Lupine alkaloid extraction and GC analysis (Fabaceae)

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Method synopsis

Comprehensive method for extracting quinolizidine and piperidine alkaloids from plant tissues using acid-base extraction followed by GC/FID and GC/MS analysis for identification and quantification.

Synthesized from method descriptions across 1 paper using this protocol.

Procedure from a recent peer-reviewed implementation

Steps below were extracted from the most recent peer-reviewed implementation of this protocol in the corpus — Pollen and vegetative secondary chemistry of three pollen-rewarding lupines (2019), American Journal of Botany. Implementations in other papers (listed below) may differ.

1. Sample preparation
   Air-dried all plant material and ground to pass through a 2 mm screen. Weighed measured quantities into 16 mL screw-top glass test tubes for extraction.
   Quantities: Measured quantities (specific amounts not specified)Duration: Until completely driedConditions: Air-dried
   Equipment: 2 mm screen, 16 mL screw-top glass test tubes
2. Alkaloid extraction
   Extracted plant material by mechanical rotation using Rugged Rotator with mixture of 1 N HCl (4.0 mL) and CHCl3 (4.0 mL) for 15 min. Centrifuged samples 5 min and removed the clean test tube containing plant material and CHCl3. Added additional 2.0 mL of 1 N HCl, repeated extraction by mechanical rotation for 15 min, centrifuged, and removed clean test tubes. Combined aqueous portions in clean test tubes. Adjusted pH to 9-10 with concentrated NH4OH and extracted the basic solution twice with CHCl3, first with 4.0 mL then with 2.0 mL. Combined and filtered solutions through anhydrous Na2SO4, then clean test tubes, and the solvent was evaporated under N2 at 60°C.
   Quantities: 4.0 mL 1N HCl + 4.0 mL CHCl3 initially, then additional 2.0 mL 1N HCl, then 4.0 mL CHCl3 followed by 2.0 mL CHCl3Duration: 15 min mechanical rotation twice, 5 min centrifugationConditions: pH adjusted to 9-10, evaporated at 60°C under N2
   Equipment: Rugged Rotator, centrifuge, anhydrous Na2SO4, N2 source
3. Sample reconstitution for analysis

Typical Equipment

• Rugged Rotator
• Shimadzu GC-2010
• Finnigan MAT GCQ
• centrifuge

Output Measurements

• alkaloid concentrations
• alkaloid identification
• alkaloid composition profiles

Papers Using This Protocol (1)

Species Studied With This Protocol

Places Where Used