臺灣博碩士論文加值系統

目的：複合式樹脂為牙科中常使用填補材料，其主要成分Bis-GMA
(bisphenylglycidyl dimethacrylate )、TEGDMA (triethylene glycol dimethacrylate)、 UDMA (urethane dimethacrylate ) 因聚合不完全與唾液的侵蝕分解導致單體釋出進入人體,造成健康危害。本研究擬開發一個以直接固相微萃取( Solid-Phase Microextraction ；SPME)吸附的方式，結合高效液相層析儀(High Performance Liquid Chromatography；HPLC)來分析唾液中樹脂釋出物，進而探討聚合時間、溫度與pH影響釋出量的關係。
方法：本研究以60 μm Polydimethylsiloxane-divinylbenzene(PDMS/DVB)纖維作為吸附介質，以直接法萃取樹脂釋出物。完成萃取後的纖維浸入HPLC的Chamber進行靜態脫附，脫附完後進行分析。
實驗室技術建立時，製備標準溶液進行條件探討。研究探討的參
數包括: 纖維種類、脫附溶劑、吸附時間、脫附時間、吸附溫度、
攪拌速度、及pH值。於不同聚合時間: 20、40、60、80、120秒；不同培養溫度: 0°C、27°C、40°C;不同pH值: 3、5、7、9、11下，以商品Tetric Ceram培養7日後分析，探討聚合時間、溫度、pH值對釋出量的影響，並探討樹脂於不同天數下釋出量的變化。
結果：1.SPME方法為：將標準品添加於未經pH值調整唾液中，裝滿4 mL vial，瓶中加入攪拌磁石，密封後於40°C、轉速200 rpm(0.18g)下，以60 μm PDMS/DVB纖維直接吸附20分鐘，再以移動相( 65 %乙腈、35 %D.I.水)靜態脫附15分鐘。2.在此萃取分析條件下，所求得的檢量線範圍為TEGDMA：0.3~30µg/mL；UDMA：0.5~50 µg/mL；Bis-GMA：0.5~50µg/mL，其相關係數(r)皆在0.995以上；偵測極限分別為TEGDMA：0.035µg/mL；UDMA：0.276 µg/mL；Bis-GMA：0.064 µg/mL。3.複合式樹脂的釋出量隨著天數增加而增加，在第7日時達到一個較高值。4.溫度與pH值皆會影響釋出，隨著溫度增加UDMA、Bis-GMA釋出量增加；隨著pH減少，釋出量隨著增加，而影響最大的是聚合時間，增加聚合時間可降低其釋出量。
結論：以Direct-SPME-HPLC的方式來分析樹脂釋出物，可省去繁雜的前處理步驟，同時其具有免溶劑、快速、方法簡便、以及不受樣本基質干擾等優點。樹脂於唾液中的釋出物快速的被酵素分解，隨即進入人體循環系統，評估樹脂釋出物對人體健康的危害，應針對釋出物分解後產物較適合。在填補完牙齒的第一周，以漱口來降低口腔溫度，沖淡口腔酸性環境，盡量少吃過熱及偏酸的食物，以減少釋出量。

