以擠壓米糠/澱粉(1：8)作為碳源，從嗜鹽菌發酵生產出來的聚羥基烷酯經過FTIR及NMR分析後，確認為聚(羥基丁酯-羥基戊酯)共聚合體(Poly(3-hydroxybutyrate-co-3-hydroxyvalerate)，PHBV)，其中羥基戊酯(HV)單元佔了9.2%。利用刮膜的方式將嗜鹽菌生產的PHBV共聚合體(HmPHBV)及從化學藥品公司提供之聚羥基丁酯單聚合體(PHB)及聚(羥基丁酯-羥基戊酯)共聚合體(HV單元佔8%，PHBV8)製作不同四環素含藥量的薄膜，然後在磷酸鹽緩衝溶液(PBS)中進行藥物釋放實驗。藉由ATR-FTIR、EDS及SEM來觀察四環素在薄膜的分佈情形，結果發現薄膜上表面及截面具有比下表面多的四環素。TGA分析中包覆四環素的PHB及PHBV8薄膜的起始裂解溫度(Tonset)比純PHB及PHBV8的Tonset增加了約20oC，而嗜鹽菌生產的HmPHBV則約提高了10oC。包覆四環素藥物的PHB、PHBV8及HmPHBV薄膜在磷酸鹽緩衝溶液(PBS)進行釋放，所有的薄膜在20小時之內達到最大藥物累積釋放量。在藥物釋放部分中，包覆四環素的PHB、PHBV8及HmPHBV薄膜皆呈現二階段釋放模式。另外利用單乳化製作包覆四環素的PHB微粒，水相中分別使用了界面活性劑Tween80及水溶性高分子(WCS)，其作用皆為幫助油相(氯仿)形成分散的油滴。使用Tween80製造PHB微粒的優點為微粒中四環素的包覆率及裝載量皆比使用WCS所製造的PHB微粒高，而使用WCS所製造的PHB微粒的優點為微粒的產率較高，若混合使用Tween80及WCS製造PHB微粒的四環素包覆率及裝載量與單獨使用WCS一樣低。原因在於四環素雖然溶於氯仿，卻大部分會擴散至水中，因此在單乳化的實驗中，PHB微粒包覆四環素的產率、包覆率及裝載量皆很低。

In this study, poly(hydroxyalkanoate) was produced by Haloferax Mediterranei. The poly(hydroxyalkanoate) was analyized by FTIR and NMR. Poly(hydroxybutyrate-co-hydroxyvalerate), PHBV (HmPHBV), confirmed the structure of the poly(hydroxyalkanoate). The HV content of the HmPHBV was only 9.2 mol%. Poly(hydroxybutyrate), PHB, and poly(hydroxybutyrate-co-hydroxyvalerate), PHBV (PHBV8), were provided by Aldrich, and the HV content of PHBV8 was 8 mol%. PHB, PHBV8 and HmPHBV membranes which were contained tetracycline were made by scraping method. The distributions of tetracycline in the membranes were investigated by FTIR, SEM and EDS. Tetracycline existed on the top and in the cross-section of the membranes was more then it existed on the bottom of the membranes. In the TGA analysis, the maxium degradation temperature of  PHB and PHBV8 membranes containg tetracycline were increased 20oC then pure membranes. And the temperature of HmPHBV containg tetracycline membranes was increased about 10oC then pure HmPHBV membrane. The drug release experiment was monitored in a buffer solution (PBS) at 37oC. Abount 20 hours, drug release amounts of all membranes were almost the largest. Releasing models of PHB and PHBV8 membranes were first-order release model, and HmPHBV membranes was t1/2 release model.Single emulsion (O/W) was used to made PHB microparticles containing tetracycline. Surfactant (Tween80) and carboxymethyl chitosan (WCS) were prevented from oil drops aggregating in water phase. Structures of PHB microparticles were observed using SEM. The adventage of using Tween80 as emulsifier was that the encapsulation efficiency and loading of PHB microparticles were higher then that of the PHB micropartilces being made by using WCS as emulsifier. But the yield of PHB microparticles using WCS as emulsifier was higher that using Tween80. If Tween80 and WCS were togegher using as emulsifier, problems of the lower encapsulation efficiency and loading of PHB microparticles still could not be solved.

