高效液相層析法檢測非核苷類反轉錄酶抑製劑 Efavirenz   全文下載

Chiral Separation of Non-Nucleoside Reverse Transcription Inhibitor Efavirenz by HPLC on Cellulose-Based Chiral Stationary Phase

本研究開發並確效一立體特異性之 HPLC 方法，使用正相纖維素衍生掌性管柱（cellulose derivatized chiral column）分析 efavirenz（EFZ）原料藥及其製劑中鏡像異構物，有機修飾劑 2-propanol、乙醇及三氟乙酸組成的移動相，優化後獲得最佳的鏡像異構物分離。(R)-EFZ 和 (S)-EFZ 的滯留時間分別為 7.5 和 9.2 min。濃度範圍 200 - 6210 ng/mL 時，檢量線呈線性，(R)-isomer之 R2 為 0.9999。偵測極限（LOD）和定量極限（LOQ）分別為 66 和 200 ng/mL。本方法精密、準確且適合活性藥物成分（API）中 (R)-EFZ 的分離和定量。分析結果經統計數據確效，可成功地應用於 EFZ 原料藥及製劑的鏡像異構物純度分析。

A stereospecific HPLC method for the separation of efavirenz (EFZ) enantiomers in bulk drug and formulations was developed and validated on a normal-phase cellulose derivatized chiral column. The effect of organic modifiers, namely, 2-propanol, ethanol and trifluoroacetic acid in mobile phase was optimized to obtain the best enantiomeric separation. The retention times of (R)-EFZ and (S)-EFZ were observed to be 7.5 and 9.2 min, respectively. The calibration curve was found to be linear over the concentration range of 200 - 6210 ng/mL with a determination coefficient (R2) of 0.9999 for the (R)-isomer. The limit of detection (LOD) and limit of quantification (LOQ) were calculated to be 66 ng/mL and 200 ng/mL, respectively. The proposed method was found to be accurate, precise and suitable for the separation and quantification of unwanted (R)-EFZ in active pharmaceutical ingredients (API). The analytical results were supported by statistical parameters. The proposed method could be successfully applied to the enantiomeric purity analysis of EFZ in bulk drug samples and formulations.