Master Degree / Yüksek Lisans Tezleri

Permanent URI for this collectionhttps://hdl.handle.net/11147/3008

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Subject: search.filters.subject.Plasma spectroscopy

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  • Master Thesis
    Identification and Detection of Cis-Platin Binding Proteins by Laser Induced Breakdown Spectroscopy
    (Izmir Institute of Technology, 2015) Kaya, İbrahim; Yalçın, Şerife; Yalçın, Şerife Hanım
    In this study, an all-optically designed laser plasma spectroscopic technique for rapid identification and detection of cisplatin-binding proteins on electrophoretic gel spots prior to molecular mass spectrometric analysis is demonstrated. For this purpose, human serum albumin, human apo transferrin and horse heart myoglobin standard proteins and protein extracts from HeLa cancer cells were subjected to; incubation with cis-platin solution for several hours. Then, non-reducing polyacrylamide gel electrophoretic separation was applied. Followed by the visualization of proteins in the gel by Coomassie Brilliant Blue staining technique protein spots on the gel were dried between two cellophane sheets and subjected to laser ablation by highly energetic laser pulses. In addition, prior to nr-SDS-PAGE separation cis-platin binding to standard proteins were monitored by ESI-MS with several measurements made in 24 hours of incubation time. Using a Nd:YAG laser at its second harmonic wavelength, 532nm, 10 Hz frequency and 10 ns pulse duration, a micro-plasma was created on dried gel spots. Resulting plasma emission light was collected with collection lenses and transferred to a spectrograph via fiber optic cable. An intensified charge coupled device (ICCD) detector enabled multielemental analysis of platinum binding protein samples. Platinum binding proteins were recognized from the prominent neutral emission line, Pt (I) at 273.3 nm, in a plasma formed by the focused laser pulses on the gel, just in the center or in the vicinity of the electrophoretic spot. Spectral emission intensity of Pt lines from LIBS data has been optimized with respect to laser energy and detector timing parameters. Optimization of LIBS experimental parameters have been studied on polyacrylamide gels soaked in cis-Pt solution for Pt signal. It has been shown that, LIBS is a suitable method for identifying Pt in proteins, in gel medium, with nanogram levels of detection capability. The technique was applied to HeLa (human cervical cancer cells) cells extract for the detection of Pt-binded HSA after standard addition of known amounts of protein.
  • Master Thesis
    Development of Inductively Coupled Plasma Spectroscopic Methods for the Determination of Metals in Beluga (delphinapterus Leucas) and Pygmy Sperm (kogia Breviceps) Whale Liver Samples
    (Izmir Institute of Technology, 2005) Parlayan, Filiz; Eanes, Ritchie C.
    Inductively couple plasma optical emission spectrometry (ICP-OES) is widely used to monitor elements in biological samples from marine organisms for ecological evaluations. Matrix effects (particularly those related to acid and salt type and concentrations) can present a barrier to the applicability of this instrumental method. To have a better understanding of these effects and to choose a suitable internal standard to correct for the signal variations, a procedure based on Principal Component Analysis (PCA) of the data from an axial-mode ICP-OES instrument with sequential detection was performed. Different from other published studies, it was found that ionic lines were more affected by matrix changes. Elements with high ionization energies and energy sums such as Cd and Zn showed a significant change for signal intensities and calculated concentrations due to the presence of acid, salt, and multielement matrix effects. It was observed that acid has a higher influence on the analyte signal as compared to the "salt-only" case. Furthermore, when several interfering elements were present in the sample, the matrix effect was either enhanced or reduced when compared with a solution containing only a single interfering element. Applicability of the proposed technique for the analysis of whale liver homogenate NIST certified material was investigated. The values of the corrected concentrations were in good agreement with the certified values, confirming the capabilities of the selected internal standards for compensation of matrix effects.