摘要
采用高效液相色谱柱Alltima^TM C18（4．6mm×150mm,4μm）上定量测定发酵生产的聚苹果酸，流动相为乙腈和0．025mol／L磷酸二氢钾混合溶液（磷酸调pH2．5）（V：V=5：95），流速1．0mL／min，检测波长210nm，柱温25℃，进样量5μL。发酵液经离心除菌体、纯沉除多糖、水解处理后进样，可以将其中的L-苹果酸完全分离定量。采用液相方法的回收率为98．70％-100．65％，RSD为0．4-1．11％。采用试剂盒的方法的回收率为96．7-97．23％，RSD为0．57-1．19％。可见，液相的精确度和试剂盒相差不大。但液相的价格相对比试剂盒要便宜很多，说明该法是测定发酵液中聚苹果酸的一种很有效的方法。
For the detection of poly （malic acid） fermentation broth, a reversed phase high performance liquid chromatography method was developed. The samples was analyzed on a column of Alltima^TM C18 （4.6 mm× 150 mm, 4 μm） with a mixture of acetonitrile and 0.025 mobil KH2PO4 buffer （pH 2.5） （5：95, v：v） used as mobile phase at the flow rate of 1.0mL/min. The detection wavelength was 210 nm; the column temperature was 25 ℃ and the volum of injection was 5 μL. Before sampling, the fermentation broth was centrifuged to remove fungis, added with organic solvent to remove polysaccharide and hydrolysed. L-malic acid were completely separated and determined in 3 minutes. The recovery and relative standard deviation （RSD, n=3） of L-malic acid by RP-HPLC method were 98.70%-100.65%, and 0.40%-1.11%, respectively. The recovery and relative standard deviation （RSD, n=3） of L-malic acid by L-malic acid kit were 96.70%-97.23% and 0.57%-1.19%, respectively. The method was proved to be a effective way to determine the poly （malic acid） in fermentation broth. Key words： Reversed phase high performance liquid chromatography; Poly （malic acid）; L-mailc acid kit
出处
《现代食品科技》
EI
CAS
北大核心
2012年第4期453-455,452,共4页
Modern Food Science and Technology
基金
天津市高等学校科技发展基金计划(ZD200703)
科技部科技型中小企业技术创新基金(11C262111200191)
天津市科技型中小企业创新基金(10ZXCXSY10900)
关键词
反相高效液相色谱法
聚苹果酸
L-苹果酸试剂盒
Reversed phase high performance liquid chromatography
Poly(malic acid)
L-mailc acid kit
