首页> 中文期刊> 《动物营养学报》 >反相高效液相色谱法测定瘤胃液中的生物胺

反相高效液相色谱法测定瘤胃液中的生物胺

         

摘要

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 t. 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.%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.

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