Chemistry Department, Faculty of Science, University of Ha'il, P.O. Box 2440, 81451, Ha'il, Saudi Arabia.
Department of Chemistry, Faculty of Applied Science, Umm Al-Qura University, 24230, Makkah, Saudi Arabia.
Sci Rep. 2023 Sep 21;13(1):15729. doi: 10.1038/s41598-023-40010-1.
A precise, Eco-friendly, and highly sensitive RP-HPLC method was employed using quality-by-design principles to concurrently identify cephalexin and cefixime residues in the manufacturing machines using a hypersil BDS C18 column (250 × 4.6 mm, 5 μm) at wavelength 254 nm. The Box-Behnken design was applied to obtain the best chromatographic conditions with the fewest possible trials. Three independent factors viz organic composition, flow rate, and pH were used to assess their effects on the responses' resolution and retention time. Overlay plot and desirability functions were implemented to predict responses of the high resolution and relatively short retention time using a mobile phase composed of acidic water: acetonitrile (85:15, v/v) at pH 4.5 adjusted by phosphoric acid with a flow rate of 2.0 mL/min. The spectral overlapping of the drugs was successfully resolved by the mean centering ratio (MCR) spectra approach at 261 nm and 298 nm for cephalexin and cefixime, respectively. Good linearity results were obtained for the suggested HPLC and MCR methods over the concentration range of (0.05-10 ppm) and (5-30 ppm) with a detection limit of 0.003, 0.004, 0.26, and 0.23 ppm, and quantitation limits of 0.008, 0.013, 0.79, and 0.68 ppm for cephalexin and cefixime, respectively, with a correlation coefficient of ≥ 0.9998 and good swab recovery results of 99-99.5%. A process capability index was accomplished for chemical and micro results, illustrating that both are extremely capable. The suggested method was effectively validated using ICH recommendations.
采用质量源于设计原则，利用反相高效液相色谱法（RP-HPLC），在 254nm 波长处，使用 Hypersil BDS C18 柱（250×4.6mm，5μm），同时对头孢氨苄和头孢克肟在生产设备中的残留量进行精确、环保、高灵敏度的检测。采用 Box-Behnken 设计，用最少的试验次数获得最佳的色谱条件。采用三因素（有机组成、流速和 pH 值），评估其对响应分辨率和保留时间的影响。采用重叠图和理想函数，预测使用酸性水：乙腈（85:15，v/v）组成的流动相，在 pH4.5 下，通过磷酸调节，流速为 2.0ml/min 的条件下，高分辨率和相对较短保留时间的响应。采用均数中心化比值（MCR）光谱法，成功地解决了药物的光谱重叠问题，头孢氨苄和头孢克肟的 MCR 光谱分别为 261nm 和 298nm。采用所建议的 HPLC 和 MCR 方法，在（0.05-10ppm）和（5-30ppm）浓度范围内，得到了良好的线性结果，头孢氨苄和头孢克肟的检测限分别为 0.003、0.004、0.26 和 0.23ppm，定量限分别为 0.008、0.013、0.79 和 0.68ppm，相关系数均≥0.9998，且棉签回收率良好，均为 99-99.5%。对化学和微生物结果进行了工艺能力指数评估，表明两者均具有极高的能力。所建议的方法经 ICH 推荐得到了有效验证。
