Leukemia risk assessment of exposure to low-levels of benzene based on the linearized multistage model.

Objectives: To assess leukemia risk in occupational populations exposed to low levels of benzene. Methods: Leukemia incidence data from the Chinese Benzene Cohort Study were fitted using the Linearized multistage (LMS) model. Individual benzene exposure levels, urinary S-phenylmercapturic acid (S-PMA) and trans, trans-muconic acid (t, t-MA) were measured among 98 benzene-exposed workers from factories in China. Subjects were categorized into four groups by rounding the quartiles of cumulative benzene concentrations (< 3, 3-5, 5-12, ≥12 mg/m3·year, respectively). The risk of benzene-induced leukemia was assessed using the LMS model, and the results were validated using the EPA model and the Singapore semi-quantitative risk assessment model. Results: The leukemia risks showed a positive correlation with increasing cumulative concentration in the four exposure groups (excess leukemia risks were 4.34, 4.37, 4.44 and 5.52 × 10-4, respectively; Ptrend < 0.0001) indicated by the LMS model. We also found that the estimated leukemia risk using urinary t, t-MA in the LMS model was more similar to those estimated by airborne benzene compared to S-PMA. The leukemia risk estimated by the LMS model was consistent with both the Singapore semi-quantitative risk assessment model at all concentrations and the EPA model at high concentrations (5-12, ≥12 mg/m3·year), while exceeding the EPA model at low concentrations (< 3 and 3-5 mg/m3·year). However, in all four benzene-exposed groups, the leukemia risks estimated by these three models exceeded the lowest acceptable limit for carcinogenic risk set by the EPA at 1 × 10-6. Conclusion: This study demonstrates the utility of the LMS model derived from the Chinese benzene cohort in assessing leukemia risk associated with low-level benzene exposure, and suggests that leukemia risk may occur at cumulative concentrations below 3 mg/m3·year.

Optimization of benzene exposure risk assessment: An integrated approach utilizing internal and external concentrations with a focus on biomarkers S-PMA & t, t-MA.

In the majority of occupational settings within China, the concentrations of benzene are observed to fall markedly below the demarcated detection thresholds. Employing traditional risk assessment models, the presence of exceptionally low airborne benzene exposure concentrations may infuse heightened degrees of uncertainty. Consequently, the necessity arises to investigate risk assessment methodologies more apt for the prevalent exposure environment among employees. In the present study, a pharmacokinetic model premised on urinary benzene metabolites (S-PMA and t, t-MA) was employed to ascertain a more precise daily airborne benzene exposure concentration per individual. This value was integrated into the linear multistage model as the 'internal exposure concentration'. In conjunction with the U.S National Environmental Protection Agency's (EPA) inhalation risk assessment model predicated on the external exposure concentration, the Singapore Ministry of Manpower's (MOM) model, and the linear multistage (LMS) model, the carcinogenic and non-carcinogenic effects of benzene were evaluated for 1781 benzene-exposed employees across 76 enterprises in Jiangsu Province. Findings suggest that in the linear multilevel model assessment, the cancer risk levels based on t, t-MA and S-PMA were higher in the printing and recording media reproduction industry, automobile manufacturing industry, general equipment manufacturing industry and the furniture manufacturing industry (median 2.842 × 10-4, 2.819 × 10-4, 2.809 × 10-4, and 2.678 × 10-4), which align more consistently with the actual benzene exposure circumstances of each industry's study participants, with overall risk levels calculated by the linear multistage model exceeding those of the EPA inhalation risk assessment model and the MOM model. This implies that the linear multistage model of internal exposure, based on the reciprocal of benzene biomarkers S-PMA and t, t-MA for airborne benzene exposure, presents enhanced sensitivity and suitability for the current occupational health risk assessment of workers. Without doubt, biomarker-based benzene exposure risk assessment emerges as the optimal choice.

Benzoates and in situ formed benzene in food supplements and risk assessment.

