TY - JOUR
T1 - An ultrasonication-assisted extraction and derivatization protocol for GC/TOFMS-based metabolite profiling
AU - Liu, Yumin
AU - Chen, Tianlu
AU - Qiu, Yunpin
AU - Cheng, Yu
AU - Cao, Yu
AU - Zhao, Aihua
AU - Jia, Wei
N1 - Funding Information:
Acknowledgements This study was financially supported by the National Basic Research Program of China (2007CB914700), National Comprehensive Technology Platforms for Innovative Drug R&D (2009ZX09301-007), the National Natural Science Foundation of China (30901997, 20775048), and the Natural Science Foundation of Shanghai in China (10ZR1414800). We thank Jiaqiang Li and Limin Sun for their help and advice.
PY - 2011/5
Y1 - 2011/5
N2 - 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.
AB - 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.
KW - Derivatization
KW - Hypertension
KW - Metabolites
KW - Metabolomics
KW - Ultrasonication
UR - http://www.scopus.com/inward/record.url?scp=79955564618&partnerID=8YFLogxK
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U2 - 10.1007/s00216-011-4880-z
DO - 10.1007/s00216-011-4880-z
M3 - Article
C2 - 21448603
AN - SCOPUS:79955564618
SN - 1618-2642
VL - 400
SP - 1405
EP - 1417
JO - Analytical and Bioanalytical Chemistry
JF - Analytical and Bioanalytical Chemistry
IS - 5
ER -