Isolation of atropine and scopolamine from plant material using liquid-liquid extraction and EXtrelut®columns.
Tropane alkaloids are toxic secondary metabolites produced by Solanaceae plants. Among them, plants from Datura genus produce significant amounts of scopolamine and hyoscyamine; the latter undergoes racemization to atropine during isolation. Because of their biological importance, toxic properties and commonly reported food and animal feed contamination by different Datura sp. organs, there is a constant need for reliable methods for the analysis of tropane alkaloids in many matrices. In the current study, three extraction and sample-clean up procedures for the determination of scopolamine and atropine in plant material were compared in terms of their effectiveness and repeatability. Standard liquid-liquid extraction (LLE) and EXtrelut®NT 3 columns were used for the sample clean-up. Combined ultrasound-assisted extraction and 24h static extraction using ethyl acetate, followed by multiple LLE steps was found the most effective separation method among tested. However, absolute extraction recovery was relatively low and reached 45-67% for atropine and 52-73% for scopolamine, depending on the compound concentration. The same method was also the most effective one for the isolation of target compounds from Datura stramonium leaves. EXtrelut®columns, on the other hand, displayed relatively low effectiveness in isolating atropine and scopolamine from such a complex matrix and hence could not be recommended. The most effective method was also applied to the extraction of alkaloids from roots and stems of D. stramonium. Quantitative analyses were performed using validated method based on gas chromatography with flame ionization detector (GC-FID). Based on the results, the importance of the proper selection of internal standards in the analysis of tropane alkaloids was stressed out.
Department of Environmental Analysis, Faculty of Chemistry, University of Gdańsk, Wita Stwosza 63, 80-308 Gdańsk, Poland.
Paula
P
Śramska
Department of Environmental Analysis, Faculty of Chemistry, University of Gdańsk, Wita Stwosza 63, 80-308 Gdańsk, Poland.
Artur
A
Maciejka
Department of Environmental Analysis, Faculty of Chemistry, University of Gdańsk, Wita Stwosza 63, 80-308 Gdańsk, Poland.
Anna
A
Topolewska
Department of Environmental Analysis, Faculty of Chemistry, University of Gdańsk, Wita Stwosza 63, 80-308 Gdańsk, Poland.
Piotr
P
Stepnowski
Department of Environmental Analysis, Faculty of Chemistry, University of Gdańsk, Wita Stwosza 63, 80-308 Gdańsk, Poland. Electronic address: lukasz.halinski@ug.edu.pl.
Łukasz P
ŁP
Haliński
D001285:Atropine / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D002849:Chromatography, Gas; D032302:Datura / Q000737:chemistry; D007692:Diatomaceous Earth / Q000737:chemistry*; D057230:Limit of Detection; D016014:Linear Models; D059625:Liquid-Liquid Extraction / Q000295:instrumentation / Q000379:methods*; D010936:Plant Extracts / Q000737:chemistry; D015203:Reproducibility of Results; D012601:Scopolamine / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D013010:Sonication
D010936:Plant Extracts; D007692:Diatomaceous Earth; C018540:Extrelut; D001285:Atropine; D012601:Scopolamine
Atropine; EXtrelut(®); Gas chromatography; Liquid-liquid extraction; Scopolamine; Tropane alkaloids