Departamento de Ingeniería Química y Tecnología Farmacéutica, Facultad de Farmacia, Universidad de La Laguna, 38200 San Cristóbal de La Laguna, Spain.
Instituto Universitario de Enfermedades Tropicales y Salud Pública de Canarias, Universidad de La Laguna, 38200 San Cristóbal de La Laguna, Spain.
Molecules. 2022 Oct 21;27(20):7141. doi: 10.3390/molecules27207141.
Drug products used for treating tuberculosis are one of the most widely reported medicines to be classified as falsified or substandard in low- and middle-income countries, representing a major hazard to health. The aim of this study was, firstly, to develop an ultra-performance liquid chromatography (UPLC) method which is able to analyze fixed combination tablets with up to four active pharmaceutical ingredients, including isoniazid, pyrazinamide, rifampicin, and ethambutol. Secondly, we aimed to optimize it through the design of experiments and multi-linear regression based on a central composite design and to validate it according to the guidelines of the International Conference on Harmonization. The application of this tools enabled the identification of the influential factors (flow rate, formic acid, and temperature) and their effects on the studied responses (retention factor and percentage for each drug) as part of the quality by design approach. The method proved to be to be linear in the range from 5.0 to 15 µg/mL for isoniazid, pyrazinamide, and rifampicin, being precise (<1%) and accurate (97−101%). In addition, the method validated for ethambutol proved to be linear from 1.4 to 4.2 µg/mL, as well as precise (0.54%) and accurate (97.3%). The method was stability indicated for all the active pharmaceutical ingredients studied and was able to detect two substandard formulations sampled on the African market.
用于治疗结核病的药物产品是在中低收入国家被报告最多的假药或劣药之一，对健康构成重大危害。本研究的目的首先是开发一种超高效液相色谱 (UPLC) 方法，该方法能够分析包含异烟肼、吡嗪酰胺、利福平、乙胺丁醇在内的四种活性药物成分的固定剂量复合片剂。其次，我们旨在通过基于中心复合设计的实验设计和多元线性回归对其进行优化，并根据国际协调会议的指南对其进行验证。该工具的应用使我们能够识别出有影响的因素（流速、甲酸和温度）及其对研究响应（每种药物的保留因子和百分比）的影响，这是质量源于设计方法的一部分。该方法在 5.0 至 15 µg/mL 范围内对异烟肼、吡嗪酰胺和利福平显示出线性，精密度（<1%）和准确度（97−101%）良好。此外，为乙胺丁醇验证的方法在 1.4 至 4.2 µg/mL 范围内也呈线性，精密度（0.54%）和准确度（97.3%）良好。该方法对所有研究的活性药物成分均显示出稳定性，并能够检测到在非洲市场上采样的两种劣药制剂。
