【摘要】：β-激動(dòng)劑(β-agonists)類手性藥物的兩個(gè)對(duì)映異構(gòu)體在藥效學(xué)、藥代動(dòng)力學(xué)及毒理學(xué)等方面都存在較大差異,建立其手性分離與檢測(cè)方法具有重要意義。毛細(xì)管電泳(Capillary electrophoresis,CE)是手性藥物分離與檢測(cè)常用的一種方法。非水毛細(xì)管電泳(Nonaqueous capillary electrophoresis,NACE)是CE的一個(gè)重要分支,在手性藥物分離方面具有獨(dú)特優(yōu)勢(shì),近年來引起了人們的關(guān)注。本文在含有三乙胺的甲醇溶液中原位合成了雙丙酮-D-甘露糖醇 硼酸絡(luò)合酸手性選擇劑,并將其應(yīng)用于NACE中,成功實(shí)現(xiàn)了7種β-激動(dòng)劑類手性藥物的分離。紫外吸收光譜(Ultraviolet spectrometry,UV)和11B-核磁共振波譜(11B-nuclear magnetic resonance spectroscopy,11B-NMR)實(shí)驗(yàn)證明了雙丙酮-D-甘露糖醇 硼酸絡(luò)合酸的生成。本文還將建立的NACE手性分離方法應(yīng)用于口服液中克侖特羅對(duì)映體的含量測(cè)定和飲用水及豬肝樣品中克侖特羅對(duì)映體的殘留量分析。全文共分六章:第一章:綜述了β-激動(dòng)劑類手性藥物分離與檢測(cè)的意義及β-激動(dòng)劑類手性藥物常用的分離與檢測(cè)方法,并對(duì)NACE在手性藥物分離檢測(cè)中的應(yīng)用進(jìn)行了介紹。第二章:在非水溶液中制備了雙丙酮-D-甘露糖醇 硼酸絡(luò)合酸,并以其為手性選擇劑,在NACE中對(duì)7種β-激動(dòng)劑進(jìn)行了分離。為了獲得良好的手性分離效果,實(shí)驗(yàn)分別考察了雙丙酮-D-甘露糖醇、硼酸和三乙胺濃度對(duì)分離效果的影響。在優(yōu)化的NACE手性分離條件下,所有7種藥物均可獲得基線分離,其中6種分離時(shí)間在12分鐘之內(nèi)。第三章:分別采用UV和11B-NMR方法考察了雙丙酮-D-甘露糖醇 硼酸絡(luò)合酸在NACE緩沖溶液中的生成情況。第四章:根據(jù)人用藥物注冊(cè)技術(shù)要求國(guó)際協(xié)調(diào)會(huì)(International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use,ICH)指導(dǎo)原則對(duì)所建立的NACE手性分離方法進(jìn)行了方法學(xué)驗(yàn)證。結(jié)果表明,5種測(cè)試藥物的兩個(gè)對(duì)映體在1.25~100.00μg mL-1范圍內(nèi)線性關(guān)系良好,相關(guān)系數(shù)(r)≥0.9992;檢測(cè)限(Limit of determination,LOD)和定量限(Limit of quantification,LOQ)分別小于1.25μg mL-1和5.00μg mL-1,加標(biāo)回收率為98.2~101.9%,精密度RSD小于0.9%(n=9)。將建立的NACE法成功應(yīng)用于氨溴特羅口服液中克侖特羅對(duì)映體的含量測(cè)定。第五章:結(jié)合選擇性陽離子電動(dòng)進(jìn)樣和掃集技術(shù),建立NACE手性富集分離方法用于飲用水及豬肝樣品中克侖特羅對(duì)映體的殘留量分析。方法學(xué)驗(yàn)證結(jié)果顯示,克侖特羅的兩個(gè)對(duì)映體在0.25~12.50 ng mL-1濃度范圍內(nèi)均具有良好的線性關(guān)系,線性相關(guān)系數(shù)(r)≥0.9984,檢測(cè)限(LOD)和定量限(LOQ)分別為0.05 ng mL-1和0.25 ng mL-1。第六章:結(jié)論與展望。
[Abstract]:The two enantiomers of 尾-agonists (尾-agonists) have great differences in pharmacodynamics, pharmacokinetics and toxicology. It is of great significance to establish chiral separation and detection methods for 尾-agonists. Capillary electrophoresis (Capillary electrophoresis,CE) is a commonly used method for separation and detection of chiral drugs. Non-aqueous capillary electrophoresis (Nonaqueous capillary electrophoresis,NACE) is an important branch of CE. It has unique advantages in chiral drug separation and has attracted much attention in recent years. In this paper, diacetone-D-mannitol / boric acid chiral selector was in situ synthesized in methanol solution containing triethylamine and applied to NACE. Seven chiral drugs of 尾-agonists were successfully separated. The formation of diacetone-D-mannitol boric acid complex acid was confirmed by UV absorption spectroscopy (Ultraviolet spectrometry,UV) and 11B-nuclear magnetic resonance (11B-nuclear magnetic resonance spectroscopy,11B-NMR). The NACE chiral separation method has also been applied to the determination of clenbuterol enantiomers in oral liquid and the determination of clenbuterol enantiomers in drinking water and pig liver samples. The thesis is divided into six chapters: chapter one: the significance of separation and detection of 尾-agonists chiral drugs and the commonly used separation and detection methods of 尾-agonists chiral drugs are reviewed, and the application of NACE in the separation and detection of chiral drugs is introduced. In chapter 2, diacetone-D-mannitol borate complex acid was prepared in non-aqueous solution and used as chiral selector, seven 尾-agonists were separated in NACE. In order to obtain a good chiral separation effect, the effects of diacetone-D-mannitol, boric acid and triethylamine concentration on the separation efficiency were investigated. Under the optimized conditions of NACE chiral separation, baseline separation was obtained for all seven drugs, six of which were separated within 12 minutes. Chapter 3: the formation of diacetone-D-mannitol borate complex acid in NACE buffer solution was investigated by UV and 11B-NMR, respectively. In chapter 4, the methodology of chiral separation of NACE is verified according to the technical requirements of (International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use,ICH (International Coordination Committee for the Registration of drugs for Human use). The results showed that there was a good linear relationship between the two enantiomers in the range of 1.25 ~ 100.00渭 g / mL-1, and the correlation coefficient (r) was 鈮，

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