离子交换色谱法结合薄层色谱-激光扫描密度法测定伏马菌素 B(1)。

Use of ion-exchange chromatography coupled with TLC-laser scanning densitometry for the quantitation of fumonisin B(1).

机构信息

Department of Biochemistry, University College of Science, Osmania University, Hyderabad 500 007, India.

出版信息

Talanta. 1999 Sep 13;50(2):381-9. doi: 10.1016/s0039-9140(99)00131-9.

Abstract

A simple TLC-Laser scanning densitometric (TLC-LSD) method was developed for the quantitation of fumonisin B(1) (FB(1)) isolated from solid media cultures (corn) and liquid media cultures of toxigenic Fusarium moniliforme strains (F. moniliforme MRC 826, F. moniliforme 4223 and F. moniliforme 2927)). FB(1) was isolated from the cultures by solvent extraction (methanol:water, 3:1) and purified in a single step by ion-exchange chromatography using Dowex-1. FB(1) in the purified extracts was detected by TLC analysis using p-anisaldehyde as a post-chromatographic derivatizing agent. The major toxin identified was FB(1) (R(f) 0.51) along with traces of FB(2) (R(f) 0.57) and FB(3) (R(f) 0.60) based on their comparison with the reference standard fumonisins. The sensitivity of the TLC-LSD method for the quantitation of FB(1) was found to be 500 ng g(-1). The linear regression analysis performed for the quantitation of FB(1) by the TLC-LSD method showed a correlation coefficient (r) value of 0.9. Spiking studies revealed the recovery of standard FB(1) (5 and 10 mug g(-1)) loaded on to Dowex-1 in the range of 87-96%. The purity of FB(1) purified from the cultures was determined by the two-dimensional TLC analysis. Two-dimensional TLC-analysis of the purified FB(1) revealed the purity to be greater than 85%. The method developed may find wide application in the environmental monitoring of the FB(1) contaminations in the various agricultural commodities and screening fumonisin producing toxigenic strains of F. moniliforme.

摘要

建立了一种简单的 TLC-激光扫描密度法（TLC-LSD），用于定量检测从固体培养基（玉米）和产毒串珠镰刀菌菌株（F. moniliforme MRC 826、F. moniliforme 4223 和 F. moniliforme 2927）的液体培养基中分离出的伏马菌素 B1（FB1）。FB1 从培养物中通过溶剂提取（甲醇：水，3:1）提取，并通过使用 Dowex-1 的离子交换色谱法在单一步骤中纯化。在纯化提取物中，通过 TLC 分析使用对茴香醛作为衍生化试剂检测 FB1。根据与参考标准伏马菌素的比较，鉴定出的主要毒素是 FB1（R(f) 0.51），以及痕量的 FB2（R(f) 0.57）和 FB3（R(f) 0.60）。TLC-LSD 方法用于定量 FB1 的灵敏度被发现为 500ng g(-1)。通过 TLC-LSD 方法对 FB1 定量进行的线性回归分析显示相关系数（r）值为 0.9。加标研究表明，标准 FB1（5 和 10 µg g(-1)）在 Dowex-1 上的加载回收率在 87-96%范围内。通过二维 TLC 分析确定从培养物中纯化的 FB1 的纯度。纯化的 FB1 的二维 TLC 分析表明纯度大于 85%。所开发的方法可能在各种农业商品中 FB1 污染的环境监测以及筛选产伏马菌素的串珠镰刀菌产毒菌株方面得到广泛应用。