高效液相色谱测定山羊瘤胃液挥发性脂肪酸条件优化的研究

Optimizing Condition of High Performance Liquid Chromatography for Detecting Volatile Fatty Acids in Goat Rumen Fluid

挥发性脂肪酸(VFA)在维持反刍动物瘤胃内环境稳定方面起着重要的作用,同时也是反映个体能量代谢和发酵类型的重要指标,准确检测瘤胃液VFA含量对反刍动物营养、瘤胃内环境、消化功能及反刍动物消化道疾病防控均具有重要的意义。为了建立高效液相色谱检测瘤胃液VFA的方法并优化色谱检测条件,本研究分别对高效液相色谱不同流动相和pH对瘤胃乙酸、丙酸和丁酸的分离检测效果进行了分析。结果表明,以甲醇或乙腈作为流动相时,均能对3种VFA进行有效分离,但两种流动相出峰时间不同。用乙腈作为流动相时,3种VFA的出峰时间均较甲醇作为流动相短。通过设置2.05和2.30两个水平的流动相pH,研究发现pH对分离效果和出峰时间影响较小。这些结果表明乙腈和甲醇均可作为瘤胃液VFA测定的流动相,在实际应用中可结合二者的价格、毒性以及检测耗时综合考虑进行选择。

Volatile fatty acids play an important role in keeping stabilization of romen environment. It re- flects status of energy meterbolism in rumen. Therefore,it is meaningful for study of ruminant nutrition, rumen environment and digestive ability accurately to measure content of VFA in rumen. To optimize con- dition of high performance liquid chromatography for detecting VFA, we compared result of separating VFA using two mobile phases of methanol and acetonitrile under two pH levels of 2.05 and 2.30. Result showed that both methanol and acetonitrile can successfully separate acetic acid, met^cetonic acid and buta- noic acid, but peak times corresponding to three organic acids is different between methanol and acetoni- trile. In addition, we found two levels of pH have little effect on result of separation. The data indicated that both methanol and acetonitrile can be used as mobile phases in detecting contents of rumen VFA,it should be determined according to its price and toxicity which,methanol or acetonitrile,is selected.