Upward trend in the use of food supplements urged the assessment of their safety. Eighty-eight liquid herbal supplements collected in Novi Sad (Serbia) in 2018 (36 samples) and 2021 (52 samples) were analysed for the presence of benzoates and sorbates (HPLC-UV) and benzene (HS-GC/MS). Benzoic acid varied from 599 to 9253 mg/kg and sorbic acid between 185 and 1658 mg/kg. The acceptable daily intake of sorbic acid was not reached, but in case of benzoic acid, it was exceeded by 5.3% of the samples. The presence of benzene was confirmed in 41.2% of benzoate preserved supplements (0.9-51.7 µg/kg). Benzene exposure revealed no health concern: maximum hazard quotients ranged from 0.39% (toddlers) to 0.84% (adolescents); minimum margins of exposure were between 35,680 (adolescents) and 77,419 (toddlers); estimates of lifetime cancer risk did not reach one extra cancer case per 100 000 persons. However, measures to mitigate benzene presence in food should be considered.

Comparing in planta accumulation with microbial routes to set targets for a cost-competitive bioeconomy.

Plants and microbes share common metabolic pathways for producing a range of bioproducts that are potentially foundational to the future bioeconomy. However, in planta accumulation and microbial production of bioproducts have never been systematically compared on an economic basis to identify optimal routes of production. A detailed technoeconomic analysis of four exemplar compounds (4-hydroxybenzoic acid [4-HBA], catechol, muconic acid, and 2-pyrone-4,6-dicarboxylic acid [PDC]) is conducted with the highest reported yields and accumulation rates to identify economically advantaged platforms and breakeven targets for plants and microbes. The results indicate that in planta mass accumulation ranging from 0.1 to 0.3 dry weight % (dwt%) can achieve costs comparable to microbial routes operating at 40 to 55% of maximum theoretical yields. These yields and accumulation rates are sufficient to be cost competitive if the products are sold at market prices consistent with specialty chemicals ($20 to $50/kg). Prices consistent with commodity chemicals will require an order-of-magnitude-greater accumulation rate for plants and/or yields nearing theoretical maxima for microbial production platforms. This comparative analysis revealed that the demonstrated accumulation rates of 4-HBA (3.2 dwt%) and PDC (3.0 dwt%) in engineered plants vastly outperform microbial routes, even if microbial platforms were to reach theoretical maximum yields. Their recovery and sale as part of a lignocellulosic biorefinery could enable biofuel prices to be competitive with petroleum. Muconic acid and catechol, in contrast, are currently more attractive when produced microbially using a sugar feedstock. Ultimately, both platforms can play an important role in replacing fossil-derived products.

Biomatrix of health risk assessment of benzene-exposed workers at Thai gasoline stations.

OBJECTIVE: This study assessed the health risk of benzene exposure among Thai gasoline station workers through biomarker detection and experience of adverse symptoms. METHODS: Trans, trans-muconic acid (tt-MA) metabolites of benzene were analyzed from spot urine sampled among gasoline station workers after shift work using HPLC-UV. Air benzene monitoring was done with an active sampler connected to a charcoal sorbent tube, and analyzed by GC-FID. The health risk was calculated by using the biomatrix of the likelihood of benzene exposure and the severity of adverse symptoms. RESULTS: The tt-MA concentration, among 235 workers, ranged from less than 10-2159 µg/g Cr, which corresponded to the air benzene concentration range of <0.1 to 65.8 ppb. In total, 32.3% of workers had a higher than acceptable risk level and there was a significant association between gasoline station work zones and the likelihood of benzene exposure as well as the health risk of workers. The health risk levels estimated from the biomarker monitoring were consistent with the risk matrix of air benzene monitoring. CONCLUSION: This tt-MA biomarker monitoring and biomatrix of health risk assessment is suggested as useful for health surveillance of gasoline station workers exposed to benzene.

Assessment of Concentrated Liquid Coffee Acceptance during Storage: Sensory and Physicochemical Perspective.

