Bioassay-directed chemical analysis in environmental research

Schuetzle, Dennis; Lewtas, Joellen

doi:10.1021/ac00124a001

Cited by 154 publications

(85 citation statements)

References 20 publications

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“…This showed that the fractionation unmasked the mutagenic potency of the compounds by separating some of the nonmutagenic cytotoxic compounds from the mutagenic compounds in the mixture. The ability of the fractionation to separate a large amount of nonmutagenic mass from mutagenic mass (Tables 7 and 8, Figure 1) is one of the goals of a successful bioassay-directed fractionation (Schuetzle and Lewtas 1986).…”

Section: Discussionmentioning

confidence: 99%

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Bioassay-directed fractionation and salmonella mutagenicity of automobile and forklift diesel exhaust particles.

DeMarini

Brooks

Warren

et al. 2004

Environ Health Perspect

118

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Many pulmonary toxicity studies of diesel exhaust particles (DEPs) have used an automobile-generated sample (A-DEPs) whose mutagenicity has not been reported. In contrast, many mutagenicity studies of DEPs have used a forklift-generated sample (SRM 2975) that has been evaluated in only a few pulmonary toxicity studies. Therefore, we evaluated the mutagenicity of both DEPs in Salmonella coupled to a bioassay-directed fractionation. The percentage of extractable organic material (EOM) was 26.3% for A-DEPs and 2% for SRM 2975. Most of the A-EOM (~55%) eluted in the hexane fraction, reflecting the presence of alkanes and alkenes, typical of uncombusted fuel. In contrast, most of the SRM 2975 EOM (~58%) eluted in the polar methanol fraction, indicative of oxygenated and/or nitrated organics derived from combustion. Most of the direct-acting, base-substitution activity of the A-EOM eluted in the hexane/dichloromethane (DCM) fraction, but this activity eluted in the polar methanol fraction for the SRM 2975 EOM. The direct-acting frameshift mutagenicity eluted across fractions of A-EOM, whereas > 80% eluted only in the DCM fraction of SRM 2975 EOM. The A-DEPs were more mutagenic than SRM 2975 per mass of particle, having 227 times more polycyclic aromatic hydrocarbon-type and 8-45 more nitroarene-type mutagenic activity. These differences were associated with the different conditions under which the two DEP samples were generated and collected. A comprehensive understanding of the mechanisms responsible for the health effects of DEPs requires the evaluation of DEP standards for a variety of end points, and our results highlight the need for multidisciplinary studies on a variety of representative samples of DEPs.

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Section: Discussionmentioning

confidence: 99%

“…Given that these factors vary for the SRM and A-DEP samples, we reasoned that the biologic activities of these DEPs were likely to be different. To assess the potential impact of these differences on mutagenicity, we evaluated the two DEP samples using a bioassay-directed fractionation (Schuetzle and Lewtas 1986) coupled with the Salmonella mutagenicity assay.…”

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confidence: 99%

Bioassay-directed fractionation and salmonella mutagenicity of automobile and forklift diesel exhaust particles.

DeMarini

Brooks

Warren

et al. 2004

Environ Health Perspect

118

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“…By examining the biological responses of the different fractions of both diesel PM extracts and biodiesel PM extracts it may be possible to identify the emission source components responsible for biological effects, in much the same way bioassay-directed chemical analysis has been used in other environmental research [30,[33][34][35][36], with possible use of the empirical data in predictive modeling. Incorporating testing of biodiesel emissions may assist in identifying the similarities and differences in the health responses (e.g.…”

Section: Discussionmentioning

confidence: 99%

Release of the Pro-Inflammatory Markers by BEAS-2B Cells Following In Vitro Exposure to Biodiesel Extracts

Swanson¹,

Kado²,

Funk³

et al. 2009

TOTOXIJ

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Biodiesel, an alkyl ester of plant oils that can be used in an unmodified diesel engine, is the first renewable diesel fuel alternative to become a commercially accepted part of our nation's energy infrastructure. For traditional diesel fuel exhaust, it has been demonstrated that the particulate matter (PM) organic components play a role in acute inflammatory reactions. However, there have been only a few cytotoxicity and mutagenicity studies on biodiesel emissions. In this study, BEAS-2B cells, a transformed human airway epithelial cell line, were exposed in vitro to the PM organic extracts from Standard Reference Material (SRM) 1975, soy ethyl ester (SEE), soy methyl ester (SME), and petroleum diesel for 24 hours. This study demonstrated that the organic extracts of biodiesel PM in an aqueous solution can increase the release of pro-inflammatory cytokines IL-8 and IL-6 by respiratory epithelial cells. On a microgram PM equivalent per ml (μg PM eq/ml) basis, exposure to biodiesel extracts was associated with a greater release of IL-8 and IL-6 relative to organic extracts of two diesel PM samples. The dose range tested was not cytotoxic. It was also noted that the solvent exchange method, which was used to prepare the aqueous exposure doses, may not be appropriate for the investigation of biodiesel extracts, though it has been used extensively in petroleum diesel research. A valuable new finding from these experiments is that the soluble organic fraction (SOF) of biodiesel PM begins to elicit a cytokine response in BEAS-2B cells at an exposure lower than petroleum diesel PM extract (approximately 40 μg PM eq/ml). However, more research is required to better characterize the potency of the organic fraction of biodiesel compared to petroleum diesel.

