Gondhale Priyanka Satish, Varghese Cheriyan Binoy
Department of Pharmaceutical Chemistry and Analysis, School of Pharmacy, Vels Institute of Science, Technology and Advanced studies (VISTAS), Tamil Nadu, India
Turk J Pharm Sci. 2022 Feb 28;19(1):35-47. doi: 10.4274/tjps.galenos.2021.77177.
Tiotropium (TIO) and formoterol fumarate (FF) combination in dry powder inhaler (DPI) dosage form used for treating asthma, bronchospasm, chronic bronchitis, emphysema and chronic obstructive pulmonary diseases. Aim to develop an analytical method for estimating emerging and advancing dry powder inhaler combination toward enhanced therapeutics for the estimation of related substances but for this it is foremost to have a sensitive, simple, robust and validated method therefore, a new reverse phase-high performance liquid chromatography (HPLC) method has been developed for the determination of related substances in FF and TIO DPI.
The analytical method was performed on schimadzu HPLC with a quaternary pump, the separation achieved using BDS Hypersil C18 (250×4.6 mm, 5 μm) column with mobile phase consisting of sodium phosphate buffer pH 3.2 and acetonitrile 1.0 mL min-1 flow rate in the gradient elution. Diluent consists of a mixture of buffer pH 3.2 and acetonitrile in the ratio of (70:30) %v/v. 30°C column temperature and photodiode-array detection detector at a wavelength 240 nm. The run time was 50 min. The Retention time of FF and TIO was found to be at 7.8 and 10.3 min respectively.
Both the analyte peaks were found to be free from interference. The method was validated as the International Council on Harmonisation guidelines, the linearity was performed on 0.015 to 1.089 ppm for TIO and 0.01 to 0.728 ppm for FF concentration with correlation coefficient of 1,000. The precision and accuracy were also performed at the limit of quantification level were within the limits. Forced degradation study was also conducted.
The recommended method for the related substance determination of FF and TIO is simple, selective, specific and precise. It also demonstrates the study of the degradation pattern. Moreover, the above developed related substance analytical method was applied to the bulk analysis and pharmaceutical dosage form for routine analysis and stability study.
噻托溴铵（TIO）与富马酸福莫特罗（FF）的干粉吸入剂剂型用于治疗哮喘、支气管痉挛、慢性支气管炎、肺气肿和慢性阻塞性肺疾病。旨在开发一种分析方法，用于评估新兴和先进的干粉吸入剂组合，以增强其治疗效果，用于相关物质的测定，但为此首先要有一种灵敏、简单、稳健且经过验证的方法，因此，已开发出一种新的反相高效液相色谱（HPLC）方法用于测定FF和TIO干粉吸入剂中的相关物质。
采用岛津HPLC仪，配备四元泵，使用BDS Hypersil C18（250×4.6 mm，5μm）色谱柱进行分离，流动相由pH 3.2的磷酸钠缓冲液和乙腈组成，流速为1.0 mL min-1，采用梯度洗脱。稀释剂由pH 3.2的缓冲液和乙腈按（70:30）%v/v的比例混合而成。柱温为30°C，采用光电二极管阵列检测器，检测波长为240 nm。运行时间为50分钟。发现FF和TIO的保留时间分别为7.8分钟和10.3分钟。
两个分析物峰均无干扰。该方法按照国际协调理事会指南进行了验证，TIO的线性范围为0.015至1.089 ppm，FF的浓度线性范围为0.01至0.728 ppm，相关系数为1.000。在定量限水平也进行了精密度和准确度测定，均在规定范围内。还进行了强制降解研究。
所推荐的用于测定FF和TIO相关物质的方法简单、选择性好、特异性强且精确。它还展示了降解模式的研究。此外，上述开发的相关物质分析方法应用于原料药分析和药物剂型的常规分析及稳定性研究。
