Multivariate optimization of a headspace solid-phase microextraction method followed by gas chromatography with mass spectrometry for the determination of terpenes in Nicotiana langsdorffii.
A simple and sensitive procedure based on headspace solid-phase microextraction and gas chromatography with mass spectrometry was developed for the determination of five terpenes (α-pinene, limonene, linalool, α-terpineol, and geraniol) in the leaves of Nicotiana langsdorffii. The microextraction conditions (extraction temperature, equilibration time, and extraction time) were optimized by means of a Doehlert design. The experimental design showed that, for α-pinene and limonene, a low temperature and a long extraction time were needed for optimal extraction, while linalool, α-terpineol, and geraniol required a high temperature and a long extraction time. The chosen compromise conditions were temperature 60°C, equilibration time 15 min and extraction time 50 min. The main analytical figures of the optimized method were evaluated; LODs ranged from 0.07 ng/g (α-pinene) to 8.0 ng/g (geraniol), while intraday and interday repeatability were in the range 10-17% and 9-13%, respectively. Finally, the procedure was applied to in vitro wild-type and transgenic specimens of N. langsdorffii subjected to abiotic stresses (chemical and heat stress). With the exception of geraniol (75-374 ng/g), low concentration levels of terpenes were measured (ng/g level or lower); some interesting variations in terpene concentration induced by abiotic stress were observed.
Department of Chemistry and Industrial Chemistry, University of Genoa, Genoa, Italy.
Francisco
F
Ardini
Marina Di
MD
Carro
Maria Luisa
ML
Abelmoschi
Marco
M
Grotti
Emanuele
E
Magi
D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D010936:Plant Extracts / Q000032:analysis* / Q000302:isolation & purification*; D030821:Plants, Genetically Modified / Q000737:chemistry* / Q000235:genetics; D052617:Solid Phase Microextraction / Q000379:methods*; D013729:Terpenes / Q000032:analysis* / Q000302:isolation & purification*; D014026:Nicotiana / Q000737:chemistry* / Q000235:genetics
D010936:Plant Extracts; D013729:Terpenes
Abiotic stress; Doehlert design; Genetic modifications; Solid-phase microextraction; gas chromatography