攜帶雞卵蛋白聚乳酸甘醇酸微粒固化過程中二氯甲烷揮發特性之探討   全文下載

Dichloromethane Evaporative Behavior during the Solidifying Process of Ovalbumin-Loaded Poly (DL Lactic-co-Glycolic Acid) Microparticles

本研究之目的為探討複乳劑溶媒萃取法製備載有雞卵蛋白聚乳酸甘醇酸微粒固化過程中二氯甲烷揮發之特性。研究中使用具頂樣器之氣相層析儀，測定在複乳劑及外相中二氯甲烷之經時變化。檢品製備時，以四氯化碳作為內部標準品，並與大量溶媒氯仿混合後加入檢品中，進行二氯甲烷含量分析。在氣相層析圖譜中，四氯化碳、二氯甲烷、氯仿之滯留時間分別為2.6、2.8、3.3分鐘，二氯甲烷之最低可定量濃度為7.3 mM，在7.5-75 mM濃度範圍線性關係良好，分析之變異係數為1.2%~3.9%。研究中製備四種分別含有0~5% 氯化鈉並加入尿素調整滲透壓為1240 mOsm／kg之微粒配方，結果顯示在四種微粒配方製備時，複乳劑中二氯甲烷變化能以零級 （0~15分鐘）及一級 （15分鐘以後） 動力學模式描述。在起始15分鐘，這些配方之零級動力學揮發速率常數為2.30~2.63 mmole／min；在15分鐘後，含5%氯化鈉配方（F4）之二氯甲烷一級動力學揮發速率常數（0.193 min-1）明顯小於其它氯化鈉含量低的配方（0.288~0.367 min-1）。此結果也說明F4雞卵蛋白載藥微粒形成過程中，表面性質與其它配方的不同，經由表面形成殼狀結構而延緩二氯甲烷由內相外流揮發之過程。由上述結果顯示在本研究中我們建立一個不需前處理即可應用於微粒固化過程中二氯甲烷含量變化分析的氣相層析分析方法，此方法也可用於評估二氯甲烷含量與載藥微粒其它配方因子相關性之研究。

The purpose of this study is to develop an analytical method for determining the evaporative behavior of dichloromethane (DCM) during the solidifying process of poly(DL lactic-co-glycolic acid) (PLGA) in the preparation of ovalbumin-loaded microparticle (OVAMP) with double emulsion solvent extraction method. The time courses of DCM levels in the external phase and total mixture were determined by a gas chromatography (GC) method combined a headspace sampler. Samples were spiked with an internal standard carbon tetrachloride, prior to mixing with a large volume of chloroform. The retention times of carbon tetrachloride, DCM and chloroform were 2.6, 2.8 and 3.3 min, respectively. The analytical method had a minimum quantitative concentration of DCM 7.3 mM and good linearity from 7.5 to 75 mM with coefficient of variations 1.2~3.9%. Four formulations containing NaCl (0~5%) and urea, an osmotic agent to adjust osmolarity at 1240 mOsm/kg, were investigated. The disappearance of DCM levels in total mixtures of four formulations were described as a function of time by zero-order (0~15 min) and first-order (after 15 min) kinetics. During the initial 15 min, these formulations had almost identical zero-order rate constants, 2.30~2.63 mmole/min. After 15 min, formulation with 5% NaCl (F4) showed a significant lower value of rate constant (0.193 min-1) in comparison with other formulations using the lower amounts of NaCl (0.288~0.367 min-1). It indicated that the surface characteristic of the F4 OVA-MP was different from other formulations by forming a crust-like structure to inhibit the efflux of DCM from the inner layer. In conclusion, we established a rapid and convenient GC approach without the need for sample pre-treatment for determining DCM levels during the solidifying process of microparticles. The analytical method could be applied to further assess the relationship between DCM level and formulation factors of drug-loaded microparticles. 本研究之目的為探討複乳劑溶媒萃取法製備載有雞卵蛋白聚乳酸甘醇酸微粒固化過程中二氯甲烷揮發之特性。研究中使用具頂樣器之氣相層析儀，測定在複乳劑及外相中二氯甲烷之經時變化。檢品製備時，以四氯化碳作為內部標準品，並與大量溶媒氯仿混合後加入檢品中，進行二氯甲烷含量分析。在氣相層析圖譜中，四氯化碳、二氯甲烷、氯仿之滯留時間分別為2.6、2.8、3.3分鐘，二氯甲烷之最低可定量濃度為7.3 mM，在7.5-75 mM濃度範圍線性關係良好，分析之變異係數為1.2%～3.9%。研究中製備四種分別含有0～5% 氯化鈉並加入尿素調整滲透壓為1240 mOsm／kg之微粒配方，結果顯示在四種微粒配方製備時，複乳劑中二氯甲烷變化能以零級 （0～15分鐘）及一級 （15分鐘以後） 動力學模式描述。在起始15分鐘，這些配方之零級動力學揮發速率常數為2.30～2.63 mmole／min；在15分鐘後，含5%氯化鈉配方（F4）之二氯甲烷一級動力學揮發速率常數（0.193 min-1）明顯小於其它氯化鈉含量低的配方（0.288～0.367 min-1）。此結果也說明F4雞卵蛋白載藥微粒形成過程中，表面性質與其它配方的不同，經由表面形成殼狀結構而延緩二氯甲烷由內相外流揮發之過程。由上述結果顯示在本研究中我們建立一個不需前處理即可應用於微粒固化過程中二氯甲烷含量變化分析的氣相層析分析方法，此方法也可用於評估二氯甲烷含量與載藥微粒其它配方因子相關性之研究。