盐酸多西环素ChP2015标准检测的紧邻主成分后杂质的制备及鉴定

目的：制备盐酸多西环素中国药典2015年版（ChP 2015）标准检测的紧邻主峰后杂质，并鉴定其结构。方法：以盐酸多西环素成盐用母液为对象，采用室温条件下半制备液相色谱分离纯化杂质；色谱条件：采用Synergi C₁₈（50 mm×250 mm，10 μm）色谱柱，以0.1%三氟乙酸水溶液（A）-0.1%三氟乙酸乙腈溶液（B）为流动相，线性梯度洗脱（A-B）：0.0 min（85∶15）→ 40.0 min（60∶40）→ 40.5 min（85∶15）→50.0 min（85∶15），流速为80 mL·min^-1； 254 nm检测收集目标组分，旋转蒸发去除有机溶剂，冷冻干燥制得目标杂质；采用盐酸多西环素ChP 2015标准中有关物质检查的高效液相色谱法确证制得物质为目标杂质并采用归一化方法测定其纯度；采用光谱综合分析等手段确证其结构。结果：制得目标杂质纯度为98.7%，结构确证为2-乙酰-2-脱氨甲酰多西环素（即EP 8.0中的盐酸多西环素杂质F），在有关物质测定用供试品溶液中添加该杂质后，多西环素后相邻峰面积显著增加。结论：盐酸多西环素ChP 2015标准检测的紧邻主峰后杂质经制备色谱分离并成功鉴定，该结论被ChP 2015采用。

Objective: To prepare and identify the doxycycline hyclate impurity eluted right after the main component described in ChP 2015. Methods: The target impurity was isolated from doxycycline hyclate mother solution by a semi-preparetive HPLC with a Synergi RP-C₁₈ column(250 mm×50 mm, 10 μm) at room temperature and gradient elution using mixtures of 0. 1% TFA in water(A) as A and 0. 1% TFA in acetonitile (B) as the mobile phase at a flow rate of 80 mL·min^-1. The liner gradient program of(A-B) was employed and set as follows:0. 0 min(85:15) →40. 0 min(60:40) →40. 5 min(85:15) →50. 0 min(85:15). The analytes were monitored at a wavelength of 254 nm. The organic solvent of the collected targent impurity solution was evaporated by rotary evaporation and then was frozen to dry. The identity of the prepared target impurity was determined by the HPLC method described in ChP 2015 for the determination of the related substances of doycycline hyclate and its purity was calculated by normalization method. The chemical structure of the target impurity was identified by organic spectroscopic methods. Results: The purity of the prepared target impurity was 98.7% and the chemical structure of this impurity was identified as 2-acetyl-2-decarbamoyl-doxycycline and was confirmed to be the impurity F of doxycycline hyclate described in EP 8.0. Conclusion: The doxycycline hyclate impurity eluted right after the main component described in ChP 2015 was successfully prepared with a semipreparetive HPLC method and identified. This result has been adopted by ChP2015.

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