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[1]目的 为了测定黄刺蛾幼虫刺毛中的组胺含量,建立了反相高压液相色谱测定组胺的方法.[2]方法采用邻苯二甲醛(OPA)与组胺的柱前衍生化反应,生成的荧光产物以高压液相色谱分离,荧光检测器测定组胺含量.色谱条件:色谱柱为ZorboxODS,流动相为甲醇—磷酸二氢钠缓冲液(v:v＝47:53,pH9.45),流速为0.7ml/min黄刺蛾幼虫-40℃低温保存,剪下刺毛,用高氯酸抽提,抽提液经衍生化反应后注入色谱柱测定,荧光检测的激发波长为360nm,测量波长为450nm.[3]结果 在0.01~10.00nmol范围内,组胺OPA衍生物的色谱峰高与组胺量之间的线性回归方程为W=6.1600H—0.0021,r＝0.9973;最低检出限为2.5pmol.回收率为93.1%,变异系数CV＝3.5%(n＝4),日内差为3.9%(n＝6).黄刺蛾幼虫刺毛的组胺含量在0.5%~0.8%之间.[4]结论 本方法灵敏、可靠,有一定实用价值.
[1] Objective In order to determine the histamine content in the thorn burr of Trichoplusia ni L., a method was established for the determination of histamine by reversed-phase high pressure liquid chromatography. [2] The method used OPA and histamine Derivatization reaction, the resulting fluorescent product was separated by high pressure liquid chromatography, fluorescence detector determination of histamine content. Chromatographic conditions: the column was ZorboxODS, the mobile phase of methanol - sodium dihydrogen phosphate buffer (v: v = 47:53 , pH9.45) at a flow rate of 0.7ml / min. The Trichoplusia ni was preserved at -40 ℃. The bristles were cut and extracted with perchloric acid. The extract was injected into the column after derivatization reaction, Excitation wavelength of 360nm, the measurement wavelength of 450nm. [3] Results in 0.01 ~ 10.00nmol range of histamine OPA derivative peak height and histamine linear regression equation between the W = 6.1600H-0.0021, r = 0.9973, the minimum detectable limit was 2.5 pmol, the recovery rate was 93.1%, the coefficient of variation was 3.5% (n = 4), and the intraday difference was 3.9% (n = 6) In the range of 0.5% ~ 0.8% [4] Conclusion The method is sensitive, reliable and has certain practical value.