动物营养学报 >

2015 , Vol. 27 >Issue 10: 3047 - 3056

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

禽营养 Poultry Nutrition

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

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  • 西北农林科技大学动物科技学院, 杨凌 712100
胡如久(1989—),男,河南信阳人,硕士研究生,动物营养与饲料科学专业。E-mail:hurujiu@163.com

收稿日期: 2015-05-06

  网络出版日期: 2015-10-15

基金资助

国家自然科学基金项目(31172223)

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

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  • College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China

Received date: 2015-05-06

  Online published: 2015-10-15

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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-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。

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

本文引用格式

胡如久, 汪菲, 李晶, 姚军虎, 杨小军 . 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[J]. 动物营养学报, 2015 , 27(10) : 3047 -3056 . DOI: 10.3969/j.issn.1006-267x.2015.10.009

Abstract

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 APEL in 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 from 4.3% to 9.7% and -5.9% to 4.1% when APEL ranged from 0.1% to 3.2%, and the standard curve of mixtures of natural and 15N-leucine showed a good linear relationship (R2=0.989).2) The values of APEL in 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 APEL in main secretory sources of endogenous amino acids.The values of APEL in main secretory sources of endogenous amino acids were increased significantly (P<0.05) with an increase of administration dose.The values of APEL among different secretory sources had no significant difference (P>0.05) at 15N-leucine administration doses of 20 mg/kg BW.However, the values of APEL in pancreas and intestinal mucosa were influenced significantly (P<0.05) by 15N-leucine administration method.In conclusion, the 15N-leucine administration doses of 20 mg/kg BW is selected to meet the requirements of GC-MS analysis, and intramuscular injection of 15N-leucine is applied to reduce harmful stress, simplify the operating procedure and reduce the cost.

Key words： single-administration method; 15N-leucine; endogenous amino acid losses; GC-MS; chicken

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