以邻苯二甲醛/N-异丁酰基-L-半胱氨酸(OPA/IBLC)为手性衍生化试剂,建立了苏氨酸手性分离和光学纯度检查的高效液相色谱法.采用HederaODS-2色谱柱(250mm×4.6mm,5µm),以0.1%醋酸铵-甲醇-乙腈(体积比70︰27︰3)为流动相,检测波长为338nm,考察了溶液pH值、试剂用量、衍生化时间及温度对衍生化反应的影响.结果表明,在该优化的色谱条件下,苏氨酸的对映体分离度为1.6,L-苏氨酸最低检测限约为0.05µg/mL.该方法灵敏度高,重现性好,可用于L-苏氨酸光学纯度的检查控制.% A new high performance liquid chromatography method for chiral separation and optical purity test of threonine has been established with ortho-phthalaldehyde/N-isobutyryl-L-cysteine (OPA/IBLC) as the pre-column derivatizing reagent. The diastereoisomers produced were separated on a Hedera ODS-2 column (250 mm×4.6 mm, 5 μm) with a mobile phase consisting of 0.1% acetate-methanol-acetonitrile (volume fraction 70:27:3) at the detection wavelength of 338 nm. The effects of solution pH, the amount of OPA/IBLC, the derivatizing time and the derivatizing temperature were optimized. The results showed that under the optimized chromatographic conditions, resolution of threonine is 1.6 and the lowest detectable limit of L-threonine is 0.05 μg/mL. The method is sensitiveandhighlyreproducibleandcan beusedinthepuritytestofL-threonine.
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