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題 名 | Method for Bile Acid Determination by High Performance Liquid Chromatography=高效液相色譜法測定膽汁 |
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作 者 | 陳紅梅; 歐陽巍; | 書刊名 | Journal of Medical Sciences |
卷 期 | 23:5 2003.10[民92.10] |
頁 次 | 頁277-279 |
分類號 | 415.121 |
關鍵詞 | 高效液相色譜法; 膽汁; Bile acid; Chromatography; Conjugated; |
語 文 | 英文(English) |
英文摘要 | Background: Several high-performance liquid chromatographic (HPLC) methods have been described for determining and measuring the major conjugated bile acids in human bile. Traditionally, in-vitro bile acid experimentation has involved the use of HPLC to determine the quantity of various tauro- and glycol- conjugates in complex mixtures of bile acids in aqueous media. Methods to separate such mixtures used in in-vitro human bile experiments have required a lengthy workup involving a (1:4; v:v) sample: isopropanol extraction followed by an evaporation and re-suspension. While these methods can produce accurate results, the extraction and evaporation processes are labor intensive, time consuming and not practical for experiments requiring intensive sampling. Methods: To generate a standard set of results, standard solutions of glycholic acid (GCA), taurodeoxycholic acid (TDCA), and glycodeoxycholic acid (GDCA) were prepared and concentrations were determined by a reversed-phase C18 HPLC column, running acetate buffer and methanol (30:70). The flow rate was 1.0 ml/min and detection was performed at 205 nm. Results: the chromatograms show adequate resolution and normal retention times and the calibration curves have correlation of determinants factors (R²) ranging from 0.9920 to 0.9895. Conclusions: The method for bile acid separation identified here eliminates the time intensive and labor intensive workup step of evaporation, allowing for an efficient workup while preserving the quality of bile acid separation and quantification. |
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