High pressure liquid chromatographic analysis of conjugated bile acids in human bile: simultaneous resolution of sulfated and unsulfated lithocholyl amidates and the common conjugated bile acids.
High pressure liquid chromatographic analysis of conjugated bile acids in human bile: simultaneous resolution of sulfated and unsulfated lithocholyl amidates and the common conjugated bile acids.
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发表时间：
1987-05
影响因子：
6.5
通讯作者：
S. Rossi;J. L. Converse;A. Hofmann
中科院分区：
文献类型：
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作者：
S. Rossi;J. L. Converse;A. Hofmann
建立了一种能同时分离人胆汁中四种胆石(硫酸和非硫酸形式的甘氨酸和牛磺酸)以及其他八种主要结合胆汁酸的反相高压液相色谱(HPLC)系统。该系统采用C18十八烷基硅烷色谱柱，用pH 5.35的磷酸甲醇缓冲液等压洗脱。相对胆汁酸浓度用200 nm处的吸光度测定。报告了人体胆汁中四种主要胆汁酸(胆酸、鹅去氧胆酸、熊去氧胆酸和脱氧胆酸)的甘氨酸和牛磺酰胺酸及其相应的非结合形式相对于鹅去氧胆酸的保留时间。甘氨酸和牛磺酸酰胺酸盐以及这些胆汁酸的非共轭形式的C23去甲基衍生物的保留时间也有报道。胆汁酸酰胺酸酯的最大吸收波长为200 nm，与(未硫化)甘氨酸和牛磺酸酰胺酸酯的最大吸光度非常相似。硫酸酯酰胆酰亚胺在200 nm处表现出摩尔吸光系数，比非硫酸酯酰胺酸甲酯的摩尔吸光度大1.4倍。未结合胆汁酸在200 nm或210 nm处的吸光度比相应的多肽结合物低20~30倍。该方法已被应用于14名健康受试者的胆汁样本，以确定人体胆汁中结合胆汁酸的模式。
A reversed phase high pressure liquid chromatography (HPLC) system capable of simultaneously separating four lithocholyl species (sulfated and unsulfated forms of lithocholylglycine and lithocholyltaurine) as well as the eight other major conjugated bile acids present in human bile is described. The system uses a C18 octadecylsilane column and isocratic elution with methanol phosphate buffer, pH 5.35. Relative bile acid concentration is determined by absorbance at 200 nm. Retention times relative to chenodeoxycholylglycine are reported for the four lithocholic acid forms, the glycine and taurine amidate of the four major bile acids present in human bile (cholic, chenodeoxycholic, ursodeoxycholic, and deoxycholic), and for their corresponding unconjugated forms. Retention times are also reported for the glycine and taurine amidates as well as the unconjugated form of the C23 norderivatives of these bile acids. Maximal absorbance of bile acid amidates is at 200 nm and is very similar for the (unsulfated) glycine and taurine amidates. Sulfated lithocholyl amidates exhibit molar absorptivities at 200 nm which are 1.4 times greater than that of non-sulfated lithocholyl amidates. Unconjugated bile acid absorbance at 200 nm or 210 nm is 20 to 30 times less than that of corresponding peptide conjugates. The method has been applied to samples of gallbladder bile obtained from 14 healthy subjects to define the pattern of conjugated bile acids present in human bile.