Meyer R, Vorster S, Dubery I A
Department of Chemistry and Biochemistry, RAU-University, P.O. Box 524, Auckland Park, 2006, South Africa.
Anal Bioanal Chem. 2004 Oct;380(4):719-24. doi: 10.1007/s00216-004-2817-5. Epub 2004 Sep 15.
Self-packed micro-tip columns containing a C18-bonded silica stationary phase, based on the same principles as solid-phase extraction methods, were used to obtain gossypol and related sesquiterpenoid aldehyde-enriched fractions. The enriched metabolite fractions were then analyzed by optimized high-performance liquid chromatography (HPLC) with a C18 column (4.6 mm x 25 cm) eluted with the binary mobile phase acetonitrile-0.1% aqueous TFA solution (80:20). This method has proven to be highly reproducible. The precision and accuracy, as %RSD and %RME values, were determined to be less than 15% for the method. The minimum detection limit of gossypol was determined to be 10 ng (absolute gossypol). Absolute recovery was greater than 94% with a standard deviation of +/-3.68%. This is a simple, fast, and cost-effective method for isolation, identification, and quantification of gossypol and related secondary metabolites. Comparative analysis of gossypol content was performed on different parts of the cotton plant (seeds, stems and leaves) of two different cultivars of Gossypium hirsutum L. (Acala(1517-70) and OR19). The results indicate that the OR19 cv naturally contains higher gossypol levels than the Acala cv. It was also found that treatment of leaves with a Verticillium dahliae-derived elicitor induced production of deoxyhemigossypol rather than gossypol.
基于与固相萃取方法相同的原理，使用装有C18键合硅胶固定相的自装微尖柱来获得棉酚和相关倍半萜醛富集馏分。然后，采用优化的高效液相色谱法（HPLC），使用C18柱（4.6 mm×25 cm），以乙腈-0.1% TFA水溶液（80:20）作为二元流动相进行洗脱，对富集的代谢物馏分进行分析。该方法已被证明具有高度的可重复性。该方法的精密度和准确度，以%RSD和%RME值表示，测定结果均小于15%。棉酚的最低检测限测定为10 ng（绝对棉酚）。绝对回收率大于94%，标准偏差为±3.68%。这是一种用于分离、鉴定和定量棉酚及相关次生代谢物的简单、快速且经济高效的方法。对陆地棉（Acala(1517 - 70)和OR19）两个不同品种的棉花植株不同部位（种子、茎和叶）的棉酚含量进行了比较分析。结果表明，OR19品种天然含有的棉酚水平高于Acala品种。还发现，用大丽轮枝菌来源的激发子处理叶片会诱导产生脱氧半棉酚而非棉酚。
