Electrospray mass spectrometry alkaloid quantification (Plantae)

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Method synopsis

Plant tissues were extracted in methanol and analyzed by electrospray mass spectrometry to identify and quantify norditerpene alkaloids. Mass spectra were recorded over 150-800 m/z range and alkaloid concentrations calculated based on relative abundance of internal standard.

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 — Norditerpene alkaloid concentrations in tissues and floral rewards of larkspurs and impacts on pollinators (2013), Biochemical Systematics and Ecology. Implementations in other papers (listed below) may differ.

1. Extract alkaloids from plant material
   A measured quantity of plant material (50 mg) was extracted in 5 ml methanol for 16 h. For nectar, reserpine (250 μg for D. barbeyi and 125 μg for D. nuttallianum) was added as internal reference. Sample was mixed then centrifuged and aliquot extracted. Sample was diluted into 1:1 methanol/1% acetic acid for analysis.
   Quantities: 50 mg plant material in 5 ml methanol, 250 μg or 125 μg reserpine for nectar samplesDuration: 16 hours extraction timeConditions: Not specified
   Equipment: Centrifuge
2. Analyze samples by mass spectrometry
   Plant extracts analyzed by electrospray mass spectrometry. For nectar, reverse-phase HPLC-esi(+) MS was used with a measured volume of nectar (10 μl) extracted in 90 μl 1:1 methanol/1% acetic acid. Mass spectra recorded for each sample over 150-800 m/z range and averaged across scans at 40% peak height.
   Quantities: 10 μl nectar in 90 μl extraction solutionDuration: Not specifiedConditions: 150-800 m/z range, 40% peak height
   Equipment: Electrospray mass spectrometer, Reverse-phase HPLC

Typical Equipment

• Electrospray mass spectrometer
• Wiley mill
• Retsch mixer mill MM301
• Centrifuge

Output Measurements

• alkaloid concentrations
• alkaloid identities
• mass spectra

Papers Using This Protocol (1)

Species Studied With This Protocol

Places Where Used