Analysis of Piperaceae germplasm by HPLC and LCMS: a method for isolating and identifying unsaturated amides from Piper spp extracts.
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 microg/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.
Scott
Ian M
IM
Biology Department, University of Ottawa, Ottawa, Ontario, K1N 6N5, Canada. iscott@science.uottawa.ca
Puniani
Evaloni
E
Jensen
Helen
H
Livesey
John F
JF
Poveda
Luis
L
Sanchez-Vindas
Pablo
P
Durst
Tony
T
Arnason
John T
JT
D000330:Aedes; D000470:Alkaloids / Q000032:analysis / Q000302:isolation & purification; D000577:Amides / Q000032:analysis* / Q000302:isolation & purification*; D000818:Animals; D052117:Benzodioxoles; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D007306:Insecticides; D007814:Larva; D013058:Mass Spectrometry / Q000379:methods*; D026321:Piperaceae / Q000737:chemistry*; D010880:Piperidines / Q000032:analysis / Q000302:isolation & purification; D010936:Plant Extracts / Q000737:chemistry*; D053284:Polyunsaturated Alkamides
D000470:Alkaloids; D000577:Amides; D052117:Benzodioxoles; D007306:Insecticides; D010880:Piperidines; D010936:Plant Extracts; D053284:Polyunsaturated Alkamides; C008922:piperine