小鼠粪便中短链脂肪酸提取与检测方法的建立及应用

Establishment and Application of Extraction and Determination Method for Short-Chain Fatty Acids in Feces of Mice

目的:建立小鼠粪便中三种短链脂肪酸(乙酸、丙酸、丁酸)的检测方法及其应用。方法:使用水提法和乙醚萃取法提取小鼠粪便中的短链脂肪酸(SCFA)通过高效液相色谱(HPLC)评价两种方法的提取效果,并建立短链脂肪酸的HPLC定量检测方法并进行方法学考察,利用上述建立的体系评价新琼寡糖对小鼠短链脂肪酸合成的影响。结果:确定了SCFA的乙醚萃取方法和基于HPLC的定量检测方法,被测组分浓度与其峰面积呈良好线性关系,各组分测定的相对标准偏差在1.31%~2.61%之间,平均加标回收率为86.6%~105.8%之间;对喂饲新琼寡糖小鼠粪便中的SCFA检测显示,较对照组乙酸和丙酸含量提高了1倍,丁酸含量提高了0.45倍。结论:建立了小鼠粪便中SCFA的提取和定量检测方法,并利用该方法证明了新琼寡糖能促进小鼠短链脂肪酸的合成。

Objective：A rapid and sensitive method was developed and applied for the extraction and determination of three short-chain fatty acid（SCFA） in feces.Methods：SCFAs were purified by aqueous or diethyl ether extraction of feces.HPLC was used to evaluate the purification method and to quantitative analyze of SCFA.At last,the method was applied to investigate the effects of neoagaro-oligosaccharides（NAOS） on SCFA production in mice.Results：Diethyl ether extraction was proved to be better for purifying of SCFA from feces.The detection results showed that acetic acid,propionic acid and n-butyric acid all had a good liner relationship,the standard deviation was 1.31%~2.61%,and the recovery rate was 86.6%~ 105.8%.After feeding of NAOS for 7 days,the feces were collected to monitor the amount of SCFA,and the results demonstrated that the concentrations of acetic acid and propionic acid were increased forl-fold by comparing with the control group,and n-butyric was increased for 0.45-fold.Conclusions：The method for extraction and quantification of SCFAs was established,and NAOS was proved can improve the synthesis of SCFAs in mice.