A simple and reliable high-performance liquid chromatography method was developed to determine the enantiomeric impurity of tenofovir disoproxil fumarate, an orally bioavailable prodrug of tenofovir, commonly used for the treatment of human immunodeficiency virus and hepatitis B. Tenofovir disoproxil and its enantiomer, were completely separated on a Chiralpak IC column (3 μm, 100 × 4.6 mm, i.d.). The chiral separation was achieved using a mobile phase containing n-hexane, ethanol, methanol, and triethylamine 65/25/10/0.1 (v/v/v/v) at a flow rate of 0.6 mL/min. Ideally, the reversal of enantiomer elution order was achieved on the Chiralpak IC column, to allow the elution of the minor enantiomeric impurity before the major component. Moreover, the proposed method was able to discriminate the active ingredient from the related substances available in the tenofovir disoproxil fumarate raw materials. These compounds were isolated and structurally elucidated by MS and nuclear magnetic resonance. Based on the spectral data, the structures of related substances were confirmed as tenofovir isoproxil monoester and fumaric acid. The high-performance liquid chromatography method was optimized by the design of experiment approach and successfully validated following the International Conference on Harmonization guideline. Proposed method was effectively applied for the quantification of enantiomeric impurity in tenofovir disoproxil fumarate raw materials.

中文翻译：

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三(3,5-二氯苯基-氨基甲酸酯)纤维素手性固定相上富马酸替诺福韦二吡呋酯对映异构体杂质的测定及其相关物质的表征

开发了一种简单可靠的高效液相色谱方法来测定富马酸替诺福韦酯的对映体杂质，替诺福韦酯是一种口服生物可利用的替诺福韦前药，常用于治疗人类免疫缺陷病毒和乙型肝炎。在 Chiralpak IC 柱（3 μm，100 × 4.6 mm，内径）上分离。使用包含正己烷、乙醇、甲醇和三乙胺 65/25/10/0.1 (v/v/v/v) 的流动相以 0.6 mL/min 的流速实现手性分离。理想情况下，在 Chiralpak IC 色谱柱上实现对映异构体洗脱顺序的逆转，以允许在主要成分之前洗脱次要对映异构体杂质。而且，所提出的方法能够将活性成分与富马酸替诺福韦二吡呋酯原料中可用的相关物质区分开来。这些化合物被分离出来并通过 MS 和核磁共振进行结构阐明。根据光谱数据，相关物质的结构被确认为替诺福韦异丙酯单酯和富马酸。高效液相色谱方法通过实验方法的设计进行了优化，并按照国际协调会议指南成功验证。提出的方法有效地应用于富马酸替诺福韦二吡呋酯原料中对映异构体杂质的定量分析。根据光谱数据，相关物质的结构被确认为替诺福韦异丙酯单酯和富马酸。高效液相色谱方法通过实验方法的设计进行了优化，并按照国际协调会议指南成功验证。提出的方法有效地应用于富马酸替诺福韦二吡呋酯原料中对映异构体杂质的定量分析。根据光谱数据，相关物质的结构被确认为替诺福韦异丙酯单酯和富马酸。高效液相色谱方法通过实验方法的设计进行了优化，并按照国际协调会议指南成功验证。提出的方法有效地应用于富马酸替诺福韦二吡呋酯原料中对映异构体杂质的定量分析。