以对甲氯基苯胺为衍生试剂,采用毛细管电泳法分析了藏红花植物细胞多精中的单精组成。对衍生条件进行了优化,并对毛细管分离条件进行了系统的研究。衍生反应在醋酸含量9.5%(v/v)、80℃下反应2h的衍生产率最大,衍生产物紫外检测波长234nm。在优化的毛细管电泳分离条件(末涂层熔融石英毛细管柱(60cm(有效长度50cm)×50μm),柱温25℃,电压20kV,使用350mmol/L硼酸电解液(pH10.21),压力(3.4475kPa)进样5s)下,基线分离了11种结构相近的醛糖(来苏糖、木糖、核糖、葡萄精、甘露糖、半乳糖、鼠李糖、纤维二糖、麦芽糖、乳糖)、酮糖(果糖)的衍生产物。应用该方法定量检测了藏红花植物细胞多糖水解物中糖的成分,各糖的回收率为94.3%-105.4%,相对标准偏差为3.3%-4.6%。
The monosaccharides in the saffron corm glycoconjugate were separated by capillary electrophoresis (CE) coupled with pre-column derivatization. 4-Methoxyaniline was used as derivatization reagent. The derivatization and CE separation conditions were investigated. The ultraviolet detection wavelength was 234 nm. The maximum yield of this derivatization reactionwas obtained under the presence of 9.5% (v/v) acetic acid at 80 22 for 2 h. An uncoated fused-silica capillary of 50μm i. d. and 50/60 cm length ( effective length/total length) was employed, and a pressure injection (3. 447 5 kPa, 5 s) was applied. The baseline separation of l 1 monosaccharides and disaccharides (lyxose, xylose, ribose, glucose, mannose, galactose,rhamnose, cellobiose, maltose, lactose, fructose) was reached at 25 ℃, 20 kV of separation voltage and with 350 mmol/L boric acid (pH 10. 21 ) as running buffer. The developed method has been successfully applied to quantitatively determine the components of saffron corm glyco- conjugate, and the results showed that the recovery of each monosaccharide was in the range of 94.3% -105.4%, the relative standard deviation was 3.3% -4.5%.