简单节杆菌发酵液中羟丙基-β-环糊精含量HPLC法分析  

HPLC Determination of HP-β-CD in Fermentation Liquid of Arthrobacter simplex

作　　者：赵清 申雁冰[1] 骆健美[1] 王敏[1] 

机构地区：[1]工业发酵微生物教育部重点实验室天津科技大学生物工程学院,天津300457

出　　处：《天津科技大学学报》2012年第5期15-17,21,共4页Journal of Tianjin University of Science & Technology

基　　金：国家自然科学基金资助项目(21076158);教育部新世纪优秀人才计划(NCET-08-0911)

摘　　要：羟丙基β环糊精(hydroxypropyl-β-cyclodextrin,HP-β-CD)的特殊结构使其能作为主体包结疏水性甾体,提高甾体的溶解度,同时作为底物/产物储存库,降低底物/产物毒性,添加到发酵液中提高了底物的转化率.利用高效液相色谱法对简单节杆菌发酵液中HP-β-CD的含量进行定量分析.采用Luna NH2100,A(4.6,mm×250,mm,5,μm)色谱柱,流动相为乙腈-水(V乙腈∶V水=60∶40),流量1.0,mL/min,RID示差折光检测器,进样量20,μL.结果表明:HP-β-CD的浓度在2～18,mmol/L的范围内线性关系良好(R=0.999,9),羟丙基-β-环糊精平均回收率(n=9)为99.35%(RSD=0.57%).通过对发酵液中羟丙基-β-环糊精的含量分析证实,该方法具有快速、操作简便、准确、精密度好等特点,适应用于发酵液中羟丙基-β-环糊精的分析.Adding hydroxypropyl-β-cyclodextrin （HP-β-CD） in fermentation liquid could enhance the biotransformation rate of steroid compounds. This effect is generally attributed to the increase of the solubility of steroids and the reduction of the toxicity of the substrate/product due to the formation of inclusion complexes with HP-β-CD. A method for the determination of hydroxypropyl-β-cyclodextrin （HP-β-CD） in fermentation liquid of Arthrobacter simplex with HPLC was developed. HPLC analysis was performed using refractive index detection and a Luna NH2 100 A （4.6 mm × 250 mm, 5 um） with acetonitrile-water （60：40 v/v）as the mobile phase at the flow rate of 1.0 mL/min. The injection volume was 20 uL. There was a good linear relationship within the range of 2- 18 mmol/L for HP-β-CD （R = 0.999 9）. The average recovery rate （n = 9） was 99.35% with 0.57% of RSD. Analyses of the content of HP-β-CD in the fermentation liquid confirmed that the method is simple, fast, accurate, and can be used for the analysis of hydroxypropyl-β-cyclodextrin in fermentation liquid.

关 键 词：高效液相色谱法 羟丙基—β-环糊精 发酵液 简单节杆菌 

分 类 号：X793[环境科学与工程—环境工程]