Hammouda Mohammed E A, El-Masry Amal A, El-Ashry Saadia M, El-Wasseef Dalia R
Department of Medicinal Chemistry, Faculty of Pharmacy, Mansoura University, Mansoura, 35516, Egypt.
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Horus University - Egypt, New Damietta, Egypt.
BMC Chem. 2023 Mar 14;17(1):17. doi: 10.1186/s13065-023-00936-z.
Simple, direct, rapid, and sensitive HPLC and spectrophotometric methods were established for simultaneous estimation of a novel combination of budesonide and azelastine (BUD/AZL) in their laboratory-prepared mixture and dosage form according to the medicinally recommended ratio 1:4.28. Budesonide is an important inhalation corticosteroid that plays a vital role in the inhibition of COVID-19 replication and cytokine production. The first chromatographic method was created for the simultaneous estimation of BUD epimers in the presence of AZL with excellent efficiency in a relatively short chromatographic run (< 9 min). The separation of BUD epimers with AZL was carried out on a C column using acetonitrile: phosphate buffer of pH 3.5 adjusted by 0.2 M orthophosphoric acid (40:60, v/v) as a mobile phase, UV detection at 230 nm and a flow rate of regulated at 2 mL/min. Besides, three spectrophotometric methods were applied for the simultaneous determination of the provided mixture adopting zero order, first order derivative, and ratio first derivative approaches. The Zero-order spectrophotometry was used for the determination of AZL in presence of BUD, where BUD shows no absorbance at 290 nm. The first derivative amplitude at 265 nm (D) (zero-crossing of AZL) and the ratio of first derivative amplitudes at 270 nm (DD) using 10.0 µg mL AZL as divisor was chosen for the simultaneous determination of BUD in the presence of AZL in the binary mixture. The proposed methods were found to be rectilinear in the concentration range of (0.4-40.0 µg mL) and (0.05-40.0 µg mL) for BUD and AZL, respectively in the HPLC method. Whereas the concentration range for AZL in the zero-order method was (1.0-35.0 µg mL) and for BUD in the first derivative and ratio derivative method was (6.0-20.0 µg mL). Validation of the suggested approaches according to the ICH criteria was performed. Furthermore, to ensure the proposed approaches' greenness, The AGREE and GAPI metrics were utilized, and the afforded results revealed an excellent greenness of the proposed approaches.
建立了简单、直接、快速且灵敏的高效液相色谱法（HPLC）和分光光度法，用于按照药用推荐比例1:4.28同时测定实验室制备的布地奈德和氮卓斯汀（BUD/AZL）混合物及其剂型中的这两种成分。布地奈德是一种重要的吸入性皮质类固醇，在抑制新冠病毒复制和细胞因子产生方面发挥着至关重要的作用。第一种色谱方法用于在存在氮卓斯汀的情况下同时测定布地奈德的差向异构体，在相对较短的色谱运行时间（<9分钟）内具有出色的效率。布地奈德差向异构体与氮卓斯汀的分离在C柱上进行，使用乙腈：由0.2 M正磷酸调节至pH 3.5的磷酸盐缓冲液（40:60，v/v）作为流动相，在230 nm处进行紫外检测，流速调节为2 mL/min。此外，采用零阶、一阶导数和比率一阶导数方法应用了三种分光光度法来同时测定所提供的混合物。零阶分光光度法用于在存在布地奈德的情况下测定氮卓斯汀，因为布地奈德在290 nm处无吸收。选择265 nm处的一阶导数幅度（D）（氮卓斯汀的零交叉点）以及使用10.0 μg/mL氮卓斯汀作为除数时270 nm处的一阶导数幅度之比（DD），用于在二元混合物中存在氮卓斯汀的情况下同时测定布地奈德。在HPLC方法中，所提出的方法在布地奈德和氮卓斯汀的浓度范围分别为（0.4 - 40.0 μg/mL）和（0.05 - 40.0 μg/mL）时呈线性。而在零阶方法中氮卓斯汀的浓度范围为（1.0 - 35.0 μg/mL），在一阶导数和比率导数方法中布地奈德的浓度范围为（6.0 - 20.0 μg/mL）。根据国际协调会议（ICH）标准对所建议的方法进行了验证。此外，为确保所建议方法的绿色度，使用了AGREE和GAPI指标，所得结果表明所建议的方法具有出色的绿色度。
