柱前衍生化RP-HPLC法分离苯乙醇胺类化合物对映体

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。

Chiral separation of phenylethanolamine compounds by pre-column derivatization high performance liquid chromatography

Objective To develop a pre-column high performance liquid chromatographic method for the separation of some phenylethanolamine compounds with 2, 3, 4, 6-tetra-O-acetyl-β-D-glucopyranosyl isothiocyanate (GITC) as derivative agent. Methods The effects of concentration of triethylamine (TEA), concentration of GITC and reaction time on derivative reaction were studied. The influences of kinds and concentration of organic modifier and pH value of buffer on chiral separation were studied. The structural factors affecting chiral derivatization and separation were disccussd. Results The separation was performed on a Diamonsil C₁₈ column (150 mm×4.6 mm, 5 μm) using 30 mmol·L^–1 ammonium acetate (pH 6.0)-acetonitrile (V∶V=50∶50) as mobile phase at a flow rate of 1.0 mL&;#8226;min^–1 and room temperature. The detection wavelength was set at 254 nm. Under the above conditions, five of the derivatized compounds were resolved with resolution larger than 4. Conclusion The method can be used to the chiral separation of phenylethanolamine enantiomers.