Smedsgaard J
Department of Biotechnology, Technical University of Denmark, Lyngby, Denmark.
J Chromatogr A. 1997 Jan 31;760(2):264-70. doi: 10.1016/s0021-9673(96)00803-5.
A simple and rapid standardized micro-scale extraction procedure has been developed to prepare extracts from fungal cultures for high-performance liquid chromatographic (HPLC) analysis. The method is based on ultrasonic extraction of three 6-mm plugs cut from a culture using 0.5 ml of solvent followed by a simple solvent change, filtration and injection. Approximately 5 min of work is involved in the extraction and work-up process and the extract can prepared for HPLC analysis within 60-70 min. The method has been used for determination of chromatographic metabolite profiles from 395 fungal isolates, including all terverticillate Penicillium species, cultivated on both Czapek Yeast Autolysate agar and Yeast Extract Sucrose agar. The concentration of the extracts proved to be sufficient to determine all secondary metabolites reported to be produced by these species using HPLC with diode array detection. These findings were confirmed by analyses of 132 pure metabolite standards.
已开发出一种简单快速的标准化微量提取程序，用于从真菌培养物中制备提取物，以进行高效液相色谱（HPLC）分析。该方法基于以下步骤：从培养物中切下三个6毫米的菌块，用0.5毫升溶剂进行超声提取，然后进行简单的溶剂更换、过滤和进样。提取和后处理过程大约需要5分钟的工作量，提取物可在60 - 70分钟内制备好用于HPLC分析。该方法已用于测定395株真菌分离物的色谱代谢物谱，这些分离物包括所有轮枝状青霉属物种，它们在察氏酵母自溶物琼脂和酵母提取物蔗糖琼脂上培养。事实证明，提取物的浓度足以使用配备二极管阵列检测的HPLC测定据报道由这些物种产生的所有次生代谢物。对132种纯代谢物标准品的分析证实了这些发现。
