Subject Area物理化学
An Alternative Method for Blood Plasma Preparation Prior to Metabonomics Study
Dong J(董军); Cai XM(蔡晓明); Zhang XL(张秀莉); Zhao LL(赵丽丽); Wang YY(王媛媛); Xue XY(薛兴亚); Liang XM(梁鑫淼)
Conference Name13th International Biotechnology Symposium & Exhibition
Conference Date2008-10-12
Conference Place中国
Alternative Title一种用于代谢组学研究的血浆制备方法
Funding Organization中国科学院、中国工程院、教育部、科技部和国家发改委
AbstractSample preparation for liquid chromatography with mass spectrometric detection is a major task in metabonomics study which significantly impacts assay throughput and data quality. Protein precipitation technology (PPT) is one of the most popular methods for plasma preparation. However, small amount proteins remain in the supernatant, which result in the performance deterioration of chromatographic column and ion suppression in MS detection. The centrifugal ultrafiltration after methanol extraction of whole plasma could be used as an alternative method for blood plasma preparation prior to metabonomics. The Ultra-performance Liquid Chromatography coupled with Q-TOF Mass Spectrometry was used to analyze the prepared sample and the Markerlynx software for processing the raw data. At first, four common using organic solvents (acetonitrile, methanol, ethanol and acetone) were compared in human plasma for protein precipitation to assess method reproducibility and number of reproducible peaks. Methanol has been demonstrated to extract more than 100 metabolites and has the best reproducibility of four organic solvent. The residual protein values pre- and post ultrafiltration after methanol PPT were 4.8 % and 0.5 % of original plasma by using the Bradford assay method. The data also showed that the numbers of peaks before and after ultrafiltration were 4648 and 4150, while the average RSD values for all metabolites were 21.3% and 16.7 %. Most of important, the column pressure was stable after 500 runs, and the sample analysis displayed excellent separation degree. These results showed that methanol protein precipitation followed by centrifugation and then ultrafiltration was an effective, concise, reproducible method, resulting in plasma metabolites containing over 4000 detected compounds peaks. References: [1] Chang, M.S., Ji, Q. Zhang, J., and El-Shourbagy, T.A. 2007. Historical Review of Sample Preparation for Chromatographic Bioanalysis: Pros and Cons. Drug Dev. Res. 68:107–133 [2] Want, E.J., O’Maille, G., Smith, C.A., Brandon, T.R., Uritboonthai, W., Qin, C., Trauger, S.A. and Siuzdak, G. 2006. Solvent-Dependent Metabolite Distribution, Clustering, and Protein Extraction for Serum Profiling with Mass Spectrometry. Anal. Chem. 78: 743-752 [3] Yin, P.Y., Zhao, X.J., Li,Q.R., Wang, J.S., Li, J.S., and Xu, G.W. 2006. Metabonomics Study of Intestinal Fistulas Based on Ultraperformance Liquid Chromatography Coupled with Q-TOF Mass Spectrometry (UPLC/Q-TOF MS). J.Proteome Res. 5: 2135-2143
WOS IDWOS:000208979400122
Citation statistics
Document Type会议论文
Corresponding AuthorLiang XM(梁鑫淼)
Recommended Citation
GB/T 7714
Dong J,Cai XM,Zhang XL,et al. An Alternative Method for Blood Plasma Preparation Prior to Metabonomics Study[C],2008:S54/2.
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