Development and Optimization of a New UPLC-UV/MS Method through DoE and MLR for Detecting Substandard Drug Products to Treat Tuberculosis

• Published in: Molecules (Basel, Switzerland) Vol. 27; no. 20; p. 7141
• Main Authors: Suárez-González, Javier, Cáceres-Pérez, Amor R, Oliva, Alexis, Santoveña-Estévez, Ana, Fariña, José B
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01.10.2022; MDPI
• Subjects: Antitubercular agents; Antitubercular Agents - chemistry; Antitubercular Agents - therapeutic use; central composite design; Chromatography, Liquid; Composition; Design; design of experiment; Design of experiments; Design optimization; Drug counterfeiting; Drug development; Ethambutol; Ethambutol - chemistry; Flow velocity; Formic acid; Formulations; Health hazards; Humans; Ingredients; Isoniazid; Isoniazid - chemistry; Isoniazid - therapeutic use; Linear models (Statistics); Linear regression models; Liquid chromatography; Mass spectrometry; Methods; multi-linear regression; Optimization; Pharmaceutical research; Pharmaceuticals; Product quality; Pyrazinamide; Pyrazinamide - chemistry; Pyrazinamide - therapeutic use; quality by design; Rifampin; Rifampin - chemistry; Rifampin - therapeutic use; Substandard Drugs; Tablets; Testing; Tuberculosis; Tuberculosis - drug therapy; Ultraviolet detectors; Spain; central composite design; quality by design; design of experiment; multi-linear regression; tuberculosis
• ISSN: 1420-3049; 1420-3049
• 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.