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GRADIENT CHROMATOFOCUSING AND REVERSED PHASE HPLC IN PROTEIN ANALYSIS AND LC-MS/MS DETERMINATION OF THIAZOLIDINEDIONE NL-1 IN MOUSE SERUM AND BRAIN

Pedada, Kiran K, Mr
http://rave.ohiolink.edu/etdc/view?acc_num=csu1390322545

Abstract Details

2013, Doctor of Philosophy in Clinical-Bioanalytical Chemistry, Cleveland State University, College of Sciences and Health Professions.
Gradient chromatofocusing (GCF) is a technique developed using linear pH gradients on weak-anion exchange HPLC columns. GCF greatly extends the capabilities of chromatofocusing by overcoming the shortcomings of the conventional chromatofocusing technique. GCF has been interfaced to mass spectrometry in protein separations. Chapter 1 demonstrates the advantage of GCF technique over reversed-phase (RP) HPLC in protein analysis. All RP protein peaks were eluted in a narrower region (40 – 60% organic in a gradient) of the gradient compared to GCF, in which the proteins peaks were distributed along the entire length of the gradient due their differences in iso-electric points. Regarding peak widths, some had narrower widths with RP, others with GCF. The median, average peak widths for both the techniques were similar, however, overall, GCF is advantageous when compared with RP technique in the separation of proteins. Chapter 2 demonstrates a new procedure for characterizing weak anion-exchange (WAX) HPLC columns. Peak figures-of-merit results from injection of the 100 µM NaNO3 standard were compared to that of injections toluene recommended by the manufacturer (20 mM toluene) has reveled that the peak width increases by 50% for the NaNO3 while the increase in toluene peak width is 30%. And, the shift in the retention time of 100 µM NaNO3 from 3.17 mins to 1.52 mins beginning and after extensive use respectively has shown that 52% of function ion-exchange sites has been lost after an extensive usage. Chapter 3, 4 and 5 demonstrates developing a quantitative LC-MS/MS method for determination of thiazolidinedione (TZD) mitoNEET ligand NL-1 in mouse serum and its pharmacokinetic applications. A sensitive LC-MS/MS assay has been developed and validated for quantification of NL-1 {5-[(3,5-di-tert-butyl-4-hydroxyphenyl)methyl]-1,3-thiazolidine-2,4-dione} in mouse serum. The method is suitable for pharmacokinetic (PK) studies of the parent drug NL-1 based on the preliminary serum results from dosed NL-1 mouse studies. The key pharmacokinetic parameters of NL-1 in its serum and brain concentration –were shown good penetration of drug into mice brain, More extensive metabolite studies should be performed as a part of future studies in order to investigate the fate of the analyte.
David Anderson , PhD (Committee Chair)
Aimin Zhou, PhD (Committee Member)
Xue-Long Sun, PhD (Committee Member)
Xiang Zhou, PhD (Committee Member)
Masaru Miyagi, PhD (Committee Member)
Analytical Chemistry; Chemistry

Recommended Citations

Citations

  • Pedada, K. K. (2013). GRADIENT CHROMATOFOCUSING AND REVERSED PHASE HPLC IN PROTEIN ANALYSIS AND LC-MS/MS DETERMINATION OF THIAZOLIDINEDIONE NL-1 IN MOUSE SERUM AND BRAIN [Doctoral dissertation, Cleveland State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=csu1390322545

    APA Style (7th edition)

  • Pedada, Kiran. GRADIENT CHROMATOFOCUSING AND REVERSED PHASE HPLC IN PROTEIN ANALYSIS AND LC-MS/MS DETERMINATION OF THIAZOLIDINEDIONE NL-1 IN MOUSE SERUM AND BRAIN . 2013. Cleveland State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=csu1390322545.

    MLA Style (8th edition)

  • Pedada, Kiran. "GRADIENT CHROMATOFOCUSING AND REVERSED PHASE HPLC IN PROTEIN ANALYSIS AND LC-MS/MS DETERMINATION OF THIAZOLIDINEDIONE NL-1 IN MOUSE SERUM AND BRAIN ." Doctoral dissertation, Cleveland State University, 2013. http://rave.ohiolink.edu/etdc/view?acc_num=csu1390322545

    Chicago Manual of Style (17th edition)

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