摘要
试验旨在建立1种基于在线柱前邻苯二醛衍生-高效液相色谱法(OPA-HPLC)测定海兰褐蛋鸡血浆及肌肉中γ-氨基丁酸(GABA)含量的分析方法。将蛋鸡血浆及匀浆肌肉样品经纯净水提取、超声处理并离心后,取上清液,经0.22μm滤膜,与4倍体积的衍生试剂进行在线柱前衍生反应。结果显示,在EP-C18色谱柱(4.6 mm×250.0 mm,5μm)、流动相乙腈∶磷酸盐缓冲液为27∶73、流速0.5 mL/min、柱温30℃、波长333 nm条件下,进行紫外检测效果最佳。GABA在8~1000 mg/L范围内线性关系良好,回归方程为y=1.115x-7.598(R^(2)=0.9996),方法检出限(LOD)为5.41 mg/L,定量限(LOQ)为18.04 mg/L。血浆中GABA的平均回收率为95.43%~101.35%,相对标准偏差(RSD)为5.08%~7.42%;肌肉中GABA的平均回收率为94.09%~100.90%,RSD为3.23%~6.88%。试验所用方法测得蛋鸡血浆中GABA含量为37.265 mg/L,胸肌中为1.064 mg/g,腿肌中为12.723 mg/g。研究表明,OPA-HPLC法操作简便、回收率高、精密度良好、特异性强,适用于蛋鸡血浆及不同部位肌肉中GABA含量的快速、批量检测。
The experiment aimed to establish an analytical method based on online pre-column o-phthaldialdehyde derivatization high-performance liquid chromatography(OPA-HPLC)for the determination ofγ-aminobutyric acid(GABA)content in plasma and muscle tissues of Hyline brown laying hens.After extracting the laying hens plasma and homogenized muscle samples with pure water,subjecting them to ultrasonic treatment and centrifugation,the supernatant was collected.This supernatant was then filtered through a 0.22μm membrane and underwent an online column reaction with four times the volume of the derivatization reagent.The results showed that optimal UV detection was achieved under the following conditions:An EP-C18 column(4.6 mm×250.0 mm,5μm),a mobile phase consisting of acetonitrile∶phosphate buffer ratio of 27∶73,a flow rate of 0.5 mL/min,a column temperature of 30℃,and a detection wavelength of 333 nm.GABA exhibited a good linear relationship in the concentration range of 8 to 1000 mg/L,with a regression equation of y=1.115x-7.598(R^(2)=0.9996).The limit of detection(LOD)was 5.41 mg/L,and the limit of quantification(LOQ)was 18.04 mg/L.The average recovery rate of GABA in plasma was 95.43%to 101.35%,with a relative standard deviation(RSD)of 5.08%to 7.42%.The average recovery rate of GABA in muscles was 94.09%to 100.90%,with RSD ranging from 3.23%to 6.88%.The GABA content in the plasma of Hyline brown laying hens was measured to be 37.265 mg/L using this method,1.064 mg/g in the chest muscle,and 12.723 mg/g in the leg muscle.The study indicates that the OPA-HPLC method is simple to operate,provides high recovery and precision,exhibits strong specificity,and is suitable for rapid and batch detection of GABA content in plasma and various muscle tissues of Hyline brown laying hens.
作者
孙云竹
赵宇
尤春雪
刘鼎阔
任超琪
赵昕
李留安
于晓雪
SUN Yunzhu;ZHAO Yu;YOU Chunxue;LIU Dingkuo;REN Chaoqi;ZHAO Xin;LI Liuan;YU Xiaoxue
出处
《饲料研究》
北大核心
2025年第18期127-132,共6页
Feed Research
基金
天津市科技局项目(项目编号:24ZYCGSN00060、23ZYCGSN00250)
天津市教委科研计划项目(项目编号:2021KJ108)
天津市青年科技人才(第三层次)项目(项目编号:QN20230301)。
关键词
在线柱前衍生
高效液相色谱法
Γ-氨基丁酸
海兰褐蛋鸡
血浆
肌肉
online pre-column derivatization
high-performance liquid chromatography
γ-aminobutyric acid
Hyline brown laying hens
plasma
muscle