目    錄

中文摘要 I
Abstract	III
目    錄	V
圖  目  錄  IX
表  目  錄  XVI
符號表	XX
第一章 緒論	1
1-1生物可分解聚酯的應用	1
1-2藥物控制釋放技術	3
1-3研究目的	4
第二章 文獻回顧	6
2-1 PHA的結構	6
2-2 PHA的發展歷史	8
2-3 PHB及PHBV的生化合成過程	9
2-4 PHB及PHBV的發酵生產	9
2-5 PHB及PHBV從微生物分離的程序	12
2-6 PHB及PHBV的性質	13
2-7 PHA在藥物釋放的應用	15
2-8四環素介紹	17
2-9四環素在藥物釋放的應用	19
2-10 PHB及PHBV與四環素類相關文獻	28
2-11水溶性幾丁聚醣	30
2-12界面活面劑與乳化作用	32
2-12-1界面活性劑的定義	32
2-12-2非離子性界面活性劑	33
2-12-3 HLB值與界面活性劑的關係	36
2-12-4乳化的原理	36
2-12-5單乳化(油/水(O/W))	38
2-12-6雙乳化(水/油/水(W1/O/W2))	39
2-12-7微胞的形成	39
2-13藥物釋放類型及釋放動力原理	42
2-13-1藥物釋放之類型	42
2-13-2 Fick’s First Law觀念	45
2-13-3零階、一階及t-1/2的藥物釋放模型	46
2-13-4時間延遲(time lag)與突釋現象(burst effect)	49
2-13-5藥物釋放之動力原理	50
第三章 實驗	60
3-1實驗藥品	60
3-2實驗儀器	61
3-3實驗步驟	63
3-3-1檢量線的製作	63
3-3-2包覆四環素之PHB及PHBV薄膜製作	64
3-3-3包覆四環素的PHB及PHBV薄膜之藥物釋放	66
3-3-4單乳化(O/W)溶劑揮發法製作包覆四環素之PHB及PHBV微粒	67
3-4紫外光光譜分析(UV)	71
3-5傅利葉紅外線光譜儀分析(FTIR-ATR)	71
3-6場發射掃描式電子顯微鏡分析(SEM)	71
3-7 X光繞射分析儀分析(XRD)	71
3-8核磁共振光譜儀分析(NMR)	71
3-9微分掃描卡計分析(DSC)	72
3-10膠體滲透層析儀分析(GPC)	72
3-11熱重分析儀分析(TGA)	72
第四章 結果與討論	73
4-1嗜鹽菌生產的PHA結構鑑定	73
4-2熱轉移性質	77
4-3水溶性幾丁聚醣的結構鑑定	79
4-4檢量線的製作	80
4-5 PHB及PHBV包覆四環素薄膜的結構與性質測定	90
4-5-1 ATR-FTIR分析	90
4-5-2 結晶構造分析	96
4-5-3熱裂解行為	101
4-5-5薄膜形態	107
4-5-6 EDS分析	142
4-5-7系統一、二、三、四薄膜在釋放後內部藥物殘餘量的測量	144
4-6系統一、二、三、四薄膜的藥物釋放動力	146
4-7單乳化(油/水(O/W))方法製備PHB包覆四環素藥物微粒	164
4-7-1 PHB微粒的製程探討	164
4-7-2 PHB包覆四環素藥物微粒的SEM分析	170
第五章 結論	178
第六章 參考文獻	180
附錄	189


圖  目  錄
圖1-1藥物釋放的理論濃度範圍	3
圖2-1 PHA的結構	6
圖2-2 PHB的結構	7
圖2-3 PHBV的結構	7
圖2-4微生物生化合成PHB及酵素解聚合的過程【3】	10
圖2-5四環素(tetracycline base)的分子結構	18
圖2-6鹽酸四環素(tetracycline hydrochloride)的分子結構	18
圖2-7聚合物接枝與藥物鍵結的示意圖【43】	22
圖2-8水溶性幾丁聚醣的結構	31
圖2-9界面活性劑的分類	33
圖2-10 D-葡萄糖轉變成D-山梨糖醇的氫化反應	34
圖2-11 Span系列的結構	34
圖2-12 Tween系列的結構	35
圖2-13界面活性劑在水油界面的排列	38
圖2-14乳液中顆粒分散的型態	38
圖2-15表面張力-濃度曲線與界面活性劑之溶解狀態	40
圖 2-16微胞的形狀【77】	40
圖2-17 Critical packing parameters(CPP=V/al) of surfactant molecules and preferred aggregate structures for geometrical packing reasons(V:圓錐部分的體積、l:圓錐最大長度、a:圓錐最大半徑)【77】	41
圖2-18儲槽式(reservoir device)藥物釋放模式	42
圖2-19整體式(monolithic device)藥物釋放模型	43
圖2-20溶蝕式(erosion type)藥物釋放模型	43
圖2-21化學反應式(chemical reactions type)釋放模型	44
圖2-22膨潤式(swelling type)藥物釋放模型	44
圖2-23滲透式(osmosis type)藥物釋放模型	45
圖2-24零階、一階及t-1/2的藥物累積釋放量與時間關係圖	47
圖2-25零階、一階及t-1/2的藥物釋放速率與時間關係圖	47
圖2-26時間延遲及突釋現象的藥物累積釋放量與時間的關係	49
圖2-27儲槽式濃度梯度圖(a)Steady-state concentration模式	51
(b)Unsteady-state(pseudo-steady-state)模式	51
圖2-28整體式濃度梯度圖(a)dissolved形態(b)dispersed形態	54
圖4-1市售PHB及PHBV8與HmPHBV的ATR-FTIR圖	74
圖4-2嗜鹽菌生產的HmPHBV的1H-NMR圖	75
圖4-3嗜鹽菌生產的HmPHBV的13C-NMR圖	75
圖4-4 PHBV結構圖位置標示	76
圖4-5 純PHB的熱示差掃描圖	78
圖4-6純PHBV8的熱示差掃描圖	78
