A kind of nanoprobe and its preparation method and application
A nanoprobe and organic matter technology, applied in the field of nanoprobes, can solve the problems of difficulty in accurately detecting albumin content, short excitation wavelength, background interference, etc., and achieve enhanced fluorescence intensity and photostability, low detection limit, and detection range. wide effect
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Embodiment 1
[0051] (1) Heptamethine dye is , according to the reaction formula Preparation of cyanine organic matter; the specific preparation steps are: the heptamethine dye (100mg, 0.14mmol) and 4-(amino)phenyl 2,2,6,2-terpyridine (100mg, 0.31mmol) in anhydrous N , stirred in N-dimethylformamide (DMF) (4mL) at 90°C for 24 hours, cooled to room temperature, and spin-dried to remove the solvent, using dichloromethane / methanol (25:1 by volume) as eluent, After separation by silica gel column chromatography, cyanine organic compounds were obtained. The structural characterization data of the cyanine organic matter is: 1H NMR (600MHz, CDCl 3 )δ8.32(d, J=14.1Hz, 2H), 8.09(d, J=8.5Hz, 2H), 7.85(dd, J=19.1, 8.2Hz, 4H), 7.56(t, J=7.6Hz, 2H), 7.43(s, 2H), 7.34(d, J=8.8Hz, 2H), 7.28(m, 9H), 7.08(d, J=7.5Hz, 1H), 6.77(s, 1H), 6.24( d,J=14.1Hz,1H),5.58(s,2H),5.24(s,1H),2.52(s,4H),2.00(s,9H),1.65(s,2H),1.19(s,2H ); The molecular weight that flight mass spectrometry records is 1023.5080, and th...
Embodiment 2
[0055] Characterize the sensitivity of the nanoprobe (PNC) that embodiment 1 makes: prepare the aqueous solution of the nanoprobe (PNC) (15 μ M) that 3mL embodiment 1 makes; Bovine serum albumin (BSA) is dissolved in deionized Prepare the sample solution to be tested (55mg / mL) with water, then add 0-1.0% bovine serum albumin in the nanoprobe aqueous solution at a volume concentration interval of 0.1%, measure the fluorescence intensity by a fluorescence spectrophotometer, and the nanoprobe The change of fluorescence spectrum with the concentration of BSA is as follows Figure 5 As shown, it shows that the fluorescence intensity of the nanoprobe solution increases with the concentration of BSA. The nanoprobe (PNC) that embodiment 1 makes is the fluorescence intensity when fluorescence wavelength is 610nm and the change relationship with the concentration of BSA is as follows Image 6 As shown, it can be seen that the two have a linear relationship, and the calculation shows th...
Embodiment 3
[0057] Characterize the specificity of the nanoprobe (PNC) prepared in Example 1: prepare the aqueous solution of 3mL nanoprobe (15 μ M), respectively starch, esterase, trypsin, transferrin, glutathione, spherical Protein, cholesterol, bovine serum albumin, human serum albumin biological sample were dissolved in deionized water respectively to prepare test sample solution (1.0mM), then each test solution of above-mentioned 5% was added in the nanoprobe solution, with Add the aqueous solution of the nanoprobe (15 μ M) of test solution as blank control experiment, adopt fluorescence spectrophotometer to measure fluorescence, the fluorescence intensity contrast situation of the PNC solution that contains different biological samples is as follows Figure 7 As shown, it can be seen that only two solutions of bovine serum albumin and human serum albumin interact with the nanoprobe to produce significant fluorescence, indicating that albumin can specifically bind to the nanoprobe to ...
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