分步酸解法快速检测发酵液中普鲁兰糖含量  

Rapid Determination of Pullulan Content in Fermentation Broth by Stepwise Acid Hydrolysis

作　　者：刘飞[1,2,3] 张林军 袁超 张金华 刁梦奇 凌沛学 陈会萍[4] 段荣帅 王凤山 LIU Fei;ZHANG Lin-jun;YUAN Chao;ZHANG Jin-hua;DIAO Meng-qi;LING Pei-xue;CHEN Hui-ping;DUAN Rong-shuan;WANG Feng-shan(School of Pharmaceutical Sciences,Shandong University,Ji'nan 250012,China;Key Laboratory of Biophar-maceuticals Postdoctoral Scientific Research Workstation,Shandong Academy of Pharmaceutical Sciences,Ji'nan250101,China;National-Local Joint Engineering Laboratory of Polysaccharide drugs,Shandong Academy ofPharmaceutical Sciences,Ji'nan 250101,China;Shandong Institute of Commerce and Technology,Ji'nan250103)

机构地区：[1]山东大学药学院,山东济南250012 [2]山东省药学科学院山东省生物药物重点实验室博士后科研工作站,山东济南250101 [3]山东省药学科学院多糖类药物发酵与精制国家地方联合工程实验室,山东济南250101 [4]山东商业职业技术学院,山东济南250103

出　　处：《药物生物技术》2021年第1期21-25,共5页Pharmaceutical Biotechnology

基　　金：山东省重点研发计划(No.2018GSF121033,No.2018GHY115009,No.2018GSF118099,No.2019GSF107040,No.2019GSF108225,No.2019GHY112009);泰山学者工程专项;山东省高等学校“青创科技计划”(No.2019KJE014)。

摘　　要：建立分步酸解快速检测发酵液中普鲁兰糖含量的方法。使用不同浓度盐酸对两个相同发酵液样品进行酸解,分别将蔗糖和普鲁兰糖降解为单糖,再利用生物传感分析仪对酸解之后样品中葡萄糖含量进行定量分析,从而利用两者之间葡萄糖含量的差值计算出发酵液中普鲁兰糖的含量。在对酸解时间、盐酸浓度、蔗糖和普鲁兰糖检测范围以及该方法精密度和准确度进行研究之后,确定了蔗糖酸解条件为:0.6 mol/L盐酸、沸水浴3 min;普鲁兰糖的酸解条件为:2 mol/L盐酸、沸水浴30 min。实验证明,此方法精密度良好,RSD为0.7743%,准确度高,回收率为97.96%~100.72%;用时较短,40 min左右即可较准确的测定发酵液中普鲁兰糖含量。实验证明此方法是一种可行的高效、快速测定发酵液中普鲁兰糖含量的方法。A step-by-step acidolysis strategy for rapid detection of pullulan in fermentation broth was proposed in this study.The main concept for strategy is that two samples with the same fermentation broth were acidified under different concentrations of hydrochloric acid,thus sucrose and pullulan were degraded into monosaccharides.And then the content of glucose in the sample after acidolysis was quantitatively analyzed by biosensor analyzer,the content of pullulan in fermentation broth was calculated by using the difference in glucose content.After investigating the acidolysis time,hydrochloric acid concentration,detection range,precision and accuracy of sucrose and pullulan,the acidolysis conditions were determined as follows:sucrose(0.6 mol/L hydrochloric acid,boiling water bath 3 min)and pullulan(2 mol/L hydrochloric acid,boiling water bath 30 min).Results confirmed that this method had high precision up to 0.7743%(RSD),high accuracy(recovery rate 97.96%~100.72%)and short time(about 40 min)and could accurately determine pullulan content in fermentation broth.Altogether,this strategy was an efficient and rapid method for the determination of pullulan in fermentation broth.

关 键 词：普鲁兰糖 酸解 生物传感分析仪 快捷检测 精密度 准确度 

分 类 号：O652.7[理学—分析化学]