Study Of Sample Preparation Method And Comparison Of Serum Versus Plasma Based On Gas Chromatography-Mass Spectrometry

The effects of different deproteinization and derivatization methods, different storage temperature and time in serum sample prepatation on metabolome were compared by gas chromatography-mass spectrometry. The difference between serum and plasma metabolome were also assessed. The relative standard derivation (RSD) of peak area and information content of profile were calculated. The principal components analysis (PCA), qualitative and quantitative analysis of serum/plasma samples and quality control samples (QC) were done. The conclutions are as follows:1) the reproducibility result demonstrated the robustness of the GC-MS system and reproducibility of preparation procedure.2) The PCA result of profile produced by four preparation methods showed that the influence of derivatization reagents on serum metabolome was great than that of deproteinization reagents.3) The results of different storage conditions in serum sample prepatation showed that sample thawed at both4℃and25℃for up to24h appeared to ensure sample stability. Similarly,48h storage of Nitrogen dried samples at4℃still seemed to keep the sample be stable. The stability of Nitrogen dried samples kept at25℃for up to48h was also assessed, with clear differences appearing after24h. Short-term storage of derivatized samples at4℃for up to48h still provided useable samples. However, apparent effect appeared after10h when kept at25℃, or after undergoing two freeze-thaw cycles when kept at4℃in a freezer.4) Thirty-eight metabolites were obtained in both serum and plasma. In addition, two metabolites were obtained in plasma. Among them, eleven metabolites showed significant difference in content of serum and plasma.