反相高效液相色谱分析克拉霉素合成工艺中区域选择性甲基化的关键中间体

Analysis of the Key Intermediates for the Regioselectivity Control in the Methylation of the Manufacture of Clarithromycin by Reversed-Phase High Performance Liquid Chromatography

采用反相高效液相色谱法对克拉霉素合成工艺中关键中间体E 2′,4″ 双(三甲基硅) 红霉素A 9 (1 异丙氧环己基)肟、Z 2′,4″ 双(三甲基硅) 红霉素A 9 (1 异丙氧环己基)肟和E 2′,4″ 双(三甲基硅) 6 甲基红霉素A 9 (1 异丙氧环己基)肟、Z 2′,4″ 双(三甲基硅) 6 甲基红霉素A 9 (1 异丙氧环己基)肟及其相关的6种工艺杂质进行了分离、定性和定量分析。色谱柱为DIKMA公司的InertsilODS 3(150mm×4 6mmi d ,5μm);流动相为乙腈 水(体积比为95∶5),流速1 5mL/min;检测波长UV205nm;柱温40℃。在进样量为6～60μg时具有良好的线性关系并可以基线分离,为研究区域选择性甲基化提供了基础。

2′,4″-O-Bis(trimethylsilyl) erythromycin A-9-O-(1-isopropoxycyclohexyl)oxime (2′,4″-TMS-EMIPCH) and 2′,4″-O-bis(trimethylsilyl)-6-O-methylerythromycin A-9-O-(1-iso-propoxycyclohexyl)oxime (2′,4″-TMS-IPCH) are the key intermediates for manufacturing clarithromycin. A qualitative and quantitative method for baseline separation of E- and Z-isomers and related process substances has been established. A DIKMA-Inertsil ODS-3 column (150 mm×4.6 mm i.d., 5 μm) was used. The column temperature was maintained at 40 ℃. The mobile phase was CH_3CN-H_2O (95∶5, v/v). The flow rate was 1.5 (mL/min) and the detection wavelength was UV 205 nm. Good linearities for E-2′,4″-TMS-EMIPCH and E-2′,4″-TMS-IPCH were obtained in the ranges of 6-60 μg (r=(0.999 4)) and 6-72 μg (r=(0.999 8)), respectively. The method described has also been demonstrated to work equally well on other 2′,4″-O-bis(trimethylsilyl)erythromycin 9-oxime hydroxyl derivatives, which provided the criterion for optimizing the protective groups at 9-oxime hydroxyl position and the study of regioselectivity of methylation at the 6-OH position.