Spiering Martin J, Davies Elizabeth, Tapper Brian A, Schmid Jan, Lane Geoffrey A
Institute of Molecular BioSciences, College of Sciences, Massey University, Palmerston North, New Zealand.
J Agric Food Chem. 2002 Oct 9;50(21):5856-62. doi: 10.1021/jf025602b.
An improved extraction and cleanup procedure for quantitative analysis of ergovaline in Neotyphodium-infected grass tissues by high-performance liquid chromatography was developed, utilizing aqueous 2-propanol-lactic acid as extraction solvent. Losses of sample material and time requirements were significantly reduced, handling procedures simplified, and ergovaline and internal standard ergotamine recovered with similar efficiency from extracts. Analyses can be carried out on very small amounts (2-5 mg of dry weight) of samples and another endophyte-alkaloid, peramine, determined in the same extracts. Calibration curves with 2-propanol-lactic acid were linear over the range 0.004-0.938 microM ergovaline (= 2-500 ng/mL) in extracts, corresponding to 0.04-10 microg/g in samples. The distribution of ergovaline in the plant was extremely heterogeneous, indicating low in-planta mobility and strong regulation of accumulation by the internal plant environment. In contrast, peramine was much more uniformly distributed. These results clearly demonstrate very large differences in the tissue specificities of ergovaline and peramine.
开发了一种改进的提取和净化程序，用于通过高效液相色谱法定量分析感染内生真菌的禾本科植物组织中的麦角缬氨酸，使用2-丙醇-乳酸水溶液作为提取溶剂。样品材料的损失和时间要求显著降低，处理程序简化，麦角缬氨酸和内标麦角胺从提取物中的回收率相似。可以对非常少量（2-5毫克干重）的样品进行分析，并在相同提取物中测定另一种内生真菌生物碱——过胺。在提取物中，2-丙醇-乳酸的校准曲线在0.004-0.938微摩尔麦角缬氨酸（=2-500纳克/毫升）范围内呈线性，相当于样品中0.04-10微克/克。麦角缬氨酸在植物中的分布极不均匀，表明其在植物体内的移动性较低，且受植物内部环境的强烈调控。相比之下，过胺的分布更为均匀。这些结果清楚地表明麦角缬氨酸和过胺在组织特异性上存在很大差异。
