Grantee Research Project Results
2007 Progress Report: Integrated Microfluidic System for Bioluminescent Bioreporting, Separations, Vibrational Spectroscopy, and Microcantilever Transducer Evaluation of Endocrine Disrupting Chemicals
EPA Grant Number: R832740Title: Integrated Microfluidic System for Bioluminescent Bioreporting, Separations, Vibrational Spectroscopy, and Microcantilever Transducer Evaluation of Endocrine Disrupting Chemicals
Investigators: Sepaniak, Michael J. , Sayler, Gary S.
Institution: University of Tennessee
EPA Project Officer: Hahn, Intaek
Project Period: November 1, 2005 through October 31, 2008 (Extended to October 31, 2009)
Project Period Covered by this Report: November 1, 2006 through October 31,2007
Project Amount: $590,240
RFA: Exposure Measurement Tools for Endocrine Disrupting Chemicals in Mixtures (2005) RFA Text | Recipients Lists
Research Category: Environmental Justice , Human Health , Safer Chemicals , Endocrine Disruptors
Objective:
The objective of this research is to develop analytical methodologies for the quantitative and qualitative measurement of mixtures of endocrine disrupting chemicals by utilizing the tools of analytical chemistry that include microcantilever arrays (MCA) for nanomechanical sensing, surface enhanced Raman spectroscopy (SERS), and chemical separations with a possible complement by biosensing microorganisms. By improving the existing technology and attacking the problem of EDC exposure and activity monitoring in realistic mixtures with an arsenal of informative tools such as MCAs, SERS, and chemical separations, one can clarify and elucidate which chemicals, and in what combinations, can mimic or inhibit endocrine signaling molecules. Speed and portability of these analytical techniques, either on separate or partially integrated platforms, will greatly facilitate screening and characterization of samples in the field.
Progress Summary:
In Year 2 of this project, we have moved beyond the scope of original task 1, using new approaches to sequester EDCs on SERS active extraction materials as well as improving the basic science of high-end lithographic SERS substrates. In original task 2, a technique of immobilizing estrogen receptors on microcantilevers (MCs) for screening of EDCs is maturing and expanding into a novel nuclear receptor protein on a MC array as nanomechanical mimics of synergistic biological systems. To address microfluidic separation and handling of EDC contaminated samples in real sample matrices, low volume preconcentration of standard and real water samples spiked with EDCs are being evaluated for response from bioreporters and laser induced fluorescence in fused silica capillaries. These samples have been prepared via syringe-loaded cartridges in preparation for the ultra-trace levels of EDCs in natural samples. In this manner, original tasks 3 and 4 are being handled in tandem with complementary studies, keeping in sight the reality that optimum performance of descriptive spectroscopy and nanomechanical response assays may be best achieved on distinct but complementary platforms. Mechanisms involving nuclear receptor proteins (e.g., PXR and CAR proteins) that protect the body from diverse harmful chemicals can also alter the ultimate physiological impact of EDCs, and the analytical consequences of this synergism are being assessed with developing MCAs.
The originally stated specific tasks of this research were:
- Test known EDCs in relevant matrices for SERS response characteristics;
- Develop EDC nanomechanical response signatures on microcantilever arrays;
- Integrate bioreporter yeast into conditions suitable for predictable function on a μfluidic platform with electrophoretic separations of EDCs; and
- Validate an integrated fluidic device that incorporates bioluminescent reporters, separations, and vibrational and nanomechanical detection of EDC-containing samples in realistic matrices.