Concentrated liquid coffees (CLCs) refer to stored extracts stable at environmental temperature, used as ingredients in the retail market. Their low chemical stability affects the sensory profile. This study was performed in two CLCs, one without additives (BIB) and another with a mix of sodium benzoate and potassium sorbate additives (SD), stored at 25 °C for one year. Quantitative-Descriptive (QDA) and discriminant analyses permitted identifying the critical sensory attributes and their evolution over time. The concentrate without additives presented an acceptance limit of 196 days (evaluated at a 50% acceptance ratio), while the additives increased the shelf life up to 226 days (38.9% improvement). The rejection was related to a decreased aroma, increased acidity, and reduced bitterness. A bootstrapped feature selection version of Partial Least Square analysis further demonstrated that reactions of 5-caffeoylquinic acid (5CQA) and 3,5-dicaffeoylquinic acid (3,5diCQA) could cause changes in the aroma at the first degradation stage. In the following stages, changes in fructose and stearic acid contents, a key indicator of acceptance for both extracts possibly related to non-enzymatic reactions involving fructose and other compounds, might affect the bitterness and acidity. These results provided valuable information to understand flavor degradation in CLCs.

Adaptation of Campylobacter field isolates to propionic acid and sorbic acid is associated with fitness costs.

AIMS: To reduce the burden of Campylobacter at different stages of the food chain, recent studies have shown the effectiveness of organic acids as a risk mitigation strategy. However, very little is known about possible adaptation responses of Campylobacter that lead to reduced susceptibility to organic acids. Here we investigated the adaptive responses of Campylobacter field isolates to organic acids and estimated the fitness costs. METHODS AND RESULTS: Exposure of two Campylobacter jejuni and one Campylobacter coli isolate to subinhibitory concentrations of propionic acid or sorbic acid resulted in twofold to fourfold increased minimal inhibitory concentration values for the adapted variants. With one exception, the decreased susceptibility was stable in at least 10 successive subcultures without selection pressure. Growth competition experiments revealed a reduced fitness of adapted variants compared to the wild-type isolates. A linear regression model allowed an estimation of the fitness cost. Growth kinetics experiments showed significantly prolonged lag phases in five of six adapted isolates while there was not a direct correlation in the maximum growth rates compared to the wild-type isolates. CONCLUSIONS: The results of the study showed that a stepwise adaptation of Campylobacter to organic acids is possible, but at the detriment of changes in growth behaviour and reduced fitness. SIGNIFICANCE AND IMPACT OF THE STUDY: The study contributes to the understanding of adaptive responses of Campylobacter to organic acids treatments, for example, as part of risk mitigation strategies.

Limited life cycle and cost assessment for the bioconversion of lignin-derived aromatics into adipic acid.

Lignin is an abundant and heterogeneous waste byproduct of the cellulosic industry, which has the potential of being transformed into valuable biochemicals via microbial fermentation. In this study, we applied a fast-pyrolysis process using softwood lignin resulting in a two-phase bio-oil containing monomeric and oligomeric aromatics without syringol. We demonstrated that an additional hydrodeoxygenation step within the process leads to an enhanced thermochemical conversion of guaiacol into catechol and phenol. After steam bath distillation, Pseudomonas putida KT2440-BN6 achieved a percent yield of cis, cis-muconic acid of up to 95 mol% from catechol derived from the aqueous phase. We next established a downstream process for purifying cis, cis-muconic acid (39.9 g/L) produced in a 42.5 L fermenter using glucose and benzoate as carbon substrates. On the basis of the obtained values for each unit operation of the empirical processes, we next performed a limited life cycle and cost analysis of an integrated biotechnological and chemical process for producing adipic acid and then compared it with the conventional petrochemical route. The simulated scenarios estimate that by attaining a mixture of catechol, phenol, cresol, and guaiacol (1:0.34:0.18:0, mol ratio), a titer of 62.5 (g/L) cis, cis-muconic acid in the bioreactor, and a controlled cooling of pyrolysis gases to concentrate monomeric aromatics in the aqueous phase, the bio-based route results in a reduction of CO2 -eq emission by 58% and energy demand by 23% with a contribution margin for the aqueous phase of up to 88.05 euro/ton. We conclude that the bio-based production of adipic acid from softwood lignins brings environmental benefits over the petrochemical procedure and is cost-effective at an industrial scale. Further research is essential to achieve the proposed cis, cis-muconic acid yield from true lignin-derived aromatics using whole-cell biocatalysts.

