Simultaneous LC-MS/MS bioanalysis of alkaloids, terpenoids, and flavonoids in rat plasma through salting-out-assisted liquid-liquid extraction after oral administration of extract from Tetradium ruticarpum and Glycyrrhiza uralensis: a sample preparation strategy to broaden analyte coverage of herbal medicines.

Publisher: Springer-Verlag Country of Publication: Germany NLM ID: 101134327 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1618-2650 (Electronic) Linking ISSN: 16182642 NLM ISO Abbreviation: Anal Bioanal Chem Subsets: MEDLINE

Original Publication: Heidelberg : Springer-Verlag, 2002-

Herbal Medicine*; Alkaloids/*blood ; Chromatography, Liquid/*methods ; Evodia/*chemistry ; Flavonoids/*blood ; Glycyrrhiza uralensis/*chemistry ; Liquid-Liquid Extraction/*methods ; Plant Extracts/*administration & dosage ; Tandem Mass Spectrometry/*methods ; Terpenes/*blood; Administration, Oral ; Animals ; Female ; Limit of Detection ; Male ; Rats ; Rats, Sprague-Dawley ; Reference Standards

Herbal medicines have historically been practiced in combinatorial way, which achieves therapeutic efficacy by integrative effects of multi-components. Thus, the accurate and precise measurement of multi bioactive components in matrices is inalienable to understanding the metabolism and disposition of herbal medicines. In this study, aiming to provide a strategy that improves analyte coverage, evaluation of six protocols employing sample pretreatment methods- protein precipitation (PPT), liquid-liquid extraction (LLE), sugaring-out-assisted liquid-liquid extraction (SULLE), and salting-out-assisted liquid-liquid extraction (SALLE)- was performed by LC-MS/MS using rat plasma and a mixture of alkaloid (evodiamine, rutaecarpine, dehydroevodiamine), terpenoid (limonin, rutaevin, obacunone), and flavonoid (liquiritin, isoliquiritin, liquiritigenin) standards isolated from Tetradium ruticarpum and Glycyrrhiza uralensis. These protocols were as follows: (1) PPT with methanol, (2) PPT with acetonitrile, (3) LLE with methyl tertiary-butyl ether-dichloromethane, (4) LLE with ethyl acetate-n-butanol, (5) SALLE with ammonium acetate, (6) SULLE with glucose. The results suggested that SALLE produced broader analyte coverage with satisfactory reproducibility, acceptable recovery, and low matrix interference. Then, sample preparation procedure of SALLE, chromatographic conditions, and mass spectrometric parameters were optimized, followed by method validation, showing that good sensitivity (LLOQ ≤ 1 ng mL ^-1 ), linearity (r ≥ 0.9933), precision (RSD ≤ 14.45%), accuracy (89.54~110.87%), and stability could be achieved. Next, the developed method was applied successfully to determine the pharmacokinetic behavior of the nine compounds in rat plasma after intragastric administration with an extract from Tetradium ruticarpum and Glycyrrhiza uralensis (Wuzhuyu-Gancao pair). Based on an extensive review and experiments, a sample preparation procedure that matches with LC-MS/MS technique and can get wider analyte coverage was outlined. The developed SALLE method is rapid, reliable, and suitable for bioanalysis of analytes with diverse polarity, which was expected to be a promising strategy for the pharmacokinetic studies of herbal medicines. Graphical abstract.
(© 2021. Springer-Verlag GmbH Germany, part of Springer Nature.)

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ZY(2018-2020)-CCCX-5002 Three-year Action Plan for the Development of Traditional Chinese Medicine of Shanghai

Keywords: Bioanalysis; Glycyrrhiza uralensis; Liquid-liquid extraction; Phase separation; Protein precipitation; Tetradium ruticarpum

0 (Alkaloids)
0 (Flavonoids)
0 (Plant Extracts)
0 (Terpenes)

Date Created: 20210731 Date Completed: 20220105 Latest Revision: 20220105

20240829

10.1007/s00216-021-03568-1

34331552