethrins < or =2 ng/cm2; piperonyl butoxide < or =17 ng/cm2), while under seats, on seats and on headrests the concentrations were up to 55.5 ng/cm2 for pyrethrins and 1162.5 ng/cm2 for piperonyl butoxide (median values). The inhaled doses for sprayers (using 100 g of spray) and persons sitting in the passenger cabin were calculated to be 17 microg for pyrethrins and 200 microg for piperonyl butoxide (maximum values). Maximum total external body doses for the applicators during spraying were 830 microg for pyrethrins and 8840 microg for piperonyl butoxide. The potential dermal dose for persons sitting in the passenger cabin was about a factor of two lower.

Pyrethroids used indoors--biological monitoring of exposure to pyrethroids following an indoor pest control operation.

A prospective epidemiological study with respect to pyrethroid exposure was carried out combining clinical examination, indoor monitoring and biological monitoring. The results of the biological monitoring are presented. Biological monitoring was performed in 57 persons before (T1) as well as 1 day (T2), 3 days (T3), 4-6 months (T4), and 10-12 months (T5) following a pest control operation (PCO) with pyrethroid containing products such as cyfluthrin, cypermethrin, deltamethrin or permethrin. Pyrethroids in blood were measured by GC-ECD. The respective metabolities cis- and trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane carboxylic acid (DCCA), cis-3-(2,2-dibromovinyl)-2,2-dimethylcyclopropane carboxylic acid (DBCA), 3-phenoxybenzoic acid (3-PBA) and fluorophenoxybenzoic acid (FPBA) were measured in urine using GC/MS. For all cases the concentrations of pyrethroids in blood were found to be below the detection limit of 5 micrograms/l before and after the PCO. With a detection limit of 0.2 microgram/l of the investigated metabolites, the percentage of positive samples were 7% for cis-DCCA, 3.5% for trans-DCCA and 5.3% for 3-PBA before PCO. One day after PCO (T2) the percentage of positive samples increased remarkably for cis-DCCA (21.5%), trans-DCCA (32.1%) and 3-PBA (25%) showing significantly increased internal doses as compared to pre-existing values. This holds also true for T3, whereas at T4 and T5 the significant increase was no more present. FPBA and DBCA concentrations were below the respective detection limit before PCO and also in most cases after PCO. In 72% of the subjects the route of pyrethroid uptake (measured by determining the DCCA isomeric ratio) was oral/inhalative and in 28% it was dermal. Based on the biological monitoring data it could be shown that appropriately performed pest control operations lead to a significant increase of pyrethroid metabolite concentration in the early phase (1 and 3 days) after pyrethroid application as compared to the pre-exposure values. However, evaluated metabolite concentrations 4-6 months after PCO did not exceed values of published background levels.

Indoor pyrethroid exposure in homes with woollen textile floor coverings.

In order to investigate human's exposure to permethrin from treated woollen textile floor coverings and possible adverse health effects, a study was carried out in 80 private homes in Hannover (Germany) equipped with woollen textile floor coverings (wool wall-to-wall carpets or woven or knotted rugs). For indoor monitoring, permethrin was determined both in house dust and on suspended particles. While permethrin concentrations in house dust (< 2 mm) were high (arithmetic mean: 53.7 mg/kg, 90th percentile 129.1 mg/kg), the permethrin concentrations in the air (suspended particles) were very low (arithmetic mean 2.8 ng/m3, 90th percentile 5.8 ng/m3, first sampling). Additional experiments demonstrate that permethrin on suspended particles result from carpet fiber abrasion (and not from an evaporation/re-condensation process). The internal exposure of the 145 inhabitants participating in the study was determined by biological monitoring (permethrin metabolites in urine). In a first sampling period almost 14% of the samples showed concentrations of the metabolite DCCA and almost 23% of the metabolite 3-PBA above the limit of detection (0.2 microgram/l). A model was developed which allows the calculation of the metabolite concentration in urine due to inhalative uptake of permethrin. Even for the worst case situation the calculated metabolite concentrations were ca. 30 times lower than the experimental results. The observed concentrations of metabolites are comparable to those of the background concentrations of the general population in Germany, suggesting that they must origin from other sources than woollen textile floor coverings. The indoor and biological monitoring data as well as the evaluation of the reported symptoms give no indication of an adverse health effect due to carpet treatment by permethrin.