Porter D W, Banks M A, Castranova V, Martin W G
Division of Animal and Veterinary Sciences, College of Agriculture and Forestry, West Virgina University, Morgantown 26506.
J Chromatogr. 1988 Nov 11;454:311-6. doi: 10.1016/s0021-9673(00)88624-0.
Taurine (2-aminoethanesulfonic acid) was quantitated by reversed-phase chromatography on a C18 Resolve column using a linear gradient of 9-11% methanol in water. Glutamine was used as the internal standard. Pre-column derivatization of the amino acid with o-phthalaldehyde allowed the detection of as little as 0.1 pmol taurine. Dual ion-exchange column chromatography was employed to remove other amino acids and metabolic precursors of taurine from the samples. Cysteic acid and cysteine sulfinic acid did not interfere with taurine analysis by the high-performance liquid chromatographic method. For sample deproteinization, boiling and picric acid precipitation were used. Recovery of taurine averaged 93.5 +/- 5.0% (means +/- standard error of the mean) from standard solutions and was not affected by the method of deproteinization. Using this procedure, plasma taurine concentrations for the rat and chick were determined to be 100.7 +/- 13.1 microM and 108.0 +/- 0.3 microM, respectively. Recovery of taurine from plasma samples averaged 97.2 +/- 4.7%.
采用反相色谱法,在C18 Resolve柱上,使用含9 - 11%甲醇的水线性梯度洗脱,对牛磺酸(2 - 氨基乙磺酸)进行定量分析。以谷氨酰胺作为内标。氨基酸与邻苯二甲醛进行柱前衍生化,可检测低至0.1皮摩尔的牛磺酸。采用双离子交换柱色谱法从样品中去除其他氨基酸和牛磺酸的代谢前体。半胱磺酸和半胱亚磺酸不干扰高效液相色谱法对牛磺酸的分析。样品脱蛋白采用煮沸和苦味酸沉淀法。牛磺酸从标准溶液中的回收率平均为93.5±5.0%(平均值±平均标准误差),且不受脱蛋白方法的影响。采用该方法,测定大鼠和雏鸡血浆中牛磺酸浓度分别为100.7±13.1微摩尔/升和108.0±0.3微摩尔/升。牛磺酸从血浆样品中的回收率平均为97.2±4.7%。