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Dissertation_Sujatha Chilakala.pdf (2.07 MB)

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DEVELOPMENT OF LIQUID CHROMATOGRAPHY-MASS SPECTROMETRIC ASSAYS AND SAMPLE PREPARATION METHODS FOR THE BIOLOGICAL SAMPLE ANALYSIS

CHILAKALA, SUJATHA
http://rave.ohiolink.edu/etdc/view?acc_num=csu1512927043412916

Abstract Details

2017, Doctor of Philosophy in Clinical-Bioanalytical Chemistry, Cleveland State University, College of Sciences and Health Professions.
The area of biosample analysis encompasses a very broad range of assays which support the clinical and nonclinical studies. Biosample analysis is used to provide a quantitative or qualitative measure of the active drug and/or its metabolite(s) in the biological matrix for the purpose of pharmacokinetics, toxicokinetics, bioequivalence, and exposure-response (pharmacokinetics /pharmacodynamics) studies. Due to the significance of pharmacological analysis, sensitive, reproducible and robust analytical methods are critically needed for pharmacological studies of the biosamples. A bioanalytical method mainly contains two components I) Sample preparation II) detection of the compound. Therefore, the main aims of this thesis are the development of quantitative and qualitative analytical methods for the target compounds using LC-MS(/MS) and development of accelerated sample preparation for high throughput sample analysis for DNA and proteins. In this dissertation, a brief review on the method rationale, workflow of the method development, sample preparation methods, instrumentations and analytical method validation, are discussed in Chapter 1. Also, research projects were discussed and the techniques used in the experiments for this thesis were reviewed. As so, chapter II and III were mainly focused on the accelerated sample preparation methods for the high throughput sample analysis of DNA and proteins respectively, where the sample preparation time was significantly reduced from hours to minutes, which are suitable for qualitative and quantitative analysis of DNA and proteins. In Chapter IV, a systematic study on the structural characterization of the model glycoprotein Human IgG was described. In chapter V successful development of LC-MS method was developed for the determination of Oxygen -18 isotope enrichment in the phosphate samples in the positional isotope exchange reactions to study the reversibility of certain enzymatic reactions was described. Successful development and validation of a new and sensitive analytical LC-MS/MS method for the determination and quantitation of incorporation rates of decitabine, an anti-cancer drug which can be applied to determine the sensitivity and responsiveness in patients treated with decitabine was described in Chapter VI.
Yan Xu, Ph.D. (Committee Chair)
David Anderson, Ph.D. (Committee Member)
Bin Su, Ph.D. (Committee Member)
Aimin Zhou, Ph.D. (Committee Member)
Petru Fodor, Ph.D. (Committee Member)
177 p.
Chemistry
DNA digestion; Protein digestion; Peptide extraction; Oxygen-labelled phosphate analysis; Decitabine incorporation; DNA hypomethylation

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Citations

  • CHILAKALA, S. (2017). DEVELOPMENT OF LIQUID CHROMATOGRAPHY-MASS SPECTROMETRIC ASSAYS AND SAMPLE PREPARATION METHODS FOR THE BIOLOGICAL SAMPLE ANALYSIS [Doctoral dissertation, Cleveland State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=csu1512927043412916

    APA Style (7th edition)

  • CHILAKALA, SUJATHA. DEVELOPMENT OF LIQUID CHROMATOGRAPHY-MASS SPECTROMETRIC ASSAYS AND SAMPLE PREPARATION METHODS FOR THE BIOLOGICAL SAMPLE ANALYSIS . 2017. Cleveland State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=csu1512927043412916.

    MLA Style (8th edition)

  • CHILAKALA, SUJATHA. "DEVELOPMENT OF LIQUID CHROMATOGRAPHY-MASS SPECTROMETRIC ASSAYS AND SAMPLE PREPARATION METHODS FOR THE BIOLOGICAL SAMPLE ANALYSIS ." Doctoral dissertation, Cleveland State University, 2017. http://rave.ohiolink.edu/etdc/view?acc_num=csu1512927043412916

    Chicago Manual of Style (17th edition)

csu1512927043412916
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This open access ETD is published by Cleveland State University and OhioLINK.