Fichtel Jörg, Köster Jürgen, Scholz-Böttcher Barbara, Sass Henrik, Rullkötter Jürgen
Institut für Chemie und Biologie des Meeres, Carl von Ossietzky Universität Oldenburg, D-26129 Oldenburg, Germany.
J Microbiol Methods. 2007 Aug;70(2):319-27. doi: 10.1016/j.mimet.2007.05.008. Epub 2007 May 21.
A high-performance liquid chromatographic method with indirect fluorescence detection has been developed for quantification of dipicolinic acid, a major constituent of bacterial endospores. After separation on a reversed-phase column, a post-column reagent of sodium acetate at 1 mol l(-1) with 50 micromol l(-1) terbium chloride was added for complexation of dipicolinic acid. Terbium monodipicolinate complexes formed were quantified by measuring the fluorescence emission maximum at 548 nm after excitation with UV light at 270 nm wavelength. Parameters of post-column complexation were optimized to achieve a detection limit of 0.5 nmol DPA l(-1), corresponding to about 10(3) Desulfosporosinus orientis endospores per ml. The method was applied to the analysis of spore contamination in tuna and for estimating the endospore numbers in marine sediments.
已开发出一种采用间接荧光检测的高效液相色谱法，用于定量测定细菌芽孢的主要成分吡啶二羧酸。在反相柱上分离后，加入1 mol l(-1)醋酸钠与50 micromol l(-1)氯化铽的柱后试剂，用于吡啶二羧酸的络合。通过在270 nm波长的紫外光激发后，测量548 nm处的最大荧光发射来定量形成的单吡啶二羧酸铽络合物。优化柱后络合参数，以实现0.5 nmol DPA l(-1)的检测限，相当于每毫升约10(3)个东方脱硫芽孢杆菌芽孢。该方法应用于金枪鱼中芽孢污染的分析以及海洋沉积物中芽孢数量的估算。
