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Nuclear magnetic resonance spectroscopy is highly sensitive for lipid-soluble metabolites

Neural Regeneration Research 2013年第22期8卷 2103-2110页

作者： Haiyang Dai Bikai Hong Zhifeng Xu Lian Ma Yaowen Chen Yeyu Xiao Renhua Wu Department of Medical Imaging Second Affiliated HospitalShantou University Medical College Department of Medical Imaging Huizhou Municipal Central Hospital Department of Radiology the First People's Hospital of Foshan Department of Pediatrics Second Affiliated HospitalShantou University Medical College Central Laboratory Shantou University

Although the water-soluble metabolite profile of human mesenchymal stem cells is known, the lipid profile still needs further investigation. In this study, methanol-chloroform was used to extract lipid-soluble metabolites and perchloric acid was used to extract water-soluble metabolites. Fur- thermore, a dual phase extraction method using methanol-chloroform and water was used to obtain both water and lipid fractions simultaneously. All metabolite extractions were analyzed on a 9.4T high-resolution nuclear magnetic resonance spectrometer. Metabolite resonance peaks were as- signed in the acquired spectra according to the chemical shift, and the extraction efficiency of dif- ferent methods was compared. Results showed that in the spectra of water-soluble extracts, major metabolites comprised low molecular weight metabolites, including lactate, acetic acid, fatty acids, threonine, glutamic acid, creatine, choline and its derivatives, while in the spectra of lipid-soluble extracts, most metabolites were assigned to fatty acids. Among the different extraction procedures, perchloric acid was more efficient in extracting water-soluble metabolites and methanol-chloroform was efficient in extracting organic components compared with the dual phase extraction method. Nuclear magnetic resonance spectroscopy showed that as low as 0.7 mg organic yield was enough to obtain clear resonance peaks, while about 6.0 mg water-soluble yield was needed to obtain rela- tively favorable spectral lines. These results show that the efficiency of extracting water and lipid fractions is higher using perchloric acid and methanol-chloroform compared with dual phase ex- traction and that nuclear magnetic resonance spectroscopy is highly sensitive for analyzing lipid-soluble extracts.

关键词： neural regeneration nuclear magnetic resonance spectroscopy mesenchymal stem cells metabo-lite profiles extraction method optimization water-soluble lipid-soluble perchloric acid metha-nol-chloroform grants-supported paper neuroregeneration

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