• Login
    View Item 
    •   Home
    • Theses and Dissertations
    • Theses and Dissertations
    • View Item
    •   Home
    • Theses and Dissertations
    • Theses and Dissertations
    • View Item
    JavaScript is disabled for your browser. Some features of this site may not work without it.

    Browse

    All of TUScholarShareCommunitiesDateAuthorsTitlesSubjectsGenresThis CollectionDateAuthorsTitlesSubjectsGenres

    My Account

    LoginRegister

    Help

    AboutPeoplePoliciesHelp for DepositorsData DepositFAQs

    Statistics

    Most Popular ItemsStatistics by CountryMost Popular Authors

    DEVELOPMENT OF NOVEL METHODS FOR THE RAPID SEPARATION OF BIOMOLECULES

    • CSV
    • RefMan
    • EndNote
    • BibTex
    • RefWorks
    Thumbnail
    Name:
    Mamunooru_temple_0225E_11315.pdf
    Size:
    2.657Mb
    Format:
    PDF
    Download
    Genre
    Thesis/Dissertation
    Date
    2013
    Author
    Mamunooru, Manasa
    Advisor
    Stanley, Robert J.
    Committee member
    Spano, Francis C.
    Nicholson, Allen W.
    Nicholson, Rhonda H.
    Department
    Chemistry
    Subject
    Chemistry
    Chemistry, Analytical
    Amino Acids
    Capillary Electrophoresis
    Dna
    Dna Damage and Repair
    Hplc
    Method Development
    Permanent link to this record
    http://hdl.handle.net/20.500.12613/1822
    
    Metadata
    Show full item record
    DOI
    http://dx.doi.org/10.34944/dspace/1804
    Abstract
    Successful methods for the separation of biomolecules like amino acids, proteins, peptides, and DNA have been developed previously using HPLC, GC, GC-MS, and CE. Recently CE has become a routine laboratory technique in the analysis of biological molecules. Even though high-resolution separations with small sample volumes is the main advantage, CE is limited by lower sensitivity detection of analytes when universal detectors like UV absorption or refractive index detectors are used. Therefore, sensitivity enhancement can be obtained by either using different detection schemes or electrophoretically based pre- or on-line concentration methods. These can be grouped into two categories. The first category includes IEF, CGF or TGF where sensitivity is achieved through equilibrium electrofocusing. In these methods, electrophoresis and bulk solution is combined in the capillary or separation column to form a null velocity point, a point at which the net velocity of the analyte is zero. Using these methods 10-10,000 fold sensitivity enhancement is achieved. The second category uses velocity gradients but not the nul velocity for the enrichment of samples. These methods include FASS, LVSS, NSM, etc., which are applied for the analysis of small molecules, and 10-10,000 fold sensitivity enhancement is reported by using these methods. In this work, first GEITP an on-line preconcentration technique is applied for the detection of amino acids (using Trp and Tyr as model analytes). This work also established the effects of different parameters on enrichment. The parameters studied include effect of current flow acceleration across capillary inner diameter, the effect of leading electrolyte (LE) concentration on current density, and the effect of applied electric fields on the current density. To explore the application of GEITP in biological fluids, optimized parameters were developed for the detection and separation of Trp and Tyr in artificial cerebrospinal fluid (aCSF). Next, GEITP was applied for enrichment and separation of physiologically relevant concentrations of chromophore-derivatized Asp and Glu in high conductivity samples like artificial cerebrospinal fluid (aCSF). It was concluded from this work that the major factors which influence the enrichment is the ratio of current density to sample conductivity. Finally, GEITP is applied as a prior step before CZE to increase the resolution between analytes without using ampholyte mixtures. In this method GEITP was combined to CZE to achieve resolution adjustment between amino acids mixture using low pressure hydrodynamic flow during CZE without changing the separation column, field strength, or electrolyte system. In this work, a rapid CE method for extraction and analysis of amino acids in planarians, labeled with 4-Fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), was developed. This method was applied to detect the changes in the levels of amino acids when planarians were fed and starved. This method can be applied to study pharmacological effects in planaria, as it can monitor different amino acid levels with respect to feeding. Finally, ssDNA photoproducts of different lengths (11-mer and 63-mer) were separated using two different matrices, a traditional C18 and a PV/DBS (PLRP-S) matrix. A faster separation (within ~ 10 mins) was achieved for a 11-mer by the PLRP-S column. A separation was achieved in the PLRP-S column for the 63-mer while there was no separation in C18 column. Baseline resolution was not achieved. Therefore, C18 can best be used for small length DNA while PLRP-S can be applied for longer length DNA, as it is more hydrophobic than C18 column. Parameters can still be optimized for a baseline separation.
    ADA compliance
    For Americans with Disabilities Act (ADA) accommodation, including help with reading this content, please contact scholarshare@temple.edu
    Collections
    Theses and Dissertations

    entitlement

     
    DSpace software (copyright © 2002 - 2023)  DuraSpace
    Temple University Libraries | 1900 N. 13th Street | Philadelphia, PA 19122
    (215) 204-8212 | scholarshare@temple.edu
    Open Repository is a service operated by 
    Atmire NV
     

    Export search results

    The export option will allow you to export the current search results of the entered query to a file. Different formats are available for download. To export the items, click on the button corresponding with the preferred download format.

    By default, clicking on the export buttons will result in a download of the allowed maximum amount of items.

    To select a subset of the search results, click "Selective Export" button and make a selection of the items you want to export. The amount of items that can be exported at once is similarly restricted as the full export.

    After making a selection, click one of the export format buttons. The amount of items that will be exported is indicated in the bubble next to export format.