Development of a measurement method to determine the ceiling exposure concentration of ortho-phthalaldehyde handling workers.

OBJECTIVES: The purpose of this research was to develop and validate an analytical method for rapid determination of the exposure of workers to ortho-phthalaldehyde (OPA) at the ceiling threshold concentration. METHODS: A 2,4-dinitrophenylhydrazine (DNPH)-silica cartridge was chosen as a sampler. OPA collected by the DNPH-silica cartridge was subsequently extracted with 5 mL of acetonitrile. A 50-µL aliquot of phosphoric acid/acetonitrile solution (2%, v/v) was added to 950 µL of the extraction solution and allowed to stand for 30 minutes at room temperature. This solution was then analyzed by high-performance liquid chromatography tandem mass spectrometry. The basic characteristics of the proposed method, such as recovery, repeatability, limit of quantification, and storage stability of the samples, were examined. RESULTS: The overall recoveries of OPA from OPA-spiked DNPH-silica cartridges were 93.6%-100.1% with relative standard deviations, representing the repeatability, of 1.5%-10.8%. The limit of quantification was 0.165 ng/sample. The recovery of OPA from DNPH-silica cartridges after 5 days of storage in a refrigerator exceeded 95%. CONCLUSIONS: The proposed method enabled the determination of the OPA concentration corresponding to the Threshold Limit Value-Ceiling of 0.1 ppb recommended by the American Conference of Governmental Industrial Hygienists, with a minimum sampling time of 18 seconds (corresponding to a sampling volume of 300 mL at 25°C and 1 atm). Thus, this method will be useful for estimating worker exposures to OPA.

Concentration determination of urinary metabolites of N,N-dimethylacetamide by high-performance liquid chromatography-tandem mass spectrometry.

OBJECTIVES: N,N-Dimethylacetamide (DMAC) is widely used in industry as a solvent. It can be absorbed through human skin. Therefore, it is necessary to determine exposure to DMAC via biological monitoring. However, the precision of traditional gas chromatography (GC) is low due to the thermal decomposition of metabolites in the high-temperature GC injection port. To overcome this problem, we have developed a new method for the simultaneous separation and quantification of urinary DMAC metabolites using liquid chromatography-tandem mass spectrometry (LC-MS/MS). METHODS: Urine samples were diluted 10-fold in formic acid, and 1-µl aliquots were injected into the LC-MS/MS equipment. A C18 reverse-phase Octa Decyl Silyl (ODS) column was used as the analytical column, and the mobile phase consisted of a mixture of methanol and aqueous formic acid solution. RESULTS: Urinary concentrations of DMAC and its known metabolites (N-hydroxymethyl-N-methylacetamide (DMAC-OH), N-methylacetamide (NMAC), and S- (acetamidomethyl) mercapturic acid (AMMA) ) were determined in a single run. The dynamic ranges of the calibration curves were 0.05-5 mg/l (r≥0.999) for all four compounds. The limits of detection for DMAC, DMAC-OH, NMAC, and AMMA in urine were 0.04, 0.02, 0.05, and 0.02 mg/l, respectively. Within-run accuracies were 96.5%-109.6% with relative standard deviations of precision being 3.43%-10.31%. CONCLUSIONS: The results demonstrated that the proposed method could successfully quantify low concentrations of DMAC and its metabolites with high precision. Hence, this method is useful for evaluating DMAC exposure. In addition, this method can be used to examine metabolite behaviors in human bodies after exposure and to select appropriate biomarkers.

Decomposition Characteristics of Toluene Vapor Using Titanium Dioxide Photocatalyst and Zeolite Thermally Sprayed on an Aluminum Fiber Filter.

Decomposition characteristics of toluene vapor by titanium dioxide photocatalyst and zeolite that are prepared by thermal spraying on an aluminum fiber filter (photocatalyst filter) were investigated. Toluene vapor was injected into a small chamber made of stainless steel, and an air cleaner equipped with the photocatalyst filter was operated. The vapor concentration in the chamber decreased exponentially. The decreasing rate of toluene vapor in the chamber depended on the initial toluene concentration, and the higher the initial vapor concentration was, the lower the decreasing rate was obtained. The decreasing rate was constant during each decomposition experiment, although the concentration decreased with time. To investigate the effect of zeolite on the reduction of the vapor concentration, we compared the decreasing rates of toluene vapor by photocatalyst filters with and without zeolite.The decreasing rate of toluene concentration using the filter without zeolite was larger than that with zeolite. The reason for this would be that photocatalyst decomposed toluene not only in air but also adsorbed in zeolite.

