Journal Article

Development of a gradient method for sulfamethoxazole, trimethoprim, isoniazid, and pyridoxine hydrochloride in rabbit plasma through QbD-driven investigation

Ansary, Akramul, K M, Premsagar, C, Bhagyalakshmi, B, Ramesh, Sola, Piyong, Pasha, T. Yunus, Dutta, Koushik Nandan, Majumder, Manish, Kumar Das, Tridib

2024

Overview

The current study developed a method for quantifying four drugs—Sulfamethoxazole, Trimethoprim, Isoniazid, and Pyridoxine—in rabbit plasma. The method uses gradient liquid chromatography based on analytical quality by design. To achieve separation, a Eclip Plus C18 (250 mm × 5 mm, 4.6 µm) column with L1 packing was used, and analytes were detected at 254 nm at ambient temperature. The optimized mobile phase consisted of 50 mM potassium dihydrogen phosphate buffer (pH 6.5) and Methanol. The concentration of Methanol was 3% (0–5 min), 15% (5–15 min), 55% (15–27 min), and 3% Methanol until the end of the 30-min runtime, and the flow rate was set at 0.95 mL/min. Control Noise Experimentation was used to screen studies, revealing that flow rate, pH, and Methanol concentration significantly affected the analytical attributes. The study identified critical attributes (resolution and asymmetric factor) and developed a quality target method profile. A central composition design was used to optimize the essential parameters. The method developed for the drugs showed peaks at retention times of 6.990 min for Isoniazid, 7.880 min for Pyridoxine, 15.530 min for Sulfamethoxazole, and 26.890 min for Trimethoprim, respectively. The method was validated with linearity in the range of 10–640 ng ml −1 , with R 2 of 0.9993, 0.9987, 0.9993, and 0.9992 for Sulfamethoxazole, Trimethoprim, Isoniazid, and Pyridoxine, respectively.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 639/638/11 / 639/638/11/872 / Acquired immune deficiency syndrome / AIDS / Ambient temperature / Animals / Bioanalytical quality by design / Chromatography / Chromatography, High Pressure Liquid - methods / Chromatography, Liquid - methods / Design of experiments / Drug development / Flow rates / Gradient elution / HIV / Human immunodeficiency virus / Humanities and Social Sciences / Infections / Isoniazid / Isoniazid - blood / Liquid chromatography / Methanol / multidisciplinary / Pharmaceutical sciences / Pyridoxine / Pyridoxine - analysis / Pyridoxine - blood / Pyridoxine hydrochloride / Quality control / Rabbits / Risk assessment / Science / Science (multidisciplinary) / Sulfamethoxazole / Sulfamethoxazole - analysis / Sulfamethoxazole - blood / Trimethoprim / Trimethoprim - analysis / Trimethoprim - blood