TWI628436B - 奈米點及用其偵測葡萄糖濃度的方法 - Google Patents
奈米點及用其偵測葡萄糖濃度的方法 Download PDFInfo
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- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 title claims abstract description 42
- 239000008103 glucose Substances 0.000 title claims abstract description 42
- 238000000034 method Methods 0.000 title claims abstract description 10
- 239000002096 quantum dot Substances 0.000 title description 2
- 239000002245 particle Substances 0.000 claims abstract description 25
- 108010015776 Glucose oxidase Proteins 0.000 claims abstract description 23
- 239000004366 Glucose oxidase Substances 0.000 claims abstract description 23
- 229940116332 glucose oxidase Drugs 0.000 claims abstract description 23
- 235000019420 glucose oxidase Nutrition 0.000 claims abstract description 23
- 239000002094 self assembled monolayer Substances 0.000 claims abstract description 19
- 239000013545 self-assembled monolayer Substances 0.000 claims abstract description 19
- -1 3-glycidoxypropyl group Chemical group 0.000 claims abstract description 8
- JGCGXJDWQQCNFF-UHFFFAOYSA-N niobium(2+);oxygen(2-) Chemical group [O-2].[Nb+2] JGCGXJDWQQCNFF-UHFFFAOYSA-N 0.000 claims abstract description 3
- 230000003647 oxidation Effects 0.000 claims abstract description 3
- 238000007254 oxidation reaction Methods 0.000 claims abstract description 3
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 20
- 230000008033 biological extinction Effects 0.000 claims description 9
- 240000007594 Oryza sativa Species 0.000 claims description 2
- 235000007164 Oryza sativa Nutrition 0.000 claims description 2
- 239000002105 nanoparticle Substances 0.000 claims description 2
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- 239000000243 solution Substances 0.000 description 20
- 230000000694 effects Effects 0.000 description 5
- UHYPYGJEEGLRJD-UHFFFAOYSA-N cadmium(2+);selenium(2-) Chemical class [Se-2].[Cd+2] UHYPYGJEEGLRJD-UHFFFAOYSA-N 0.000 description 4
