Baker Mostafa M, Belal Tarek S
Pharco Pharmaceuticals Co., Methodology Department, Alexandria, Egypt.
University of Alexandria, Faculty of Pharmacy, Pharmaceutical Analytical Chemistry Department, Elmessalah, Alexandria 21521, Egypt.
J AOAC Int. 2018 Jul 1;101(4):993-1000. doi: 10.5740/jaoacint.16-0439. Epub 2017 Sep 1.
This work presents a simple, sensitive, and generic HPLC-diode-array detection method for the simultaneous determination of six drugs prescribed for the treatment of open-angle glaucoma and ocular hypertension. The investigated drugs include brimonidine tartarate (BMN), acetazolamide (AZA), brinzomaide (BZA), dorzolamide HCl (DZA), levobunolol HCl (LVB), and timolol maleate (TIM). Efficient chromatographic separation was achieved using a Thermo Hypersil BDS C18 column (4.6 × 250 mm, 5 µm) with a mobile phase consisting of phosphate buffer pH 5 and acetonitrile in a ratio of 78 + 22. The flow rate was 1 mL/min, and quantification was based on measuring peak areas at 298 nm for TIM and 254 nm for the other drugs. Peaks were perfectly resolved, with retention times at 3.06, 3.87, 4.53, 5.78, 7.31, and 10.78 min for BMN, AZA, DZA, TIM, LVB, and BZA respectively. The developed method was validated according to International Conference on Harmonization guidelines with respect to system suitability, linearity, ranges, accuracy, precision, robustness, and LODs and LOQs. The proposed method showed good linearity in the ranges of 2-80, 2.5-100, 2.5-100, 5-200, 3.75-150, and 1.75-70 µg/mL for BMN, AZA, DZA, TIM, LVB, and BZA, respectively. LODs were 0.20-1.01 μg/mL for the analyzed compounds. Applicability of the proposed method to real-life situations was assessed through the analysis of five different pharmaceutical formulations, and satisfactory results were obtained.
本研究提出了一种简单、灵敏且通用的高效液相色谱 - 二极管阵列检测方法，用于同时测定六种用于治疗开角型青光眼和高眼压症的药物。所研究的药物包括酒石酸溴莫尼定（BMN）、乙酰唑胺（AZA）、布林佐胺（BZA）、盐酸多佐胺（DZA）、盐酸左布诺洛尔（LVB）和马来酸噻吗洛尔（TIM）。使用Thermo Hypersil BDS C18柱（4.6×250 mm，5 µm），以pH 5的磷酸盐缓冲液和乙腈按78 + 22的比例作为流动相，实现了高效的色谱分离。流速为1 mL/min，定量基于在298 nm处测量TIM的峰面积以及在254 nm处测量其他药物的峰面积。峰得到了完美分离，BMN、AZA、DZA、TIM、LVB和BZA的保留时间分别为3.06、3.87、4.53、5.78、7.31和10.78分钟。根据国际协调会议指南，对所开发的方法在系统适用性、线性、范围、准确性、精密度、稳健性以及检测限和定量限方面进行了验证。所提出的方法在BMN、AZA、DZA、TIM、LVB和BZA的浓度范围分别为2 - 80、2.5 - 100、2.5 - 100、5 - 200、3.75 - 150和1.75 - 70 µg/mL时显示出良好的线性。所分析化合物的检测限为0.20 - 1.01 μg/mL。通过分析五种不同的药物制剂评估了所提出方法在实际情况中的适用性，并获得了满意的结果。
