硫酸特布他林的质量控制方法研究

发布时间：2018-07-21 20:01

【摘要】：硫酸特布他林为选择性β2受体激动剂,用于支气管哮喘、慢性支气管炎和其它伴有支气管痉挛的肺部疾病,也可用于预防早产和胎儿窒息,具有选择性高、副作用少等特点,在国内外已得到广泛应用。本课题针对现行硫酸特布他林质量控制的不足,就影响其质量的关键指标——残留溶剂、工艺杂质与降解产物以及含量分别进行研究,为硫酸特布他林的质量评价和控制提供了新的方法和参考。在全面分析硫酸特布他林合成过程中所用溶剂的基础上,首次建立了一种同时测定硫酸特布他林中甲醇、乙醇、丙酮、叔丁基甲基醚、乙酸乙酯、四氢呋喃、苯、醋酸、甲苯和氯化苄共10种残留溶剂的顶空气相色谱法;建立了超高效液相色谱高分辨质谱联用(UHPLC-Q-TOF-MS/MS)法,用于硫酸特布他林工艺杂质与降解产物的结构解析和鉴定;建立了高效液相色谱(HPLC)法,用于硫酸特布他林的含量测定。第一部分顶空气相色谱法同时测定硫酸特布他林中10种残留溶剂目的:建立测定硫酸特布他林中甲醇、乙醇、丙酮、叔丁基甲基醚、乙酸乙酯、四氢呋喃、苯、醋酸、甲苯和氯化苄共10种残留溶剂的顶空气相色谱法。方法:采用Agilent DB-624UI(30 m×0.32 mm,1.8μm)毛细管柱,柱温采用程序升温,起始柱温为40°C,保持3 min,以10°C/min升至90°C,保持1 min,再以40°C/min升至200°C,保持5 min。FID检测器,检测器温度为250°C,进样口温度为200°C。载气为氮气,流速为1.0mL/min;分流比为15:1。顶空进样,顶空平衡温度为90°C,顶空时间为30 min。定量环温度为100°C,传输线温度为110°C。压力平衡时间为1min,进样时间为1 min。结果:10种溶剂完全分离;在考察的浓度范围内线性关系良好,相关系数r均不小于0.9990,平均回收率为95.7%-102.4%,仪器精密度、重复性、稳定性、检测限和定量限均符合要求。3批供试品中甲醇的质量分数分别为0.035%、0.043%、0.043%,乙醇的质量分数分别为0.033%、0.034%、0.034%,丙酮的质量分数分别为0.00059%、0.00062%、0.00062%,叔丁基甲基醚、乙酸乙酯、四氢呋喃、苯、醋酸、甲苯、氯化苄均低于检测限(lod)。结论:建立的顶空气相色谱法简单、准确、重复性好,可用于硫酸特布他林中残留溶剂的测定。第二部分硫酸特布他林工艺杂质与降解产物的uhplc-q-tof-ms/ms分析目的:采用超高效液相色谱高分辨质谱联用技术(uhplc-q-tof-ms/ms)对硫酸特布他林工艺杂质与降解产物进行结构解析和鉴定。方法:对硫酸特布他林质谱裂解规律进行研究,然后将硫酸特布他林供试品分别进行酸、碱、氧化、高温、高湿、光照降解及长期试验,采用uhplc-q-tof-ms技术对获得的14种降解样品进行分析。色谱条件:采用phenomenexlunac18色谱柱(2.0mm×150mm,3μm),流动相为甲醇-10mmol/l甲酸铵(用甲酸调ph至3.2),梯度洗脱,流速为0.3ml/min,进样量为5μl。质谱条件:采用duospraytm离子源正离子模式,全扫描采集,采用动态背景扣除和信息依赖采集触发二级碎片的采集,每一循环采集12个最强峰的二级质谱图。根据测得的各降解产物及其碎片离子的准确质量数和可能分子组成,以硫酸特布他林质谱裂解规律为基础,比较降解产物与硫酸特布他林以及降解产物碎片离子之间的质量数变化,对其进行结构鉴定。结果:共鉴定出17个杂质,包括14个新的降解产物和3个已知杂质,其中3个已知杂质既是工艺杂质,也是降解产物。结论:硫酸特布他林在氧化、碱性水解和光照(碱性溶液)条件下不稳定;容易发生反应的官能团为叔丁胺基、苄基碳和苯环。本研究首次对降解产物可能的降解途径进行了推断。第三部分硫酸特布他林的含量测定方法改进目的:对硫酸特布他林现有含量测定方法进行改进,建立高效液相色谱(hplc)法用于硫酸特布他林的含量测定。方法:选用agilentzorbaxsb-c18色谱柱(4.6mm×150mm,5μm);流动相为30mmol/l甲酸铵(用甲酸调ph至3.0)-甲醇(体积比为90:10);流速为1.0ml/min;检测波长为276nm;进量样为10μl;柱温为室温。结果:方法的线性范围为0.01-0.40 mg/mL(r=1.000)。回收率为99.2%,RSD为0.8%(n=6)。仪器精密度、重复性、稳定性、检测限和定量限均符合要求。3批供试品中硫酸特布他林的含量分别为100.5%、98.9%、99.2%。结论:与现有电位滴定法和离子对高效液相色谱法(IP-HPLC)比较,改进的HPLC法更为简单、快速、经济,并可以从液相色谱图上直观反映样品中主要杂质的变化,可用于硫酸特布他林的含量测定。
[Abstract]:Terbutaline sulfate, a selective beta 2 receptor agonist, is used in bronchial asthma, chronic bronchitis and other pulmonary diseases with bronchial spasm. It can also be used to prevent premature and fetal asphyxia. It has high selectivity and less side effects. It has been widely used at home and abroad. This subject is aimed at the quality control of current terbutaline sulfate. A new method and reference for the quality evaluation and control of terbutaline sulfate is provided for the quality evaluation and control of terbutaline sulfate, which is the key index which affects its quality, such as residual solvents, process impurities and degradation products and their content respectively, and a new method is established on the basis of the comprehensive analysis of the solvent used in the process of terbutaline sulfate synthesis. A headspace gas chromatographic method for the determination of 10 residual solvents of methanol, ethanol, acetone, tert butyl methyl ether, ethyl acetate, tetrahydrofuran, benzene, acetic acid, toluene and benzyl chloride in terbutaline sulfate, and a high resolution liquid chromatography high resolution mass spectrometry (UHPLC-Q-TOF-MS/ MS) method for the synthesis of impurities and degradation products in terbutaline sulphate process was established. Structure analysis and identification; a high performance liquid chromatography (HPLC) method was established for the determination of the content of terbutaline sulphate. The first part, headspace gas chromatography, was used for the simultaneous determination of 10 residual solvents in terbutaline sulphate. The determination of methanol, ethanol, acetone, tert butyl methyl ether, ethyl acetate, tetrahydrofuran, benzene, acetic acid, methyl in terbutaline sulfate was established. The headspace gas chromatography of 10 kinds of residual solvents of benzene and benzyl chloride. Method: using Agilent DB-624UI (30 m x 0.32 mm, 1.8 m) capillary column, the column temperature was programmed heating, the starting column temperature was 40 C, the 3 min was kept, 10 degree C/min to 90 C, 1 min, and then 40 [degree C/min] to 200 degrees C, and the detector temperature was 250 degrees. The sample mouth temperature is 200 C. nitrogen, the flow rate is 1.0mL/min, the flow ratio is 15:1. headspace sample, the headspace equilibrium temperature is 90 C, the headspace is 30 min. quantitative ring temperature 100 C, the transmission line temperature is 110 degree C., the pressure balance time is 1min, the injection time is 1 min. results: 10 solvents are completely separated; linear in the range of investigation is linear. The relationship is good, the correlation coefficient r is not less than 0.9990, the average recovery is 95.7%-102.4%, instrument precision, repeatability, stability, detection limit and quantitative limit all meet the requirements of the.3 batch test products, the mass fraction of methanol is 0.035%, 0.043%, 0.043%, and the mass fraction of ethanol is 0.033%, 0.034%, 0.034%, and the acetone's mass fraction is 0. respectively. 