An ultrasonication-assisted extraction and derivatization protocol for GC/TOFMS-based metabolite profiling

• Published in: Analytical and bioanalytical chemistry Vol. 400; no. 5; pp. 1405 - 1417
• Main Authors: Liu, Yumin, Chen, Tianlu, Qiu, Yunpin, Cheng, Yu, Cao, Yu, Zhao, Aihua, Jia, Wei
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer-Verlag 01.05.2011; Springer
• Subjects: Aged; Analytical Chemistry; Biochemistry; blood serum; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Chromatographic methods and physical methods associated with chromatography; derivatization; epidemiological studies; Exact sciences and technology; Female; Food Science; Gas chromatographic methods; Gas Chromatography-Mass Spectrometry - economics; Gas Chromatography-Mass Spectrometry - methods; glucosamine; homocysteine; Human; Humans; hypertension; Hypertension - blood; Hypertension - metabolism; Laboratory Medicine; Male; mass spectrometry; Mathematical analysis; Metabolites; metabolomics; Metabolomics - economics; Metabolomics - methods; Middle Aged; Monitoring/Environmental Analysis; Original Paper; patients; Profiling; reference standards; Samples; sampling; Serum - metabolism; Sonication; sorbitol; Spectrometric and optical methods; Statistical analysis; Statistical methods; Time Factors; unsaturated fatty acids; Hypertension; Metabolomics; Ultrasonication; Metabolites; Derivatization; Sonication; Glucosamine; Metabolite; Time; Organic acids; Plant; Accuracy; Sample preparation; Efficiency; Serum; Extraction; Carbohydrate; Peak; Ultrasound; Human; Vegetals; Sample; Use; Marker; Carbonyl; Statistical method; Gas chromatography; Time of flight method; Aminoacid; Animal; Reference material; Unsaturated fatty acid; Large scale; Limit; Mass spectrometry; Comparative study
• ISSN: 1618-2642; 1618-2650; 1618-2650
• DOI: 10.1007/s00216-011-4880-z

Conventional chemical derivatization of metabolites in biological specimens is time-consuming, which limits the throughput and efficiency of metabolite profiling using a gas chromatography/time-of-flight mass spectrometry (GC/TOFMS) platform. We report an ultrasonication-assisted protocol which reduces the derivatization time from hours to about 30 min and significantly enhances the derivatization efficiency prior to a GC/TOFMS analysis. The protocol was evaluated using 40 compounds representing different classes of human metabolites, and demonstrated good analytical precision and accuracy. In comparison with the conventional method, the new protocol was able to increase the intensity of most of the identified peaks (71.0%) in the GC/TOFMS chromatograms of human serum samples. The detected compounds with increased intensity include most amino acids, keto-containing organic acids, carbonyl-containing carbohydrates, and unsaturated fatty acids. We applied this protocol in a metabolomic study of human serum samples obtained from 34 patients diagnosed with hypertension and 29 age- and gender-matched healthy subjects. Metabolite markers associated with hypertension, including glucosamine, d -sorbitol, 1-stearoylglycerol, and homocysteine, were identified and validated by statistical methods and use of reference standards. Our work highlights the potential of this novel approach for the large-scale metabolite profiling of samples generated from plant, animal, and clinical and epidemiological studies.