The fingerprint chromatogram of AZC was established through high-performance liquid chromatography (HPLC) employing a Waters® system equipped with a Waters® 2998 photodiode array detector. Separation was achieved using Purospher® STAR, C-18 encapped (5µM), LiChro-CART® 250 − 4.6, and HPLC-Cartridge, Sorbet Lot No. HX255346 (Merck, Germany).
In this study, the mobile phase gradient comprised 100% methanol (solvent A) (Fisher Scientific, USA) and 2.5% acetic acid (solvent B) (Fisher Scientific, USA) in deionized (DI) water. The gradient elution was conducted at a flow rate of 1.0 mL/min, with the following gradient profile: 0–17 min, 70% A; 18–20 min, 100% A; 20.5–25 min, 10% A. Before administration, the sample underwent filtration (0.45 μm, Millipore), and a 20 µL aliquot of the sample was directly utilized. The chromatogram was assessed at 280 nm using a UV detector, and data analysis was carried out employing EmpowerTM3 [22 (link)].
In this study, the mobile phase gradient comprised 100% methanol (solvent A) (Fisher Scientific, USA) and 2.5% acetic acid (solvent B) (Fisher Scientific, USA) in deionized (DI) water. The gradient elution was conducted at a flow rate of 1.0 mL/min, with the following gradient profile: 0–17 min, 70% A; 18–20 min, 100% A; 20.5–25 min, 10% A. Before administration, the sample underwent filtration (0.45 μm, Millipore), and a 20 µL aliquot of the sample was directly utilized. The chromatogram was assessed at 280 nm using a UV detector, and data analysis was carried out employing EmpowerTM3 [22 (link)].
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