Assessment of environmental tobacco smoke exposure in children with asthmatic symptoms by questionnaire and cotinine concentrations in plasma, saliva, and urine.

To validate a detailed questionnaire for assessment of environmental tobacco smoke (ETS) exposure by the biomarker cotinine in various media, a population-based study in the urban area of Malmö, Sweden was performed in children aged 8-13 years with and without asthmatic symptoms. There were strong correlations between urinary and saliva cotinine concentrations and also, though to a lesser extent, between these media and plasma. Even a detailed questionnaire gave only a rough picture of the ETS exposure, as indicated by the biomarkers. In a multivariate model, the most significant questionnaire-derived predictor of the cotinine levels was the maternal smoking habits; other questionnaire variables gave only a minimal explained variance. Children with a history of asthmatic symptoms had statistically significantly lower median cotinine levels in urine and saliva compared to referent children, most likely because of the antismoking information to their parents. This should be considered in epidemiological studies of ETS risks.

Long-term use of nicotine chewing gum and mercury exposure from dental amalgam fillings.

In experimental studies, chewing gum has been shown to increase the release rate of mercury vapor from dental amalgam fillings. The aim of the present study was to investigate the influence of long-term frequent chewing on mercury levels in plasma and urine. Mercury levels in plasma (P-Hg) and urine (U-Hg), and urinary cotinine were examined in 18 subjects who regularly used nicotine chewing gum, and in 19 referents. Age and number of amalgam surfaces were similar in the two groups. Total mercury concentrations in plasma and urine were determined by means of cold vapor atomic absorption spectrometry. Urinary cotinine was determined by gas chromatography-mass spectrometry. The chewers had been using 10 (median) pieces of gum per day for the past 27 (median) months. P-Hg and U-Hg levels were significantly higher in the chewers (27 nmol/L and 6.5 nmol/mmol creatinine) than in the referents (4.9 nmol/L and 1.2 nmol/mmol creatinine). In both groups, significant correlations were found between P-Hg or U-Hg on the one hand and the number of amalgam surfaces on the other. In the chewers, no correlations were found between P-Hg or U-Hg and chewing time per day or cotinine in urine. Cotinine in urine increased with the number of pieces of chewing gum used. The impact of excessive chewing on mercury levels was considerable.

Determination of toluenediamine isomers by capillary gas chromatography and chemical ionization mass spectrometry with special reference to the biological monitoring of 2,4- and 2,6-toluene diisocyanate.

The determination of 2,3-, 3,4-, 2,6-, 2,4- and 2,5-toluenediamine (TDA) in hydrolysed human urine and blood plasma was studied by GC-MS. The TDA isomers as their perfluoro-fatty acid anhydride derivatives were investigated. Chemical ionization with ammonia and isobutane as reagent gas and monitoring both positive and negative ions are studied. Negative ion monitoring using ammonia and the TDA pentafluoropropionic anhydride (PFPA) derivatives were chosen owing to the low detection limits and good separations of the isomers studied. The ions monitored were m/z 394 and 374 corresponding to the (M-20)- and (M-40)- ions and the m/z = 397 and 377 ions of the tri-deuterium-labelled TDA used as an internal standard. The performance of 2,4-, 2,5- and 2,6-TDA-PFPA in the ion source was studied by varying the ammonia pressure, temperature and electron energy. A 1-ml volume of human urine was added to 1.5 ml of 6 M HCl containing 0.5 micrograms/l of each of the trideuterated 2,6- and 2,4-TDA and the solution was hydrolysed at 100 degrees C overnight. TDA was extracted into 2 ml of toluene by the addition of 5 ml of saturated NaOH solution. Derivatization was performed in toluene by the addition of 10 microliters of PFPA. The excesses of the reagent and acid formed were removed by extraction with 1 M phosphate buffer solution (pH 7.5). Analyses of 2,6-, 2,4- and 2,5-TDA-spiked human urine (0.2-2.5 micrograms/l) were performed. The correlation coefficients were 0.999 (n = 6). The precision (R.S.D.) for human urine spiked at 1 micrograms/l was 1.6% for 2.6-TDA, 3,5% for 2,4-TDA and 3.2% for 2,5-TDA (n = 10). The detection limit, defined as twice the signal-to-noise ratio, was 1-5 fg injected, corresponding to less than 0.05 micrograms/l of TDA in human urine or plasma.

