Nuclear magnetic resonance spectroscopy is highly sensitive for lipid-soluble metabolites.
Nuclear magnetic resonance spectroscopy is highly sensitive for lipid-soluble metabolites.
复制标题
DOI：
10.3969/j.issn.1673-5374.2013.22.010
复制
发表时间：
2013-08-05
影响因子：
6.1
通讯作者：
Wu R
中科院分区：
文献类型：
--
作者：
Dai H;Hong B;Xu Z;Ma L;Chen Y;Xiao Y;Wu R
尽管人间充质干细胞的水溶性代谢物谱已知，但脂质谱仍需要进一步研究。本研究采用甲醇-氯仿提取吡啶溶性代谢物，采用高氯酸提取水溶性代谢物。此外，采用甲醇-氯仿和水的双相萃取方法同时获得水和脂质组分。所有代谢物提取物均在 9.4T 高分辨率核磁共振波谱仪上进行分析。根据化学位移在获得的光谱中分配代谢物共振峰，并比较不同方法的提取效率。结果表明，水溶性提取物谱中，主要代谢物为低分子量代谢物，包括乳酸、乙酸、脂肪酸、苏氨酸、谷氨酸、肌酸、胆碱及其衍生物，而脂溶性提取物谱中，大部分代谢物为脂肪酸。在不同的提取方法中，与双相提取方法相比，高氯酸提取水溶性代谢物的效率更高，甲醇-氯仿提取有机成分的效率更高。核磁共振波谱显示，低至0.7 mg的有机收率就足以获得清晰的共振峰，而约6.0 mg的水溶性收率则需要获得相对良好的谱线。这些结果表明，与双相提取相比，使用高氯酸和甲醇-氯仿提取水和脂质组分的效率更高，并且核磁共振波谱对于分析脂溶性提取物高度敏感。
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 pid-soluble metabolites and perchloric acid was used to extract water-soluble metabolites. Furthermore, 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 assigned in the acquired spectra according to the chemical shift, and the extraction efficiency of 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 relatively 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 extraction and that nuclear magnetic resonance spectroscopy is highly sensitive for analyzing lipid-soluble extracts.
登录
查看更多内容
DOI：
10.12659/msm.882112
发表时间：
2011-12
期刊：
Medical science monitor : international medical journal of experimental and clinical research
影响因子：
--
作者：
Walecki J;Barcikowska M;Ćwikła JB;Gabryelewicz T
通讯作者：
Gabryelewicz T
影响因子：
2.6
作者：
Loewenbrueck, Kai F.;Fuchs, Beate;Storch, Alexander
通讯作者：
Storch, Alexander
影响因子：
2.9
作者：
Martínez-Bisbal, MC;Martí-Bonmatí, L;Celda, B
通讯作者：
Celda, B
影响因子：
2.9
作者：
Le Belle, JE;Harris, NG;Bhakoo, KK
通讯作者：
Bhakoo, KK
影响因子：
3.3
作者：
Tyagi, RK;Azrad, A;Salomon, Y
通讯作者：
Salomon, Y