Quantification of loline alkaloids

Lolines were extracted from 50 mg of freeze-dried, ground plant samples using a solution of 40% methanol/5% ammonia and 1,2-dichloroethane containing 54.8 ng mL⁻¹ 4-phenylmorpholine as internal standard. Plant extracts were centrifugated, and supernatants transferred to glass GC vials via a 20 µm filter for analysis. The analysis was conducted using a GC flame ionization detector (GC2010Plus, Shimadzu Corporation, Japan) and separation was achieved on a ZB-5 capillary column (30 m × 0.32 mm × 0.25 µm film). More information can be found in Bastías et al. (2018a). The detection limit was 25 µg g⁻¹ DW.