Hinojo M J, Medina A, Valle-Algarra F M, Gimeno-Adelantado J V, Jiménez M, Mateo R
Depto. Microbiologia y Ecologia, Facultad de Biologia, Universitat de Valencia, Dr. Moliner 50, E-46100 Burjassot, Valencia, Spain.
Food Microbiol. 2006 Apr;23(2):119-27. doi: 10.1016/j.fm.2005.03.006.
Fumonisin B1 (FB1) and fumonisin B2 (FB2) are the main members of a family of mycotoxins produced by various fungal species belonging to the Gibberella fujikuroi complex. The present work shows the results of a comparative study of various clean-up and derivatization procedures for analysis of fumonisins in rice cultures. Fumonisins were extracted from rice with acetonitrile/water (50/50, v/v). For clean-up, three solid-phase extraction procedures were assayed (C18 cartridge, SAX cartridge, and a combination of both). Two reagents (o-phthaldialdehyde and 4-fluoro-7-nitro-benzofurazan) were studied comparatively for formation of fluorescent derivatives. The separation was carried out by LC using a fluorescence detector. The best procedure for analysis of fumonisins in rice involved clean-up with C18 cartridge and derivatization with o-phthaldialdehyde. The limit of detection was 0.010 mg kg(-1) for both toxins. In the 10-500 mg kg(-1) spiking level range, the recovery rates for FB1 and FB2 in rice varied from 94.6% to 103.6% and from 96.3% to 101.9%, respectively. The optimized analytical method for determination of fumonisins in rice was applied to the study of FB1 and FB2 production by four isolates of the G. fujikuroi species complex in rice cultures carried out at different temperatures and water activities to establish the influence of strain and environmental conditions on fumonisin production in this cereal. In general, fumonisin production was the highest at 20 degrees C and lowest at 37 degrees C. Four of the five assayed water activity (aw) values (0.97, 0.98, 0.99, and 1.0) did not affect significantly fumonisin accumulation but fumonisins were not detected in cultures when aw was 0.96.
伏马菌素B1(FB1)和伏马菌素B2(FB2)是由藤仓赤霉复合体中的多种真菌产生的一类霉菌毒素的主要成员。本研究展示了对水稻培养物中伏马菌素进行分析时,各种净化和衍生化程序的比较研究结果。伏马菌素用乙腈/水(50/50,v/v)从水稻中提取。为了净化,测试了三种固相萃取程序(C18柱、SAX柱以及两者的组合)。比较研究了两种试剂(邻苯二甲醛和4-氟-7-硝基苯并呋咱)用于形成荧光衍生物的情况。采用荧光检测器通过液相色谱进行分离。分析水稻中伏马菌素的最佳程序是用C18柱净化并用邻苯二甲醛衍生化。两种毒素的检测限均为0.010 mg kg⁻¹。在10 - 500 mg kg⁻¹的加标水平范围内,水稻中FB1和FB2的回收率分别为94.6%至103.6%和96.3%至101.9%。将优化后的水稻中伏马菌素测定分析方法应用于对藤仓赤霉物种复合体的四个分离株在不同温度和水分活度下进行的水稻培养物中FB1和FB2产生情况的研究,以确定菌株和环境条件对这种谷物中伏马菌素产生的影响。总体而言,伏马菌素产量在20℃时最高,在37℃时最低。所测定的五个水分活度(aw)值中的四个(0.97、0.98、0.99和1.0)对伏马菌素积累没有显著影响,但当aw为0.96时,培养物中未检测到伏马菌素。