Thermal reconditioning characteristics of a respirator cartridge for organic vapors using humid air as the desorption gas.

OBJECTIVES: Thermal reconditioning characteristics of organic vapors from a respirator cartridge were studied by introducing humid air into a cartridge that had adsorbed organic vapors in order to develop a thermal reconditioning method. METHODS: Five different organic vapors (methanol, 2-propanol, acetone, dichloromethane and methyl acetate), most of which have relatively weak adsorption affinity to charcoal, were used in this study. Adsorption was carried out at a temperature of 25 degrees C. The relative humidity of the adsorption air with organic vapor was 50%. When the vapor concentration in the downstream of the respirator exceeded the breakthrough concentration, that is, the occupational exposure limits in Japan, the vapor supply was stopped. Then, desorption was started by introducing clean humid air from opposite side of the cartridge under a heated condition. When the desorbed vapor concentration fell below the limit of quantification, desorption process was ended and the next adsorption cycle was started after the temperature had returned to room temperature. This adsorption - desorption cycle was repeated more than three times. The desorption temperature was 65 degrees C and the relative humidity of desorption air was 20%, 50% or 70% at 25 degrees C. RESULTS: When the relative humidity was 20%, the breakthrough times of regenerated cartridges were shorter than that of a new one, but no difference was observed in the breakthrough curves when the relative humidity was greater than 50%. CONCLUSION: The results suggest that the thermal reconditioning of respirator cartridges using humid air is possible for these vapors.

Biological effect of carbon graphite whisker in rat lung by long-term Inhalation.

Carbon graphite whisker (CGW) was used in a 1-year inhalation study in male Wistar rats and its biological effect was observed until the 1-year clearance period. The inhalation study was conducted at 2.6 +/- 0.5 mg/m(3) (equivalent to 44.5 +/- 15.0 fibers/mL) for 6 hours a day, 5 days a week for 1 year. There were no differences in survival rate between the exposure and control groups during this examination; however, the body weights were significantly different at the end of the 1-year clearance. The lung weight at 3 days and 1 year after the end of exposure was not significantly different in both groups. The deposited amount of CGW was 6.83 +/- 0.75 mg at 3 days post-exposure; the deposition rate was 17.6%. Only around 30% of the total deposited CGW was cleared during the 1-year clearance period. The geometric means of CGW in the lung, i.e. CMD (count median diameter) and CML (count median length), hardly changed, and the clearance was delayed. In the histopathological examination, there was mild fibrosis in all exposed rats irrespective of the clearance period. One adenoma was observed in a single animal at 3 days post-exposure, while no adenomas were observed in the exposure group after the 1-year clearance. Epithelial hyperplasia was found in some animals.

Effects of 1-bromopropane, a substitute for chlorofluorocarbons, on BDNF expression.

1-Bromopropane (1-BP) has been widely used as an alternative to ozone-depleting chlorofluorocarbons in various industries. Although the neurotoxicity of 1-BP has been recently reported, there is little information about the effect of 1-BP on the cells in brain by experimental approach. Here we studied the effect of 1-BP on brain-derived neurotrophic factor (BDNF) expression in astrocytes in vitro. The BDNF mRNA level was remarkably decreased by 1-BP in a human astrocytoma cell line, U251, and in mouse primary astrocytes. The DNA-binding and specific reporter activity of cAMP response element-binding transcription factor (CREB), which is one of the key molecules regulating BDNF expression, were reduced by 1-BP in U251 and/or mouse primary astrocytes. Additionally, protein kinase A (PKA) activity was suppressed by 1-BP in U251. These results suggest that BDNF expression was affected by 1-BP through at least PKA.

Hypothalamo-pituitary-adrenal gland axis in mice inhaling toluene prior to low-level long-term exposure to formaldehyde.