- 229910000420 cerium oxide Inorganic materials 0.000 description 4
- DRVWBEJJZZTIGJ-UHFFFAOYSA-N cerium(3+);oxygen(2-) Chemical group [O-2].[O-2].[O-2].[Ce+3].[Ce+3] DRVWBEJJZZTIGJ-UHFFFAOYSA-N 0.000 description 4
- 238000006460 hydrolysis reaction Methods 0.000 description 4
- BMMGVYCKOGBVEV-UHFFFAOYSA-N oxo(oxoceriooxy)cerium Chemical compound [Ce]=O.O=[Ce]=O BMMGVYCKOGBVEV-UHFFFAOYSA-N 0.000 description 4
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- MFAWEYJGIGIYFH-UHFFFAOYSA-N 2-[4-(trimethoxymethyl)dodecoxymethyl]oxirane Chemical compound C(C1CO1)OCCCC(C(OC)(OC)OC)CCCCCCCC MFAWEYJGIGIYFH-UHFFFAOYSA-N 0.000 description 1
- 239000004593 Epoxy Substances 0.000 description 1
- ROZSPJBPUVWBHW-UHFFFAOYSA-N [Ru]=O Chemical group [Ru]=O ROZSPJBPUVWBHW-UHFFFAOYSA-N 0.000 description 1
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- 150000001412 amines Chemical class 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 238000007098 aminolysis reaction Methods 0.000 description 1
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- RUDFQVOCFDJEEF-UHFFFAOYSA-N oxygen(2-);yttrium(3+) Chemical group [O-2].[O-2].[O-2].[Y+3].[Y+3] RUDFQVOCFDJEEF-UHFFFAOYSA-N 0.000 description 1
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- 229910001925 ruthenium oxide Inorganic materials 0.000 description 1
- WOCIAKWEIIZHES-UHFFFAOYSA-N ruthenium(iv) oxide Chemical group O=[Ru]=O WOCIAKWEIIZHES-UHFFFAOYSA-N 0.000 description 1
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- BPSIOYPQMFLKFR-UHFFFAOYSA-N trimethoxy-[3-(oxiran-2-ylmethoxy)propyl]silane Chemical compound CO[Si](OC)(OC)CCCOCC1CO1 BPSIOYPQMFLKFR-UHFFFAOYSA-N 0.000 description 1
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Abstract
一種用以偵測葡萄糖濃度之奈米點，係包含：一氧化矽核；一自組裝單層，包含一3-環氧丙氧基丙基，該自組裝單層以一共價鍵結合該氧化矽核；及一葡萄糖氧化酶粒子，以一共軛鍵結合該自組裝單層之3-環氧丙氧基丙基。本發明另關於以該奈米點偵測葡萄糖濃度的方法。
Description
本發明係關於一種奈米點，特別係關於一種用以偵測葡萄糖濃度的奈米點，本發明另關於以該奈米點偵測葡萄糖濃度的方法。
