大鼠肾组织中小分子代谢物的GC-MS分析

摘要

Objective:To establish a method to determine the metabolites in rat kidney tissues by gas chromatography-mass spectrometry (GC-MS) combined with chemometric techniques. Methods:Metabolites were separated and identiifed on HP-5MS column (30 m × 0.25 μm × 0.25 mm). The initial column temperature was 100℃lasting 3 min, and then programmed at 8℃/min to 300℃, maintaining at this temperature for 6 min. hTe internal standard was heptadecanoic acid. hTe grinded kidney tissue was exacted by methanol. hTe supernatant was dried by nitrogen. Atfer the oximation and derivation, the supernatant was analyzed by GC-MS. hTe overlapped peaks were resolved into pure chromatogram and mass spectra with chemometric techniques. Qualitative analysis was performed by comparing the obtained pure mass spectra with those in NIST mass spectra database and certiifcated by the standards and the references. hTe internal method was used for semi-quantitation. Results:A total of 53 compounds were identiifed. hTe main constitutions in the kidney tissue were amino acids, saccharides, fatty acids and urea. Conclusion:hTe combination of methods is rapid and accurate for the analysis of metabolites in the kidney tissue, which provides more information for further study of metabonomics in kidney tissues.%目的：采用气相色谱-质谱法(gas chromatography-mass spectrometry，GC-MS)结合化学计量学分析测定大鼠肾组织中小分子代谢物。方法：色谱柱为HP-5MS石英毛细管柱(30 m ×0.25μm ×0.25 mm)，初始柱温为100℃，保持3 min，程序升温为8℃/min至300℃，保持6 min。肾脏组织绞碎后加甲醇萃取，十七酸为内标，离心分离得上清液，氮气吹干，再用甲氧胺吡啶肟化、衍生化试剂衍生后进行GC-MS分析，对重叠色谱峰采用化学计量学分辨，得各化合物的纯色谱与光谱曲线，通过质谱库检索定性化合物，同时结合标准品并参考文献确定化合物结构，用内标法对各成分进行半定量。结果：共鉴定出53种成分，肾组织中主要代谢物成分为尿素、氨基酸、糖类与脂肪酸。结论：GC-MS辅以化学计量学法相比单用GC-MS分析，结果更准确可靠，将为采用代谢组学技术研究肾脏病变分子标记物提供基础。