气相色谱法测定猪结肠内容物中短链脂肪酸含量

A Determination Method Based on Gas Chromatography for Analysis of Short-chain Fatty Acids in Colonic Contents of Piglet

目的：为了准确测定猪结肠内容物中蛋白质代谢物短链脂肪酸含量以研究猪结肠蛋白质代谢情况,本文拟利用气相色谱建立一种更准确快速分析猪结肠内容物中短链脂肪酸含量的方法。方法：实验选用体重相近的21日龄断奶的杜长大三元杂交仔猪,单栏饲养。试猪麻醉后放血致死,迅速剖开腹腔,收集结肠末端内容物,保存于-80℃。收集到的结肠末端内容物用偏磷酸预处理,利用毛细管气相色谱法程序升温测定猪结肠内容物中短链脂肪酸含量。结果：结肠内容物经偏磷酸预处理3 h后,在程序升温、0.8 m L/min载气流量的气相色谱条件下,内容物中乙酸、丙酸、异丁酸、丁酸、异戊酸和戊酸得到有效分离,其回收率为93%~113%,相对标准偏差为0.46%~0.70%,变异系数小于1%。结论：此方法具有操作简便、快速、准确的优点,是测定动物肠道内容物中短链脂肪酸较为理想的方法。

Objective： This study was conducted to establish a more accurate method for analysis the short-chain fatty acids in colonic contents of piglet using gas chromatography in order to accurately measure the short-chain fatty acid content in the colonic contents to study the situation of intestinal protein metabolism. Methods： The experiment was conducted with similar weight 21 day weaned piglets of Duroc × Landrace × Yorkshire hybrid, single bar feeding. After anesthesia bleeding to death, the abdominal cavity was quickly opened, the end of the colon contents were collected and stored at -80 ℃. Samples were treated with metaphosphoric acid and then capillary gas chromatography was used for determination of short-chain fatty acid with a temperature programmed. Results： The results showed that temperature programmed, 0.8 mL/min of carrier gas flow rate can effectively separate acetic acid, propionic acid,isobutyric acid, butyric acid, isovaleric acid and pentanoic acid, the recoveries were 93 %~113 %, the relative standard deviations were 0.46 % ~ 0.70 % and the variation coefficient is less than 1 %. Conclusions： This method proved to be not only stable, practical, fast and easy to operate, but also better in repetition and accuracy, and would be an ideal way to determinate the short-chain fatty acids in colonic contents of piglet.