肌酸酐 (Creatinine) 是肌肉中肌酸代謝的產物，為診斷腎功能的重要因子，血液或尿液肌酸酐的濃度通常在臨床上是相當重要的生理指標。本研究乃是製備對肌酸酐具模印記憶的材料，同時，使此材料具有螢光可調節性，即可依肌酸酐被吸附量與螢光強度變化比例的關係進行對肌酸酐分子具專一性之螢光式檢測。芴 (Fluorene) 的聚合物聚芴，其導電性和電致發光已廣泛的被用於研究有機發光二極體之應用，本研究即想延伸其特性，以芴的衍生物為功能性單體，合成具螢光性與辨識性吸附之模版高分子。
首先先合成螢光功能性單體，以2-acetylfluorene為反應物，2-acetylfluorene由還原反應形成2-(α-hydroxyethyl)fluorene，再行脫水反應合成2-vinylfluorene (2-VF)，並以傅利葉轉換紅外線光譜儀 (Fourier transform infrared spectroscopy, FT-IR) 與氫核磁共振光譜 (1Hnuclear magnetic resonance, 1H NMR) 進行分子結構之鑑定，再以螢光光譜儀 (FL spectrophotometer) 進行3D螢光圖譜測定以決定2-VF之最佳激發波長與放射波長。
模版高分子之合成則是以肌酸酐為模版分子，加入功能性單體methylacrylic acid (MAA)、自行合成之螢光單體2-vinylfluorene (2-VF)、交聯劑ethylene glycol dimethacrylate (EGDMA) 以及起始劑azobisisobutyronitrile (AIBN) 一起進行熱聚合反應。此模版材料 (MIP) 在10 mg/dL肌酸酐溶液中，對肌酸酐的平均吸附量為4.26 ± 0.25 mg/g；而非模版材料 (NIP) 則為1.39 ± 0.14 mg/g，因此可計算其模印因子 (imprinting factor) 為3.08 ± 0.32，此乃是以HPLC進行肌酸酐成份分析之結果。若以螢光檢測，則螢光模版高分子吸附肌酸酐後的螢光變化率為13.12 ± 0.91 %，而非模版高分子的螢光變化率為5.08 ± 0.27 %，則其模印因子為2.58 ± 0.16；且在不同肌酸酐濃度下以MIP對肌酸酐的平均吸附量對應螢光變化率所作的校正圖，具有良好的線性關係，顯示此模版材料具有以螢光檢測方式檢測未知樣本中肌酸酐濃度的可行性。因此不論以液相層析或螢光分析方式，皆可達到良好的模印效果。單體與交聯劑的比例是製備螢光模版高分子重要的條件，且以模印因子做為比較良劣的指標。螢光模版高分子在選擇性吸附感測實驗中，在肌酸酐/肌酸雙成份溶液中的選擇率相當良好，達2.78 ± 0.13；而在螢光模版高分子的干擾性吸附測試，在小牛血清肌酸酐濃度為10 mg/dL時，其螢光變化率為9.72 ± 1.26 %，肌酸酐平均吸附量為2.42 ± 0.43 mg /g MIP，皆呈現良好的辨識效果。
綜合前述，於本研究中已確認以螢光單體製備螢光模版高分子材料的可行性，且在對肌酸酐吸附檢測上，不論在專一性、選擇性上皆有良好的效果，顯示此螢光模版高分子以螢光檢測方式進行肌酸酐吸附檢測的可用性已確立。

Creatinine is a metabolite from muscle. Its concentration in serum or urine is an important physiological index in clinics and is essential to the diagnosis of renal functions. In this research, we intend to prepare molecularly imprinted polymer (MIP) that could provide specific cavity for the binding of creatinine as well as the fluorescence quenching via the binding of creatinine. Polyfluorenes, with electro-conductivity and electroluminescence, have long been applied to the organic light-emitting diodes. Consequently, this study further extends fluorene’s properties, by the utilization of a fluorene derivative as the fluorescence monomer, for the preparation of creatinine imprinted polymer. In this way, the MIP thus prepared would be able to own the fluorescence property and also recognize creatinine via specifically binding with creatinine.
First of all, the fluorescent monomer, 2-vinylfluorene (2-VF) is synthesized. By reduction, 2-acetylfluorene forms 2-(α-hydroxyethyl)fluorene. 2-(α-Hydroxyethyl)fluorene further reacts to produce 2-VF by dehydration. The molecular structure could be identified by Fourier transform infrared spectroscopy (FTIR) and 1H nuclear magnetic resonance (1H NMR). Three-dimensional fluorescence spectrum is analyzed to determine the best excitation and emission wavelengths for 2-VF measurement.
In the following, 2-VF (fluorescence monomer), methylacrylic acid (MAA, functional monomer), ethylene glycol dimethacrylate (EGDMA, crosslinker), and azobisisobutyro- nitrile (AIBN, initiator) were mixed together in the presence of creatinine template. The binding capacity of the MIP and the NIP are 4.26 ± 0.25 and 1.39 ± 0.14 mg/g, respectively. The ratio, defined as the imprinting factor, is 3.08 ± 0.32.The proportions of the corresponding fluorescence intensity change of MIP and NIP upon the binding the creatinine are 13.12 ± 0.91% and 5.08 ± 0.27% respectively. Thus, the imprinting factor of fluorescence detection is calculated to be 2.58 ± 0.16. Besides, the binding capacity of MIP for creatinine under different creatinine concentration against proportion of fluorescence intensity change appears to be an excellently linear correlation for valid calibration. Both HPLC and fluorescence analysis confirm the imprinting effect of the MIP. Hence, the detection of the creatinine concentration is indeed feasible by the fluorescent MIP proposed in this work. The composition of prepolymerization solution for the preparation of MIP is also investigated. The molar ratio of 2-VF, MAA, and EGDMA is considered as an essential factor affecting the performance of MIP. The imprinting factor is used to judge and compare the performance of the MIPs. Additionally, the imprinting effect and the selectivity w.r.t. creatinine from the creatinine mixtures are also evaluated. Selectivity of MIP for creatinine from creatinine/creatine mixture is 2.78 ± 0.13. Creatinine was also spiked into bovine calf serum (BCS) for the interference investigation of serum creatinine analysis by the MIP. The proportion of fluorescence intensity change caused by the binding of creatinine is 9.72 ± 1.26 % and the average binding capacity is 2.42 ± 0.43 mg /g MIP.
Concluded from the above, it is confirmed that the fluorescent MIP thus prepared is feasible for the clinical detection of serum creatinine. Its specific binding ability towards creatinine is also studied and proved in this work.

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