Assessment of sodium benzoate and potassium sorbate preservatives in some products in Kashan, Iran with estimation of human health risk.

The purpose was to assess of sodium benzoate (SB) and potassium sorbate (PS) preservatives in 103 samples of cake, toast bread, tomato paste, mayonnaise sauce, carbonated soft drink and Olovieh salad in Kashan, by spectrophotometry and high performance liquid chromatography (HPLC) methods. The chronic daily intake (CDI), target hazard quotient (THQ) and hazard index (HI) of SB and PS for Iranian population were calculated. The results showed that SB and PS were not detected in the tomato paste samples. SB and PS concentrations for all samples were less than regulatory limits except for PS in one cake sample (3.57%). CDI and THQ of PS for mayonnaise sauce, Olovieh salad and cake products, except toast bread, were less than the acceptable daily intakes (ADIs) and one, respectively. While HI value of PS for the selected products was more than one, indicating that the non-carcinogenic risk represent a threat to consumers. THQ and HI values of SB for mayonnaise sauce and carbonated soft drink products were more than one through consumption of these products, indicating considerable non-carcinogenic risk. Therefore, the results highlighted the importance of a more attentive monitoring of these preservatives by the public and food health authorities in Iran.

Safety assessment of sodium acetate, sodium diacetate and potassium sorbate food additives.

Cytotoxicity and genotoxicity of sodium acetate (SA), sodium diacetate (SDA), and potassium sorbate (PS) was tested on Human Umbilical Vein Endothelial Cells (HUVEC). Cytotoxicity was investigated by MTT assay and flow cytometry analysis, while genotoxicity was evaluated using DNA fragmentation and DAPI staining assays. The growth of treated HUVECs with various concentrations of SA, SDA and PS decreased in a dose-and time-dependent manner. The IC50 of 487.71, 485.82 and 659.96 µM after 24 h and IC50 of 232.05, 190.19 and 123.95 µM after 48 h of treatment were attained for SA, SDA and PS, respectively. Flow cytometry analysis showed that early and late apoptosis percentage in treated cells was not considerable. Also neither considerable DNA fragmentation nor DNA smear was observed using DAPI staining and DNA ladder assays. Overall, it can be concluded that the aforementioned food additives can be used as safe additives at low concentration in food industry.

Assessment of exposure to mixture pollutants in Mexican indigenous children.

The aim of the present work was to complete an exposure assessment in three Mexican indigenous communities using the community-based health risk assessment, which is the first step in the CHILD framework. We used 1-hydroxypyrene (1-OHP) as an exposure biomarker to polycyclic aromatic hydrocarbons (PAHs) and trans, trans-muconic acid (t,t-MA) as an exposure biomarker to benzene, persistent organic pollutants (POPs), lead, manganese, arsenic, and fluoride. Anthropometric measurements were also taken. In these communities, high percentages of children with chronic malnutrition were found (28 to 49 %) based on their weight and age. All communities showed a high percentage of children with detectable levels of four or more compounds (70 to 82 %). Additionally, our results showed that in indigenous communities, children are exposed to elevated levels of certain environmental pollutants, including manganese with 17.6, 16.8, and 7.3 µg/L from SMP, TOC, and CUA, respectively. Lead and HCB levels were similar in the indigenous communities (2.5, 3.1, and 4.2 µg/dL and 2.5, 3.1, and 3.7 ng/mL, respectively). 1-OHP and t,t-MA levels were higher in TOC (0.8 µmol/mol of creatinine, 476 µg/g of creatinine, respectively) when compared with SMP (0.1 µmol/mol of creatinine, 215.5 µg/g of creatinine, respectively) and CUA (0.1 µmol/mol of creatinine, 185.2 µg/g of creatinine, respectively). DDE levels were 30.7, 26.9, and 9.6 ng/mL in CUA, SMP, and TOC, respectively. The strength of this study is that it assesses exposure to pollutants with indications for the resultant risk before an intervention is made by the CHILD program to manage this risk in the indigenous communities. Considering the large number of people, especially children, exposed to multiple pollutants, it is important to design effective intervention programs that reduce exposure and the resultant risk in the numerous indigenous communities in Mexico.

