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目的以2,3,4,6-四-O-乙酰基-β-D-葡萄糖异硫氰酸酯(GITC)为手性衍生化试剂,建立苯乙醇胺类化合物对映体RP-HPLC分离分析方法。方法采用RP-HPLC法。考察了衍生化反应中碱化试剂和衍生化试剂的浓度等反应条件对衍生化产率的影响,并考察流动相的组成和pH值等因素对生成的非对映异构体分离的影响。讨论了化合物的分子结构对手性衍生化及衍生后的非对映异构体色谱分离的影响。结果在三乙胺和GITC的浓度分别为10和5 mmol.L-1的乙腈溶液中,室温下反应20 min后,有5个苯乙醇胺化合物转化成相应的非对映异构体的硫脲衍生物。在色谱条件为:Diamonsil C18色谱柱(150 mm×4.6 mm,5μm),30 mmol.L-1醋酸铵(pH6.0)-乙腈(体积比为50∶50)为流动相,检测波长254 nm,流速1.0 mL.min-1,室温下,5个苯乙醇胺化合物对映体衍生化后非对映异构体的分离度达到4以上。结论该方法可作为苯乙醇胺类化合物对映体分离的方法之一。
OBJECTIVE To establish RP-HPLC separation of enantiomers of phenylethanolamines with 2,3,4,6-tetra-O-acetyl-β-D-glucose isothiocyanate (GITC) as chiral derivatization reagent method. Methods RP-HPLC method. The effects of reaction conditions such as the concentration of alkalizing agent and derivatizing reagent on the derivatization reaction were investigated. The effects of mobile phase composition and pH on the separation of diastereoisomers were also investigated. The influence of molecular structure of compounds on chiral derivatization and derivatization of diastereomer chromatographic separation was discussed. Results In the acetonitrile solution with triethylamine and GITC concentration of 10 and 5 mmol.L-1 respectively, after 20 min reaction at room temperature, 5 phenylethanolamine compounds were converted into the corresponding diastereomers of thiourea derivative. The chromatographic conditions were: Diamonsil C18 column (150 mm × 4.6 mm, 5 μm), 30 mmol·L -1 ammonium acetate (pH 6.0) -acetonitrile (50:50 by volume) , The flow rate was 1.0 mL.min-1. The enantiomers of five phenylethanolamine compounds were separated and their diastereoisomers were more than 4 at room temperature. Conclusion This method can be used as one of the enantiomer separation methods for phenylethanolamines.