臺灣博碩士論文加值系統

本論文開發一新型之肌酸酐螢光生物檢測方法，能快速檢測檢品中肌酸酐之濃度且具備低成本等優勢。於高鹼性環境下，肌酸酐會與3,5-二硝基苯甲酸(3,5-Dinitrobenzoic acid )複合後，反應物會吸收波長400 nm紫外光並放出波長491 nm之螢光，並由發出螢光強度可推算出肌酸酐之濃度。因此本論文利用微型光譜儀結合三維平移台及雷射二極體建立一套螢光感測系統，並針對不同濃度之肌酸酐量測。其肌酸酐量測範圍分別為12.5

In this thesis, we have developed a novel fluorescent bioassay for creatinine which has advantages of fast detection and low cost. Creatinine will be reacting with 3,5-Dinitrobenzoic acid under the highly alkaline condition. The complexes have fluorescent characteristic that absorb excitation light at 400 nm and release fluorescence at 491 nm. By detecting the fluorescence, we can find out the concertation of creatinine. Thus we used laser diode (LD), spectrometer and 3-D translation stage to construct this fluorescent bio-system. The results show that this fluorescent bio-system have good linearity from 5 μmol/L to 300 μmol/L (R2 = 0.9934) and the limit of detection (LOD) is 5 μmol/L. The accuracy and precision of intra-day ranged from 81.82 to 99.75 % and 7.33 to 10.11 %, respectively. The accuracy and precision of inter-day ranged from 86.75 to 87.85% and 3.08 to 9.39 %, respectively. Also, compared to National Laboratory Animal Center and our bio-system, the mice serum test results show that the correlations (R) are 0.80.
In addition, we use the fluorescent bioassay for D-lactate that was previously developed by our labs. To construct a fluorescent bio-system for creatinine and D-lactate assaying. The results show that this fluorescent bio-system have good linearity from 5 μmol/L to 125 μmol/L (R2 = 0.998) and the limit of detection (LOD) is 5 μmol/L. The accuracy and precision of intra-day ranged from 96.22 to 108.01 % and 3.94 to 52.67 %, respectively. The accuracy and precision of inter-day ranged from 95.13 to 101.24 % and 7.65 to 23.86 %, respectively. Also, compared to high performance liquid chromatography (HPLC) and our bio-system, the mice serum test results show that the correlations (R) are 0.69. From this, this fluorescent bio-system provides a rapid, accurate, low cost and convenient for creatinine and D-lactate determination in biological sample.

摘要 II
ABSTRACT IV
誌 謝 VI
目錄 VII
圖目錄 XI
表目錄 XIII
第一章 緒論 1
1.1 肌酸酐之簡介 1
1.2 肌酸酐與D-乳酸之相關性 2
1.3 肌酸酐之生物感測器介紹 4
1.4 研究動機 10
1.5 研究流程 11
第二章 實驗設計 12
2.1 肌酸酐螢光生物感測系統之設計 12
2.1.1 肌酸酐螢光感測系統之搭設 12
2.1.2 肌酸酐螢光感測原理 14
2.1.3 肌酸酐標準品之調配 14
2.1.4 肌酸酐螢光檢測試劑之調配 15
2.2肌酸酐實驗藥品與儀器設備 15
2.2.1 實驗藥品 15
2.2.2 實驗儀器 15
2.3 肌酸酐螢光感測系統之標準品實驗分析 16
2.3.1 螢光與激發光譜分析 16
2.3.2 肌酸酐標準品之時序分析 16
2.3.3 不同濃度之3,5-二硝基苯甲酸與氫氧化鋰之實驗分析 17
2.3.4 肌酸酐標準品溶液之檢量線分析 17
2.4 肌酸酐螢光感測系統之大鼠血清實驗分析 18
2.4.1 大鼠血清之去蛋白方法 18
2.4.2 大鼠血清之肌酸酐時序分析 19
2.4.3 大鼠血清之肌酸酐檢量線分析 19
2.4.4 大鼠血清之肌酸酐的準確度(Accuracy)與精密度(Precision)試驗分析 19
2.4.5 大鼠血清之肌酸酐測定 20
第三章 肌酸酐與D-乳酸螢光生物感測系統 21
3.1 D-乳酸與肌酸酐螢光生物感測系統之設計 21
3.1.1 肌酸酐與D-乳酸螢光感測系統之搭設 21
3.1.2 D-乳酸螢光感測原理 22
3.1.3 甘胺酸-聯胺緩衝液(Glycine-Hydrazine buffer, G-H buffer)之調配 23
3.1.4 D-乳酸標準品之調配 23
3.2 D-乳酸實驗藥品與儀器設備 23
3.2.1 實驗藥品 23
3.2.2 實驗儀器 24
3.3 D-乳酸螢光感測系統之標準品實驗分析 24
3.3.1 螢光與激發光譜分析 24
3.3.2 D-乳酸標準品之時序分析 25
3.3.3 D-乳酸標準品溶液之檢量線分析 25
3.4 D-乳酸螢光感測系統之大鼠血清實驗分析 26
3.4.1 大鼠血清之去蛋白方法 26
3.4.2 大鼠血清之肌酸酐時序分析 26
3.4.3 大鼠血清之肌酸酐的準確度(Accuracy)與精密度(Precision)試驗分析 27
3.4.4 大鼠血清之D-乳酸測定 27
第四章 肌酸酐與D-乳酸生物感測系統之標準品檢測 28
4.1 肌酸酐標準品之確效性實驗分析 28
4.1.1 螢光與激發光譜分析 28
4.1.2肌酸酐之反應時序分析 29
4.1.3不同濃度之3,5-二硝基苯甲酸與氫氧化鋰之實驗分析 30
4.1.3 肌酸酐標準品溶液之檢量線分析 32
4.2 D-乳酸標準品之確效性實驗分析 35
4.2.1 螢光與激發光譜分析 35
4.2.2 D-乳酸標準品之反應時序分析 36
4.2.3 D-乳酸標準品之檢量線分析 37
第五章 大鼠血清之肌酸酐與D-乳酸檢測結果 39
5.1 大鼠血清檢品之檢測確效性分析–肌酸酐 39
5.1.1 大鼠血清檢品之時序分析 39
5.1.2 大鼠血清之檢量線實驗分析 40
5.1.3 大鼠血清之準確度和精密度實驗分析 41
5.2 大鼠血清檢品之確效性分析–D-乳酸 42
5.2.1 大鼠血清檢品之時序分析 42
5.2.2 大鼠血清之準確度和精密度實驗分析 43
5.3 大鼠血清之肌酸酐與D-乳酸測定分析 44
第六章 結論 46
第七章 未來展望 49
參考文獻 51
附錄 56

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