2008 Progress Report: Integrated Microfluidic System for Bioluminescent Bioreporting, Separations, Vibrational Spectroscopy, and Microcantilever Transducer Evaluation of Endocrine Disrupting Chemicals

EPA Grant Number: R832740
Title: 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 - Knoxville
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, 2007 through October 31,2008
Project Amount: $590,240
RFA: Exposure Measurement Tools for Endocrine Disrupting Chemicals in Mixtures (2005) RFA Text |  Recipients Lists
Research Category: Economics and Decision Sciences , 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:

Task 1. With advanced substrates, test known EDCs in relevant matrices for SERS response characteristics.
Surveys of SERS responses of many EDCs have been made that are distinctive but inadequate sensitivity and cost issues have limited success. We have address this with a novel approaches to nanofabrication (most notably nanotransfer printing) of substrates and with integrating with μ-fluidics & on-line concentrators.

Task 2. Develop EDC nanomechanical response signatures on functionalized microcantilever arrays.
Nanomechanical sensing using MCAs with conventional (organic coatings) responsive phases lacked desired sensitivity and selectivity. This issue has been successful addressed in years 2 & 3 with endocrine receptor proteins as selective and sensitive responsive phases. Both estrogen and thyroid receptor phases have been shown to function with the nanomechanical sensing approach for screening studies.

Task 3. Integrate bioreporter elements and conditions onto μfluidic platforms with electrophoretic separations (including extraction/concentration) and detection of EDCs. Capillary electrophoretic separations of representative EDCs have been performed and coupled with fluorescence detection and SERS. A novel lab-on-achip platform is being pursued in Year 3. Bioreporting elements (receptor proteins) have been included in MCA systems (see above) and we are developing MCAs with multiple receptors. An extensive bioluminescence bioreporting screening study has been conducted by the Co-PI’s group.

Task 4. Validate integrated μ−fluidic devices that incorporates combinations of these analytical components for EDC-containing samples. Integration has progressed relatively slowly do to status of 1-3. However, validation of integrated systems has been made with separations with SERS, bioreporter elements (receptor proteins) with nanomechanics on MCs, and integrating extraction/concentration with these techniques.

Future Activities:

  • Task 1. Pursue advanced better performing substrates created by sophisticated nano-lithographic techniques, as well as integrating micro-pre-concentration to enhance detection sensitivity.
  • Task 2. Further optimization of receptor protein immobilization protocols and research the creation of true arrays with multiple types of receptors to simultaneously screen both EDC and TDC as well as PXR.
  • Task 3. Couple these techniques more intimately, both with conventional separation capillaries and on-chip. In addition, sample pre-concentration protocols that have been developed and will be integrated.
  • Task 4. We will perform an analytical validation of the performance of different integrated techniques (see above) in the analysis of EDC mixtures.
  • 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 final report as well as final report upon completion of no-cost extension.


Journal Articles on this Report : 17 Displayed | Download in RIS Format

Other project views: All 53 publications 30 publications in selected types All 30 journal articles
Type Citation Project Document Sources
Journal Article 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)
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  • Abstract from PubMed
  • Abstract: Sage-Abstract
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  • Journal Article Abu-Hatab NA, Oran JM, Sepaniak MJ. Surface-enhanced Raman spectroscopy substrates created via electron beam lithography and nanotransfer printing. ACS Nano 2008;2(2):377-385. R832740 (2008)
    R832740 (Final)
  • Abstract from PubMed
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  • Abstract: ACS-Abstract
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  • Other: ACS-Full Text PDF
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  • Journal Article 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)
  • Abstract from PubMed
  • Full-text: ScienceDirect-Full Text HTML
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  • Abstract: ScienceDirect-Abstract
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  • Other: ScienceDirect-Full Text PDF
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  • Journal Article Bhandari D, Walworth MJ, Sepaniak MJ. Dual function surface-enhanced Raman active extractor for the detection of environmental contaminants. Applied Spectroscopy 2009;63(5):571-578. R832740 (2008)
    R832740 (Final)
  • Abstract from PubMed
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  • Journal Article 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)
  • Abstract from PubMed
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  • Abstract: ACS-Abstract
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  • Journal Article 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 (2008)
    R832740 (Final)
  • Abstract from PubMed
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  • Abstract: ACS-Abstract
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  • Journal Article Connatser RM, Cochran M, Harrison RJ, Sepaniak MJ. Analytical optimization of nanocomposite surface-enhanced Raman spectroscopy/scattering detection in microfluidic separation devices. Electrophoresis 2008;29(7):1441-1450. R832740 (2008)
    R832740 (Final)
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  • Abstract: Wiley-Abstract
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  • Journal Article De Jesus MA, Giesfeldt KS, Oran JM, Abu-Hatab NA, Lavrik NV, Sepaniak MJ. Nanofabrication of densely packed metal-polymer arrays for surface-enhanced Raman spectrometry. Applied Spectroscopy 2005;59(12):1501-1508. R832740 (2008)
  • Abstract from PubMed
  • Abstract: IngentaConnect-Abstract
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  • Journal Article Dutta P, Chapman PJ, Datskos PG, Sepaniak MJ. Characterization of ligand-functionalized microcantilevers for metal ion sensing. Analytical Chemistry 2005;77(20):6601-6608. R832740 (2008)
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  • Abstract: ACS Publications-Abstract
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  • Journal Article Dutta P, Sanseverino J, Datskos PG, Sepaniak MJ. Nanostructured cantilevers as nanomechanical immunosensors for cytokine detection. NanoBiotechnology 2005;1(3):237-244. R832740 (2008)
  • Abstract: SpringerLink-Abstract
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  • Journal Article 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)
  • Abstract from PubMed
  • Full-text: National Tsing Hua University-Full Text PDF
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  • Abstract: RSC Publishing-Abstract
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  • Journal Article 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)
  • Abstract from PubMed
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  • Abstract: ScienceDirect-Abstract
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  • Journal Article 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)
  • Abstract: Wiley-Abstract
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  • Journal Article Sanseverino J, Eldridge ML, Layton AC, Easter JP, Yarbrough J, Schultz TW, Sayler GS. Screening of potentially hormonally active chemicals using bioluminescent yeast bioreporters. Toxicological Sciences 2009;107(1):122-134. R832740 (2008)
    R832740 (Final)
  • Abstract from PubMed
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  • Abstract: OUP-Abstract & Full Text HTML
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  • Journal Article Senesac LR, Dutta P, Datskos PG, Sepaniak MJ. Analyte species and concentration identification using differentially functionalized microcantilever arrays and artificial neural networks. Analytica Chimica Acta 2006;558(1-2):94-101. R832740 (2008)
  • Full-text: ScienceDirect-Full Text HTML
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  • Abstract: Science Direct
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  • Journal Article 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)
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  • Abstract: ACS-Abstract
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  • Journal Article 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)
  • Abstract from PubMed
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  • Abstract: ACS-Abstract
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  • Supplemental Keywords:

    MEMS, preconcentration, nuclear receptor protein sensing, microcantilever arrays, SPE-SERS, RFA, Health, Scientific Discipline, Health Risk Assessment, Endocrine Disruptors - Environmental Exposure & Risk, endocrine disruptors, Microbiology, Biochemistry, Endocrine Disruptors - Human Health, endocrine disruptor screening program, microcantilever transducer evaluation, bioavailability, EDCs, endocrine disrupting chemicals, exposure studies, bioluminescent testing

    Relevant Websites:

    http://www.chem.utk.edu/~sepaniak/ Exit

    Progress and Final Reports:

    Original Abstract
  • 2006 Progress Report
  • 2007 Progress Report
  • Final Report