AUTOMATIC RECORDING APPARATUS FOR USE IN THE CHROMATOGRAPHY OF AMINO ACIDS
AUTOMATIC RECORDING APPARATUS FOR USE IN THE CHROMATOGRAPHY OF AMINO ACIDS
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DOI：
10.1021/ac60139a006
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发表时间：
1958-01-01
影响因子：
7.4
通讯作者：
MOORE, S
中科院分区：
文献类型：
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作者：
SPACKMAN, DH;STEIN, WH;MOORE, S
氨基酸的定量测定是由一种能自动记录离子交换柱流出液的茚三酮色值的仪器进行的，它使测定更简单、更迅速。将不含空气的流入缓冲液以恒定速率泵送通过磺化聚苯乙烯树脂柱。流出物与由第二泵输送的茚三酮试剂的毛细管流相遇。通过将试剂和流出液的混合物通过浸没在沸水浴中的螺旋形聚四氟乙烯毛细管管来显色。当所得溶液流过2 mm孔径的圆柱形玻璃池时，在570和440 m/i下连续测量所得溶液的吸光度。对于0.1 - 3.0 jumoles的每种氨基酸，记录曲线上的峰可以以100±3%的精度进行积分。蛋白质或肽的水解产物可以在少于24小时内分析。血浆、尿液和哺乳动物组织的更复杂的混合物特征可以在2天内分析。该仪器原理上适用于各种色谱柱流出液中茚三酮阳性组分的检测。
Quantitative determination of amino acids is made simpler and more rapid by an instrument for automatically recording the ninhydrin color value of the effluent from ion exchange columns. The influent buffer, freed of air, is pumped at a constant rate through a column of sulfonated poly-styrene resin. The effluent is met by a capillary stream of ninhydrin reagent delivered by a second pump. The color is developed by passing the mix-ture of reagent and effluent through a spiral of capillary Teflon tubing im-mersed in a boiling water bath. The absorbance of the resulting solution is measured continuously at 570 and 440 m/i as it flows through a cylindrical glass cell of 2-mm. bore. The peaks on the recorded curves can be inte-grated with a precision of 100±3% for loads from 0.1 to 3.0 jumoles of each amino acid. A hydrolyzate of a protein or peptide may be analyzed in less than 24 hours. The more complex mixtures characteristic of bloodplasma, urine, and mammalian tissues can be analyzed in 2 days. The instrument is appli-cable in principle to detection of nin-hydrin-positive constituents in the efflu-ent from various types of chromatograph columns.