Analysis of the reactivity of [14C]toluene diisocyanate (TDI) in an isolated, perfused lung model.

An isolated, perfused, guinea pig lung model was used to investigate the molecular events which occur when a 14C-labeled TDI vapor reaches the airways. Exposure concentrations of 0.2 and 0.7 ppm were tested. Perfusate composition included: Krebs Ringer buffer only, as well as buffer containing either guinea pig serum albumin, human serum albumin, or diluted guinea pig plasma. Radioactivity was detected in the perfusate within minutes of exposure, and following a delay, increased linearly. The rate of uptake was dependent on TDI concentration and the composition of the perfusate. Biochemical characterization of the state of the 14C-labeled material in the perfusate was performed. The distribution between low and high molecular weight reaction products was determined by molecular sieve fractionation and varied as a function of perfusate composition but no variability was observed as a function of time during the 45 min of exposure. An increase in nucleophile concentration in the perfusate was associated with both a higher percentage of conjugated products (from 15% with buffer only to 45% with diluted guinea pig plasma) and an increase in the rate of TDX uptake (from 0.5 microns Eq/min with buffer alone to 0.1 micrograms Eq/min with diluted GPSA as perfusate at 0.7 ppm). GC-MS analysis of the samples for free TDA, before and after acid hydrolysis, showed that the low molecular weight product(s), which represented from 55-85% of the circulating radioactivity, was composed of hydrolyzable and non-hydrolyzable conjugates and metabolites with approximately 4% of the label associated with free TDA. Although the distribution between high and low molecular weight species varies, this result is analogous to the findings from in vivo studies and suggests that the isolated, perfused lung (IVPL) system may be a useful tool in investigating the molecular mechanisms of isocyanate-induced disease and metabolic activity of the lung.

Acute respiratory disorder, rhinoconjunctivitis and fever associated with the pyrolysis of polyurethane derived from diphenylmethane diisocyanate.

OBJECTIVES: A case is described of complex reactions associated with exposure to diphenylmethane diisocyanate (MDI), with some immunologic observations. METHODS: Medical history, clinical examinations, and analyses of immunologic parameters and the 4,4'-MDI-related amine 4,4'-diaminodiphenylmethane (MDA) in hydrolyzed serum and urine were used. RESULTS: The patient, a mechanic whose medical history suggested repeated attacks of a work-related pulmonary or systemic disease, was examined because of acute respiratory disorder, rhinoconjunctivitis, and a late systemic reaction after exposure to polyurethane pyrolysis products, including 4,4'-MDI (air level 15 micrograms.m3). Spirometry showed a partly reversible obstructive dysfunction, and a skin-prick test was positive versus isocyanates conjugated with human serum albumin (HSA). MDA was detected in hydrolyzed serum (5.6 ng.ml) and urine (1.6 micrograms.g creatinine-1). In serum, there were specific immunoglobulin (Ig) G (IgG1 and IgG4) and IgE antibodies to 4,4'-MDI-HSA and other isocyanates (phenylisocyanate, toluene diisocyanate, p-toluene monoisocyanate, hexamethylene diisocyanate) conjugated with HSA, a very high total IgE, a raised total IgG, and moderate neutrophilia and eosinophilia. The specific antibodies declined, but were still increased five years later. Furthermore, the values of circulating immune complexes were high. In vitro, the circulating immune complexes in serum increased after the addition of 4,4'-MDI-HSA. The patient had anti-Clq antibodies, which probably accounted for part of the circulating immune complexes. CONCLUSIONS: The reactions associated with MDI exposure (in combination with exposure to pyrolysis products) had features compatible with immediate hypersensitivity and with a complement-mediated immune-complex reaction.

