Beil D, Kinder H, Paschke A, Steinhart H, Vieluf D, Behrendt H, Ring J
Department of Food Chemistry, University of Hamburg, Germany.
J Chromatogr Sci. 1998 Jun;36(6):284-6. doi: 10.1093/chromsci/36.6.284.
A reversed-phase high-performance liquid chromatographic method with gradient elution for the separation of the mediator substances histamine and the leukotrienes C4 (LTC4), D4 (LTD4), and E4 (LTE4) is described. The detection occurs fluorimetrically after automated precolumn derivatization with o-phthaldialdehyde. All components are chromatographically separable. Because of the different excitation and emission wavelengths, only the most important biological active mediators histamine and LTC4 are determinable in one parallel chromatographic run. The method is examined by linearity and precision tests and is applicable to biological sample matrices like cell supernatants of human basophils enriched by Percoll-density gradient centrifugation and stimulated for mediator release by anti-IgE. The established method is suitable to separate the mediators from other matrix components. The determination limit for histamine is 55.0 micrograms/L and that for LTC4 16.0 micrograms/L, referring to the reference solutions. Therefore, a fast, economical method for the common determination of the most important mediators histamine and LTC4 is established. This method is also suitable for high sample amounts in routine medical analysis.
描述了一种采用梯度洗脱的反相高效液相色谱法，用于分离介质物质组胺和白三烯C4（LTC4）、D4（LTD4）及E4（LTE4）。检测是在通过邻苯二甲醛进行自动柱前衍生化后采用荧光法进行。所有组分在色谱上均可分离。由于激发和发射波长不同，在一次平行色谱运行中仅能测定最重要的生物活性介质组胺和LTC4。该方法通过线性和精密度测试进行了检验，适用于生物样品基质，如通过Percoll密度梯度离心富集并经抗IgE刺激释放介质的人嗜碱性粒细胞的细胞上清液。所建立的方法适用于从其他基质成分中分离介质。以参考溶液计，组胺的测定限为55.0微克/升，LTC4的测定限为16.0微克/升。因此，建立了一种快速、经济的方法用于同时测定最重要的介质组胺和LTC4。该方法也适用于常规医学分析中的大量样品。
