Sarvin Boris, Fedorova Elizaveta, Shpigun Oleg, Titova Maria, Nikitin Mikhail, Kochkin Dmitry, Rodin Igor, Stavrianidi Andrey
Chemistry Department, Lomonosov Moscow State University, 119991 Moscow, Russia.
Biology Department, Lomonosov Moscow State University, 119234 Moscow, Russia.
J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Mar 30;1080:64-70. doi: 10.1016/j.jchromb.2018.02.012. Epub 2018 Feb 13.
In this paper, the ultrasound assisted extraction method for isolation of steroidal glycosides from D. deltoidea plant cell suspension culture with a subsequent HPLC-MS determination was developed. After the organic solvent was selected via a two-factor experiment the optimization via Latin Square 4 × 4 experimental design was carried out for the following parameters: extraction time, organic solvent concentration in extraction solution and the ratio of solvent to sample. It was also shown that the ultrasound assisted extraction method is not suitable for isolation of steroidal glycosides from the D. deltoidea plant material. The results were double-checked using the multiple successive extraction method and refluxing extraction. Optimal conditions for the extraction of steroidal glycosides by the ultrasound assisted extraction method were: extraction time, 60 min; acetonitrile (water) concentration in extraction solution, 50%; the ratio of solvent to sample, 400 mL/g. Also, the developed method was tested on D. deltoidea cell suspension cultures of different terms and conditions of cultivation. The completeness of the extraction was confirmed using the multiple successive extraction method.
本文开发了一种超声辅助提取方法，用于从三角叶薯蓣植物细胞悬浮培养物中分离甾体糖苷，并随后进行高效液相色谱-质谱测定。通过双因素实验选择有机溶剂后，采用拉丁方4×4实验设计对以下参数进行优化：提取时间、提取溶液中有机溶剂浓度以及溶剂与样品的比例。结果还表明，超声辅助提取方法不适用于从三角叶薯蓣植物材料中分离甾体糖苷。使用多次连续提取法和回流提取法对结果进行了复核。超声辅助提取法提取甾体糖苷的最佳条件为：提取时间60分钟；提取溶液中乙腈（水）浓度50%；溶剂与样品的比例400 mL/g。此外，所开发的方法在不同培养期和培养条件的三角叶薯蓣细胞悬浮培养物上进行了测试。使用多次连续提取法确认了提取的完整性。