Urinary cotinine in children and adults during and after semiexperimental exposure to environmental tobacco smoke.

Urinary cotinine (U-cotinine) as a biomarker of environmental tobacco smoke exposure was evaluated in 14 children (age 4-11 y) and in 7 adults who were exposed to environmental tobacco smoke at an air nicotine level of 110 mg/m3 for 2 h in a bus. Nicotine in air and U-cotinine were measured by gas chromatography/mass spectrometry before, during, and after the experiment. U-cotinine rose rapidly to a maximum after a median of 6 h following the end of exposure; remained at an apparent plateau for half a day; and then decreased exponentially, with a mean half-time of 19 h (95% confidence interval 18-20 h; no significant difference between children and adults). The maximum U-cotinine was higher in the children (mean = 22 mg/l) than in the adults (13 mg/l; p = .005); decreased with age among the children (r = -.74; p = .002); and increased as the estimated inhaled nicotine dose increased. Therefore, the findings of the present study showed that young children had higher U-cotinine than adults at the same experimental environmental tobacco smoke exposure, probably because they had a higher relative nicotine dose because of a higher relative ventilation rate, and possibly also because of metabolic differences; the elimination rate did not differ. The long half-time makes U-cotinine a good biomarker of environmental tobacco smoke exposure; the time of sampling is not very critical. Dilution-adjusted concentrations should be employed, and in children, preferably by density correction. A certain urinary cotinine level indicates a lower environmental tobacco smoke exposure in a small child than in an adult.

Exposure to environmental tobacco smoke in the household and urinary cotinine excretion, heavy metals retention, and lung function.

The relationship between urinary levels of cotinine (U-cotinine) and arsenic (U-As), blood levels of cadmium (B-Cd), blood levels of lead (B-Pb), lung function, and questionnaire data on smoking habits were studied in 107 parents and their 46 children (7-10 y of age). There was a statistically significant relationship between the reported amount of tobacco smoked and U-cotinine levels. Nonsmokers who were married to persons who smoked had three times higher U-cotinine levels than nonsmokers whose spouses did not smoke. There was a significant association between the number of parents who smoked in the family and the U-cotinine levels of children. If only one parent smoked, maternal smoking was of greater importance than paternal smoking. There was also an association between U-cotinine and B-Cd. A study of lung function in the children revealed that vital capacity and functional residual capacity (corrected for sex, age, and height) increased as the number of parents who smoked increased. Therefore, the present study showed that (1) U-cotinine was a useful index of active smoking and environmental tobacco smoke exposure in adults and children, (2) U-cotinine was associated with the blood concentration of cadmium, and (3) environmental tobacco smoke exposure was associated with changes in lung function of children.

Method for the biological monitoring of hexahydrophthalic anhydride by the determination of hexahydrophthalic acid in urine using gas chromatography and selected-ion monitoring.

A method for the determination of hexahydrophthalic acid, a metabolite of hexahydrophthalic anhydride, in human urine has been developed. The urine was worked-up by liquid-solid extraction, esterified with boron trifluoride-methanol, and analysed by capillary gas chromatography and selected-ion monitoring. Hexadeuterium-labelled hexahydrophthalic acid was used as the internal standard. The precision was 4% at 0.7 microgram/ml and 5% at 0.07 microgram/ml. The recovery of the acid for the overall method was 101% at 0.07 micrograms/ml of urine (with a coefficient of variation of 4%) and 95% at 0.7 microgram/ml (coefficient of variation 2%). The limit of detection was 20 ng/ml urine.

Determination of 4,4'-methylenedianiline in hydrolysed human urine using liquid chromatography with UV detection and peak identification by absorbance ratio.