Objective：Dental composite resin materials are widely used for fixing teeth. The main components are BisGMA (bisphenylglycidyl dimetha-crylate)、TEGDMA (triethylene glycol dimethacrylate)、UDMA (urethane dimethacrylate ). These resin monomers may be released due to incomplete polymerization and have cytotoxicity and genotoxicity to human. The purpose of this study is to develop an analytical method for determining the resin monomers
in saliva using solid phase microextraction followed by high performance liquid chromatography (HPLC) analysis.
Method：The 60 μm Polydimethylsiloxane-divinylbenzene(PDMS/DVB)fiber through direct extraction was tested to be the most suitable material for sample extraction. The SPME fiber was inserted into the injection chamber of HPLC and analyzed. The optimum extraction parameters investigated are pH、extraction temperature, stirring speed, extraction time、desorption time、desorption solvent.
The effect of polymeric time (20、40、60、80、120s), temperature (0°C、27°C、40°C) and pH(3、5、7、9、11) to releasing amount are investigated. The test sample were polymerized for 120 s and stored in ethanol and saliva solution in room temperature with no adjustment pH for 12 h, 24 h, 3, 5, 7 or 10 days to investigate influence of immersing solution and time.
Results：1. The SPME method was placing magnetic stirring-bar in an 4 mL vial, stirring 200 rpm at 40 °C, PDMS/DVB fiber was used to perform direct solid-phase microextraction for 20 min and mobile phase(65 %acetonitrile:35 %D.I. water) desorption for 15 min.
2. For the quality control calibration, the concentration ranges of TEGDMA, UDMA, and Bis-GMA were 0.3~30 µg/mL, 0.5~50 µg/mL, 0.5~50 µg/mL. The correlation coefficients(r) were all above 0.995. The method detection limit（MDL）for TEGDMA, UDMA, and Bis-GMA were 0.035 µg/mL , 0.276 µg/mL and 0.064 µg/mL respectively.
3. Results showed a maximum concentration of monomers after 7 days almost.
4. Under longer polymeric time, can reducing the amount of releasing. With higher temperature and lower pH, releasing will increase.
Conclusion：The analytical method of eluted composite resins in whole saliva by Direct-SPME-HPLC can be reduced complicated pretreated procedure, as well as having the advantage of solvent-free,rapidity, simplicity, and avoid interfering with a complex matrix.The enzyme of saliva will react with releasing fast and into blood circulation.

目 錄
目次 頁次
第壹章 序論 1
第一節 研究背景 1
第二節 研究目的 4
第三節 研究內容 4
第四節 研究假設 4
第貳章 文獻探討 5
第一節 填補牙材的演化 5
第二節 複合式樹脂及聚合 6
第三節 單體釋出、唾液分解與健康危害 9
第四節 物化性及毒性 11
(一) TEGDMA 11
(二) UDMA 11
(三) Bis-GMA 11
(四)分解產物與毒性 11
第五節 傳統萃取唾液中釋出物方法 13
第六節 固相微萃取(Solid-Phase Microextraction) 14
(一) 直接固相微萃取 14
(二) 頂空固相微萃取 15
(三) 固相微萃取裝置 16
(四) 固相微萃取操作程序 16
(五) 影響固相微萃取效率的因素 16
(六) 分配係數 21
(七) 固相微萃取法於生物檢體的應用 22
第叁章 材料設備與方法 30
第一節 儀器設備與試藥 30
(一) 試藥 30
(二) 設備 31
第二節 儀器參數條件 34
(一) HPLC條件 34
(二) LC-MS條件 35
第三節 研究架構 36
(一) HPLC分析方法的建立 36
(二)直接固相微萃取方法建立 36
(三)分配係數 39
(四)分析品質管制 39
(五)真實樣本培養 40
第肆章 結果與討論 48
第一節 HPLC分析 48
第二節 固相微萃取方法建立 49
(一) 固相微萃取纖維的選擇 49
(二) 萃取時間 49
(三) 脫附溶劑 49
(四) 脫附時間探討 50
(五) 萃取溫度探討 50
(六) 攪拌速度對萃取的影響 51
(七) 樣本pH值探討 51
第三節 分配係數 52
第四節 分析品質管制 53
(一) 檢量線建立 53
(二) 方法偵測極限(MDL) 53
(三) 分析之精密度與準確度 53
第五節 真實樣本培養 54
(一) 市售複合式樹脂 54
(二) LC-MS-MS質譜 54
(三) 聚合時間探討 54
(四) 溫度探討 55
(五) pH值探討 55
第六節 討論 56
(一) 固相微萃取 56
(二) 複合式樹脂 59
第伍章 結論與建議 104
(一) 結論與建議 104
(二) 未來發展 105
參考文獻 106

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