We studied the change in the hypothalamo-pituitary-adrenal gland (HPA) axis upon adding prior toluene inhalation to our previous formaldehyde inhalation experiments to determine whether short term exposure to relatively high levels of toluene triggers multiple chemical sensitivity (MCS). Data come from immunocytochemical, morphometrical and RT-PCR measurements. Four groups of adult female mice were exposed to differing concentrations (0, 80, 400, and 2,000 ppb) of formaldehyde for 16 hr/day, 5 days/week for twelve weeks, after the mice were exposed intranasally to 500 ppm toluene per mouse for 6 hr/day, for 3 days. We found that the number of corticotropin releasing hormone (CRH)-immunoreactive (ir) neurons was up-regulated according to the amount of formaldehyde as well as inhalation of formaldehyde alone in our previous experiment. The proportion of adrenocorticotropin hormone (ACTH)-ir cells increased according to the formaldehyde concentration, though there was no significant difference between the 400 and 2,000 groups. The number of ACTH-ir cells was higher in the 400 group than in the other groups (0, 80, and 2,000). Expression of ACTH-mRNA was also up-regulated according to the quantity of formaldehyde. The sinusoid in the anterior pituitary showed more dilatation in the 400 and 2,000 groups than in the control group, especially in the 2,000 group. We propose that exposure to toluene prior to inhalation of formaldehyde has no effect on the HPA axis and as a trigger of MCS, although greater sinusoid dilatation was found in the anterior pituitary gland at higher concentrations of formaldehyde.

Inhalation of low-level formaldehyde enhances nerve growth factor production in the hippocampus of mice.

OBJECTIVE: The effects of low-level formaldehyde (FA) inhalation on the amount of nerve growth factor (NGF) in the hippocampus of immunized mice were studied. METHODS: Evaluation of NGF in the hippocampus was performed by ELISA and RT-PCR. RESULTS: Exposure to 80 and 400 ppb FA significantly increased the brain NGF levels in the immunized mice. Evaluation of the NGF levels in the hippocampus of immunized mice showed that 400 ppb FA significantly increased the NGF content. The RT-PCR evaluation also showed higher concentrations of hippocampal NGF mRNA in the mice exposed to 80 and 400 ppb FA with immunization. CONCLUSION: Exposure of immunized mice to low levels of FA significantly increases NGF levels in the hippocampus.

Effect of prolonged exposure to low concentrations of formaldehyde on the corticotropin releasing hormone neurons in the hypothalamus and adrenocorticotropic hormone cells in the pituitary gland in female mice.

We examine the effect on the hypothalamus-pituitary-adrenal gland (HPA) axis of prolonged exposure to low levels of formaldehyde in female C3H/He mice, using immunocytochemical and RT-PCR methods. Two groups of female mice were exposed to differing concentrations (0, 80, 400, 2000 ppb) of formaldehyde inhalation for 16 h/day, 5 days/week, for 12 weeks. The corticotropin releasing hormone (CRH)-immunoreactive (ir) neurons in the hypothalamus were then examined, together with the adrenocorticotropin hormone (ACTH)-ir cells and ACTH mRNA in the pituitary. One group comprised sham control mice. The other group was made allergic by injection of ovalbumin (OVA) and alum prior to exposure to formaldehyde, since most sick building syndrome (SBS) sufferers are women with allergic disease. These animals were further exposed to aerosolized OVA as a booster four times during the exposure period. Our results showed a dose-dependent increase in the number of CRH-ir neurons in the non-allergy (NAG) group. A similar pattern was found in ACTH-ir cells and ACTH mRNA. The allergy (AG) model group showed an increase in basal levels of all markers of HPA activity. Moreover, the AG mice appeared to respond to the lowest concentration of formaldehyde, and all indices of HPA activity were reduced at the highest concentrations of formaldehyde. These results relate to an important clinical issue and also have implications in the broader area of HPA regulation. We conclude that our experimental system may be a suitable animal model for SBS and/or multiple chemical sensitivity (MCS).

Pathology and mechanism of lung toxicity following inhalation of hair spray in rats.

