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Analysis of Piperaceae germplasm by HPLC and LCMS: a method for isolating and identifying unsaturated amides from Piper spp extracts
- Scott, I.M., Puniani, E., Jensen, H., Livesey, J.F., Poveda, L., Sanchez-Vindas, P., Durst, T., Arnason, J.T.
- Journal of agricultural and food chemistry 2005 v.53 no.6 pp. 1907-1913
- germplasm, black pepper, pesticidal plants, insecticidal properties, Georgecraigius atropalpus, plant extracts, amides, high performance liquid chromatography, botanical insecticides, bioassays, plant genetic resources, larvae, mass spectrometry, wild relatives, Piper nigrum
- A method for extraction and high performance liquid chromatography-mass spectrometer (HPLC-MS) analysis of the medicinally important genus Piper (Piperaceae) was developed. This allows for a rapid and accurate measure of unsaturated amides, or piperamides, in black pepper, Piper nigrum L., and in wild species from Central America. Reflux extraction provided the highest recovery of piperine (>80%) from leaf and peppercorn material. HPLC analysis using a binary gradient of acetonitrile and water separated the major amide peaks between 5 and 12 min. Atmospheric pressure chemical ionization (APCI)-MS improved the detection limit to 0.2 ng, 10-fold below the 2 ng limit of the HPLC-diode array detector (DAD) based on linear standard curves between 0.1 and 250 microgram/mL (R2 = 0.999). The HPLC-MS method identified pellitorine, piperylin, 4,5-dihydropiperlonguminine, piperlonguminine, 4,5-dihydropiperine, piperine, and pipercide. The biological activity of six Costa Rican Piper species assessed by mosquito larval bioassays correlated well with piperamide content.