Arakawa Y, Tachibana S
Anal Biochem. 1986 Oct;158(1):20-7. doi: 10.1016/0003-2697(86)90582-8.
A convenient method for the routine measurement of histamine (HA) in biological samples was developed. This method does not require any preliminary purification or concentration of HA, and features high sensitivity, specificity, and reliability. The method consists of the direct application of the acid-deproteinized sample to high-performance liquid chromatography on a sulfonated polystyrene column with detection by means of a postcolumn fluorogenic reaction with o-phthaladehyde. The detection limit was found to be 0.1 pmol (signal-to-noise ratio = 3). The coefficient of variation for measurements of 10 pmol of standard histamine was 1.1%. Each chromatography takes only 10 min and therefore more than 50 samples can be measured in a day. The high sensitivity of the method allows it to be applied even to samples of very low HA concentration such as human plasma without any procedure for concentration of the sample, and further, only 0.1 ml of the sample is necessary for determination. The method was applied to compare the HA levels of the whole blood and plasma of man and various animals. Applications of the method to the supernatant of rat peritoneal mast cell incubates and to extracts of mouse brain and stomach are also described.
开发了一种用于常规测量生物样品中组胺（HA）的简便方法。该方法无需对HA进行任何初步纯化或浓缩，具有高灵敏度、特异性和可靠性。该方法包括将酸脱蛋白后的样品直接应用于磺化聚苯乙烯柱上的高效液相色谱，并通过与邻苯二甲醛的柱后荧光反应进行检测。检测限为0.1 pmol（信噪比 = 3）。10 pmol标准组胺测量的变异系数为1.1%。每次色谱分析仅需10分钟，因此一天内可测量50多个样品。该方法的高灵敏度使其即使应用于HA浓度非常低的样品（如人体血浆）也无需对样品进行任何浓缩程序，而且测定仅需0.1 ml样品。该方法用于比较人和各种动物全血和血浆中的HA水平。还描述了该方法在大鼠腹膜肥大细胞培养上清液以及小鼠脑和胃提取物中的应用。
