柱前衍生高效液相色谱法测定酵母培养物中甘露聚糖的含量  

Determination of mannan in yeast cultures by HPLC with pre-column derivatization

作　　者：张相飞[1] 刘明 李文辉[3] 

机构地区：[1]青岛农业大学化学与药学院,山东青岛266109 [2]北京中农弘科生物技术有限公司,北京102206 [3]廊坊市农业局,河北廊坊065000

出　　处：《饲料工业》2017年第16期60-62,共3页Feed Industry

摘　　要：采用1-苯基-3-甲基-5-吡唑啉酮(PMP)为衍生化试剂,建立了反相高效液相色谱测定酵母培养物中甘露聚糖的含量方法。样品酵母培养物经均质处理,其中的多糖在酸性条件下水解为单糖,PMP衍生后,以0.1 mol/l pH值5.5乙酸铵缓冲液乙腈(V/V,7030)为流动相,反相色谱柱分离,流速1.0 ml/min,紫外波长250 nm检测。结果表明,甘露糖浓度在1.0~100 mg/l范围内与对应峰面积呈线性关系(R2=0.99)。甘露聚糖添加回收率为96.3%~102.6%,相对标准偏差2.5%,甘露聚糖最低检出限为0.11 mg/l。该方法适用于酵母培养物中甘露聚糖含量的分析检测。A high performance liquid chromatographic method was developed for the determination of mannan in yeast cultures by using 1-phenyl-3-methyl-5-pyrazolone （PMP） as the derivatization reagent. After the yeast cultures were processed by homogenizer, the polysaccharides in the yeast cultures were hydrolyzed into monosaccharides under an acid condition. After being derived by PMP, the samples were separated on reversed-phase C 18 column by using 0.1 mol/l pH5.5 ammonium acetate buffer solution-acetonitrile （V/V, 70 ： 30）as the mobile phase. The UV detector was operated at 250 nm. The flow velocity was 1.0 ml/min. The results showed that the good linearity relationship （R^2=0.99） was obtained between the concentration of mannose in the range of 1.0 mg/l to 100 mg/l and its peak area. The recovery rate was in the range of 96.3% -102.6% and the relative standard deviation （RSD） was 2.5%. The limit of detection （LOD） was 0.11 mg/l for marman. This method can he used for the determination of mannan in yeast cultures.

关 键 词：酵母培养物 甘露聚糖 PMP衍生化法 高效液相色谱法 

分 类 号：TQ926.3[轻工技术与工程—发酵工程]