Robitaille L, Hoffer L J
McGill Nutrition and Food Science Centre, McGill University, Montréal, Qué., Canada.
Can J Physiol Pharmacol. 1988 May;66(5):613-7. doi: 10.1139/y88-095.
A simple and rapid high performance liquid chromatographic technique is described for the separation and quantitation of plasma branched chain amino acids. After addition of a norleucine internal standard, plasma samples are acidified with acetic acid, and amino acids are separated from proteins and other plasma components by passage of the acidified plasma through an ion exchange resin. The ammonium hydroxide eluate from the resin is dried, phenylisothiocyanate derivatives are prepared, and the amino acids are separated on a Waters reverse-phase "Pico-Tag" column with an ultraviolet detector set at 254 nm. In addition to the branched chain amino acids (leucine, valine, and isoleucine), aspartate, glutamate, serine, threonine, alanine, and methionine are quantitated with high precision and accuracy, as verified by quantitative recovery and comparison with an automatic amino acid analyzer. The advantages of the method are its simplicity, speed, stability of derivatives, high reproducibility, low per-sample cost, and the use of a simple fixed-wavelength ultraviolet detector.
本文描述了一种简单快速的高效液相色谱技术，用于分离和定量血浆支链氨基酸。加入正亮氨酸内标后，血浆样品用乙酸酸化，通过将酸化的血浆通过离子交换树脂，使氨基酸与蛋白质和其他血浆成分分离。树脂的氢氧化铵洗脱液干燥后，制备苯异硫氰酸酯衍生物，氨基酸在 Waters 反相“Pico-Tag”柱上分离，紫外检测器设置在 254 nm。除了支链氨基酸（亮氨酸、缬氨酸和异亮氨酸）外，天冬氨酸、谷氨酸、丝氨酸、苏氨酸、丙氨酸和蛋氨酸也能高精度和准确地定量，通过定量回收率以及与自动氨基酸分析仪的比较得到验证。该方法的优点是简单、快速、衍生物稳定性好、重现性高、每个样品成本低，并且使用简单的固定波长紫外检测器。
