采用高效毛细管电泳法测定人体皮肤中的组氨酸和尿刊酸异构体。

Determination of histidine and urocanic acid isomers in the human skin by high-performance capillary electrophoresis.

作者信息

Hermann K, Abeck D

机构信息

Technische Universitat Munchen, Klinik und Poliklinik fur Dermatologie und Allergologie, Munich, Germany.

出版信息

J Chromatogr B Biomed Sci Appl. 2000 Nov 10;749(1):41-7. doi: 10.1016/s0378-4347(00)00376-5.

DOI:10.1016/s0378-4347(00)00376-5

PMID:11129077

Abstract

Histidine was baseline separated from histamine, 1-methylhistamine and cis- and trans-urocanic acid using high-performance capillary electrophoresis (HPCE) on a fused-silica column (50 cm x 75 microm) with 0.05 M NaH2PO4 buffer, pH 5.0, and 12 kV. The detection limit of histidine, trans- and cis-urocanic acid was 10(-6) M at a wavelength of 214 nm. The detection limit of the urocanic acid isomers was slightly enhanced to 5 x 10(-7) M at 267 nm. The transformation of the trans-urocanic acid standard in vitro into the cis-isomer was dependent on the time of exposure and the energy of the light source. UVB light induced a significantly faster conversion than UVA light. The HPCE method was used for the characterization and measurement of histidine and urocanic acid in human skin eluates. The concentrations of histidine, trans- or cis-urocanic acid in ethanol washes from the skin of healthy, non-allergic volunteers were 2.22+/-0.40 x 10(-5) 0.96+/-0.26 x 10(-5) and 1.04+/-0.30 x 10(-5) M, respectively, (mean+/-SEM, n=8). The results obtained by HPCE correlated well with data obtained by HPLC. Correlation coefficients of r2=0.981, r2=0.814 and r2=0.956 were found for histidine, trans- and cis-urocanic acid, respectively.

摘要

使用高效毛细管电泳（HPCE），在一根熔融石英柱（50 cm×75 微米）上，以 0.05 M 磷酸二氢钠缓冲液（pH 5.0）和 12 kV 的条件，组氨酸与组胺、1-甲基组胺以及顺式和反式尿刊酸实现了基线分离。在 214 nm 波长下，组氨酸、反式和顺式尿刊酸的检测限为 10⁻⁶ M。在 267 nm 波长下，尿刊酸异构体的检测限略有提高，为 5×10⁻⁷ M。反式尿刊酸标准品在体外转化为顺式异构体取决于暴露时间和光源能量。UVB 光诱导的转化比 UVA 光显著更快。HPCE 方法用于表征和测量人皮肤洗脱液中的组氨酸和尿刊酸。来自健康、非过敏志愿者皮肤的乙醇洗脱液中，组氨酸、反式或顺式尿刊酸的浓度分别为 2.22±0.40×10⁻⁵、0.96±0.26×10⁻⁵ 和 1.04±0.30×10⁻⁵ M（平均值±标准误，n = 8）。HPCE 获得的结果与 HPLC 获得的数据相关性良好。组氨酸、反式和顺式尿刊酸的相关系数分别为 r² = 0.981、r² = 0.814 和 r² = 0.956。