用于原料药及制剂中替诺福韦和恩曲他滨含量测定的反相高效液相色谱法的建立与验证

Development And Validation Of Rp-Hplc Method For The Estimation Of Tenofovir And Emtricitabine In Bulk And Pharmaceutical Dosage Form.

作者信息

Sahoo Biswa Mohan, Rao Pallepogu Venkateswara, Rao Naidu Srinivasa

机构信息

Roland Institute of Pharmaceutical Sciences, Berhampur-760010, Odisha, India.

Vikas College of Pharmacy, Vissannapeta, Krishna Dist-521215, Andhra Pradesh, India.

出版信息

Curr Drug Res Rev. 2023 Jun 2. doi: 10.2174/2589977515666230602151222.

DOI:10.2174/2589977515666230602151222

PMID:37278041

Abstract

BACKGROUND

A novel, simple, efficient, rapid, and precise reverse-phase high-performance liquid chromatography (RP-HPLC) method was developed for the estimation of Tenofovir and Emtricitabine in the bulk and pharmaceutical dosage form. The currently developed method was subsequently validated according to ICH guidelines in terms of linearity, accuracy, precision, the limit of detection, the limit of quantification, robustness, etc. Methods: The separation of the selected drugs was optimized after several trials including change of mobile phase and its composition, stationary phase, flow rate, column temperature, etc. The separation was performed by using an Inertsil ODS C18 column (250 mm x 4.6 mm, 5μ) and UV absorption was measured at 231 nm. Methanol: Acetonitrile: Water was selected as the mobile phase in the ratio of 50:20:30 (V/V/V) at a flow rate of 1 mL/min. As per International Conference on Harmonization (ICH) Q2 R1 guidelines, several validation parameters were evaluated which include specificity, linearity, precision, accuracy, the limit of detection (LOD), and the limit of quantitation (LOQ).

RESULTS

The acceptable degree of linearity range was found to be 40-100 µg/mL. The standard solution exhibited retention times of 3.06 minutes and 5.07 minutes for Tenofovir and Emtricitabine respectively. The LOD and LOQ obtained were 0.05µg/ml and 0.02µg/mL, 15µg/mL, and 0.08µg/mL for Tenofovir and Emtricitabine respectively. The percent recovery was found to be 98 to 102%.

CONCLUSION

Hence, the proposed method is simple, selective, and specifically meets the requirements of ICH guidelines for the validation of the analytical method.

摘要

背景

开发了一种新颖、简单、高效、快速且精确的反相高效液相色谱（RP-HPLC）方法，用于测定原料药和药物剂型中的替诺福韦和恩曲他滨。随后，根据国际人用药品注册技术协调会（ICH）指南，对当前开发的方法在线性、准确性、精密度、检测限、定量限、耐用性等方面进行了验证。方法：在进行了多次试验后，对所选药物的分离条件进行了优化，包括流动相及其组成、固定相、流速、柱温等的改变。使用Inertsil ODS C18柱（250 mm×4.6 mm，5μm）进行分离，并在231 nm处测量紫外吸收。选择甲醇：乙腈：水作为流动相，比例为50:20:30（V/V/V），流速为1 mL/min。根据国际协调会议（ICH）Q2 R1指南，评估了几个验证参数，包括特异性、线性、精密度、准确性、检测限（LOD）和定量限（LOQ）。