摘要
本实验建立了高效液相色谱法测定饲料中维生素B_2的含量。采用了国标法和优化法两种方法进行测定。国标法:按照GB/T14701-2002的方法进行。优化方法:采用Agilent-1260高效液相色谱仪,ZORBAX SB-C_(18)(4.6 mm×150 mm,5-Microm)色谱柱;流动相A液:1.1003 g庚烷磺酸钠和0.0502 g EDTA及25 m L冰乙酸、5 m L三乙胺溶液,用冰乙酸、三乙胺调节pH值至3.4;B液为甲醇;A∶B=85∶15;检测波长:265 nm;柱温:35℃;流速:1.0 m L/min;进样量:0.5μL。结果表明:国标法的保留时间为70 min左右,优化法的保留时间为9 min,且优化法的相关系数、重复性和精密度都满足要求。可见优化法测定所需时间更短,更具有快速、精确测定等优点。
In this experiment,the content of vitamin B2 in feed was determined by high performance liquid chromatography （HPLC）.Two methods of national standard method and optimization method were adopted.The national standardmethod： according to GB/T 14701-2002.Optimization method：using high performance liquid chromatograph Agilent-1260, ZORBAX SB-C18 （4.6 mm×150 mm,5-Microm）chromatographic column;mobile phase for A liquid： 1000 mL contained 1.1003 g heptane sulfonic acid sodium,0.0502 g EDTA and 25 mL ice acetic acid,5 mL triethylamine solution,with ice acetic acid and triethylamine to adjust pH value of 3.4;B liquid methanol ; A：B （85：15） ;detection wavelength of 265 nm; column temperature 35 ℃; the flow rate and sample volume： 1.0 mL/min,0.5 μL.Results showed that the retention time of national standard method for 70 minutes,optimization method of retention time was about 9 minutes,and the correlation coefficient,repeatability and precision of optimization method meet the requirements.The results indicated that the required time of optimization was shorter,more had the advantages of fast and accurate determination.
出处
《中国饲料》
北大核心
2016年第20期38-40,44,共4页
China Feed
关键词
高效液相色谱法
维生素B2
色谱条件优化
high performance liquid chromatography
Vitamin B2
chromatographic condition optimization