量子點(quantum dot)為一種奈米級的半導體材料，由於其特別的光學性質，常被應用於作為生物偵測的螢光探針(fluorescent probe)。舉例而言，CdSe/ZnS量子點可以搭配葡萄糖氧化酶粒子的使用，藉由葡萄糖氧化酶粒子作用所產生的過氧化氫分子對該CdSe/ZnS量子點進行一螢光消光作用(fluorescence quenching)，即可以得知一待測溶液中的葡萄糖濃度。
然而，當該葡萄糖氧化酶粒子固定於如上述的CdSe/ZnS量子點時，容易造成該葡萄糖氧化酶粒子失去正常的酵素活性，因而影響該CdSe/ZnS量子點偵測葡萄糖濃度的靈敏度及精確度，因此，仍需要提供一種用以偵測葡萄糖的奈米點，進而可以靈敏地、精確地偵測該待測溶液中的葡萄糖濃度。
為解決上述問題，本發明提供一種奈米點，係可以靈敏地、精確地偵測一待測溶液中的葡萄糖濃度者。
本發明另提供一種以前述之奈米點偵測葡萄糖濃度的方法。
本發明之用以偵測葡萄糖濃度之奈米點，係包含：一氧化矽
核；一自組裝單層，包含一3-環氧丙氧基丙基，該自組裝單層以一共價鍵結合該氧化矽核；及一葡萄糖氧化酶粒子，以一共軛鍵結合該自組裝單層之3-環氧丙氧基丙基；據此，本發明之用以偵測葡萄糖濃度之奈米點係藉由該葡萄糖氧化酶粒子可以將該待測溶液中的葡萄糖氧化形成一過氧化氫分子，該過氧化氫分子續可以對該奈米點進行一螢光消光作用，因而可以藉由偵測該奈米點受該螢光消光作用所造成的螢光強度之變化來靈敏地、精準地偵測該待測溶液中的葡萄糖濃度，為本發明之功效。
基於相同的技術概念之下，本發明之偵測葡萄糖的方法，係包含：提供如前述之奈米點；提供一待測溶液，該待測溶液包含葡萄糖；使該待測溶液中的葡萄糖受該奈米點的葡萄糖氧化酶粒子作用形成一過氧化氫分子；使該過氧化氫分子對該奈米點進行一螢光消光作用；及偵測該奈米點受該螢光消光作用所造成的螢光強度之變化；據此，本發明之偵測葡萄糖的方法係藉由該奈米點的使用，可以靈敏地、精準地偵測該待測溶液中的葡萄糖濃度，為本發明之功效。
其中，係偵測該奈米點於497nm之波長下的螢光強度之變化；藉此可以靈敏地、精準地偵測該待測溶液中的葡萄糖濃度。
1‧‧‧奈米點
11‧‧‧氧化矽核
12‧‧‧自組裝單層
13‧‧‧葡萄糖氧化酶粒子
第1圖：本發明之一實施例的奈米點之示意圖。
第2a圖：本發明之一實施例的奈米點之共價鍵結合氧化矽之自組裝單層的原子力顯微鏡粒徑分布圖(AFM particle size distribution)。
第2b圖：本發明之一實施例的奈米點之共價鍵結合氧化矽之自組裝單層的穿透式顯微鏡分析結果。
第3a圖：本發明之一實施例的奈米點之表面增強拉曼散射光譜分析結果。
第3b圖：本發明之一實施例的奈米點之光致螢光光譜分析結果。
第4圖：本發明之一實施例的奈米點與葡萄糖反應後之光致螢光光譜分析結果。
為讓本發明之上述及其他目的、特徵及優點能更明顯易懂，下文特舉本發明之實施例，並配合所附圖式，作詳細說明如下：請參照第1圖所示，本發明之一實施例的奈米點1，大抵可以包含：一氧化矽核11、一自組裝單層12及一葡萄糖氧化酶粒子13。
詳而言之，該氧化矽核11及該自組裝單層12係可以藉由使一3-環氧丙氧基丙基三甲氧基矽烷(3-glycidoxypropyl trimethoxysilane)經一水解縮合反應(hydrolysis and condensation)所獲得，使該自組裝單層可以包含一3-環氧丙氧基丙基(3-glycidoxypropyl group)，且該自組裝單層12以一共價鍵(covalent bond)結合該氧化矽核11。於本實施例中，係於350℃、常壓之環境下，使該3-環氧丙氧基丙基三甲氧基矽烷進行該水解縮合反應90分鐘，續冷卻至室溫後，即可以獲得以該共價鍵結合該氧化矽核11之自組裝單層12，其pH值約為7。請參照第2a、2b圖所示，該共價鍵結合該氧化矽11之自組裝單層12的粒徑約為3.1±0.3nm。
續請參照第1圖所示，該葡萄糖氧化酶粒子13係可以經由一胺解反應(aminolysis reaction)，使該葡萄糖氧化酶粒子13之胺基(amino group)以一共軛鍵(conjugated bond)結合該自組裝單層12之3-環氧丙氧基丙基，如此可以使該葡萄糖氧化酶粒子13穩固地固定於該自組單層12；又，由於該葡萄糖氧化酶粒子13必須於pH值介於4~7.5之間時，才會具有正常的酵素活性，因而可以使該葡萄糖氧化酶粒子13仍維持有正常的酵素活性，亦無需要EDC/NHS活化劑促進該葡萄糖氧化酶粒子13與該自組裝單層12的鍵結，可以避免繁瑣的製備流程損耗該葡萄糖氧化酶粒子
