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用羟丙基-β-环糊精(HP-β-CD)作为手性流动相添加剂,在UFCN(220×4.6 mm×5μm)氰基柱上通过反相高效液相色谱实现了4-氯扁桃酸(4-Cl MA)对映体的拆分。考察了水相HP-β-CD浓度,NaH_2PO_4浓度,p H,流动相中甲醇的比例及柱温等因素对分离度的影响。确定的最优色谱条件为:水相(5 mmol/L HP-β-CD,8 mmol/L NaH_2PO_4,p H 2.8),V水相/V甲醇=95:5,柱温25℃,流速1 m L/min。此条件下4-Cl M A对映体的分离度可达1.80。同时在此色谱条件下考察了3-氯扁桃酸(3-Cl MA),2-氯扁桃酸(2-Cl MA)与扁桃酸(MA)对映体的色谱拆分结果。结果发现在此条件下,3-Cl M A对映体分离度为1.51,而2-Cl M A与M A完全没有分开。对方法进行了验证,(S),(R)-4-Cl MA两对映体低、中、高浓度日内和日间精密度的RSD均小于1.1%;(S),(R)-4-Cl M A两对映体回收率分别为101.0%和100.6%;(S),(R)-4-Cl M A两对映体检出限与定量限分别为0.028,0.030 mg/L与0.093,0.098 mg/L。
By HPLC on a UFCN (220 × 4.6 mm × 5 μm) column with HP-β-cyclodextrin (HP-β-CD) as a chiral mobile phase additive, Resolution of mandelic acid (4-Cl MA) enantiomer. The effects of factors such as HP-β-CD concentration, NaH 2 PO 4 concentration, p H, mobile phase methanol and column temperature on the resolution were investigated. The optimum chromatographic conditions were as follows: aqueous phase (5 mmol / L HP-β-CD, 8 mmol / L NaH 2 PO 4, p H 2.8), V aqueous phase / V methanol = 95: m L / min. Under this condition, the resolution of 4-Cl M A enantiomer can reach 1.80. The chromatographic resolution of 3-chloro-mandelic acid (3-Cl MA), 2-chloromandelic acid (2-Cl MA) and mandelic acid (MA) was also investigated under this chromatographic conditions. As a result, it was found that under this condition, the resolution of 3-Cl M A enantiomer was 1.51, and 2-Cl M A did not completely separate from M A. The RSDs of intra-day and inter-day precision of the (S), (R) -4-Cl MA enantiomers were all less than 1.1% -Cl MA was 101.0% and 100.6%, respectively. The detection limits and quantification limits of (S), (R) -4-Cl MA enantiomers were 0.028,0.030 mg / L and 0.093,0.098 mg / L.