Benzoic and sorbic acid in soft drink, milk, ketchup sauce and bread by dispersive liquid-liquid microextraction coupled with HPLC.

Benzoic acid and sorbic acid are widely used for food preservation. These preservatives are generally recognised as safe. The aim of this study was to determine the level of benzoic and sorbic acid in food samples that are usually consumed in Iran. Therefore, 54 samples, including 15 soft drinks, 15 ultra-high-temperature milk, 15 ketchup sauces and 9 bread samples, were analysed by high-performance liquid chromatography with UV detection. Benzoic acid was detected in 50 (92.5%) of the samples ranging from 3.5 to 1520 µg mL⁻¹, while sorbic acid was detected in 29 (50.3%) samples in a range of 0.8 and 2305 µg mL⁻¹. Limits of detection and limits of quantification for benzoate were found to be 0.1 and 0.5 µg mL⁻¹, respectively, and for sorbate 0.08 and 0.3 µg mL⁻¹, respectively. The results showed that benzoic acid and sorbic acid widely occur in food products in Iran.

Sorbate and benzoate in Turkish retail foodstuffs.

From 2008 to 2011, surveys were conducted to determine the levels of benzoic and sorbic acids and their respective salts in 983 retail food samples which included sauces, vegetable and fruit preparations, flavoured syrups, food supplements, cereals, bakery products, jelly, synthetic cream, sprays, mustards, jam and preserves, molasses, chewing gum, confectionery, non-alcoholic beverages, tea, wine, vinegar, brine and beers. The analysis involved methanol extraction of the foodstuff and direct determination by HPLC with UV detection. Quality assurance was employed with each batch of samples. Accuracy was ensured through regular participation in proficiency tests. Over this four-year period, a total of 23 samples (2.3%), some syrups, tomato sauces and fruit contained individual or combined levels of sorbic and benzoic acids above regulatory limits. Unauthorised use of benzoic acid was also detected in a syrup sample, bakery products and fruit preserves.

Assessment of occupational exposure to benzene, toluene and xylenes in urban and rural female workers.

OBJECTIVES: This is the first research study to compare among female, non-smoker workers: (a) the exposure to benzene, toluene and xylenes (BTXs) in urban air during work in the street (traffic policewomen, TP) vs. work in vehicles (police drivers, PD); (b) the exposure to BTXs in urban environments (in street and in car) vs. rural environments (roadwomen, RW); (c) the values of blood benzene, urinary trans, trans muconic acid (t,t-MA) and urinary S-phenylmercapturic acid (S-PMA) in urban areas (in street and in car) vs. rural areas. METHODS: Passive personal samplings and data acquired using fixed monitoring stations located in different areas of the city were used to measure environmental and occupational exposure to BTXs during the work shift in 48 TP, 21 PD and 22 RW. In the same study subjects, blood benzene, t,t-MA and S-PMA were measured at the end of each work shift. RESULTS: Personal exposure of urban workers to benzene seemed to be higher than the exposure measured by the fixed monitoring stations. Personal exposure to benzene and toluene was (a) similar among TP and PD and (b) higher among urban workers compared to rural workers. Personal exposure to xylenes was (a) higher in TP than in PD and (b) higher among urban workers compared to rural workers. Blood benzene, t,t-MA and S-PMA levels were similar among TP and PD, although the blood benzene level was significantly higher in urban workers compared to rural workers. In urban workers, airborne benzene and blood benzene levels were significantly correlated. CONCLUSIONS: Benzene is a human carcinogen, and BTXs are potential reproductive toxins at low dose exposures. Biological and environmental monitoring to assess exposure to BTXs represents a preliminary and necessary tool for the implementation of preventive measures for female subjects working in outdoor environments.

