建立樣品電動進樣堆積之毛細管電泳分析法對兩種類固醇Budesonide與Fluticasone propionate之同時分析研究及其應用

Simultaneous determination of two steroids of budesonide and fluticasone propionate by capillary electrophoresis coupling with electrokinetic stacking injection

石祐安(You-An Shih)

指導教授 ： 陳素惠

高雄醫學大學/藥學院/藥學研究所/碩士(2014年)

摘要

布地奈德及氟替卡松丙酸酯屬於化學合成性的類固醇，能應用在發炎性疾病的治療上。論文中會先介紹類固醇的疾病治療與相關副作用，並且介紹可用來分析中性分析物之新興毛細管電泳法---非水毛細管電泳、重覆大體積進樣搭配微胞破裂法之毛細管電泳法及常見之微胞電動層析法與毛細管區帶電泳，進而介紹本論文應用之電動進樣堆積毛細管電泳法，再將五種分析方法做比較。
本方法係在有效長度30公分之熔矽毛細管內充滿硼酸鹽緩衝溶液(28 mM, pH 9.6)內含9%甲醇、80 mM十二烷基羥硫酸鹽，再以電壓進行取樣 (15 kV, 5 min)，通電後產生之電滲透流將分析物帶入毛細管後並且同時進行線上樣品濃縮的方式來提高毛細管電泳法對吸入劑中布地奈德及氟替卡松丙酸酯之偵測感度，最後用15 kV的電壓將待測物分離，並於波長254 nm下偵測之，內部標準品為Trazodone。
本實驗加以探討了影響分離的因素，包括：硼酸鹽緩衝溶液的濃度和酸鹼值、十二烷基羥硫酸鹽的濃度以及緩衝溶液中甲醇的含量。此方法之線性範圍為40-500 ng/mL，偵測極限皆為20 ng/mL。最後應用在普米克及輔舒酮二種市售乾粉吸入劑之單一劑量均一度測試，其含量規範皆符合美國藥典第三十五版之規定。

關鍵字

布地奈德 ； 毛細管電泳法 ； 電動進樣 ； 氟替卡松丙酸酯 ； 類固醇

並列摘要

Budesonide and fluticasone propionate were chemical synthetic steroids, and they could be applied to treat inflammatory diseases. First, this study would introduce the therapies and adverse effects of glucocorticoids. Second, we compared electrokinetic stacking injection (EKI) with four different analytical methods of capillary electrophoresis for neutral analytes including non-aqueous capillary electrophoresis (NACE), repetitive large volume sample injection combined sweeping MEKC (rLVSS-MEKC), micellar electrokinetic capillary chromatography (MEKC) and capillary zone electrophoresis (CZE).
Before sample loading, a background electrolyte was filled with an uncoated fused silica capillary of effective length 30 cm. Electrokinetic injection (15 kV, 300 s) was used to introduce neutral sample. The separation condition for budesonide and fluticasone propionate was performed in electrolyte solutions containing borate buffer (28 mM, pH 9.6) with 80 mM sodium dodecyl sulfate (SDS) and 9% methanol (MeOH). The analytes would stack at the interface of the sample matrix and separation buffer micelle zones. The separation was performed at 15 kV and detected at 254 nm. We used trazodone as an internal standard.
In the research, some analytical factors were investigated including the concentrations and pH values of borate buffer, the concentrations of SDS and the percentages of methanol. The linear ranges of the method for the determination of budesonide and fluticasone propionate in commercial samples were over a range of 40-500 ng/mL. The limit of detection was 20 ng/mL (S/N=3). Both analytical results of content uniformity of budesonide formulation (Pulmicort turbuhalerR) and fluticasone propionate formulation (Flixotide accuhalerR) were within the criterion of USP 35.

並列關鍵字

Budesonide ； Capillary electrophoresis ； Electrokinetic injection ； Fluticasone propionate ； Steroid

參考文獻

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