Розробка методик контролю якості лікарських препаратів з бензидаміну гідрохлоридом з позицій «зеленої хімії»

Abstract

У дисертації висвітлено питання розробки та валідації аналітичних методик контролю якості оромукозних препаратів з бензидаміну гідрохлоридом за принципами «зеленої хімії». Розроблено «дерево рішень» щодо вибору аналітичних методик з огляду на їх відповідність принципам «зеленої хімії», за результатами його імплементації обґрунтовано методологію досліджень. Розроблено методику визначення супровідних домішок у препаратах бензидаміну гідрохлориду спреї та розчині методом ВЕРХ, проведено дослідження деградації АФІ в стресових умовах. Запропоновано підходи до ідентифікації бензидаміну та його метаболіту в присутності НПЗЗ. Розроблено методики одночасного визначення АФІ (бензидаміну гідрохлориду) та консерванта (метилпарагідроксибензоату) методами ВЕРХ та ГХ. Визначено оптимальні умови пробопідготовки та хроматографування, проведено їх валідацію. Розроблено методику визначення залишкових кількостей АФІ на поверхні обладнання методом абсорбційної спектрофотометрії. Розроблено методику одночасного визначення бензидаміну, ментолу та метилпарабену в оригінальній комбінованій лікарській формі методом ГХ. Розроблені методики відповідають принципам «зеленої хімії» та є коректними за результатами валідаційних досліджень.

Диссертация посвящена разработке и валидации аналитических методик контроля качества оромукозних препаратов на основе бензидамина гидрохлорида по принципам «зеленой химии». Разработано «дерево решений» по выбору аналитических методик с учетом их соответствия принципам «зеленой химии», по результатам его имплементации обоснована методология исследований. Разработана методика определения сопутствующих примесей в препаратах бензидамина гидрохлорида спрей и раствор методом ВЭЖХ, проведено исследование деградации АФИ в стрессовых условиях. Разработаны методики одновременного определения АФИ (бензидамина гидрохлорида) и консерванта (метилпарагидроксибензоата) методами ВЭЖХ и ГХ. Определены оптимальные условия пробоподготовки и хроматографирования, проведена их валидация. Разработана методика определения остаточных количеств АФИ на поверхности оборудования методом абсорбционной спектрофотометрии. Разработана методика одновременного определения бензидамина, ментола и метилпарабена в оригинальной комбинированной лекарственной форме методом ГХ. Разработанные методики соответствуют принципам «зеленой химии» и являются корректными по результатам валидационных исследований

The dissertation is devoted to the development and validation of analytical methods of quality control of oromucosal preparations with benzydamine hydrochloride according to the principles and approaches of "green chemistry". The development of the "decision tree" was performed on the basis of assessing the degree of use of organic compounds and solvents by various analytical methods, as well as the direct ability of these methods to simultaneously determine multiple analytes, ie to "built-in selectivity" of analytical methods. As result of “decision tree” implementation methodology of investigations was substantiated. To develop a method for the related substances of benzydamine hydrochloride in oromucosal spray and solution, the method of liquid chromatography was used as the most effective technique for the separation of related substances. A GraceAltima C18 chromatographic column was selected for separation. As the mobile phase, a buffer solution based on sodium perchlorate, with a pH of 3.0, and triethylamine as an ion-pair additive was selected. Stress tests were performed to determine potential degradation products. The main degradation product, benzydamine N-oxide, was determined by mass spectrometric determination in tandem with HPLC. The validation characteristics of the method were selected and investigated taking into account the requirements of SPhU and ICH Q2. The characteristics of linearity, precision, accuracy and robustness are determined in accordance with the specified criteria and are within their limits. The development of a method for the simultaneous determination of benzydamine hydrochloride and methylparaben was carried out according to the further implementation of the green chemistry approach. At first step, HPLC was used to simultaneously determine the components. Wavelengths of 320 and 254 nm were used to detect benzydamine and methylpararben. A mixture of acetonitrile buffer with pH 3.0 consisting of sodium perchlorate, perchloric acid and triethylamine was chosen as the mobile phase. The specificity of the method was proved by the absence of the effect of placebo peaks on the studied components, as well as the lack of coelution between benzydamine and methylparaben. The correlation coefficient of the regression line R2 for both analytes was more than 0.999. The requirements for the suitability criteria of the chromatographic system were established. The robustness of the method was tested by slightly varying the parameters of the method, namely, the flow rate and pH of the mobile phase. When developing a method for the simultaneous determination of benzydamine and methylparaben by gas chromatography, the possibility of elution of both analytes simultaneously on a gas chromatographic column was evaluated. To do this, it was necessary to solve the problem of transferring benzydamine hydrochloride to the base to reduce the boiling point. Weakly polar chloroform was used as an extractant. Benzydamine impurity A was used as an internal standard, which is not a product of possible decomposition of the molecule and accordingly will not change its concentration during sample preparation. The gradient parameters of the method were selected so as to simultaneously determine both components. The method of absorption spectrophotometry was used to develop a method for determining the residual amounts of API on the surface of the equipment. The maximum allowable amounts of API that can be transferred to another drug during the technological process due to cross-contamination were taken into account. It was determined that the maximum allowable amount of benzydamine hydrochloride on the surface of the equipment should not exceed 10 ppm. Determination of API residues is proposed to be performed at a wavelength of 310 nm, because this is the wavelength where the maximum absorption is observed at a given concentration. The specificity of the method was proved by the lack of effect of placebo on the signal of the desired substance. The correlation coefficient in determining the linearity of the method was 0.999. The coefficient of variation of the definition did not exceed 1%. To enhance the antibacterial effect of benzydamine oromucosal spray it was proposed the addding of essential oil into the composition of the spray. Oil screening was performed by comparative studying of the antimicrobial action of benzydamine solutions after adding of essential oils as eucalypt, lavender and peppermint respectively. Peppermint essential oil has been shown the highest enhance of antibacterial effect of benzydamine solution. To quantify the components of the original combination spray, the possibility of further implementation of the "green chemistry" approach was considered, which would allow the simultaneous determination of API, preservative and menthol as marker component of essential oil. The method developed before for the simultaneous determination of benzydamine and methylparaben by gas chromatography was taken as a basis. The determination was performed after preliminary extraction components with chloroform. The calculation of the quantitative content of the components was performed through the relationship to the internal standard. Sufficient separation of all analytes from drug matrix components was shown. The coefficient of linearity for all detectable components was more than 0.999. The correctness and reproducibility of the quantification were established. RSD% for all components does not exceed 0.25% for five parallel injections. The reliability of the procedure was studied by varying the parameters of the method, namely the initial temperature, the velocity of the mobile phase and the sample volume. The proposed methods are implemented by JSC "Farmak" into the internal specification for the control of the finished dosage forms.