Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden. takashi.yoshitake @ ki.se
Skin Pharmacol Physiol. 2012;25(2):65-72. doi: 10.1159/000330899. Epub 2011 Oct 5.
To develop a sensitive and selective liquid-chromatographic method for the determination of histamine in microdialysis samples from guinea pig skin following allergenic provocation.
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.
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%.
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.
建立一种灵敏、选择性的高效液相色谱荧光检测法，用于测定过敏激发后豚鼠皮肤微透析样品中的组胺。
该新的荧光衍生化方法基于组胺的 2 个氨基与 2 分子 4-（1-芘基）丁酰氯（PBC）之间的分子内激基缔合物反应，生成相应的二芘标记衍生物。
PBC 衍生的组胺在 20 分钟内得到分离，组胺的检测限（信噪比=3）为 0.6 fmol/20 μl 进样量。豚鼠皮肤微透析样品中组胺的基础细胞外浓度为 20.6±1.7 fmol/10 μl。皮下给予组胺释放剂化合物 48/80（3 mg/kg）可使皮肤透析液中细胞外组胺水平升高约 860%，而致敏豚鼠静脉内给予卵清蛋白（2 mg/kg）可使细胞外组胺水平升高约 3030%。
该测定豚鼠皮肤微透析样品中组胺的新方法可用于抗组胺药物的临床前研究以及各种皮肤制剂（如透皮给药系统）的变应原性评价。
