一次注射^(15)N-亮氨酸示踪法检测鸡内源氨基酸损失量适宜参数的研究  

^(15)N-Leucine Single-Administration Method for Measuring Endogenous Amino Acid Losses in Chicken:Optimization of Main Parameters

作　　者：胡如久[1] 汪菲[1] 李晶[1] 姚军虎[1] 杨小军[1] 

机构地区：[1]西北农林科技大学动物科技学院,杨凌712100

出　　处：《动物营养学报》2015年第10期3047-3056,共10页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：国家自然科学基金项目(31172223)

摘　　要：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30 mg/kg BW)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M TBSTFA衍生物在GC-M S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30 mg/kg BW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20 mg/kg BW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20 mg/kg BW可满足GC-M S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。The current study was conducted to optimize parameters of the^15N-leucine single-administration method for measuring endogenous amino acid losses in chicken. Experiment 1 was designed to provide suitable parameters of gas chromatography-mass spectrometry（ GC-MS） for determing^15N-enrichment of leucine（ APEL）and ascertain an appropriate scope of APELin samples. Experiment 2 was carried out to optimize administration parameters of^15N-leucine,using 48 Avian roosters to study the effects of administration methods（ intramuscular injection and intravenous injection） and doses（ 10,20 and 30 mg / kg BW） of^15N-leucine on^15N-enrichment in main secretory sources of endogenous amino acids. The results showed as follows： 1） the biggest characteristic fragment ion peaks of MTBSTFA derivatives of leucine in GC-MS is m / z = 200. Precision and accuracy of measurement were poor when APEL〈0. 1% in samples; measurement precision and accuracy respectively ranged from4. 3% to 9. 7% and-5. 9% to 4. 1% when APELranged from 0. 1% to 3. 2%,and the standard curve of mixtures of natural and^15N-leucine showed a good linear relationship（ R^2= 0. 989）. 2） The values of APELin main secretory sources of endogenous amino acids were greater than 0. 1% for^15N-leucine administration doses of 20 and 30 mg / kg BW. The experimental birds administrated^15N-leucine through intravenous injection showed harmful stress including congestion and extravasation at the injection sites. 3） There were not administration methods and doses interaction effects（ P〈0. 05） of APELin main secretory sources of endogenous amino acids. The values of APELin main secretory sources of endogenous amino acids were increased significantly（ P〈0. 05） with an increase of administration dose. The values of APELamong different secretory sources had no significant difference（ P〉0. 05） at^15N-leucine administration doses of 20 mg / kg BW. However,the values of APELin pancreas and intestinal mucosa were influenced significantly（ P�

关 键 词：一次注射法 ^15N-亮氨酸 内源氨基酸损失量 GC-M S 公鸡 

分 类 号：S852.2[农业科学—基础兽医学]