Rönnberg A L, Hansson C, Håkanson R
Anal Biochem. 1984 Jun;139(2):338-44. doi: 10.1016/0003-2697(84)90014-9.
A high-performance liquid chromatographic (HPLC) method for the determination of histamine in tissues, based on precolumn derivation with o-phthalaldehyde, is described. Trichloroacetic acid extracts of rat brain, but not of rat stomach or of rat peritoneal mast cells, had to be cleaned-up by a chromatographic step before HPLC. The extracts were allowed to react with o-phthalaldehyde at pH 12.5 and -20 degrees C for 12 h, followed by acidification to pH 2.0. HPLC was performed on a reverse-phase column with isocratic elution using sulfuric acid in methanol as solvent system. A fluorescence detection system was used; excitation was set at 353 nm and emission was read at 451 nm. One chromatographic run was completed in 20 min. The detection limit with the conventional procedure was 1.5 ng histamine per sample, with a scaled-down procedure it was 250 pg per sample. With extracts of rat gastric mucosa the within-run variation was 2.7% and the day-to-day variation 4.5%.
描述了一种基于邻苯二甲醛柱前衍生化测定组织中组胺的高效液相色谱（HPLC）方法。大鼠脑的三氯乙酸提取物，而非大鼠胃或大鼠腹膜肥大细胞的提取物，在进行HPLC分析前必须经过色谱步骤净化。提取物在pH 12.5和-20℃条件下与邻苯二甲醛反应12小时，随后酸化至pH 2.0。使用硫酸甲醇溶液作为溶剂系统，在反相柱上进行等度洗脱的HPLC分析。采用荧光检测系统；激发波长设定为353nm，发射波长读取为451nm。一次色谱运行在20分钟内完成。常规方法的检测限为每个样品1.5ng组胺，缩小规模的方法检测限为每个样品250pg。对于大鼠胃黏膜提取物，批内变异为2.7%，日间变异为4.5%。
