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一次注射15N-亮氨酸示踪法检测鸡内源氨基酸损失量适宜参数的研究

DOI: 10.3969/j.issn.1006-267x.2015.10.009

Keywords: 一次注射法,15N-亮氨酸,内源氨基酸损失量,GC-MS,公鸡

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Abstract:

本研究旨在优化一次注射15N-亮氨酸(15N-Leu)示踪法检测鸡内源氨基酸(AA)损失量的适宜参数。试验一,研究气相色谱-质谱联用仪(GC-MS)测定Leu的15N丰度(APEL)的适宜参数,确定适宜的丰度测定范围;试验二,以48只艾维茵公鸡为对象,研究15N-Leu的引入方式(肌肉注射和静脉注射)和剂量(10、20和30mg/kgBW)对内源AA主要分泌源中APEL的影响,旨在优化15N-Leu的引入参数。结果显示:1)Leu的MTBSTFA衍生物在GC-MS上的最大特征碎片离子峰为质荷比(m/z)=200;当样品APEL<0.1%时,测定精准度(13.1%)和准确度(13.8%)较差;当样品APEL为0.1%~3.2%时,测定精准度和准确度分别为4.3%~9.7%和-5.9%~4.1%,所得到的同位素稀释曲线良好,R2为0.989;2)当15N-Leu引入剂量为20和30mg/kgBW时,内源AA主要分泌源中APEL>0.1%;静脉注射的试鸡在注射部位出现淤血、外渗现象;3)15N-Leu引入方式和剂量对内源AA主要分泌源的APEL不存在交互作用(P>0.05);各分泌源的APEL均随引入剂量的增大而显著增加(P<0.05),当引入剂量为20mg/kgBW时,各分泌源间的APEL无显著差异(P>0.05);引入方式显著影响胰腺和肠黏膜的APEL(P<0.05)。以上结果表明,15N-Leu引入剂量为20mg/kgBW可满足GC-MS分析要求,同时选取肌肉注射可减少动物应激、简化操作程序,并降低成本。

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