圖4-7純HmPHBV的熱示差掃描圖	79
圖4-8水溶性幾丁聚醣的ATR-FTIR圖	80
圖4-9四環素在磷酸鹽緩衝溶液(PBS)中不同時間(照光)的UV圖	81
圖4-10四環素在磷酸鹽緩衝溶液(PBS)中不同時間(不照光)的UV圖	82
圖4-11四環素在磷酸鹽緩衝溶液(PBS)中不同標準溶液的UV圖	83
圖4-12四環素在磷酸鹽緩衝溶液(PBS)中的檢量線圖	83
圖4-13四環素在水中不同標準溶液的UV圖	85
圖4-14四環素在水中的檢量線圖	85
圖4-15四環素在溶有PHB的氯仿中的UV圖	86
圖4-16四環素在溶有PHB的氯仿中的檢量線圖	87
圖4-17不同比例標準溶液求水/氯仿的分散係數K值	88
圖4-18 Tween80在水中的UV圖	89
圖4-19純四環素(粉末)ATR-FTIR圖	91
圖4-20系統一的PHB(含藥物)薄膜上表面ATR-FTIR分析	92
圖4-21系統一的PHB(含藥物)薄膜下表面ATR-FTIR分析	92
圖4-22系統二的PHB(含藥物)薄膜上表面ATR-FTIR分析	93
圖4-23系統二的PHB(含藥物)薄膜下表面ATR-FTIR分析	93
圖4-24系統三的PHBV8(含藥物)薄膜上表面ATR-FTIR分析	94
圖4-25系統三的PHBV8(含藥物)薄膜下表面ATR-FTIR分析	94
圖4-26系統四的HmPHBV(含藥物)薄膜上表面ATR-FTIR分析	95
圖4-27系統四的HmPHBV(含藥物)薄膜下表面ATR-FTIR分析	95
圖4-28系統一的PHB(含藥物)薄膜上表面的XRD圖	97
圖4-29系統一的PHB(含藥物)薄膜下表面的XRD圖	97
圖4-30系統二的PHB(含藥物)薄膜上表面的XRD圖	98
圖4-31系統二的PHB(含藥物)薄膜下表面的XRD圖	98
圖4-32系統三的PHBV8(含藥物)薄膜上表面的XRD圖	99
圖4-33系統三的PHBV8(含藥物)薄膜下表面的XRD圖	99
圖4-34系統四的HmPHBV(含藥物)薄膜上表面的XRD圖	100
圖4-35系統四的HmPHBV(含藥物)薄膜下表面的XRD圖	100
圖4-36系統一的PHB(含藥物)薄膜的熱重量損失圖	102
圖4-37系統二的PHB(含藥物)薄膜的熱重量損失圖	102
圖4-38系統三的PHBV8(含藥物)薄膜的熱重量損失圖	103
圖4-39系統四的HmPHBV (含藥物)薄膜的熱重量損失圖	103
圖4-40系統一的PHB(含藥物)薄膜的微分圖	104
圖4-41系統二的PHB(含藥物)薄膜的微分圖	104
圖4-42系統三的PHBV8(含藥物)薄膜的微分圖	105
圖4-43系統四的HmPHBV(含藥物)薄膜的微分圖	105
圖4-44純四環素粉末	107
圖4-45純PHB、PHBV8及HmPHBV薄膜上表面SEM圖	110
圖4-46純PHB、PHBV8及HmPHBV薄膜下表面SEM圖	111
圖4-47純PHB、PHBV8及HmPHBV薄膜截面SEM圖	112
圖4-48 PHB薄膜(含藥物)在藥物釋放前的上表面圖	113
圖4-49系統一的PHB(含藥物)薄膜在藥物釋放前的上表面圖	114
圖4-50系統二的PHB(含藥物)薄膜在藥物釋放前的上表面圖	115
圖4-51 PHB(含藥物)薄膜在藥物釋放前的下表面圖	117
圖4-52系統一的PHB(含藥物)薄膜在藥物釋放前的下表面圖	118
圖4-53系統二的PHB(含藥物)薄膜在藥物釋放前的下表面圖	119
圖4-54 PHB(含藥物)薄膜在藥物釋放前的截面圖	121
圖4-55系統一的PHB(含藥物)薄膜在藥物釋放前的截面圖	122
圖4-56系統二的PHB(含藥物)薄膜在藥物釋放前的截面圖	123
圖4-57系統一的PHB(含藥物)薄膜在藥物釋放後的上表面圖	124
圖4-58系統二的PHB(含藥物)薄膜在藥物釋放後的上表面圖	125
圖4-59系統一的PHB(含藥物)薄膜在藥物釋放後的下表面圖	126
圖4-60系統二的PHB(含藥物)薄膜在藥物釋放後的下表面圖	127
圖4-61系統一的PHB(含藥物)薄膜在藥物釋放後的截面圖	128
圖4-62系統二的PHB(含藥物)薄膜在藥物釋放後的截面圖	129
圖4-63系統三的PHBV8(含藥物)薄膜在藥物釋放前的上表面圖	130
圖4-64系統三的PHBV8(含藥物)薄膜在藥物釋放前的下表面圖	131
圖4-65系統三的PHBV8(含藥物)薄膜在藥物釋放前的截面圖	132
圖4-66系統三的PHBV8(含藥物)薄膜在藥物釋放後的上表面圖	133
圖4-67系統三的PHBV8(含藥物)薄膜在藥物釋放後的下表面圖	134
圖4-68系統三的PHBV8(含藥物)薄膜在藥物釋放後的截面圖	135
圖4-69系統四的HmPHBV(含藥物)薄膜在藥物釋放前的上表面圖	136
圖4-70系統四的HmPHBV(含藥物)薄膜在藥物釋放前的下表面圖	137
圖4-71系統四的HmPHBV(含藥物)薄膜在藥物釋放前的截面圖	138
圖4-72系統四的HmPHBV(含藥物)薄膜在藥物釋放後的上表面圖	139
圖4-73系統四的HmPHBV(含藥物)薄膜在藥物釋放後的下表面圖	140
圖4-74系統四的HmPHBV(含藥物)薄膜藥物釋放後的截面圖	141
圖4-75系統一的薄膜在不同時間的藥物累積釋放量	150
圖4-76系統二的薄膜在不同時間的藥物累積釋放量	151
圖4-77系統三的薄膜在不同時間的藥物累積釋放量	152
圖4-78系統四的薄膜在不同時間的藥物累積釋放量(Mt)	153
圖4-79系統一的薄膜在不同時間的藥物累積釋放百分比	154
圖4-80系統二的薄膜在不同時間的藥物累積釋放百分比	155
圖4-81系統三的薄膜在不同時間的藥物累積釋放百分比	156
圖4-82系統四的薄膜在不同時間的藥物累積釋放百分比	157
圖4-83系統一的PHB(含藥物)薄膜的藥物對數式釋放動力模式	160
