Wild medicinal plant secondary metabolite profiling via HPLC-UV (Apiaceae)

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Method synopsis

Field collection of Ligusticum porteri roots followed by methanol extraction and HPLC-UV analysis to quantify five secondary metabolites. Method adapted from previously published work by Rivero et al. (2012).

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 — Chemotypic variation in oshá (<i>Ligusticum porteri</i>) in Colorado, USA (2018), J Appl. Res. Med. Aromat. Plants. Implementations in other papers (listed below) may differ.

1. Field collection of plant specimens
   Whole roots were collected from four replicate plants in each population. Plants were identified using morphological criteria outlined in Weber and Wittmann's Colorado Flora. A voucher specimen from one population (Lower Mt. Herman) was deposited in the herbarium at Colorado College (COCO) in Colorado Springs, CO, USA.
   Quantities: Four replicate plants per population, eleven total populationsDuration: Collections occurred during the second week of August 2015 for 2015 samplesConditions: Field collections in subalpine and montane zones
   Equipment: GPS for location marking
2. Light environment measurement
   Light environment was measured as photosynthetically active radiation (PAR) at plant height between 10:00 and 15:00 h using a light meter and sensor. Populations were classified as understory (<40% full-sun PAR) or meadow (>40% full-sun PAR).
   Quantities: Light measurements taken at each plant locationDuration: Measurements between 10:00 and 15:00 hoursConditions: Daytime measurements during peak solar radiation hours
   Equipment: LI-250 A & LI-191R light meter and sensor, LI – COR, Lincoln, NE
3. Root extract preparation
   Roots were stored at 4°C until crude methanol extraction was made in late August of 2015 for roots collected in August 2015, or at -20°C for roots from 2012. Methanol extracts were stored in a freezer at -20°C. Crude extracts were filtered with a sterile 0.45 μm cellulose-acetate syringe filter prior to HPLC analyses.

Typical Equipment

• GPS
• LI-250 A & LI-191R light meter
• Agilent 1100 HPLC
• Restek Ultra II C18 column

Output Measurements

• compound concentrations in ppm
• retention times
• peak areas

Papers Using This Protocol (1)

Species Studied With This Protocol

Places Where Used