GC-MS with ethyl chloroformate derivatization for comprehensive analysis of metabolites in serum and its application to human uremia.
An optimized method based on GC-MS with ethyl chloroformate derivatization has been developed for the comprehensive analysis of endogenous metabolites in serum. Twenty-two reference standards and serum samples were used to validate the proposed method. The correlation coefficient was higher than 0.9900 for each of the standards, and the LOD varied from 125 to 300 pg on-column. The analytical equipment exhibited good repeatability (RSD<10%) for all of the standards. Both the repeatability and the within-48-h stability of the analytical method were satisfactory (RSD<10%) for the 18 metabolites identified in the serum samples. Mean recovery was acceptable for the 18 metabolites, ranging from 70% to 120% with RSDs of less than 10%. Using the optimized protocol and a subsequent multivariate statistical technique, complete differentiation was achieved between the metabolic profile of uremic patients and that of age- and sex-matched normal subjects. Significantly decreased levels of valine, leucine, and isoleucine and increased levels of myristic acid and linoleic acid were observed in the patient group. This work demonstrated that this method is suitable for serum-based metabolic profiling studies.
Tao
Xiumei
X
School of Pharmacy, Shanghai Jiao Tong University, Shanghai, 200240, China.
Liu
Yumin
Y
Wang
Yihuang
Y
Qiu
Yunping
Y
Lin
Jingchao
J
Zhao
Aihua
A
Su
Mingming
M
Jia
Wei
W
D005563:Formic Acid Esters / Q000737:chemistry*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D007532:Isoleucine / Q000097:blood; D007930:Leucine / Q000097:blood; D019787:Linoleic Acid / Q000097:blood; D019814:Myristic Acid / Q000097:blood; D012015:Reference Standards; D015203:Reproducibility of Results; D014511:Uremia / Q000097:blood* / Q000175:diagnosis / Q000378:metabolism*; D014633:Valine / Q000097:blood
D005563:Formic Acid Esters; D007532:Isoleucine; C007658:ethyl chloroformate; D019814:Myristic Acid; D019787:Linoleic Acid; D007930:Leucine; D014633:Valine