摘要
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。
The enantioseparation of 4-chloromandelic acid（ 4-ClMA） was performed by reverse phase high performance liquid chromatography with hydroxypropyl- β-cyclodextrin（ HP- β-CD） as chiral mobile phase additive on a cyanopropyl column（UF-CN： 220 × 4.6 mm × 5 μm）.The effects of aqueous phase PH,the concentration of hydroxypropyl- β-cyclodextrin and NaH_2PO_4 buffer,the amount of organic modifier methanol and column temperature on retention were investigated.The satisfactory resolution of nearly 1.80 for 4-ClMA enantiomers were achieved at 25℃ using a mobile phase consisting of a mixture of aqueous buffer（ pH of 2.8,8 mmol/L sodium dihydrogen phosphate and 5 mmol/L HP- β-CD） and methanol with a volumetric ratio of 95：5 at a flow rate of 1.0 m L/min.The chiral separation of 3-chloromandelic acid（ 3-ClMA）,2-chloromandelic acid（ 2-Cl MA）,mandelic acid（ MA） were also investigated in the same chromatographic condition.The results appeared that the resolution for 3-Cl MA enantiomers was 1.51,while 2-ClMA and MA were not separated.Finally,the developed analytical method for 4-ClMA was validated by examining the inter and intra precision（ lower than 1.1%,for lower,moderate and higher concentration）,the recoveries（ 100.98% for（ S）- 4-Cl MA and100.6% for（ R）- 4-ClMA）,the limits of detection（ 0.028 mg/L for（ S）- 4-Cl MA and 0.030 mg/L for（ R）- 4-ClMA） and the limits of quantification（ 0.093 mg/L for（ S）- 4-Cl MA and 0.098 mg/L for（ R）- 4-ClMA）.
出处
《分析试验室》
CAS
CSCD
北大核心
2016年第6期700-704,共5页
Chinese Journal of Analysis Laboratory
关键词
羟丙基-Β-环糊精
手性流动相添加剂
氰基柱
4-氯扁桃酸
拆分
Hydroxypropyl-β-cyclodextrin
Chiral mobile phase additive
Cyanopropyl column
4-Chloromandelic acid
Resolution
