Determination of Histamine in Microdialysis Samples from Guinea Pig Skin by High-performance Liquid Chromatography with Fluorescence Detection
Determination of Histamine in Microdialysis Samples from Guinea Pig Skin by High-performance Liquid Chromatography with Fluorescence Detection
复制标题
高效液相色谱荧光检测法测定豚鼠皮肤微透析样品中的组胺
DOI：
10.1159/000330899
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发表时间：
2012
影响因子：
2.7
通讯作者：
M.Yamaguchi
中科院分区：
文献类型：
--
作者：
T.Yoshitake;S.Ijiri;S.Yoshitake;K.Todoroki;H.Yoshida;J.Kehr;H.Nohta;M.Yamaguchi
目的：建立一种灵敏、选择性高的液相色谱法测定豚鼠皮肤微透析样品中组胺的含量。方法：基于组胺的2个氨基与2分子4-（1-芘）丁酰氯（PBC）之间的分子内准分子形成反应，生成相应的联芘标记衍生物。结果：组胺的PBC衍生物在20 min内分离，组胺的检测限（信噪比= 3）为0.6 fmol/20 µl进样体积。豚鼠皮肤微透析液中组胺的基础细胞外水平为20.6 ± 1.7 fmol/10 µl。皮下施用组胺释放剂化合物48/80（3 mg/kg）使皮肤透析液中的细胞外组胺水平增加约860%，而卵清蛋白攻击（2 mg/kg i. v.）结论：该方法可用于抗组胺药物的临床前研究和皮肤制剂如透皮给药系统的过敏性评价。
Aim:To develop a sensitive and selective liquid-chromatographic method for the determination of histamine in microdialysis samples from guinea pig skin following allergenic provocation.Methods:The novel fluorescence derivatization method is based on an intramolecular excimer-forming reaction between 2 amino moieties of histamine and 2 molecules of 4-(1-pyrene)butanoyl chloride (PBC) yielding the corresponding dipyrene-labeled derivative.Results:The PBC derivative of histamine was separated within 20 min, and the detection limit (signal-to-noise ratio = 3) of histamine was 0.6 fmol/20 µl volume injected. The basal extracellular levels of histamine in guinea pig skin microdialysates were 20.6 ± 1.7 fmol/10 µl. Subcutaneous administration of histamine liberator compound 48/80 (3 mg/kg) increased the extracellular histamine levels in the skin dialysates by about 860%, whereas ovalbumin challenge (2 mg/kg i.v.) in the sensitized guinea pigs increased the extracellular histamine levels by about 3,030%.Conclusion:The novel technique for histamine determination in microdialysis samples from the guinea pig skin may be utilized in preclinical research of antihistaminergic drugs and evaluation of allergenic properties of various dermal preparations such as transdermal drug delivery systems.