RP-HPLC法测定长期盐胁迫下盐爪爪和盐穗木中的甜菜碱(英文)

Determination of Glycinebetaine in Kalidium foliatum and Halostachys caspica Chronical Treatment with NaCl by RP-HPLC

通过对影响甜菜碱提取和测定的各种因素进行研究,建立了一种准确、快速的测定长期盐胁迫下盐爪爪和盐穗木中甜菜碱的HPLC方法.样品采用超声波甲醇提取,并依次通过强阴阳离子交换树脂（Dowex 50×8和Dowex 50×2）进行纯化,得到的样品采用反相C18柱进行分离,色谱条件：流动相为50 mmol/L、pH 4.5的KH2PO4溶液;波长200 nm下紫外测定,流速0.7 mL/min;此方法的检出限为0.02μg/mL,平均回收率为97.2%.通过对0～500 mmol/L NaCl胁迫下的盐爪爪和盐穗木中甜菜碱进行测定,甜菜碱的含量均呈现先增加后减少的趋势,盐爪爪在NaCl浓度为200 mmol/L时达到最大,而盐穗木在NaCl浓度为300 mmol/L时达到最大.

A series of studies was conducted to establish a methodology for the accurate and efficient determination of glycinebetaine in Kalidium foliatum and Halostachys caspica chronically cultured with different concentration of NaCl. The final methodology involves an extraction step in which the sample was extracted with ultrasonic and ion-exchange resin（Dowex 50 × 8,Dowex 50 × 2）. An HPLC method using an ODS-C18 column and UV detection for the determination of glycinebetaine in plant tissue extracts was described. The mobile phase was kept constant at pH 4.5 and 50 mmol/L KH2PO4 in water,and eluted compounds are detected by UV absorbance （200 nm）. The flow rate was maintained at 0.7 mL/min. This assay was very accurate with a lower detection limit of 0.02 μg/mL. Recovery assays done with standard glycinebetaine and wild Kalidium foliatum resulted in a consistent recovery of 97. 2%. Glycinebetaine content was quantified in K. Foliatum and H. Caspica cultured with different concentration of NaCl chronically and the analysis according to their current from 0 to 500 mmol/L NaCl was also discussed in this paper.