117828223

520 M. Daszykowski et al.

High-Performance Liąuid Chromatography with Diodę-Array

Detection (HPLC-DAD)

HPLC was performed with a Gyncotek (Macclesfield, UK) liąuid chromato-graph eąuipped with the Gyncotek Giną 50 autosampler, the Gyncotek P 580A LPG pump, the Gyncotek UVD 340U diode-array detector (DAD), and Chromeleon Dionex v. 6.4 software for data acąuisition and processing. Compounds were separated on a 250 mm * 4.6 mm i.d., 5-pm particie, RP-18 column (Varian, Harbor City, CA, USA; #1215-9307) with acetonitrile-water 55:45 (v/v), containing a tracę amount of glacial acetic acid, as iso-cratic mobile phase. The flow ratę was 0.6 mL min^-1. The quantitative com-position of the mobile phase was specially developed for the purpose of this analysis).

Development of the ASE procedurę used for plant extraction followed by HPLC-DAD acąuisition of the sagę fingerprints has been described in detail elsewhere [32].

Head-Space Gas Chromatography with Mass Spectrometric

Detection (HS-GC-MS)

Each sagę species (1 g) was weighed, powdered in a porcelain mortar, and placed in a 10-mL glass vial stoppered with a Teflon-faced silicone rubber septum. Three replicates of each sample were processed identically. HS-GC-MS analysis was performed with a Tracę 2000 gas chromatograph, an MS Tracę mass detector (both from ThermoQuest, Waltham, MA, USA), and a CTC Analytics autosampler (Combi PAL, Basel, Switzerland), used in the head-space modę. Temperaturę and time of head-space desorption were, respectively, 70°C and 15 min. The head-space phase (0.5 mL) was in-troduced into a 30 m * 0.25 mm i.d. capillary column coated with a 0.25-pm film of DB-5 (Agilent Technologies, Pało Alto, CA, USA; formerly J&W Sci-entific, Folsom, CA, USA). Helium (p = 100 kPa) was used as carrier gas. The column temperaturę was maintained at 40°C for 3 min, then pro-grammed at 8° min^-1 to 150°C, which was maintained for 15 min. The injec-tor temperaturę was 150°C.

The mass spectrometer was fitted with an El source operated at 70 eV. Identification of individual compounds was based on comparison of mass spectra obtained from sample components with those from standards or available from the NIST (National Lnstitute of Standards and Technology, Gaithersburg, MD, USA) software library. Retention times of peaks from the Salina samples were also compared with those of known components of other essential oils. For this purpose we used pine oil, peppermint oil, euca-