圖4-84系統二的PHB(含藥物)薄膜的藥物對數式釋放動力模式	161
圖4-85系統三的PHBV8(含藥物)薄膜的藥物對數式釋放動力模式	161
圖4-86系統四的HmPHBV(含藥物)薄膜的藥物對數式釋放動力模式	162
圖4-87不同轉速(10000、14000、18000rpm)對乳化的影響	167
圖4-88不同溶劑揮發溫度(40、50oC)對乳化的影響	168
圖4-89不同Tween80量(0.0234、0.0468、0.1875g)對乳化的影響	169
圖4-90 Tween80系列PHB微粒(Tw005~Tw02)的結構形態	172
圖4-91 WCS系列PHB微粒(CS005~CS02)的結構形態	174
圖4-92 Tween80/WCS作為界面活性劑所獲得的PHB微粒的實驗結果	176
附錄圖-1 Tween80在水中的UV光譜圖	190
附錄圖-2 Tween80在水中的檢量線圖	190
附錄圖-3不同轉速、溶劑揮發溫度及時間所製成的PHB微粒	206

表  目  錄

表2-1各種不同側基的PHA	6
表2-2可生產PHB的微生物	7
表2-3 PHA發展簡述	8
表2-4可溶解PHB的溶劑溶解度參數比較表【27】	14
表2-5 Span系列的R基結構及HLB值	35
表2-6 Tween系列的R基結構及HLB值	35
表2-7界面活性劑的HLB值和水溶性的關係	37
表2-8 HLB值與界面活性劑用途之關係	37
表3-1 系統一的薄膜代號	65
表3-2 系統二的薄膜代號	65
表3-3系統三的薄膜代號	66
表3-4系統四的薄膜代號	66
表3-5 Tween80系列的樣品代號	68
表3-6 WCS系列的樣品代號	69
表3-7 Tween80與WCS系列的樣品代號	70
表4-1 NMR分析結果	76
表4-2 PHB及PHBV的分子量值	77
表4-3四環素在磷酸鹽緩衝溶液(PBS)中的不同標準溶液	82
表4-4四環素溶於水中的不同標準溶液	85
表4-5四環素在含有PHB的氯仿溶液中的標準溶液	86
表4-6不同比例標準溶液求水/氯仿的分散係數K值	88
表4-7系統一的PHB(含藥物)薄膜的起始裂解溫度( Tonset)及600oC時殘餘的焦碳含量(char yield)，升溫速率為20oC/min	106
表4-8系統二的PHB(含藥物)薄膜的起始裂解溫度( Tonset)及600oC時殘餘的焦碳含量(char yield)，升溫速率為20oC/min	106
表4-9系統三的PHBV8(含藥物)薄膜的起始裂解溫度( Tonset)及600oC時殘餘的焦碳含量(char yield)，升溫速率為20oC/min	106
表4-10系統四的HmPHBV(含藥物)薄膜的起始裂解溫度( Tonset)及600oC時殘餘的焦碳含量(char yield)，升溫速率為20oC/min	107
表4-11薄膜中的C原子個數比	143
表4-12薄膜中的O原子個數比	143
表4-13薄膜中的N原子個數比	143
表4-14系統一薄膜在釋放後內部藥物殘餘量的測量	144
表4-15系統二薄膜在釋放後內部藥物殘餘量的測量	144
表4-16系統三薄膜在釋放後內部藥物殘餘量的測量	145
表4-17系統四薄膜在釋放後內部藥物殘餘量的測量	145
表4-18系統一、二、三、四不同釋放百分比	149
表4-20系統一的兩段釋放動力模式n及R值	162
表4-21系統二的兩段釋放動力模式n及R值	163
表4-22系統三的兩段釋放動力模式n及R值	163
表4-23系統四的兩段釋放動力模式n及R值	163
表4-24文獻中水相及油相比例的比較表	165
表4-25轉速、溶劑揮發溫度及時間的影響	166
表4-26 Tween80作為界面活性劑所獲得的PHB微粒的實驗結果	173
表4-27 WCS作為界面活性劑所獲得的PHB微粒的實驗結果	175
表4-28 Tween80及WCS系列微球的實驗結果	177
附錄表-1 Tween80在水中的不同標準溶液	189
附錄表-2藥物累積釋放量的計算式	191
附錄表-3系統一實驗紀錄	195
附錄表-4系統二實驗紀錄	195
附錄表-5系統三實驗紀錄	195
附錄表-6系統四實驗紀錄	195
附錄表-7系統一(HB1及HB2)實驗記錄	196
附錄表-8系統一(HB3及HB4)實驗記錄	197
附錄表-9系統二(HB01及HB02)實驗記錄	198
附錄表-10系統二(HB03及HB04)實驗記錄	199
附錄表-11系統三(HV01及HV02)實驗記錄	200
附錄表-12系統三(HV03及HV04)實驗記錄	201
附錄表-13系統四(Hm01及Hm02)實驗記錄	202
附錄表-14系統四(Hm03及Hm04)實驗記錄	203
附錄表-15四環素在水相損失的重量(Tween80系列)	206
附錄表-16微粒中四環素的重量(Tween80系列)	207
附錄表-17四環素在水相損失的重量(WCS系列)	207
附錄表-18微粒中四環素的重量(WCS系列)	207
附錄表-19四環素在水相損失的重量(Tween80及WCS系列)	208
附錄表-20微粒中四環素的重量(Tween80及WCS系列)	208

參考文獻

[1] Wendy Amass, Allan Amass, Brian Tighe. A review of biodegradable polymers: Uses, current developments in the synthesis and characterization of biodegradable polyesters, blends of biodegradable polymers and recent advances in biodegradation studies. Polymer International 1998;47:89-144.