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“…Diesel exhaust is believed to consist of thousands of organic constituents and is a major cause of urban pollution (Bayona et al 1988;Draper 1986;Erdinger et al 2005;Mestres 2005;Morville 2006;Ona 2006;Schuetzle 1983;Schuetzle & Lewtas 1986;Seidel et al 2006). A number of studies on the adverse effects of diesel exhaust particles (DEP) on human health, including lung cancer (McClellan 1987;Ichinose et al 1997), allergic rhinitis (Muranaka 1986;Takafuji et al 1987), bronchial asthma-like disease (Sagai et al 1996;Miyabara et al 1998), and disruption of endocrine function, have been reported (Yoshida et al 1999;Watanabe & Oonuki 1999;Tsukue et al 2001Tsukue et al , 2002.…”

Section: Introductionmentioning

confidence: 99%

Isolation and identification of new vasodilative substances in diesel exhaust particles

Seki

Noya

Mikami

et al. 2009

Environ Sci Pollut Res

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Goal, Scope, and Background We recently developed a new isolation method for diesel exhaust particles (DEP), involving successive extraction with H 2 O, sodium bicarbonate, and sodium hydroxide, in which the sodium hydroxide extract was found to consist of phenolic components. Analysis of the extract revealed that vasodilative active nitrophenols are in DEP in significantly higher concentration than those estimated by an earlier method involving a combination of solvent extraction and repeated chromatography. These findings indicated that our new procedure offers a simple, efficient and reliable method for the isolation and identification of bioactive substances in DEP. This encouraged us to extend our work to investigating new vasodilatory substances in the sodium bicarbonate extract. Materials and Methods DEP were collected from the exhaust of a 4JB1-type engine (ISUZU Automobile Co., Tokyo, Japan). GC-MS analysis was performed with a GCMS-QP2010 instrument (Shimadzu, Kyoto, Japan).Results DEP dissolved in 1-butanol was successively extracted with water, sodium bicarbonate, and then aqueous sodium hydroxide. The sodium bicarbonate extract was neutralized and the resulting mixture of acidic components was subjected to reverse-phase (RP) column chromatography followed by RP-HPLC with fractions assayed for vasodilative activity. This led to the identification of telephthalic acid, p-hydroxybenzoic acid, isophthalic acid, phthalic acid, 3-hydroxy-4-nitrobenzoic acid, 4-hydroxy-3-nitrophenol, and 1,4,5-naphthalene tricarboxylic acid as components of DEP. DiscussionThe sodium bicarbonate extract was rich in the acidic components. Repeated reverse-phase chromatography resulted in the successful isolation of several acidic substances including the new vasodilative materials, 4-hydroxy-3-nitrobenzoic acid and 3-hydroxy-4-nitrobenzoic acid. Seidel et al. 2006). A number of studies on the adverse effects of diesel exhaust particles (DEP) on human health, including lung cancer (McClellan 1987;Ichinose et al. 1997), allergic rhinitis (Muranaka 1986;Takafuji et al. 1987), bronchial asthma-like disease (Sagai et al. 1996;Miyabara et al. 1998), and disruption of endocrine function, have been reported (Yoshida et al. 1999;Watanabe & Oonuki 1999;Tsukue et al. 2001Tsukue et al. , 2002. While these studies have furnished epidemiologically important results, they have been done without knowledge of the bioactive entities in DEP, making it difficult to estimate the effect of individual components on human health. In our earlier studies on the isolation and identification of bioactive compounds contained in DEP, we have isolated aromatic substances including nitrophenols (Mori et al. 2003a;Taneda et al. 2004a) that showed strong vasodilatory activity by a method involving successive solvent extractions, followed by repeated column chromatography. This multi-step isolation procedure, however, resulted in poor fractionation with impaired accuracy and reproducibility. Therefore, it was crucial to develop a more efficient and ...

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Bioassay-directed chemical analysis in environmental research

Cited by 154 publications

References 20 publications

Bioassay-directed fractionation and salmonella mutagenicity of automobile and forklift diesel exhaust particles.

Bioassay-directed fractionation and salmonella mutagenicity of automobile and forklift diesel exhaust particles.

Release of the Pro-Inflammatory Markers by BEAS-2B Cells Following In Vitro Exposure to Biodiesel Extracts

Isolation and identification of new vasodilative substances in diesel exhaust particles

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