Simultaneous determination of sesquiterpenes and pyrrolizidine alkaloids from the rhizomes of Petasites hybridus (L.) G.M. et Sch. and dietary supplements using UPLC-UV and HPLC-TOF-MS methods.
UPLC-UV and HPLC-TOF-MS methods have been developed for the analysis of major sesquiterpenes and pyrrolizidine alkaloids from rhizomes of Petasites hybridus (L.) G.M. et Sch. (Family, Asteracea) and dietary supplements claiming to contain P. hybridus. The best results were obtained with Acquity UPLC™ HSS T3 (100 mm × 2.1 mm, I.D., 1.8 μm) column system using a gradient elution with a mobile phase consisting of ammonium formate (50mM) and acetonitrile (0.05% formic acid) at a constant flow rate of 0.25 mL/min via UPLC-UV. The newly developed method was validated according to the ICH guidelines with respect to specificity, linearity, accuracy and precision. The limits of detection were found to be 5 μg/mL and 0.1 μg/mL for pyrrolizidine alkaloids and sesquiterpenes, respectively by UPLC-UV and 0.001 and 0.01 μg/mL, respectively using HPLC-TOF-MS. The methods were successfully used to analyze different P. hybridus market products, as well as to distinguish between two other Petasites species. The total content of petasins was found to be in the range of 0.02-11.6 mg/dosage form for 15 dietary supplements and no petasins were detected in an additional six dietary supplements. Additionally, pyrrolizidine alkaloids, which are considered to be toxic for the liver, were detected in seven dietary supplements. The amount of petasin in seven dietary supplements was found to be within limits of label claim and no pyrrolizidine alkaloids were detected. HPLC-mass spectrometry coupled with electrospray ionization (ESI) interface method is described for the identification and confirmation of sesquiterpenes and pyrrolizidine alkaloids from plant extracts and dietary supplements that claim to contain P. hybridus as well as different species of Petasites.
National Center for Natural Products Research, Research Institute of Pharmaceutical Sciences, School of Pharmacy, The University of Mississippi University, MS 38677, USA. bavula@olemiss.edu
Bharathi
B
Avula
Yan-Hong
YH
Wang
Mei
M
Wang
Troy J
TJ
Smillie
Ikhlas A
IA
Khan
D002021:Buffers; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000592:standards; D002853:Chromatography, Liquid / Q000592:standards; D019587:Dietary Supplements / Q000032:analysis* / Q000592:standards; D057230:Limit of Detection; D036901:Petasites / Q000737:chemistry* / Q000145:classification; D011763:Pyrrolizidine Alkaloids / Q000032:analysis*; D011786:Quality Control; D012015:Reference Standards; D015203:Reproducibility of Results; D027343:Rhizome / Q000737:chemistry*; D012717:Sesquiterpenes / Q000032:analysis*; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000592:standards; D013056:Spectrophotometry, Ultraviolet / Q000592:standards
D002021:Buffers; D011763:Pyrrolizidine Alkaloids; D012717:Sesquiterpenes; C418727:petasin