摘要
应用薄层色谱(TLC)和高效液相色谱(HPLC)技术相结合,建立苜蓿皂苷的分离检测方法以甲醇为溶剂,超声波辅助萃取苜蓿皂苷;萃取液首先经TLC分离(薄层板规格5 cm×20 cm),以氯仿-甲醇-水(65:35:10,下层)做展开荆,于254 nm处观察并收集不同比移值的分离组分;分离后的组分再次用HPLC进行检测,色谱柱为Venusil Mp C18(2)(4.6 mm×250 mm),在室温下以甲醇-水(15:85)为流动相,流速为0.15 mL/min、0.2 mL/min,以及甲醇-水(20:80)做流动相,流速为0.3 mL/min,检测波长为200 nm。结果表明:经薄层层析后分离得5种皂苷化合物,比移值分别为Ⅰ(0.08)、Ⅱ(0.18)、Ⅲ(0.49)、Ⅳ(0.63)、Ⅴ(0.80);分别将5种皂苷化合物经高效液相色谱进一步分离,共获得单体苜蓿皂苷13种。薄层层析与高效液相色谱技术相结合可用于苜蓿皂苷单体的分离检测,在本实验条件下检测到苜蓿提取物中的13种皂苷类化合物:
A method of determination for alfalfa saponins by TLC and HPLC was established. Alfalfa saponins were extracted with methanol by ultrasonic wave; Alfalfa saponins were separated by TLC （silica gel plate 5 cm×20 cm）with the developing system of chloroform： methanol： water （ 65：35：10, the under layer）, the silica gel plate was observed at 254 nm and collected the separate samples; All the separate samples were tested by HPLC. Venusil Mp C18（2） （4.6 mm × 250mm） column was used with mobile phase ： methanol - water （15 ： 85） at a flow rate of 0.15 mL/min, 0.2 mL/min and methanol - water （20：80）at a flow rate of 0.3 mL/min, detecting wavelength： 200 nm, column temperature： room temperature. The results showed that five alfalfa saponins compounds were obtained by TLC, the Rf values were I （0.08）, II（0.18）, III （0.49）, IV（0.63） and V （0.80）, respectively; In conclusion, thirteen single alfalfa saponins were totally detected by HPLC. Conclusion： Thirteen alfalfa saponins compound were detected under the condition of the experiment; TLC combined HPLC can be used for determination of alfalfa saponins.
出处
《中国畜牧杂志》
CAS
北大核心
2015年第21期58-62,共5页
Chinese Journal of Animal Science
基金
国家科技支撑计划(2013BAD10B04-5)
河南省高校科技创新团队(131RSTHN007)
国家自然科学基金资助项目(31172241)
