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邱哥新蟲 (初入文壇)
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[求助]
材料制備 文獻翻譯
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| Synthesis of meso-5,10,15,20-Tetrakis(4-N-methylpyridiniumyl)porphyrinato Manganese(III) Pentachloride (Mn−PyP). Mn−PyP was synthesized according to literature methods (Scheme S1, Supporting Information).20 Briefly, porphyrin tetratosylate (101 mg, 0.074 mmol) and manganese(II) chloride tetrahydrate (16 mg, 0.081 mmol) were added to 15 mL of methanol. The mixture was refluxed with stirring under N2 atm. UV−vis absorption spectroscopy was used to monitor the reaction, and it was stopped when the Soret band completely shifted from 422 to 463 nm. Subsequently, the tosylate anion was exchanged by chloride ion on 5 g anion-exchange resin (DOWEX 1 × 8−200) for 24 h with slow stirring. The solution was filtered, and the resin was washed with methanol. The methanol solution was concentrated to 1−2 mL, and the product was precipitated by the addition of Et2O. The precipitate was filtered, washed with Et2O, dried, and dissolved in 4 mL of water. Finally, the manganese salt was allowed to precipitate at 4 °C, and the aqueous phase was filtered and then evaporated. After drying under vacuum, Mn−PyP was obtained as a purple solid. The product was characterized by electrospray ionization mass spectrometry (ESI-MS) (Figure S1). The ESI-MS analysis provided a m/z value of 182.9, whereas the calculated m/z value for [C44H36MnN8]4+ was 182.8, which confirmed the identity and purity of the probe. Assay Procedures. The VEGF luminol CL assay was performed at an ambient temperature. Briefly, 980 μL of Tris− HCl solution (20 mM, pH 9.0) containing 50 nM VEGF165 aptamer 1 (or 80 nM VEGF165 aptamer 2), 10 nM Mn−PyP, 25 μM luminol, 2 mM H2O2, 0.05% Triton X-100, and VEGF of different concentrations were mixed. Then, 20 μL of H 2O2 stock solution (100 mM) was quickly added. The CL signal at 1 min was collected by the MCDR-A system. The photomultiplier tube (PMT) voltage was set at 500 V. The biological fluid analysis was conducted in the sample mixtures containing 0.1%, 1% human serum, or 1% human serum with albumin and IgG removed (Figures S9−S11). The procedures were the same as described above. |
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jiangguofeng
金蟲 (著名寫手)
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| 內(nèi)消旋5,10,15,20 -四（4 - N -甲基吡啶基）卟啉錳(III)五氯化物( Mn - PyP)的合成。Mn - PyP是根據(jù)文獻方法（圖S1,支持信息）合成的。20 簡單地說就是,將卟啉四對甲苯磺酸酯( 101毫克，0.074毫摩爾）和氯化錳四水合物( 16毫克, 0.081毫摩爾）加到15毫升甲醇中�；旌衔镉贜2氣壓下攪拌回流。用紫外-可見吸收光譜法監(jiān)控反應(yīng),當(dāng)索瑞氏光譜帶完全地由422移至463nm時停止反應(yīng)。隨后,緩慢攪拌的同時于５克陰離子交換樹脂（道威克斯1?8 - 200)上用氯離子對甲苯磺酸鹽陰離子進行交換24小時。過濾溶液,用甲醇洗滌樹脂。將甲醇溶液濃縮至1-２毫升，添加Et2O使產(chǎn)物析出。濾出沉淀物用Et2O洗滌、干燥再溶于４毫升水中。最后,于4℃使錳鹽沉淀,過濾水相然后蒸發(fā)。 真空干燥后得到紫色固體Mn-PyP。 產(chǎn)物通過電噴霧質(zhì)譜( ESI - MS)進行表征(圖S1)。電噴霧質(zhì)譜分析給出[C44H36MnN8]4+的m/Z值為182.9,而計算的m／Z值為182.8，這就證明了試樣的同一性和純度。 VEGF魯米諾CL含量測定于常溫中進行。簡言之,將含有50nM的VEGF165適配體1(或80nM的VEGF165適配體2）、10 nM的Mn-PyP、25μΜ的魯米諾、2mM的H2O2、0.05%的曲拉通X-100和不同濃度的VEGF的980μL的鹽酸溶液（20mM, pH值9.0)進行混合。然后, 將20μL的H 2O2儲備溶液( 100 mM)迅速地加進。通過MCDR - A系統(tǒng)收集于1分鐘的CL信號。光電倍增管(PMT)電壓設(shè)置為500 V。使用含有0.1%、1%人體血清或1%除去白蛋白和IgG的人體血清的混合物樣品進行生物流體分析（圖S9 - S11）。具體程序如上所述。 |
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