A liquid chromatographic method using multi-wavelength UV detection (258 and 285 nm) is presented for the determination of 4,4'-methylenedianiline (MDA) in hydrolysed human urine. The method is based on hydrolysis under strongly acidic conditions followed by derivatization with pentafluoropropionic anhydride. The perfluoro fatty acid amide derivative formed was analysed on a bonded octadecylsilyl column using isocratic elution with acetonitrile-water (67:33, v/v) as mobile phase. The overall recovery for urine samples containing 115 micrograms/l of MDA was 97 +/- 3%. The calibration graph was linear in the investigated range (12-122 micrograms/l) with a correlation coefficient of 0.998. The precision was 2.3% for urine samples containing 122 micrograms/l and the detection limit was 8 micrograms/l. The chromatograms were evaluated using a combination of retention time data and absorbance ratio by the simultaneous monitoring of the wavelengths 285 and 258 nm. The absorbance ratio (285/258 nm) was virtually constant (0.28 +/- 0.04) in the range 78-10,000 micrograms/l. The precision for the absorbance ratio was 6.1% for urine samples containing 124 micrograms/l and the lowest amount of MDA to give an absorbance ratio was 50 micrograms/l. The procedure for the hydrolysis of urine spiked with MDA and N,N'-diacetyl-MDA and urine from skin-exposed workers was studied under strongly acidic, weakly acidic and basic conditions. MDA was found in hydrolysed urines from skin-exposed epoxy resin workers in the concentration range 8-700 micrograms/l.

Determination of 4,4'-methylenediphenyldianiline (MDA) and identification of isomers in technical-grade MDA in hydrolysed plasma and urine from workers exposed to methylene diphenyldiisocyanate by gas chromatography-mass spectrometry.

Gas chromatography-mass spectrometry using chemical ionization with ammonia as reagent gas monitoring both positive and negative ions was applied. Negative-ion monitoring using ammonia and the pentafluoropropionic anhydride (PFPA) derivatives were chosen owing to low detection limits and good separation for the isomers studied. Technical-grade methylenediphenyldiioscyanate (MDI) was analysed and three isomers, 4,4'-, 2.4'- and 2,2'-methylenediphenyldianiline (MDA), were determined in addition to methylated MDA. Plasma and urine from an exposed worker were hydrolysed and analysed and the MDA isomers were identified in the biological samples.

Trace analysis of airborne 1,6-hexamethylenediisocyanate and the related aminoisocyanate and diamine by glass capillary gas chromatography.

A capillary gas chromatographic method was developed for the analysis of complex air mixtures of 1,6-hexamethylenediisocyanate, 1,6-hexamethyleneaminoisocyanate and 1,6-hexamethylenediamine. The method is based on derivatization in the sampling step of the reactive isocyanate groups to corresponding urethane groups by the alkaline ethanolic solvent and a subsequent derivatization of remaining amino groups to amide groups with heptafluorobutyric acid anhydride. The overall procedure, including sampling, gave a linear response at air concentrations of 3-300 micrograms/m3 for 1,6-hexamethylenediisocyanate with a precision of ca. 4% at 15 micrograms/m3 and a detection limit of ca. 0.2 microgram/m3 using nitrogen selective detection. In a field measurement of air concentrations in welding work on lacquered metal parts at a motor-car workshop, concentrations of 1,6-hexamethylenediisocyanate above 600 micrograms/m3 were found. Also 1,6-hexamethyleneaminoisocyanate and 1,6-hexamethylenediamine were found at concentrations of the order of 15% of the 1,6-hexamethylenediisocyanate concentration.

Determination of cotinine in urine using glass capillary gas chromatography and selective detection, with special reference to the biological monitoring of passive smoking.

