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Application of MALDI-Orbitrap-MS in the Lipidomic Analysis of Renal Cell Carcinoma Samples
Authors: Jirásko Robert | Holčapek Michal | Vrána David | Študent Vladimír | Melichar Bohuslav
Year: 2015
Type of publication: ostatní - přednáška nebo poster
Page from-to: nestránkováno
Titles:
Language Name Abstract Keywords
eng Application of MALDI-Orbitrap-MS in the Lipidomic Analysis of Renal Cell Carcinoma Samples The aim of this work was the optimization and validation of MALDI-Orbitrap mass spectrometry in the analysis of renal cell carcinoma (RCC) samples. RCC is the most common type of kidney cancer in adults and it accounts for approximately 3% of adult malignancies and 90-95% of kidney neoplasms. Early diagnosis markedly increases RCC cure rates. The clarification of cancer biochemistry or discovery of cancer biomarkers for early cancer diagnosis requires detailed analytical characterization of tumor and normal tissue samples using reliable analytical techniques. In the present study, the ultrahigh-resolution MALDI mass spectrometer LTQ Orbitrap XL (Thermo Fisher Scientific, Waltham, MA, USA) equipped with the nitrogen UV laser (337 nm, 60 Hz) was used for the analyses of RCC samples. The instrument was operated in negative-ion mode in the normal mass range (m/z 350 - 2000). 9-Aminoacridine was selected as the best matrix candidate based on the previous comparison of various matrices and nanostructured materials with regard to best reproducibility of uniform spotting and number of detected lipids. More than 500 lipids from 18 lipid classes were found in RCC samples with the help of high mass accuracy and tandem mass spectra measurements and were added to our RCC lipid database. The nonendogenous internal standards consisted of either shorter fatty acid chains (12:0, 14:0) or odd-carbon fatty acyls were used per each lipid class and added to the RCC sample before the extraction. The method was validated for linearity, dynamic range, LOD, LOQ, matrix effect, inter/intra-day precision, accuracy and recovery rate for each lipid class. Finally, the mass spectrometry imaging of tumor and autologous normal tissues was performed and compared with H&E histological staining. The results indicate the potential of MALDI-Orbitrap-MS for the quantitative lipidomic analysis of RCC samples and open the way for the investigation of lipidomic biomarkers for early RCC diagnosis. Renal cell carcinoma; Lipidomic analysis; MALDI; Mass spectrometry; Orbitrap

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