13。於本實施例中，係於20~30℃之溫度下，將2mg之葡萄糖氧化酶粒子13溶解於10μL之磷酸緩衝溶液中，續混合590μL之以該共價鍵結合該氧化矽核11之自組裝單層12，持續震盪30分鐘後即可以獲得該奈米點1。
經由上述流程所獲得之奈米點1，其激發波長(excitation wavelength)為400nm，且於發射波長(emission wavelength)為497nm時具有最大螢光強度(fluorescent intensity)，因此即可以透過此一特性來偵測一待測溶液中的葡萄糖濃度，詳如下述。
本實施例之奈米點1在應用於偵測葡萄糖濃度時，係可以於該待測溶液中加入該奈米點1，使該待測溶液中的葡萄糖受該奈米點1之葡萄糖氧化酶粒子13的氧化作用，而形成一過氧化氫分子(H2O2)，該過氧化氫分子續可以對該奈米點1進行一螢光消光作用(quenching)，因而可以藉由偵測該奈米點1受該螢光消光作用所造成的螢光強度之變化來換算該待測溶液中的葡萄糖濃度。
為證實該奈米點1確實能夠應用於偵測葡萄糖之濃度，遂進行以下試驗：
(A)奈米點1的性質
本試驗係分析該奈米點1之表面增強拉曼散射光譜(surface-enhanced Raman scattering spectrum)，其結果如第2a圖所示，第A1組之共價結合該氧化矽核11之自組裝單層12分別於1268、1475及1489cm-1處具有波峰(環氧環振動，epoxide ring vibration)，第A2組之葡萄糖氧化酶粒子3則分別於1258、1672及1470cm-1處具有波峰，且分別代表第III個醯胺基(amide III)、第I個醯胺基(amideI)及CH2/CH3構型(CH2/CH3 deformation)，而二者經該胺解反應所形成之奈米點1的1268、1475及1489cm-1處的波峰則消失了(第A3組)。
又，請參照第2b圖所示，利用波長為400nm之激發光對該奈米點1進行光致螢光光譜(photoluminescence spectrum)分析，並記錄400~700nm之間的發射光譜，其結果顯示該奈米點1於497nm之波長下具有最大螢光強度。
(B)葡萄糖濃度之偵測
本試驗係分別混合不同濃度之葡萄糖(濃度介於8~800μM，各1μL)及該奈米點1，於室溫下震盪20分鐘後，進行光致螢光光譜分析，其結果如第3圖所示，隨著該待測溶液中的葡萄糖濃度增加，該奈米點1於497nm之最大螢光強度有下降之趨勢。另將該奈米點1於497nm之波長下的螢光強度之變化相對該待測溶液中的葡萄糖濃度繪製關係圖，當該待測溶液中的葡萄糖濃度介於88~400μM之間時，二者呈正相關(R2=0.99)。
綜上所述，本發明之用以偵測葡萄糖濃度之奈米點係藉由該葡萄糖氧化酶粒子可以將該待測溶液中的葡萄糖氧化形成該過氧化氫分子，該過氧化氫分子續可以對該奈米點進行該螢光消光作用，因而可以藉由偵測該奈米點受該螢光消光作用所造成的螢光強度之變化來靈敏地、精準地偵測該待測溶液中的葡萄糖濃度，為本發明之功效。
再者，本發明之偵測葡萄糖的方法係藉由該奈米點的使用，可以靈敏地、精準地偵測該待測溶液中的葡萄糖濃度，為本發明之功效。
雖然本發明已利用上述較佳實施例揭示，然其並非用以限定本發明，任何熟習此技藝者在不脫離本發明之精神和範圍之內，相對上述實施例進行各種更動與修改仍屬本發明所保護之技術範疇，因此本發明之保護範圍當視後附之申請專利範圍所界定者為準。
Claims (3)
- 一種用以偵測葡萄糖濃度之奈米點，係包含：一氧化矽核；一自組裝單層，包含一3-環氧丙氧基丙基，該自組裝單層以一共價鍵結合該氧化矽核；及一葡萄糖氧化酶粒子，以一共軛鍵結合該自組裝單層之3-環氧丙氧基丙基。
- 一種偵測葡萄糖濃度的方法，係包含：提供如申請專利範圍第1項所述之奈米點；提供一待測溶液，該待測溶液包含葡萄糖；使該待測溶液中的葡萄糖受該奈米點的葡萄糖氧化酶粒子作用形成一過氧化氫分子；使該過氧化氫分子對該奈米點進行一螢光消光作用；及偵測該奈米點受該螢光消光作用所造成的螢光強度之變化。
- 如申請專利範圍第2項所述之偵測葡萄糖濃度的方法，其中，係偵測該奈米點於497nm之波長下的螢光強度之變化。
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| US15/622,592 US20180327808A1 (en) | 2017-05-09 | 2017-06-14 | Nanodot and method for detecting glucose concentration thereof |
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