反相HPLC色谱法测定盐芥和菠菜中的甜菜碱含量

QUANTITATIVE DETERMINATION OF BETAINE IN TH. SALSUGINEA.O.E.SCH ULZ AND SPINACH BY REVERSE PHASE HPLC

本实验利用反相HPLC色谱法测定了盐芥和菠菜中的甜菜碱含量,实验分别以菠菜和盐芥的叶组织为材料,用去离子水提取甜菜碱.提取液经浓缩后依次通过阳离子交换树脂柱和阴离子交换树脂柱进行纯化,用NH4 OH洗脱阳离子交换树脂柱,洗脱液收集浓缩干后用磷酸盐流动相溶解.样品用C1 8反相色谱柱进行分离,流动相为5 0mmol/L的KH2 PO4 溶液(pH 4.5 6) ,色谱条件为0 .7ml/min ,192nm波长测定;此色谱条件能使甜菜碱与其它组分很好的分离,甜菜碱加样回收率达到90 %～98% .经测定,盐芥叶片中的甜菜碱含量为1.3 67mg/g ,菠菜叶片中的甜菜碱含量为1.65 6mg/g .

The experiment applies reverse phase HPLC to detect be ta ine levels in leaf of spinach and Th.salsuginea.O.E.Schulz.The extracts were pas sed through a 10 ml column of 201×7 in series with a second column of a 10 ml 7 32.Bound betaine was recovered from 732 with 10 ml of 2 mol/L NH 4OH and the el uate was evaporated to dryness at 60～65 ℃ in vacuum.Separation was perform ed o n a LC column eluted with 50 mmol/L KH 2PO 4(pH 4.56).Detection was performed by UV absorbance at 192 nm.The recovery rate of betaine in tissue extracts range d from 90% to 98%.Betaine concentrations in leaf of spinach is 1.656 mg/g fresh w eight (FW).Betaine level in leaf of Th.salsuginea.O.E.Schulz was inc reased to 1.367 mg/g FW.