[2] Gerhart Braunegg, Gilles Lefebvre, Klaus F. Genser. Polyhydroxyal-
kanoates, biopolyesters from renewable resources: Physiological and engineering aspects (Review ariticle). Journal of Biotechnology 1998;65:127-161.
[3] Colin.W.Pouton, Saghir Akhtar. Biosynthetic polyhydroxyalkanoates and their potential in drug. Advanced Drug Delivery Reviews 1996;18: 133-162.
[4] Dunja-Manal Abou-Zeid, Rolf-Joachim Müller, Wolf-Dieter Deck-wer. Degradation of natural and synthetic polyesters under anaerobic conditions. Journal of Biotechnology 2001;86:113-126.
[5] Yong-Bin Yan, Qiong Wu, Ri-Qing Zhang. Dynamic accumulation and degradation of poly(3-hydroxyalkanoate)s in living cells of Azotobacter vinelandii UWD characterized by 13C NMR. FEMS Microbiology Letters 2000;193:269-273.
[6] Yoshie N., Nakasato K., Fujiwara M., Kasuya K., Abe H., Doi Y., Inoue Y.. Effect of low molecular weight additives on enzymatic degradation of poly(3-hydroxybutyrate). Polymer 2000;41:3227-3234.
[7] Seymour R.A., Heasman P.A.. Pharmacological control of periodon-
tal disease. II Antimicrobial agents (Review paper). Journal of Dentistry 1995;23:5-14.
[8] 梁文權.生物藥劑學與藥物動力學(第二版). 人民衛生出版社 
2000.
[9] Richard A. Gross, Christopher DeMello, Robert W. Lenz, Helmut Brandl, R. Clinton Fuller. Biosynthesis  and characterization of poly
(β-hydroxyalkanoates) produced by Pseudomonas oleovorans. Macromolecules 1989;22:1108-1115.
[10] Manfred Zinn, Bernard Witholt, Thomas Egli. Occurrence synthesis and application of bacterial polyhydroxyalkanoate. Advanced Drug    Delivery Reviews 2001;53:5-21.
[11] Byrom D.. Polymer synthesis by microorganisms: Technology and economics. Tibtech 1987;5:247-250.
[12] Griffin G. J. L.. Chemistry and technology of biodegradable polym-ers. London: Chapman & Hall 1994;48-89.
[13] Arthur C. Eschenlauer, Sandra K. Stoup, Friedrich Srienc, David A. Somers. Production of heteropolymeric polyhydroxyalkanoate in Eschericha coli from a single carbon source. International Journal of Biological Macromolecules 1996;19:121-130.
[14] Klaassien J. Ganzeveld, Annechien van Hagen, Martin H. van Agteren, Wim de Koning, Anton J.M. Schoot Uiterkamp. Upgrading of organic waste: production of the copolymer poly-3-hydroxy-butyrate-co-valerate by Ralstonia eutrophus with organic waste as sole carbon source. Journal of Cleaner Production 1999;7:413–419.
[15] Tatiana Volova , Ekaterina Shishatskaya, Viktor Sevastianov, Sergei
Efremov, Olga Mogilnaya. Results of biomedical investigations of PHB and PHB/PHV fibers. Biochemical Engineering Journal 2003;16: 125–133.
[16] Sudesh K., Abe H., Doi Y.. Synthesis, structure and properties of polyhydroxyalkanoates: Biological polyesters. Prog. Polym. Science. 2000;25:1503-1555.
[17] Tsuneo Yamane. Yield of poly-D(-)-3-hydroxybutyrate from various carbon sources: A theoretical study. Biotechnology and Bioengin-eering 1993;41:165-170.
[18] Tsuneo Yamane, Masaaki Fukunaga, Yong Woo Lee. Increased PHB productivity by high-cell-density fed-batch culture of Alcalig-enes latus, a growth-associated  PHB producer. Biotechnology and Bioengineering 1995;50:197-202.
[19] Wendlandt K.-D., Jechorek M., Helm J., Stottmeister U.. Production of PHB with a high molecular mass from methane. Polymer Degradation and Stability 1997;59:191-194.