In order to elucidate the pathology and mechanism of lung toxicity induced by chronic hair spray inhalation, male Wister rats 9 wk of age were exposed to a uniform concentration of hair spray for up to 12 wk using a jet nebulizer. The aerosol concentration to which the rats were exposed was about 7 g/m(3). Differential cell counts in bronchoalveolar lavage fluid (BALF), the expression of several cytokines from alveolar macrophage using the reverse-transcription polymerase chain reaction (RT-PCR) method, and histopathologic evaluation using a computer-aided graphic analyzer (IBAS) were conducted 1, 4, 8, and 12 wk after exposure. Over the passage of time, neutrophils and macrophages increased in BALF, and neutrophils infiltrated in the lung interstitium from the peribronchial interstitium to the alveolar septum. Alveolar macrophages showed increased expression of both the mRNA of tumor necrosis factor (TNF)-alpha and the mRNA of the chemokines macrophage inflammatory protein 2 (MIP2) and cytokine-induced neutrophil attractant (CINC). From these findings, chronic inhalation of hair spray is considered to induce at first intra-alveolar accumulation and activation of alveolar macrophages, followed by recruitment of neutrophils in the lung through the expression of proinflammatory cytokine, CINC, and MIP2, which cause predominantly neutrophilic inflammation in the lung.

[Method for installing an effective smoking room and the effectiveness of real-time monitoring].

We investigated the methodology for installing effective smoking rooms in workplaces. It is absolutely necessary to install exhaust ventilation in smoking rooms. There are two bases for deciding the exhaust ventilation rate. The most important is to eliminate the leakage of environmental tobacco smoke (ETS) from the smoking room. An airflow rate of more than 0.2 m/s at the opening of the smoking room is required by the Guidelines for Smoking Control in Workplaces (Ministry of Labour, Welfare and Health) to eliminate the leakage. This ventilation rate is decided by multiplying the opening area by 0.2 m/s. The second important point is to keep the concentration of ETS in the smoking room less than control concentration (0.15 mg/m3). This ventilation rate is decided by dividing the rate of generation of ETS by the control concentration. It is confirmed that an effective smoking room can be installed by following these guidelines. We used real-time monitoring to evaluate the leakage of ETS from the smoking room and the ETS concentration in the smoking room before and after the improvement. It is concluded that real-time monitoring of ETS is a useful method for evaluating the effectiveness of the smoking room.

Pulmonary effects and clearance after long-term inhalation of potassium octatitanate whiskers in rats.

The pulmonary effects of long-term inhalation of potassium octatitanate whisker (PT1), one of the durable man-made fibers (MMFs), were examined in rats. Male Wistar rats were exposed to PT1 by inhalation for 6 h/day, 5 days/wk for 1 yr. The daily average exposure concentration of PT1 aerosol was 2.2 +/- 0.7 mg/m3 (111 +/- 34 fiber/ml) during the exposure. Rats were sacrificed at 3 days, 6 mo, and 12 mo after 1 yr of inhalation exposure. The amount of deposited PT1 in rat lungs (lung burden) was 2.4 +/- 0.7 mg and the deposition fraction was 7.2% at 3 days after 1 yr. The clearance of inhaled PT1 after 1-yr inhalation was prolonged so that the biological half-life time (BHT) was difficult to estimate. The histopathological findings showed that mild fibrotic changes were observed around the macrophages that had engulfed the PT1 in the 3-day, 6-mo, and 12-mo rat sacrifice groups. As for pulmonary tumors, no malignant tumors were observed, although 2 adenomas at 6 mo and 1 adenoma and 1 squamous metaplasia at 12 mo after the exposure were found in the rat lungs.

c-jun mRNA expression and profilin mRNA amplification in rat alveolar macrophages exposed to volcanic ash and sulfur dioxide.

Local residents exposed to heavy falls of ash discharged by Mt. Sakurajima, an active volcano, have been reported to develop acute and chronic inflammation of the respiratory tract. The present study aimed to determine the primary cause of this inflammation using an experimental model. Wistar rats were exposed for 5 days (4 h/d) to air containing 100 mg/m3 volcanic ash (mass median aerodynamic diameter, 4.3 microm; geometric standard deviation, 1.7) with or without 1.5 ppm sulfur dioxide (SO2). The lungs were then lavaged, and mRNA was extracted from alveolar macrophages and assessed by reverse transcription-polymerase chain reaction (RT-PCR). In the lavage fluid, no change in cellularity or increase in the content of tumor necrosis factor (TNF)-alpha was detected. However, at 1 h following exposure, 80% of macrophages were seen to have phagocytosed the volcanic ash. This percentage was unchanged at 24 h after exposure. Profilin mRNA content of the macrophages was elevated, and c-jun mRNA was expressed. Alveolar macrophages exposed to volcanic ash and SO2, therefore, are likely to have some inflammatory and fibrogenic potential.

