高效液相色谱柱后衍生法在线测定海带淀粉的相对分子质量  

Online Determination of Laminarin Relative Molecular Weight Using HPLC-Post-column Derivatization System

作　　者：韩平[1] 郑立[1] 刘涛[2] 崔志松[1] 郭秀春[1] 田黎[1] 王小如[1] 

机构地区：[1]国家海洋局第一海洋研究所海洋生态研究中心,山东青岛266061 [2]中国海洋大学海洋生命学院,山东青岛266100

出　　处：《分析测试学报》2009年第9期1066-1068,1073,共4页Journal of Instrumental Analysis

基　　金：国家自然科学基金资助项目(20602009);国家"863计划"资助项目(2007AA09Z435);国家海洋局公益资助项目(200805039);青岛市发展计划资助项目(06-2-2-16-jch)

摘　　要：建立了HPLC-柱后衍生-紫外可见检测器在线测定海带淀粉相对分子质量的方法。以10μ/TSK G3000PW300mm×7.5mmi.d.凝胶柱为色谱柱,0.02%NaN3-0.1mol.L-1NaNO3溶液为色谱柱流动相,流速为0.5mL.min-1;以pH8.0的0.01%刚果红-0.1mol.L-1磷酸缓冲液为柱后流动相,流速0.5mL.min-1;柱后衍生温度为60℃;二极管阵列检测器于550nm检测。多糖相对分子质量标准曲线的线性范围为505～133800u,相关系数(R2)为0.9993;海带淀粉的检出限(S/N=3)为30mg.L-1,保留时间的相对标准偏差为1.89%(n=6)。该方法能够实现在线检测,无需对海带多糖粗提物进行分离纯化。A novel method was established to determine the relative ＇molecular weight of laminarin in kelp using HPLC with post-column derivatization and UV - Vis detector. 10℃/TSK G3000 PW 300 mmx 7.5 mm i. d. column was used with a mobile phase of 0. 02% NaN3 -0. 1 mol · L-1 NaNO3 with a flow rate of 0. 5 mL ·min- 1 The post column mobile phase was 0. 01% congo red -0. 1 mol · L-1 phosphate buffer（ pH 8.0） with a flow rate of 0. 5 mL · min-1 at 60 ℃. The signals were obtained by diode array detector（DAD） at the wavelength of 550 nm. The calibration curve of relative molecular weight of standard polysaccharide was linear in the range of 505 - 133 800 u with a correlation coeffi- cient of 0. 999 3. The detection limit （ S/N = 3 ） of laminarin was 30 mg· L- 1 and the relative stand- ard deviation （n = 6） of retention time was 1.89% . The method could applied in the online determi- nation of laminarin from crude polysaccharide extraction without any seperation.

关 键 词：海带淀粉 相对分子质量 高效液相色谱 柱后衍生 刚果红 

分 类 号：O675.7[理学—化学] O636.12