title: High-speed homogenization coupled with microwave-assisted extraction followed by liquid chromatography-tandem mass spectrometry for the direct determination of alkaloids and flavonoids in fresh Isatis tinctoria L. hairy root cultures.
abstract: A new, simple and efficient analysis method for fresh plant in vitro cultures-namely, high-speed homogenization coupled with microwave-assisted extraction (HSH-MAE) followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS)-was developed for simultaneous determination of six alkaloids and eight flavonoids in Isatis tinctoria hairy root cultures (ITHRCs). Compared with traditional methods, the proposed HSH-MAE offers the advantages of easy manipulation, higher efficiency, energy saving, and reduced waste. Cytohistological studies were conducted to clarify the mechanism of HSH-MAE at cellular/tissue levels. Moreover, the established LC-MS/MS method showed excellent linearity, precision, repeatability, and reproducibility. The HSH-MAE-LC-MS/MS method was also successfully applied for screening high-productivity ITHRCs. Overall, this study opened up a new avenue for the direct determination of secondary metabolic profiles from fresh plant in vitro cultures, which is valuable for improving quality control of plant cell/organ cultures and sheds light on the metabolomic analysis of biological samples. Graphical Abstract HSH-MAE-LC-MS/MS opened up a new avenue for the direct determination of alkaloids and flavonoids in fresh Isatis tinctoria hairy root cultures.
authors.affiliation: State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin, 150040, China.
authors.forename: Jiao
authors.initials: J
authors.lastname: Jiao
authors.forename: Qing-Yan
authors.initials: QY
authors.lastname: Gai
authors.forename: Lin
authors.initials: L
authors.lastname: Zhang
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Meng
authors.initials: M
authors.lastname: Luo
authors.forename: Yuan-Gang
authors.initials: YG
authors.lastname: Zu
authors.forename: Yu-Jie
authors.initials: YJ
authors.lastname: Fu
mesh_terms: D000470:Alkaloids / Q000032:analysis*; D002853:Chromatography, Liquid / Q000379:methods*; D005419:Flavonoids / Q000032:analysis*; D031219:Isatis / Q000166:cytology / Q000378:metabolism*; D008872:Microwaves; D018517:Plant Roots / Q000166:cytology*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000470:Alkaloids; D005419:Flavonoids