反相高效液相色谱法测定瘤胃液中的生物胺

王东升, 霍文婕, 朱伟云, 毛胜勇

doi:10.3969/j.issn.1006-267x.2011.12.018

动物营养学报 >

2011 , Vol. 23 >Issue 12: 2165 - 2169

DOI: https://doi.org/10.3969/j.issn.1006-267x.2011.12.018

饲料检测

反相高效液相色谱法测定瘤胃液中的生物胺

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南京农业大学动物科技学院,南京 210095

王东升(1987―),男,山东潍坊人,硕士研究生,从事奶牛营养研究。E_mail: wangdongshengdk2@163.com

收稿日期: 2011-06-16

网络出版日期: 2011-12-14

基金资助

国家重点基础研究发展计划(2011CB100801)

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Determination of Biogenic Amines in Rumen Fluid by Reverse Phase-High Performance Liquid Chromatography

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College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China

Received date: 2011-06-16

Online published: 2011-12-14

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摘要

本文旨在应用反相高效液相色谱法建立瘤胃液中生物胺的测定方法。样品经0.5 mol/L盐酸提取后,用丹磺酰氯柱前衍生,流动相为乙腈和0.1 mol/L乙酸铵溶液;采用梯度洗脱,流速为1.0 mL/min;紫外检测器波长254 nm,柱温30 ℃。应用该方法,研究高精料和低精料饲粮条件下瘤胃液中5种生物胺浓度的差异。结果表明:采用该方法,色胺、腐胺、甲胺、组胺和酪胺5种生物胺能在40 min内得到良好地分离,回收率为88.46％~98.49％,相对标准偏差小于10％。在高精料饲粮条件下,饲喂前山羊瘤胃液中的酪胺和甲胺浓度显著高于低精料饲粮(P<0.05),但组胺、色胺和腐胺浓度无显著性差异(P>0.05)。结果提示,该方法可用于瘤胃液中的生物胺的分析,饲喂前山羊瘤胃液中的酪胺和甲胺浓度受饲粮精粗比的影响。

关键词： 反相高效液相色谱法; 生物胺; 瘤胃液

本文引用格式

王东升, 霍文婕, 朱伟云, 毛胜勇 . 反相高效液相色谱法测定瘤胃液中的生物胺[J]. 动物营养学报, 2011 , 23(12) : 2165 -2169 . DOI: 10.3969/j.issn.1006-267x.2011.12.018

Abstract

This study was conducted to establish a method of simultaneously detecting concentration of biogenic amine in rumen fluid by reverse phase-high performance liquid chromatography (RP-HPLC). Samples were extracted by hydrochloric acid (0.5 mol/L), and then derived using dansyl chloride, the mobile phase was a gradient elution program with a binary mixture of acetonitrile and 0.1 mol/L ammonium acetate and the flow rate was 1 mL/min; the wavelength of UV detector was 254 nm, and the column temperature was 30 ℃. The method was then used to investigate the effect of diet forage-to-concentrate ratio on concentrations of the five biogenic amines in rumen fluid of goats. The results showed that a good separation of tryptamine, putrescine, methylamine, histamine and tyramine was seen within 40 min, and the average recovery were ranged from 88.46% to 98.49%, and the relative standard deviations were less than 10%; Compared with goats fed low concentrate diet, tryptamine and histamine concentrations in rumen fluid of goats fed high concentrate diet were significantly higher (P<0.05), while no significant changes were observed on putrescine, histamine and tyramine concentrations (P>0.05). The results indicate that this method presented here can be used to determine biogenic amine concentrations in rumen fluid, and concentrations of biogenic amine in rumen fluid of goats can be affected by diet forage-to-concentrate ratio.

Key words： reverse phase-high performance liquid chromatography; biogenic amine; rumen fluid

参考文献

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