[20] Cho Kyung-Suk, Ryu Hee Wook, Park Chang-Ho, Goodrich Philip R.. Utilization of Swine Wastewater as a Feedstock for the Production of Polyhydroxyalkanoates by Azotobacter vinelandii UWD. Journal of Bioscience and Bioengineering 2001;91:129-133.
[21] Hahn Sei Kwang, Chang Yong Keun, Kim Beom Soo, Chang Ho Nam. Communication to the editor optimization of microbial poly(3-hydroxybutyrate) recovery using dispersions of sodium hypochlorite solution and chloroform. Biotechnology and Bioengineering 1994;44:256-261.
[22] Chen Yinguang, Yang Haizhen, Zhou Qi, Chen Jian, Gu Guowei. Cleaner recovery of poly(3-hydroxybutyric acid) synthesized in Alcaligenes eutrophus. Process Biochemistry 2001;36:501-506.
[23] Chen Yinguang, Chen Jian, Yu Cairong, Du Guocheng, Lun Shiyi. Recovery of poly-3-hydroxybutyrate from Alcaligenes Eutrophus by surfactant-chelate aqueous system. Process Biochemistry 1999;34:153-157.
[24] Chen Yinguang, Xu Qing, Yang Haizhen, Gu Guowei. Effects of cell fermentation time and biomass drying strategies on the recovery of poly-3-hydroxyalkanoates from Alcaligenes Eutrophus using a surfactant-chelate aqueous system. Process Biochemistry 2001;36:773-779.
[25] Holmes P. A., Lim G. B., US. 4910145 (1990).
[26] Terada Mikio, Marchessault R. H.. Determination of solubility para-meters for poly(3-hydroxyalkanoates). International Journal of Biological Macromolecules 1999;25:207-215.
[27] 林達鎔.溶劑與高分子聚合物溶解理論.化工技術2003;第11卷第4期:236-244.
[28] Valentin Henry E., Berger Pierre A., Gruys Kenneth J., Maria Fil-omena de Andrade Rodrigues, Alexander Steinbüchel, Minhtein Tran, Jawed Asrar. Biosynthesis and characterization of poly(3-hydroxy-4-pentenoic acid). Macromolecules 1999;32:7389-7395.
[29] Zhang Qiqing, Wang Chun. Polyhydroxybutyrate produced from cheap resources. I. crystallization and melting behavior. Journal of Applied Polymer Science 1994;54:515-518.
[30] Savenkova L., Gercberg Z., Bibers I., Kalnin M.. Effect of 3hy-droxy valerate content on some physical and mechanical properties  of polyhroxyalkanoates produced by Azotobacter chroococcum. Process Biochemistry 2000;36:445-450.
[31] Li Si-Dong, He Ji-Dong, Yu Peter H., Cheung Man Ken. Thermal degradation of poly(3-hydroxybutyrate) and poly(3-hydroxy-butyrate-co-3-hydroxyvalerate) as studied by TG, TG–FTIR, and Py–GC/MS.  Journal of Applied Polymer Science 2002;89:1530-1536.
[32] Chen Cheng, Fei Bin, Peng Shuwen, Zhuang Yugang, Dong Lisong, Feng Zhiliu. Nonisothermal crystallization and melting behavior of poly(3-hydroxybutyrate) and maleated poly(3-hydroxybutyrate).  European Polymer Journal 2002;38:1663-1670.
[33] Wang Zhioxiong, Itoh Yoshiaki, Hosaka Yoshifumi, Kobayashi Ich-iro, Nakano Yoshihisa, Maeda Isamu, Umeda Fusako, Yamakawa Junji, Kawase Masaya, Yagi Kiyohito. Novel transdermal drug delivery system with polyhydroxyalkanoate and starburst polyamidoamine dendrimer. Journal of  Bioscience and Bioengin-eering 2003;95:541-543.
[34] Wang Zhixiong, Itoh Yoshiaki, Hosaka Yoshifumi, Kobayashi Ichiro, Nakano Yoshihisa, Maeda Isamu, Umeda Fusako, Yamakami Junji, Nishimine Mari, Suenobu Tomoyoshi, Fukuzumi Shunichi, Kawase Masaya, Yagi Kiyohito. Mechanism of enhancement effect of dendrimer on transdermal drug permeation through polyhydroxyalk-anoate matrix. Journal of Bioscience and Bioengineering 2003;96:537-540.
[35] Yagmurlu M. Firat, Korkusuz Feza, Gürsel Ihsan, Korkusuz Petek, Örs Ülken, Hasirci Vasif. Sulbactam-cefoperazone polyhydroxy-butyrate-cohydroxyvalerate(PHBV) local antibiotic delivery system: In vivo effectiveness and biocompatibility in the treatment of implant-related experimental osteomyelitis. John Wiley & Sons Inc. 1999;494-503.
[36] Teresa Eligio, Jacques Rieumont, Rubén Sánchez. Characterization of chemically modified poly(3-hydroxy-alkanoates) and their performance as matrix for hormone release. Die Angewandte Makromolekulare Chemie 1999;270:69-75.
[37] Martin M.A., Miguens F.C., Rieumont J., Sanchez R.. Tailoring of the external and internal morphology of poly-3-hydroxy butyrate microparticles. Colloids and Surfaces B: Biointerfaces 2000;17:111-116. 
[38] Rege Pankaj R., Garmise Robert J., Block Lawrence H.. Spray-dried 
chitinosans Part II: In vitro drug release from tablets made from spray-dried chitinosans. International Journal of Pharmaceutics 2003;252:53–59.