00059%, 0.00062%, 0.00062%, tert butyl methyl ether, ethyl acetate, tetrahydrofuran, benzene, acetic acid, toluene, benzyl chloride are lower than the detection limit (LOD). Conclusion: the set up headspace gas chromatography is simple, accurate, reproducible, and can be used for the determination of residual solution in terbutaline sulphate. The second part of the terbutaline process and the uh of the degradation products Plc-q-tof-ms/ms analysis objective: to analyze and identify the structure of impurities and degradation products of terbutaline sulphate by super high performance liquid chromatography (uhplc-q-tof-ms/ms) and high resolution mass spectrometry (high resolution mass spectrometry). Method: the fragmentation of terbutaline sulfate mass spectrometry was studied, and then acid, alkali and oxidation were carried out in terbutaline sulfate. High temperature, high humidity, light degradation and long term test, 14 kinds of degradation samples were analyzed by uhplc-q-tof-ms technology. The chromatographic conditions were: using phenomenexlunac18 column (2.0mm x 150mm, 3 mu m), the flow phase was methanol -10mmol/l formate (pH to 3.2), gradient elution, velocity of 0.3ml/min, and the sampling amount of 5 u mass mass spectrometry. Using duospraytm ion source positive ion mode, full scan collection, dynamic background deduction and information dependent acquisition trigger two stage debris collection, each cycle to collect 12 strongest peaks of two level mass spectrogram. According to the accurate mass number and possible molecular composition of each degradation product and its fragment ions, the pyrolysis of terbutaline sulfate mass spectrometry On the basis of the law, the mass changes between the degradation products and the terbutaline sulfate and the fragments of the degradation products were compared and the structure was identified. Results: 17 impurities were identified, including 14 new degradation products and 3 known impurities, of which 3 known impurities were both process impurities and degradation products. Conclusion: sulphuric acid terbuterol The forest is unstable under the condition of oxidation, alkaline hydrolysis and illumination (alkaline solution); the functional groups that are easy to react are tertiary Ding Anji, benzyl carbon and benzene ring. This study is the first time to infer the possible degradation pathways of the degradation products. The third part of the method for the determination of the content determination method of terbutaline sulphate: the present content measurement of terbutaline sulfate The method is improved, and a high performance liquid chromatography (HPLC) method is established for the determination of the content of terbutaline sulphate. Methods: agilentzorbaxsb-c18 column (4.6mm x 150mm, 5 mu m), mobile phase 30mmol/l formate (pH to 3) - methanol (volume ratio 90:10); flow rate 1.0ml/min; detection wavelength of 276nm; intake sample of 10 mu L; column temperature Room temperature. Results: the linear range of the method is 0.01-0.40 mg/mL (r=1.000). The recovery rate is 99.2%, RSD is 0.8% (n=6). The instrument precision, repeatability, stability, detection limit and quantitative limit are all conformed to the requirement of.3 batch, the content of terbutaline sulfate is 100.5%, 98.9%, 99.2%. conclusion: with the current potentiometric titration method and the ion pair high performance liquid phase Compared with the chromatography (IP-HPLC), the improved HPLC method is simpler, faster, economical, and can directly reflect the changes of the main impurities in the sample from the liquid chromatography, and can be used for the determination of the content of terbutaline sulfate.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R927

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