Exposure assessment of food preservatives (sulphites, benzoic and sorbic acid) in Austria.

An exposure assessment was performed to estimate the potential intake of preservatives in the Austrian population. Food consumption data of different population groups, such as preschool children aged 3-6 years, female and male adults aged 19-65 years were used for calculation. Levels of the preservatives in food were derived from analyses conducted from January 2007 to August 2010. Dietary intakes of the preservatives were estimated and compared to the respective acceptable daily intakes (ADIs). In the average-intake scenario, assuming that consumers randomly consume food products that do or do not contain food additives, estimated dietary intakes of all studied preservatives are well below the ADI for all population groups. Sulphite exposure accounted for 34%, 84% and 89% of the ADI in preschool children, females and males, respectively. The mean estimated daily intake of benzoic acid was 32% (preschool children), 31% (males) and 36% (females) of the ADI. Sorbic acid intakes correspond to 7% of the ADI in preschool children and 6% of the ADI in adults. In the high-intake scenario assuming that consumers always consume food products that contain additives and considering a kind of brand loyalty of consumers, the ADI is exceeded for sulphites among adults (119 and 124%, respectively). Major contributors to the total intake of sulphites were wine and dried fruits for adults. Mean estimated dietary intakes of benzoic acid exceeded the ADI in all population groups, 135% in preschool children, 124% in females and 118% of the ADI in males, respectively. Dietary intakes of sorbic acid are well below the ADI, accounting for a maximum of 30% of the ADI in preschool children. The highest contributors to benzoic and sorbic acid exposure were fish and fish products mainly caused by high consumption data of this large food group, including also mayonnaise-containing fish salads. Other important sources of sorbic acid were bread, buns and toast bread and fruit and vegetable juices.

Biomarkers of internal dose for the assessment of environmental exposure to benzene.

The urinary excretion of t,t-muconic acid (t,t-MA), S-phenylmercapturic acid (SPMA) and urinary benzene and the influence of a smoking habit and of exposure to urban traffic on the urinary excretion of these biomarkers were investigated in 137 male adults from the general population. All subjects were not occupationally exposed to benzene and resident in two cities in Puglia (Southern-Italy). Environmental exposure to benzene was measured using passive personal samplers. The biomarkers t,t-MA, SPMA and urinary benzene were determined in urine samples collected from each subject at the end of the environmental sampling. The percentage of cases above the limit of detection was higher for SPMA and urinary benzene in smokers than in non-smokers, and for airborne benzene and urinary benzene in subjects exposed to urban traffic. Airborne benzene was correlated with the time spent in urban traffic during the environmental sampling. Among the biomarkers, urinary benzene was found to be correlated with airborne benzene only in non-smokers, and with the time spent in urban traffic, both in smokers and non-smokers considered together, and in non-smokers only. Finally, multiple regression analysis showed that the urinary excretion of all the biomarkers was dependent on the number of cigarettes smoked per day and, for urinary benzene, also on the time spent in urban traffic. In conclusion, urinary benzene seems to be a more valid biomarker than t,t-MA and SPMA to assess environmental exposure to extremely low concentrations of benzene. Cigarette smoking prevailed over traffic exhaust fumes in determining the internal dose of benzene.

Genotoxic monitoring and benzene exposure assessment of gasoline station workers in metropolitan Bangkok: sister chromatid exchange (SCE) and urinary trans, trans-muconic acid (t,t-MA).