A capillary gas chromatographic (GC) method using selected-ion monitoring (SIM) was developed for the analysis of cotinine (C.A.S. No. 486-56-6) in human urine. The method is based on basic extraction of cotinine from 2 ml of urine into dichloromethane. After evaporation of the dichloromethane solution to dryness, 100 microliters of toluene were added, prior to GC-mass spectrometric (MS) analysis. Trideuterated cotinine (C.A.S. No. 97664-65-8) was used as the internal standard. More than 1000 automatic chromatographic analyses were made without column degradation. Molecular ions (M) of cotinine and trideuterated cotinine, (m/e = 176 and 179), were monitored in the electron impact (EI) mode and m/e = 177 (M + 1) and m/e = 180 (M + 1) in the chemical ionization (CI) mode with isobutane. The correlation coefficient with SIM and EI was 0.998 (5-20 ng/ml) and with CI was (0.2-2 ng/ml). For thermionic specific detection the correlation coefficient was 0.998 (10-510 ng/ml). Only capillary columns with an apolar bonded stationary phase film thickness of 1 micron showed sufficient inertness for cotinine analysis at the sub ng/ml level. The relative standard deviations for 5 and 20 ng/ml were 5.2 and 3.5% respectively (n = 12) using EI. Spiked urine samples from six non-smokers (5 ng/ml) showed a relative standard deviation of 5%. The overall recovery (25 ng/ml) was 100 +/- 4%. The minimum detectable concentration, using SIM, was ca. 2 ng/ml in the EI mode and ca. 0.2 ng/ml in the CI mode. The half-time for cotinine was ca. 18 h for both active smokers and non-smokers.

Determination of 4,4'-methylenedianiline in hydrolysed human urine by micro liquid chromatography with ultraviolet detection.

4,4'-Methylenedianiline was determined in human urine by micro liquid chromatography with ultraviolet detection. The combination of a thorough work-up and the high mass sensitivity of micro liquid chromatography gave the method very high sensitivity. Derivatization with pentafluoropropionic anhydride enhanced the resolution of the 4,4'-dimethylenedianiline peak. The detection limit, defined as blank plus three times the standard deviation of the blank, was 2 nmol/l of urine, for 10-microliters injection volumes. The detection limit, defined as three times the noise, was about ten times better. The within- and between-assay coefficients of variation were 4 and 6%, respectively, for samples containing 40 nmol/l. The method was applied for the monitoring of excreted 4,4'-methylenedianiline in urine, during epicutaneous skin hypersensitivity testing (patch testing).

Determination of methylmercury in human blood using capillary gas chromatography and selected-ion monitoring.

A capillary gas chromatographic method, using selected-ion monitoring in the electron-impact mode, was developed for the analysis of methylmercury (MeHg) in human blood. The samples, spiked with the internal standard propylmercury bromide (PropHgBr), were, after addition of sodium bromide and cupric sulfate, extracted with toluene. The organic phase was extracted with an ethanol-water solution of sodium thiosulfate. After addition of sodium bromide solution, the ethanol-water phase was extracted with toluene. Methylated derivatives (MeHgCH2Br and PropHgCH2Br) were formed by the addition of a diethyl ether solution of diazomethane. The chromatographic properties of the derivatives were much better than those of the non-methylated compounds. The m/z 215 fragment of MeHgCH2Br and the molecular ion m/z 338 of PropHgCH2Br were monitored. The calibration graphs, with a linear correlation coefficient of 0.992 (n = 12) in the 1-5 micrograms/l concentration range, passed through the origin. The detection limit for MeHg in human blood was ca. 0.5 microgram/l. Analysis of spiked blood samples at concentrations of about 2 and 10 micrograms/l gave a relative standard deviation of 4.2 and 5.5%, respectively (n = 10).

Determination of piperazine in working atmosphere and in human urine using derivatization and capillary gas chromatography with nitrogen- and mass-selective detection.

A reliable routine method is presented for the determination of piperazine down to the sub-ppm level in aqueous solutions and in urine. The method includes a two-phase derivatization procedure with ethyl- or isobutyl chloroformate as the reagent, followed by a capillary gas chromatographic determination using nitrogen- or mass selective detection. The addition of ammonia ensured a quantitative recovery. Detection limits for piperazine in urine were ca. 20 ng/ml using nitrogen-selective and ca. 1 ng/ml with mass-selective detection. The calibration plots were linear in the investigated range, 100-10,000 ng/ml with nitrogen-selective and 30-3000 ng/ml with mass-selective detection. The precision was ca. 6% at a concentration of 300 ng/ml. Acid anhydrides were investigated as alternative reagents in the two-phase derivatization procedure, and heptafluorobutyric acid anhydride in aqueous solutions gave approximately 100% recovery. However, in urine the recoveries of the investigated acid anhydride derivatives were unsatisfactory.

