Scott Ian M, Puniani Evaloni, Jensen Helen, Livesey John F, Poveda Luis, Sanchez-Vindas Pablo, Durst Tony, Arnason John T
Biology Department, University of Ottawa, Ottawa, Ontario, K1N 6N5, Canada.
J Agric Food Chem. 2005 Mar 23;53(6):1907-13. doi: 10.1021/jf048305a.
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.
开发了一种用于提取和高效液相色谱 - 质谱仪（HPLC-MS）分析药用重要属胡椒属（胡椒科）的方法。这使得能够快速准确地测定黑胡椒（Piper nigrum L.）以及中美洲野生种中不饱和酰胺或胡椒酰胺的含量。回流提取从叶片和胡椒粒材料中获得了最高的胡椒碱回收率（>80%）。使用乙腈和水的二元梯度进行HPLC分析，在5至12分钟之间分离出主要的酰胺峰。大气压化学电离（APCI）-MS将检测限提高到0.2 ng，基于0.1至250μg/mL之间的线性标准曲线（R2 = 0.999），比HPLC - 二极管阵列检测器（DAD）的2 ng检测限低10倍。HPLC-MS方法鉴定出了异缬草素、胡椒酯碱、4,5 - 二氢胡椒碱、胡椒碱、4,5 - 二氢胡椒碱、胡椒碱和哌啶酮。通过蚊虫幼虫生物测定评估的六种哥斯达黎加胡椒属物种的生物活性与胡椒酰胺含量密切相关。