[39] Santhosh Kumar T.R., Bai Mary Vasantha, Krishnan Lissy K.. A freeze-dried fibrin disc as a biodegradable drug release matrix. Biologicals 2004;32:49–55.
[40] Peter B. Petratos, Jie Chen, Diane Felsen, Dix P. Poppas. Local Pharmaceutical Release from a New Hydrogel Implant. Journal of Surgical Research 2002;103:55–60.
[41] Yang Libo, Eshraghi Jamshid, Fassihi Reza. A new intragastric delivery system for the treatment of Helicobacter pylori associated gastric ulcer: In vitro evaluation. Journal of Controlled Release 1999;57:215–222.
[42] Domingues Z.R., Cortés M.E., Gomes T.A., Diniz H.F., Freitas C.S., Gomes J.B., Faria A.M.C., Sinisterra R.D.. Bioactive glass as a drug delivery system of tetracycline and tetracycline associated with β-cyclodextrin. Biomaterials 2004;25:327–333.
[43] Murugan R., Ramakrishna S.. Coupling of therapeutic molecules onto surface modified coralline hydroxyapatite. Biomaterials 2004;25:3073–3080.
[44] Kim Hae-Won, Knowles Jonathan C., Kim Hyoun-Ee. Hydroxy-apatite/poly(ε-caprolactone) composite coatings on hydroxyapatite porous bone scaffold for drug delivery. Biomaterials 2004;25:1279–1287.
[45] Pataro A.L., Franco C.F., Santos V.R., Cortés M.E., Sinisterra R.D.. 
Surface effects and desorption of tetracycline supramolecular complex on bovine dentine. Biomaterials 2003;24:1075–1080.
[46] Lin D.M., Kalachandra S., Valiyaparambil J., Offenbacher S.. A polymeric device for delivery of anti-microbial and anti-fungal drugs in the oral environment: Effect of temperature and medium on the rate of drug release. Dental Materials 2003;19:589–596.
[47] Lev E. Bromberg, Debra K. Buxton, Phillip M. Friden. Novel peri-odontal drug delivery system for treatment of periodontitis.   Journal of Controlled Release 2001;71:251–259.
[48] David S. Jones, A. David Woolfson, Andrew F. Brown, Wilson A. Coulter, Cathy McClelland, Christopher R. Irwin. Design, charac-terisation and preliminary clinical evaluation of a novel mucoadhesive topical formulation containing tetracycline for the treatment of periodontal disease. Journal of Controlled Release 2000;67:357–368.
[49] Esposito E., Carotta V., Scabbia A., Trombelli L., D’ntona P.  Mene-gatti E., Nastruzzi C.. Comparative analysis of tetracycline-containing dental gels: Poloxamer- and monoglyceride-based formulations. International Journal of Pharmaceutics 1996;142:9-23.
[50] Kelly H.M., Deasy P.B., Ziaka E., Claffey N.. Formulation and pre-liminary in vivo dog studies of a novel drug delivery system for the treatment of periodontitis. International Journal of Pharmaceutics 2004;274:167–183.
[51] Roskos K.V., Fritzinger B.K., Rao S.S., Armitage G.C., Heller J.. Development of a drug delievery system for the treatment of periodonatal disease based on bioerodible poly(ortho esters). Biomaterials 1995;16:313-317.
[52] Schwach-Abdellaoui K., Monti A., Barr J., Heller J., Gurny R.. Optimization of a novel bioerodible device based on auto-catalyzed poly(ortho esters) for controlled delivery of tetracycline to periodontal pocket. Biomaterials 2001;22:1659-1666. 
[53] Park Yoon Jeong, Nam Kyung Hee, Ha Soo Jeong, Pai Chul Min, Chung Chong Pyoung , Lee Seung Jin. Porous poly(L-lactide) membranes for guided tissue regeneration and controlled drug delivery: Membrane fabrication and characterization. Journal of Controlled Release 1997;43:151–160.
[54] Sendil Dilek, Gürsel Ihsan, Wise Donald L., Hasırcı Vasif. Anti-biotic release from biodegradable PHBV microparticles.  Journal of Controlled release 1999;59:207-217.
[55] Sendil Dilek, Gürsela Ihsan, Hasirci Vasif. Preparation of PHBV foams and investigation of their potential for drug release. Turk J Med. Sci. 2000;30:9-14. 
[56] Zhao Zhi-Ping, Wang Zhi, Wang Shi-Chang. Formation, charged characteristic and BSA adsorption behavior of carboxymethyl chitosan/PES composite MF membrane. Journal of Membrane Science 2003;217:151-158.
[57] Chen Lingyun, Du Yumin, Zeng Xiaoqing. Relationships between the molecular structure and moisture-absorption and moisture-retention abilities of  carboxymethyl chitosan II. Effect of degree deacetylation and carboxymethylation. Carbohydrate Research 2003;338:333-340.
[58] Liu Xiao Fei, Guan Yun Lin, Yang Dong Zhi, Li Zhi, Yao Kang De. Antibacterial action of chitosan and carboxymethylated chitosan.  Journal of Applied Polymer Science 2001;79:1324-1335.
[59] Chen Xi-Guang, Park Hyun-Jin. Chemical characteristics of O-car-boxymethyl chitosans related to the preparation conditions. Carbohydrate Polymers 2003;53:355-359.