- Integrated microfluidics systems for advanced EDC analysis
- Multiplexing of SERS nanoparticle activity evaluations with several analytes nearly simultaneously on a single, multichannel microfluidic platform (70% complete)
- SERS substrate improvements using rational design with electron beam lithography (50% complete) 2
- Low volume extraction and concentration for separations as well as investigation of alternative extraction materials to directly serve as SERS substrates (60% complete)
- Bioreporter compatibility analysis (80% complete)
- Functionalized MC arrays as nanomechanical mimics of synergistic systems
- Optimization studies to enhance microcantilever biosensor response sensitivity. (80% complete)
- Move toward optimized assays of EDCs using a range of receptors; estrogen receptor proteins, thyroid proteins and others with more subtle connections to the endocrine signaling system, including the complex relationship of PXR triggers. (50% complete)
- Assess nuclear receptor mediated responses with multiple receptor phases integrated into a single microcantilever array (MCA) as a surrogate for total living systems. (just initiated)
Future Activities:
Year 3 Activities Will Focus on the Following:
- Continue to develop techniques such as high efficiency preconcentration and advanced nanofabricated SERS substrates to mitigate the detriments of inadequate sensitivity for a wide range of EDCs
- Nanomechanical transduction to sense bioaffinity detection events – goal is the creation and testing of differential MCAs having different bioaffinity receptors on different MCs for broad analyte screening and fundamental studies
- Optimize μfluidic chip-based separations of EDCs for several EDC mixtures, utilizing advanced preconcentration techniques and multiple mobile phase regimes if necessary
- Prepare and submit at least 3 manuscripts for peer-review; present at least two talks at meetings such as EAS, FACSS, PittCon, EPA annual reporting conference
- Prepare and submit year 3 report as well as final report upon completion of no-cost extension
Journal Articles on this Report : 9 Displayed | Download in RIS Format
Other project views: | All 56 publications | 32 publications in selected types | All 32 journal articles |
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Abu-Hatab NA, John JF, Oran JM, Sepaniak MJ. Multiplexed microfluidic surface-enhanced Raman spectroscopy. Applied Spectroscopy 2007;61(10):1116-1122. |
R832740 (2007) R832740 (2008) R832740 (Final) |
Exit |
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Archibald R, Datskos P, Devault G, Lamberti V, Lavrik N, Noid D, Sepaniak M, Dutta P. Independent component analysis of nanomechanical responses of cantilever arrays. Analytica Chimica Acta 2007;584(1):101-105. |
R832740 (2006) R832740 (2007) R832740 (2008) R832740 (Final) |
Exit Exit Exit |
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Chapman PJ, Vogt F, Dutta P, Datskos PG, Devault GL, Sepaniak MJ. Facile hyphenation of gas chromatography and a microcantilever array sensor for enhanced selectivity. Analytical Chemistry 2007;79(1):364-370. |
R832740 (2006) R832740 (2007) R832740 (2008) R832740 (Final) |
Exit Exit Exit |
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Chapman PJ, Long Z, Datskos PG, Archibald R, Sepaniak MJ. Differentially ligand-functionalized microcantilever arrays for metal ion identification and sensing. Analytical Chemistry 2007;79(18):7062-7068. |
R832740 (2007) |
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Dutta P, Hill K, Datskos PG, Sepaniak MJ. Development of a nanomechanical biosensor for analysis of endocrine disrupting chemicals. Lab on a Chip 2007;7(9):1184-1191. |
R832740 (2007) R832740 (2008) R832740 (Final) |
Exit Exit |
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Hill K, Dutta P, Zareba A, Eldridge ML, Sepaniak MJ. Morphological and chemical optimization of microcantilever surfaces for thyroid system biosensing and beyond. Analytica Chimica Acta 2008;625(1):55-62. |
R832740 (2007) R832740 (2008) R832740 (Final) |
Exit Exit Exit |
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Oran JM, Hinde RJ, Abu-Hatab NA, Retterer ST, Sepaniak MJ. Nanofabricated periodic arrays of silver elliptical discs as SERS substrates. Journal of Raman Spectroscopy 2008;39(12):1811-1820. |
R832740 (2007) R832740 (2008) R832740 (Final) |
Exit |
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Wellman AD, Sepaniak MJ. Magnetically-assisted transport evanescent field fluoroimmunoassay. Analytical Chemistry 2006;78(13):4450-4456. |
R832740 (2006) R832740 (2007) R832740 (2008) R832740 (Final) |
Exit Exit Exit |
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Wellman AD, Sepaniak MJ. Multiplexed, waveguide approach to magnetically assisted transport evanescent field fluoroassays. Analytical Chemistry 2007;79(17):6622-6628. |
R832740 (2007) R832740 (2008) R832740 (Final) |
Exit Exit Exit |
Supplemental Keywords:
MEMS, preconcentration, nuclear receptor protein sensing, microcantilever arrays, SPE-SERS;, RFA, Scientific Discipline, Health, Health Risk Assessment, Endocrine Disruptors - Environmental Exposure & Risk, Microbiology, endocrine disruptors, Biochemistry, Endocrine Disruptors - Human Health, endocrine disruptor screening program, microcantilever transducer evaluation, bioavailability, bioluminescent testing, EDCs, endocrine disrupting chemicals, exposure studiesProgress and Final Reports:
Original AbstractThe perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.