|Detail of the metabolite profile
||Hydrolysis non-polar gas chromatography mass spectroscopy (GC-MS)
|Description for analysis method
||A portion of 0.5 mL organic extract was dried under N2 and hydrolysed by adding 0.5 ml 1.25 M HCl in MeOH and incubating for 4 h at 50 ??C. Following hydrolysis, HCl and solvent were evaporated under nitrogen. The sample was resuspended in 30 uL pyridine and derivatized through the addition of 20 uL of a commercial derivatization solution containing MSTFA+1%TMCS (Pierce Biotechnology, Rockford, IL, USA). The sample was incubated for 1 h at 50 ??C to allow though derivatization. The sample was equilibrated to room temperature, transferred to a 200 uL glass insert, and analysed using an Agilent 6890 GC coupled to a 5973 MSD scanning from m/z 50-650. Samples were injected at a 1:1 split ratio, and the inlet and transfer line were held at 280 ?C. Separation was achieved with a temperature programme of 80 ??C for 2 min, then ramped at 5 ??C/min to 315 ??C and held for 12 min, a 60 m DB-5MS column (J&W Scientific, 0.25 mm ID, 0.25 lm film thickness) and a constant flow of 1.0 ml/min.
|Species / Cultivar / Tissue
||Humulus lupulus / / Trichome
|Description for biological material
||Female cones were mashed in liquid nitrogen and glandular trichomes were separated from cones. The trichomes were shifted into a glass vial through a fine metal mesh. The samples were lyophilized prior to analysis.
||Don Sik Yang, Lloyd W. Sumner. Metabolic Profiling of trichomes from Potato Leafhopper susceptible and resistant alfalfa lines. (unpublished)
Plant Biology Division
The Samuel Roberts NOBLE Foundation, Inc.
Ardmore, OKLAHOMA, U.S.A - 73401.
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