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UPLC-Q-TOF/MS based Untargeted Metabolite and Lipid Analysis on Premature Ovarian Insufficiency Plasma Samples.

Author(s):

Yasemin Taşcı, Rahime Bedir Fındık, Meryem Kuru Pekcan, Ozan Kaplan and Mustafa Çelebier*  

Abstract:


Background: Metabolomics is one of the main areas to understand cellular process at molecular level by analyzing metabolites. In recent years metabolomics has been emerged as key tool to understand molecular basis of disease, find diagnostic and prognostic biomarkers, and develop new treatment opportunities and drug molecules.

Objective: In this study, an untargeted metabolite and lipid analysis were performed to identify potential biomarkers on premature ovarian insufficiency plasma samples. 43 POI subject plasma samples were compared with 32 healthy subject plasma samples.

Methods: Plasma samples were pooled and extracted using chloroform:methanol:water (3:3:1 v/v/v) mixture. Agilent 6530 LC/MS Q-TOF instrument equipped with ESI source was used for analysis. A C18 column (Agilent Zorbax 1.8 μM, 50 x 2.1 mm) was used for separation of metabolites and lipids. XCMS, an “R software” based freeware program, was used for peak picking, grouping and comparing the findings. Isotopologue Parameter Optimization (IPO) software was used in order to optimize XCMS parameters. The analytical methodology and data mining process were validated according to the literature.

Results: 83 metabolite peaks and 213 lipid peaks were found to be in semi-quantitatively and statistically different (fold change >1.5, p <0.05) between the POI plasma samples and control subjects.

Conclusion: According to the results, two groups were successfully separated through principal component analysis. Among the peaks, phenyl alanine, decanoyl-L-carnitine, 1-palmitoyllysophosphatidylcholine and PC(O-16:0/2:0) were identified through auto MS/MS and matched with human metabolome database and proposed as plasma biomarker for POI and monitoring the patients in treatment period.

Keywords:

Liquid Chromatography Mass Spectrometry, Ultra Performance Liquid Chromatography, Metabolomics, Lipidomics, Premature Ovarian Insufficiency

Affiliation:

University of Health Sciences, Zekai Tahir Burak Women’s Health Research Hospital, University of Health Sciences, Zekai Tahir Burak Women’s Health Research Hospital, University of Health Sciences, Zekai Tahir Burak Women’s Health Research Hospital, Hacettepe University Faculty of Pharmacy Department of Analytical Chemistry, Hacettepe University Faculty of Pharmacy Department of Analytical Chemistry



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