National Center for Natural Products Research, Research Institute of Pharmaceutical Sciences, School of Pharmacy, The University of Mississippi University, MS 38677, USA.
J Pharm Biomed Anal. 2012 Nov;70:53-63. doi: 10.1016/j.jpba.2012.05.021. Epub 2012 Jun 5.
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.
已开发出 UPLC-UV 和 HPLC-TOF-MS 方法，用于分析杂种千里光（L.）G.M.和 Sch.（家族，菊科）根茎中的主要倍半萜和吡咯里西啶生物碱，以及声称含有杂种千里光的膳食补充剂。使用 Acquity UPLC™HSS T3（100mm×2.1mm，内径，1.8μm）柱系统，在 UPLC-UV 中使用由甲酸铵（50mM）和乙腈（0.05%甲酸）组成的流动相进行梯度洗脱，以恒定流速 0.25mL/min，可获得最佳结果。新开发的方法根据 ICH 指南进行了专属性、线性、准确性和精密度验证。通过 UPLC-UV 检测到吡咯里西啶生物碱和倍半萜的检测限分别为 5μg/mL 和 0.1μg/mL，而通过 HPLC-TOF-MS 检测分别为 0.001μg/mL 和 0.01μg/mL。该方法成功地用于分析不同的杂种千里光市场产品，并区分其他两种千里光属物种。在 15 种膳食补充剂中，发现总 petasins 含量在 0.02-11.6mg/剂量形式之间，而在另外 6 种膳食补充剂中未检测到 petasins。此外，在 7 种膳食补充剂中检测到吡咯里西啶生物碱，其被认为对肝脏有毒。在 7 种膳食补充剂中发现 petasin 的量在标签声称的范围内，且未检测到吡咯里西啶生物碱。描述了与电喷雾电离（ESI）接口联用的 HPLC-质谱法，用于鉴定和确认来自植物提取物和声称含有杂种千里光的膳食补充剂以及不同种千里光的倍半萜和吡咯里西啶生物碱。
