Kakiyama Genta, Hosoda Atsuko, Iida Takashi, Fujimoto Yasuo, Goto Takaaki, Mano Nariyasu, Goto Junichi, Nambara Toshio
Department of Chemistry, College of Humanities & Sciences, Nihon University, Sakurajousui, Tokyo 156-8550, Japan.
J Chromatogr A. 2006 Aug 25;1125(1):112-6. doi: 10.1016/j.chroma.2006.05.037. Epub 2006 Jun 13.
A direct method for the separation and quantification of a series of bile acid acyl glycosides using high-performance liquid chromatography coupled to an evaporative light scattering detector (HPLC-ELSD) is described. Complete separation of each of 15 bile acid acyl 24-alpha-glucosides and their 24-beta-anomers and 24-beta-galactosides was achieved by the stepwise gradient elution mode on a C18 column using a mixture of acetonitrile-methanol (8:2, v/v) and 1% aqueous acetic acid as the mobile phase. 24-beta-Galactosides were always eluted faster than the corresponding 24-beta-glucosides, which eluted after the corresponding 24-alpha-anomers. Calibration curves of different 24-beta-galactosides were linear over a range of 0.2-40 nmol of injected amount and the detection limits (S/N > 3) were from 0.08 to 0.1 nmol. The present HPLC-ELSD method may provide an insight into the separation and quantification of the biologically interesting neutral bile acids.
描述了一种使用高效液相色谱与蒸发光散射检测器（HPLC - ELSD）联用的直接方法，用于分离和定量一系列胆汁酸酰基糖苷。通过在C18柱上采用乙腈 - 甲醇（8:2，v/v）和1%乙酸水溶液的混合物作为流动相的逐步梯度洗脱模式，实现了15种胆汁酸酰基24 - α - 葡萄糖苷及其24 - β - 端基异构体和24 - β - 半乳糖苷的完全分离。24 - β - 半乳糖苷的洗脱速度总是比相应的24 - β - 葡萄糖苷快，而24 - β - 葡萄糖苷在相应的24 - α - 端基异构体之后洗脱。不同24 - β - 半乳糖苷的校准曲线在进样量0.2 - 40 nmol范围内呈线性，检测限（S/N > 3）为0.08至0.1 nmol。目前的HPLC - ELSD方法可能为具有生物学意义的中性胆汁酸的分离和定量提供见解。