Biological monitoring of hexamethylene diisocyanate by determination of 1,6-hexamethylene diamine as the trifluoroethyl chloroformate derivative using capillary gas chromatography with thermoionic and selective-ion monitoring.

A GC method using a novel derivatization reagent, 2',2',2-trifluoroethyl chloroformate (TFECF), for the derivatization of primary and secondary aliphatic amines with the formation of carbamate esters is presented. The method is based on a derivatization procedure in a two-phase system, where the carbamate ester is formed. The method is applied to the determination of 1,6-hexamethylene diamine (HDA) in aqueous solutions and human urine, using capillary GC. Detection was performed using thermionic specific detection (TSD) and mass spectrometry (MS)-selective-ion monitoring (SIM) using electron-impact (EI) and chemical ionization (CI) with ammonia monitoring both positive (CI)+ and negative ions (CI)-. Quantitative measurements were made in the chemical ionization mode monitoring both positive and negative ions. Tetra-deuterium-labelled HDA (TDHDA; H2NC2H2(CH2)4C2H2NH2) was used as the internal standard for the GC-MS analysis. In CI+ the m/z 386 and the m/z 390 ions corresponding to the [M + 18]+ ions (M = molecular ion) of HDA-TFECF and TDHDA-TFECF were measured; in CI- the m/z 267 and the m/z 271 ions corresponding to the [M - 101]- ions. The overall recovery was found to be 97 +/- 5% for a HDA concentration of 1000 micrograms/l in urine. The minimal detectable concentration in urine was found to be less than 20 micrograms/l using GC-TSD and 0.5 micrograms/l using GC-SIM. The overall precision for the work-up procedure and GC analysis was ca. 3% (n = 5) for 1000 micrograms/l HDA-spiked urine, and ca. 4% (n = 5) for 100 micrograms/l. The precision using GC-SIM for urine samples spiked to a concentration of 5 micrograms/l was found to be 6.3% (n = 10).

Air and biological monitoring of toluene diisocyanate in a flexible foam plant.

Comparative air measurements of toluene diisocyanate (TDI) were performed in a 5.6 m3 standard atmosphere and at a TDI flexible foam plant. Air samples were collected in midget impinger flasks containing 9-(N-methyl-amino-methyl)-anthracene (MAMA) in toluene and on 13-mm glass-fiber filters impregnated with MAMA and glycerol analyzed by LC-UV and with filter-tape instruments. In the laboratory study the average amounts of the TDI-MAMA derivatives determined were higher for filters compared to impingers when tested at concentrations between 16 and 150 micrograms/m3 (n = 29). At the TDI foaming plant the amount of TDI-MAMA collected on the filters compared with impingers showed higher TDI values at low concentrations and lower values at higher concentrations. The same was seen for the filter-tape measurements, but for two samples at very low concentrations the response was much lower. The average air concentration was 29.8 micrograms/m3 (12.5-79.9; n = 12). The highest exposure peak measured was approximately 3 mg TDI/m3. 2,4- and 2,6-toluene diamine (TDA) in urine (U-TDA) and in plasma (P-TDA) from four exposed workers and one volunteer were determined after strong acid hydrolysis as their pentafluoro-propionic anhydride derivatives using gas chromatography-mass spectrometry. The ions monitored were the M-20 ions (M = molecular weight) of the TDA and trideuterium labeled TDA as the internal standard. The P-TDA among the workers varied between 1-38 micrograms/L and between 7-24 micrograms/L for 2,4- and 2,6-TDA, respectively. The individual plasma levels among the workers over the 3-day periods varied between 7-73%. For the volunteer, P-TDA reached a maximum about 24 hours after the last exposure. The half-time of P-TDA for the volunteer was about 10 days. The urine levels (U-TDA) varied greatly with time and exposure. High peaks were found during or shortly after the exposure. No clear correlation between air levels of TDI measured with the filter-tape instruments and levels of TDA in hydrolyzed urine and plasma was seen, but the U-TDAMax followed the exposure in time as measured with the filter-tape instruments.

