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suchanghui木虫 (小有名气)
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求助药学实验过程翻译！写文章用，一定要专业！
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Hydrolysis The stock solutions of complexes 1–5 (2 mM) were prepared in methanol. Then 5 mL of the solution (10 mL for complex 2) was diluted to 200 mL with deionized water in a quartz cuvette and the UV-vis spectra was recorded by scanning from 200 to 500 nm at certain time intervals at 37 1C. The wavelengths corresponding to the LMCT band at ca. 330–336 nm were selected for the kinetic study. The time-dependent absorbance were fitted using Origin 8.0 (OriginLab Corporation, US) to give the first order rate constant k,and half reaction time t1/2was calculated by the formula as follows: A = Ce-kt + A0；t1/2 = ln2/k； where A is the absorbance, and A0 and C are constants. The species before and after hydrolysis for 1 h were analysed by LC-MS as described below. High performance liquid chromatography (HPLC) A Rheodyne sample injector, an Agilent Eclipse XDB-C18 reversed phase column (150×4.6 mm, 5um, USA) and an Agilent 1200 series HPLC system were used to analyse the hydrolytic mixtures of complexes 1–5. The mobile phases were water containing 0.1% TFA (A), and acetonitrile containing 0.1% TFA (B). The gradient was increased from 10% to 80% B for 20 min, with a flow rate of 1 mL min-1. DNA interaction Hoechst 33342 (20 uM) and CT DNA (200 uM) were dissolved in Tris-HCl buffer (pH = 7.2) and incubated at ambient temperature for 4 h. Then complex 1 or 3 (0–16 mM) was added to this solution to make the final concentrations from 0 to 160 uM. After incubating for 12 h, the resulting mixture was measured on an F-4500 fluorescence spectrophotometer (HITACHI) with excitation wavelength at 370 nm and emission spectra from 400 to 650 nm. A modified Stern–Volmer plot was employed to evaluate the affinity of complexes towards DNA. Different F0/F values recorded at 490 nm with different concentrations of complex 1 or 3 was fitted using Origin 8.0 (OriginLab Corporation, US) and Ksv was calculated by equations as follows: F0/F = 1 + Ksv[Q] where F0 and F are the fluorescence intensities of the Hoechst–CT DNA complex recorded before and after adding complex 1 or 3, respectively. [Q] is the concentration of complex 1 or 3. |
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