建立了高效阴离子交换色谱-脉冲安培电化学检测法同时测定生物转化样品中海藻糖、葡萄糖和麦芽糖的分析方法。选用 CarboPacTM10色谱柱（250 mm×2 mm）对分离条件进行优化，使用标准样品测定了线性范围和工作曲线，柱温为30℃，流速为0.30 mL／min，以氢氧化钠溶液和醋酸钠溶液为流动相进行梯度洗脱，脉冲安培法进行检测。研究结果表明，该方法可在15 min内实现海藻糖生物转化液中3种糖的快速定量分析。海藻糖、葡萄糖和麦芽糖峰面积与质量浓度的线性关系良好，检出限为0.010~0.100 mg／L。将此方法用于酶法制备海藻糖的检测，加标回收率为89.40％~103.2％。在生物转化样品中检测到海藻糖浓度为101.084 g／L，转化率达到了50.5％。该方法灵敏度高，简便快速，可应用于海藻糖制备样品中各种成分的分离和定量检测。
An analytical method for the determination of trehalose,maltose,and glucose in biotransformation samples was developed by using high performance anion exchange chroma-tography coupled with pulsed ampere detection( HPAEC-PAD). The analysis was performed on a CarboPacTM 10 column( 250 mm × 2 mm ) with the gradient elution of NaOH-NaAc as the mobile phase. The column temperature was set at 30 ℃,the flow rate was 0. 30 mL/min. The results showed that trehalose,maltose,and glucose in biotransformation system were com-pletely separated and determined in 15 min. The linear ranges and the working curves were determined by using standard samples. The correlation coefficients of three kinds of carbohy-drates were over 0. 999 8 . The detection limits( LODs)were 0. 010-0. 100 mg/L. Under the optimized separation conditions,the recoveries of saccharides in the transformation system at three different spiked levels ranged from 89. 4% to 103. 2%. In biotransformation system,50 IU trehalose synthase were added into 200 g/L maltose for reaction of 8 h at 37 ℃,pH 8. 0. Under the above conditions,the concentration of trehalose in biotransformation sample was 101. 084 g/L,and the conversion rate of trehalose reached 50. 5%. The method can be applied to deter-mine the composition in the transformation system with the advantages of simplicity and con-venience.
参考文献
| [1] | Schiraldi C;Di Lernia I;De Rosa M.[J].Trends Biotechnol,200220(10):420. |
| [2] | 李群;袁勤生.[J].中国生化药物杂志,199516(05):231. |
| [3] | 吴秀丽;岳明;丁宏标.[J].微生物学通报,200936(07):1067. |
| [4] | 缪静.[J].微生物学通报,200431(05):96. |
| [5] | 周俊;刘景汉;欧阳锡林.[J].中国实验血液学杂志,200412(06):837. |
| [6] | 王兰;肖冬光;张正.[J].工业微生物,200232(02):18. |
| [7] | 王兰;肖冬光.[J].生物技术,200212(03):27. |
| [8] | 赵华;何聪芬;秦允荣.[J].吉首大学学报(自然科学版),200930(01):79. |
| [9] | 牟世芬;于泓;蔡亚岐.[J].色谱,200927(05):667. |
| [10] | Yu H;Ding Y S;Mou S F.[J].J Chromatogr A,2002966(1/2):89. |
| [11] | 李静;李仁勇;梁立娜.[J].分析化学,201240(09):1415. |
| [12] | 张磊;周光明;熊建飞.[J].食品科学,201233(08):159. |
| [13] | 周洪斌;熊治渝;李平.[J].色谱,201331(11):1093. |
| [14] | 潘广文;刘绿叶;叶明立.[J].色谱,201230(04):395. |
| [15] | 丁洪流;李灿;金萍.[J].色谱,201331(08):804. |
| [16] | 魏泱;丁明玉.[J].色谱,200119(03):226. |
| [17] | 许丽;周光明;余娜.[J].日用化学工业,201242(01):77. |
- 下载量()
- 访问量()
- 您的评分：
-
10%
-
20%
-
30%
-
40%
-
50%