Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Beni-Suef University, Alshaheed Shehata Ahmed Hegazy St., Beni-Suef 62514, Egypt.
J Chromatogr Sci. 2021 Apr 21;59(5):425-431. doi: 10.1093/chromsci/bmaa132.
Two simple, sensitive and validated chromatographic methods were developed for quantitative determination of bromhexine hydrochloride (BHX) in presence of its major impurities, impurity B (IMB) and impurity C (IMC), as specified by British Pharmacopoeia. First method (I) was high-performance thin layer chromatography-densitometry at which the chromatographic separation was performed using silica gel plates and developing system consisted of hexane:acetone:ammonia solution (9:0.5:0.08, by volume) with ultraviolet scanning at 240 nm and linearity was achieved in the ranges of 0.40-10.00, 0.20-2.00 and 0.20-2.00 μg/band of BHX, IMB and IMC, respectively. Also, second chromatographic method (II) was high-performance liquid chromatography (HPLC) where the separation was carried out on C18 column at isocratic mode at flow rate 1.5 mL/min. The mobile phase consisted of methanol:water (90:10, v/v) adjusted to pH 2.5 with O-phosphoric acid and temperature was adjusted to 40°C. The scanning wavelength was 240 nm. The chromatographic run time was 6 min. Linearity of this method was achieved in the ranges of 4.00-40.00, 0.20-10.00 and 0.50-10.00 μg/mL for BHX, IMB and IMC, respectively. The validation of these chromatographic methods was made according to International Conference on Harmonization guidelines. These methods were successfully applied for determination of BHX in its pharmaceutical formulation. Also, statistical comparison was attained between the developed methods and the reported HPLC method using Student's t-test and F-test, and the obtained results showed that there was not any significant difference between them concerning with accuracy and precision.
两种简单、灵敏且经验证的色谱方法被开发出来，用于定量测定溴己新盐酸盐（BHX），同时也能测定其主要杂质 B（IMB）和 C（IMC），这是英国药典规定的。第一种方法（I）是高效薄层色谱-密度法，其中的色谱分离是在硅胶板上进行的，展开系统由正己烷：丙酮：氨溶液（9:0.5:0.08，体积比）组成，在 240nm 处进行紫外扫描，线性范围分别为 BHX、IMB 和 IMC 的 0.40-10.00、0.20-2.00 和 0.20-2.00μg/带。此外，第二种色谱方法（II）是高效液相色谱法（HPLC），其中分离是在 C18 柱上以等度模式进行的，流速为 1.5mL/min。流动相由甲醇：水（90:10，v/v）组成，用 O-磷酸调节 pH 值至 2.5，温度调整至 40°C。扫描波长为 240nm。色谱运行时间为 6 分钟。该方法的线性范围分别为 BHX 的 4.00-40.00、0.20-10.00 和 0.50-10.00μg/mL，IMB 和 IMC 的 4.00-40.00、0.20-10.00 和 0.50-10.00μg/mL。这些色谱方法的验证是根据国际协调会议的指导方针进行的。这些方法成功地应用于其药物制剂中 BHX 的测定。此外，通过学生 t 检验和 F 检验对开发的方法和报道的 HPLC 方法进行了统计比较，结果表明，它们在准确性和精密度方面没有显著差异。