[60] Inez M. van der Lubben, J. Coos Verhoef, Gerrit Borchard, Hans E. Junginger. Chitosan and its derivatives in  mucosal drug and vaccine delivery. European Journal of Pharmaceutical Sciences 2001;14:201-207.
[61] Chen Sung-Ching, Wu Yung-Chin, Mi Fwu-Long, Lin Yu-Hsin, Yu Lin-Chien, Sung Hsing-Wen. A novel pH-sensitive hydrogel com-posed of N,O-carboxymethyl chitosan and alginate cross-linked by genipin for protein drug delivery. Journal of Controlled Release 2004;96:285-300.
[62] Chen Lingyun, Tian Zhigang, Du Yumin. Synthesis and pH sensi-tivity of carboxymethyl chitosan-based polyampholyte hydrogels for protein carrier matrices. Biomaterials 2004;25:3725-3732.
[63] 趙承琛.界面科學基礎.復文書局1993;74-118.
[64] 陳玫樺,鄭建新.界面活性劑在畜用疫苗佐劑之應用.化工技術 
1997;第五卷第十一期:144-152.
[65] 林江珍,顏孝欽,黃世吉.非離子型界面活性劑應用於精油乳化之原理.化工技術1998;第六卷第四期:164-174.
[66] 官常慶.化妝品用界面活性劑.化工技術1997;第五卷第十一期:118-123.
[67]洪耀釧,孫逸民,莊尊仁,王惠民,王豐益.有機化學第11版.歐亞書局有限公司&學銘圖書有限公司.2004;492-493.
[68] Palla Byron J., Shah Dinesh O.. Stabilization of high ionic strength slurries using the synergistic effects of a mixed surfactant system.  Journal of Colloid and Interface Science 2000;223:102–111.
[69] 莊晟榜,阮若屈.Tween系列界面活性劑對微生物降解碳氫化合物之研究.第七屆生化工程研討會論文集 2002.
[70] 李嘯風,陳志榮,李浩然,劉迪霞,韓世鈞. Span80-Tween-菜油-水乳化體系中最佳HLB值與乳化劑總用量的關係.CJI 2000;第二卷第四期.
[71] Md. Emdadul Haque, Akhil Ranjan Das, Satya Priya Moulik. Mixed Micelles of Sodium Deoxycholate and Polyoxyethylene Sorbitan Monooleate (Tween 80). Journal of Colloid and Interface Science 1999;217:1–7.
[72] Anna Hillgren, Jan Lindgren, Maggie Aldén. Protection mechanism 
of Tween 80 during freeze–thawing of a model protein, LDH. International Journal of Pharmaceutics 2002;237:57–69.
[73] Philip Sherman. Emulsion Science. London and New York: Acad-emic Press Inc. 1968.
[74] Rosen J. Milton. Surfactants and interfacial phenomena. New York: John Wiley & Sons:224-250.
[75] Rosca Iosif Daniel, Watari Fumio, Uo Motohiro. Microparticle for-mation and its mechanism in single and double emulsion solvent evaporation. Journal of Controlled Release 2004;99:271–280.
[76] 沈宗禮.制放技術與微粒包覆.高立圖書有限公司1980;127-142.
[77] Jőnsson Bo, Lindman Bjőrn, Holmberg Krister, Kronberg Bengt. Surfactant and polymer in aqueous solution. New York: John Wiley & Sons 1998:33-37, 82-85.
[78] 曹恒光,連大成.淺談微乳液.物理雙月刊 2001;第二十三卷四期
:488-493.
[79] Winston Ho W.S., Sirkar Kamalesh K.. Membrane Handbook 2. New York: Van Nostrand Reinhold 1992:915-935.
[80] Fan L.T., Singh S.K. Controlled Release: A Quantitative Treatment. Germany:Springer Verlag 1989:0-166.
[81] Culler E.L.. Diffusion: Mass Transfer in Fluid Systems Second Edi-tion. America: Cambridge University Press 1997:467-478.
[82]顏曉楓.利用pH-stat控制策略進行高密度培養Haloferax mediterranei生產poly-β-hydroxybutyrate之研究,大同大學生物工程研究所碩士論文:中華民國九十年七月.
[83] 行政院國家科學委員會補助專題研究計劃成果報告,生物可分解塑膠PHA的結構分析,摻合及應用(I),計劃編號:NSC-90-2621-Z-  
032-002.
[84] Norma Galego, Chavati Rozsa, Rubén Sánchez, Juan Fung, Analıá Vázquez, Julio Santo Tomás. Characterization and application of poly(b-hydroxyalkanoates) family as composite biomaterials. Poly-mer Testing 2000;19:485–492.
[85] Conway B.R., Eyles J.E., Alpar H.O.. A comparative study on the immune responses to antigens in PLA and PHB microspheres. Journal of Controlled Release 1997;49:1-9.
[86] Nicole Nihant, Chantal Schugens, Christian Grandfils, Robert Jero-me, Philippe Teyssie. Polylactide microparticles prepared by double emulsion-evaporation II. effect of the poly(lactide-co-glycolide) composition on the stability of the primary and secondary emulsions. Journal of colloid and interface science 1995;173:55-65.
[87] Karin Frauke Pistel, Armin Breitenbach, Regina Zange-Volland, Th-omas Kissel. Brush-like branched biodegradable polyester, part III Protein release release from microspheres of poly(vinyl alcohol)-graft-poly(D,L-lactic-co-glycolic acid). Journal of Controlled Release 2001;73:7-20.