聚乙二醇1000维生素E琥珀酸酯中单双酯的制备及5种成分含量测定

Preparation of Monoester and Diester from D-α-tocopheryl Polyethylene Glycol 1000 Succinate and Determination of Five Components

关键词:  聚乙二醇1000维生素E琥珀酸酯  单酯  双酯  制备液相色谱法  高效液相色谱法  结构确证

Key words:  D-α-tocopheryl polyethylene glycol 1000 succinate  monoester  diester  preparative HPLC  HPLC  structural identification

摘要:

目的 分离制备聚乙二醇1000维生素E琥珀酸酯中的单酯与双酯，并采用HPLC-UV、HPLC-ELSD测定聚乙二醇1000维生素E琥珀酸酯中的单酯、双酯、维生素E琥珀酸酯、α-生育酚及聚乙二醇1000的含量。方法 制备液相色谱分离采用DAC-HB50 C₁₈动态轴向压缩色谱柱（50 mm×250 mm，10 μm），以水（A）-乙醇（B）为流动相，梯度洗脱，流速75 mL·min^-1，检测波长285 nm，收集目标组分旋蒸去除溶剂制得单酯、双酯，并确证其结构。含量测定色谱条件：Waters XBridge C₈色谱柱（4.6 mm×250 mm，5 μm），0.1%甲酸溶液为流动相A，乙腈-异丙醇（1∶1）为流动相B，梯度洗脱，流速1.0 mL·min^-1，检测波长285 nm。HPLC测定制得的单双酯纯度及单酯、双酯、维生素E琥珀酸酯、α-生育酚含量；HPLC-ELSD测定聚乙二醇1000含量，采用相同色谱柱与流动相，进行不同的梯度洗脱，ELSD漂移管温度90.4℃，载气流速2.3 L·min^-1。结果 制得单、双酯纯度分别为99.1%，99.8%。各成分分离良好，在相应的范围内线性关系良好，相关系数r≥0.999 5；平均回收率为98.2%~99.7%。结论 制备液相色谱法可制得满足检测需求的单、双酯对照品；2种检测方法经验证，适用于聚乙二醇1000维生素E琥珀酸酯中单酯、双酯、维生素E琥珀酸酯、α-生育酚及聚乙二醇1000的含量测定。

Abstract:

OBJECTIVE To prepare monoester and diester from D-α-tocopheryl polyethylene glycol 1000 succinate (TPGS) using a preparative HPLC method, and to establish determination methods of monoester, diester, vitamin E succinate, α-tocopheryl and polyethylene glycol 1000 in TPGS. METHODS A preparative HPLC method with a DAC-HB50 C₁₈dynamic axis compress chromatographic column (50 mm×250 mm, 10 μm) was developed using the mobile phase of water (A)-ethanol (B) in a gradient mode at a flow rate of 75 mL·min^-1. Chromatograms were observed by a UV detector with wavelength set to 285 nm. Collected target product solution was evaporated by rotary evaporation. The target products were identified. An HPLC method with a Waters XBridge C₈ column (4.6 mm×250 mm, 5 μm) for the determination of prepared components was developed using a gradient elution of 0.1% formic acid solution and acetonitrile-isopropanol (1:1) at a flow rate of 1.0 mL·min^-1. Chromatograms were observed by a UV detector with wavelength set to 285 nm. The HPLC method was used for determination of monoester, diester, vitamin E succinate and α-tocopheryl in TPGS. An HPLC-ESLD method for the determination of polyethylene glycol 1000 in TPGS was developed with the same column and mobile phase but different gradient elution of HPLC method. Chromatograms were observed by an ELSD detector with drift tube temperature of 90.4℃ and gas flow rate of 2.3 L·min^-1. RESULTS The purity of prepared monoester and diester were 99.1% and 99.8%. The methods were validated and they were found that good resolution, linearity (r ≥ 0.999 5) within the certain range of five components and average recoveries of 98.2%-99.7% were achieved. CONCLUSION The preparative HPLC method is used successfully for the preparation of monoester and diester. Two validated detective HPLC methods can be used for detection of monoester, diester, vitamin E succinate, α-tocopheryl and polyethylene glycol 1000 in TPGS.