Biological monitoring of isocyanates and related amines. II. Test chamber exposure of humans to 1,6-hexamethylene diisocyanate (HDI).

Five male subjects were exposed to 1,6-hexamethylene diisocyanate (HDI) atmospheres for 7.5 h. The exposures were performed in an 8 m3 stainless steel test chamber, and the HDI atmospheres were generated by a gas-phase permeation method. HDI in air was determined by an HPLC method utilizing the 9-(N-methylaminomethyl)-anthracene reagent, and by a continuous monitoring device (MDA 7100). The average air concentration was ca 25 micrograms/m3, and the inhaled dose of HDI for the different subjects was estimated at ca 100 micrograms. The related amine 1,6-hexamethylene diamine (HDA) was after acid hydrolysis of urine and plasma, determined as a heptafluorobutyric derivative, by glass capillary gas-chromatography and selected ion monitoring (SIM), in a chemical ionization mode using ammonia as reagent gas. The cumulated urinary excretion of HDA during 28 h was 8.0 to 14 micrograms, which corresponds to ca 11 to 21% of the inhaled dose of HDI. The urinary level of HDA, in samples collected immediately after the end of the exposures, was on average 0.02 mmol/mol creatinine (range 0.01-0.03 mmol/mol creatinine). The urinary elimination was rapid, and half-time (t 1/2), for the concentration of HDA in urine, showed an average of 1.2 h (range 1.1-1.4 h). No specific IgE and IgG antibodies to HDI were detected before and after provocation; nor were spirometry or bronchial reactivity changed immediately and 15 h after provocation. Analysis of HDA in hydrolysed urine, as a marker of short-time exposure to HDI, is proposed.

Chromatographic determination of amines in biological fluids with special reference to the biological monitoring of isocyanates and amines. IV. Determination of 1,6-hexamethylenediamine in human urine using capillary gas chromatography and selective ion monitoring.

A capillary gas chromatographic (GC) method was developed for the determination of 1,6-hexamethylenediamine (HDA) in hydrolysed human urine. The method was based on a derivatization procedure with heptafluorobutyric anhydride. The amides formed were determined using capillary GC with selected ion monitoring in the chemical ionization mode with ammonia as reagent gas. The overall recovery was 34% for a concentration of 100 micrograms/l of HDA in urine. The minimum detectable concentration in urine was below 0.5 microgram/l. The precision of the method was 5% (n = 9). Deuterium-labelled HDA [H2NC2H2(CH2)4C2H2NH2] was used as the internal standard. A male subject was exposed to hexamethylene diisocyanate (HDI) for 7.5 h in a test chamber. The average air concentration of HDI was ca. 30 micrograms/m3, which corresponds to ca. 85% of the threshold limit value in Sweden (35 micrograms/m3). The half time of urinary levels of HDA was ca. 1.4 h and more than 90% of the urinary elimination was completed within 4 h after the exposure. The amount of HDA excreted in urine was ca. 10 micrograms, corresponding to ca. 10% of the estimated inhaled dose of HDI.

Passive smoking and childhood asthma. Urinary cotinine levels in children with asthma and in referents.

Passive exposure to tobacco smoke was assessed in children with asthma (age 3-15) and in referents. There was statistically significantly (P less than 0.0005) higher excretion of the nicotine metabolite, cotinine, in the urine of 49 children with asthma (geometric mean 10 ng/ml) compared with 77 referents (4.8 ng/ml). Maternal smoking was statistically significantly more prevalent among the asthmatics than among the referents (relative risk = RR = 2.6, 95% C1 = 1.2-5.3). In conclusion, the exposure to environmental tobacco smoke in asthmatic children was higher than among healthy children, indicating that passive smoking may be a predisposing and/or aggravating factor for childhood asthma.