Early warning of the potential of mutagens or carcinogens caused by benzene exposure that might occur in gasoline station workers can be achieved by examining 2 major biomarkers: sister chromatid exchange (SCE) and trans, trans-muconic acid (t,t-MA), a urinary metabolite of benzene. The main objective of this study was to assess benzene exposure and monitor the genotoxic effect of gasoline station workers in Bangkok, Thailand. Blood and urine samples were collected from 33 gasoline station workers, working in Pathumwan district area, central Bangkok, Thailand, for SCE and t,t-MA analysis, from April to June 2009. Control samples were collected from 30 office workers and students in the same area at the same period. Our results indicated significantly higher frequencies of SCE in gasoline exposed workers were than in controls (p<0.01), independent of gender. Urinary t,t-MA and t,t-MA/creatinine levels of gasoline exposed workers were also significantly higher than the control groups (p<0.05) were significantly higher in women than men workers (p<0.01). Calculated chromosomal damage relative risk (RR) of gasoline station workers was 3.00 (95% CI = 1.81 - 4.98, p<0.001) compared to controls. The gasoline exposed workers had potentially higher risk of chromosomal damage and cancer development because of direct contact to benzene.

Benzene exposure in childhood: Role of living environments and assessment of available tools.

Benzene is a widespread air pollutant and a well-known human carcinogen. Evidence is needed regarding benzene intake in the pediatric age group. We investigated the use of urinary (u) trans,trans-muconic acid (t,t-MA), S-phenylmercapturic acid (SPMA), and unmodified benzene (UB) for assessing exposure to low concentrations of environmental benzene and the role of living environment on benzene exposure in childhood. u-t,t-MA, u-SPMA, u-UB and u-cotinine were measured in urine samples of 243 Italian children (5-11 years) recruited in a cross-sectional study. Analytical results were compared with data obtained from questionnaires about participants' main potential exposure factors. u-UB, u-t,t-MA and u-SPMA concentrations were about 1.5-fold higher in children living in urban areas than in those in the rural group. Univariate analyses showed that u-UB was the only biomarker able to discriminate secondhand smoke (SHS) exposure in urban and rural children (medians=411.50 and 210.50 ng/L, respectively); these results were confirmed by the strong correlation between u-UB and u-cotinine in the SHS-exposed group and by multivariate analyses. A regression model on u-SPMA showed that the metabolite is related to residence area (p<0.001), SHS exposure (p=0.048) and gender (p=0.027). u-UB is the best marker of benzene exposure in children in the present study, and it can be used as a good carcinogen-derived biomarker of exposure to passive smoking, especially related to benzene, when urine sample is collected at the end of the day. In addition, it is important to highlight that SHS resulted the most important contributor to benzene exposure, underlining the need for an information campaign against passive smoking exposure.

Negative and positive ion mode LC/MS/MS for simple, sensitive analysis of sorbic acid.

Sorbic acid (SA: CH(3)-CH=CH-CH=CH-COOH) is one of the widely used food preservatives, although there have been some reports of its toxic activity, for example, on DNA and skin cells. In order to examine the effects of SA on mammalian tissues, we have developed a highly sensitive analytical method using LC/MS/MS with positive and negative ion mode electrospray ionization (ESI). In a previous study, we found that a nonacidic eluent offers better ionization efficiency than acids or their ammoniun salts. However, optimal results could not be obtained because the anion form of SA is poorly retained on a conventional reversed phase column. To resolve this problem, we chose a new type of column and used high-resolution mass spectrometry and positive ion mode analysis. There have only been a few reports using these methods in the positive mode, for example derivatized SA, because acid compounds such as SA are usually used in the negative ion mode. However, a new type of low-carbon-content and polar-endcapped C18 phase column was developed for better separation of SA from the matrix. High-resolution selected reaction monitoring (SRM) gave the best signal to noise ratio in normal-resolution SRM. In the positive ion mode, the CH(3)OH-0.05% HCOOH/0.1% CH(3)COOH eluent system yielded the best ionization efficiency. We propose a highly sensitive and simple analysis using a two-ion-mode ESI SRM method. Such systems should allow quantification of the amount of SA in or around the cells, without the need for pretreatment such as solid phase extraction.