[Measurement of emission rate of formaldehyde and volatile organic compounds(VOCs) from building materials by the small chamber method].

A small chamber for measuring the emission rate of formaldehyde gas and volatile organic compounds(VOCs) from building materials was developed, and formaldehyde and VOCs (toluene and xylene) concentrations emitted from particle boards(PB), medium density fiber-boards(MDF) and wall papers were measured. The emission rates measured by the chamber method were compared with the values by the desiccator method, several modified desiccator methods and the perforator method. The emission rate of formaldehyde increased with time and it became constant after about 20 hours. On the other hand, the emission rate of VOCs decreased with time. A good correlation was obtained on the emission rate of formaldehyde between the chamber method and the conventional desiccator method. The correlation coefficient between the perforator method and the chamber method was also high, but the correlation was low for surface treated materials.

[The effect of formaldehyde exposure on cytokine production in murine alveolar macrophages].

The vapor of formaldehyde has been reported to represent a potential health hazard, resulting in respiratory dysfunction. To determine the potential role of alterations in the alveolar macrophages induced by inhalation of formaldehyde, we studied the cytokine production of murine alveolar macrophages. Mice were exposed at 0, 2.5, 5, 10 ppm of formaldehyde for 16 hrs daily, respectively. Durations of inhalation were 7, 14, 21 and 28 days. Immediately after the formaldehyde exposure, murine alveolar macrophages were harvested from the mice. 18 hrs after LPS stimulation, the cytokines (IL-4, IL-10, IFN-gamma, TNF-alpha) were measured in the supernatant of cultured murine alveolar macrophages using the ELISA method. No levels of formaldehyde exposure changed these cytokine productions. Either our dose or duration of formaldehyde exposure might not have been enough to produce significant changes of cytokine production of murine alveolar macrophages. Further experiments with low but longer formaldehyde exposure may be needed to understand the effect of formaldehyde on cytokine production.

Development of a new respirator for organic vapors with a breakthrough detector using a semiconductor gas sensor.

A method for determining breakthrough of organic vapors in a respirator cartridge was developed. A thick film semiconductor gas sensor was used as a breakthrough detector. Air containing organic vapor was introduced into the cartridge, and an output signal from the sensor inserted in the downstream flow of the cartridge was recorded on an IC card. Simultaneously, the breakthrough curve was obtained by measuring the vapor concentration at downstream from the respirator cartridge with a gas chromatograph (GC) equipped with a flame ionization detector. When the breakthrough was almost completed, the data recorded in the card were transferred to a personal computer and the change in the output signal from the sensor was compared with the breakthrough curve obtained by the GC. Twelve organic solvents including aromatic hydrocarbons, chlorinated hydrocarbons, acetates, alcohols, ketones, and aliphatic hydrocarbons were tested under low (20%-25%) and high (70%-80%) relative humidity ranges. The sensitivity of the sensor for chlorinated hydrocarbons such as 1,1,1-trichloroethane was relatively low, especially when the relative humidity was high, but the rise time of the sensor output signal was almost the same as or earlier than the breakthrough time by the GC. Based on the experimental results, a new respirator for organic vapors that can detect the end of service life was developed.

Personal exposure level and environmental ethylene oxide gas concentration in sterilization facilities of hospitals in Japan.

Personal and environmental (stationary) ethylene oxide (EO) gas concentrations in gas sterilization facilities were measured at six workplaces in five hospitals. An ethylene oxide gas monitor (3M #3551) was used for both personal and stationary samplings. A gas detector tube was also used for instantaneous sampling. In most workplaces, the personal exposure levels of EO were below the detection limit of the gas monitor. Most of the time-weighted average (TWA) concentrations by the stationary sampling were below the threshold limit value of EO (TLV-TWA = 1 ppm), but in one workplace, more than 4 ppm of EO were detected in front of the sterilizer in a clean room during a 24-hour measurement, although all the personal exposure levels were below the detection limit. Method of aeration after the sterilization was very important for reducing the EO exposure. The EO gas concentrations in two workplaces where sufficient aeration was carried out were below the detection limit in all the stationary samples. In one workplace where insufficient aeration was performed, EO was detected from 16 of 17 stationary samples, and more than 90-200 ppm of EO was determined by the gas detector tube near the worker's face at the moment when the door of the sterilizer was opened and the sterilized materials were removed.