| The formation of complexes consisting of natural medical effective components and metal ions, has stronger pharmacological activities, which has been proven by modern pharmacological studies. Orientin is a Chinese medicine which has effective components of total flavonoids monomer, belonging to the flavonoid glycosides. The orientin has anti-inflammatory, antiviral, anti-oxidation, anti-tumor effect in vivo and in vitro and has protective effect on ischemic myocardial injury. The study of orientin-metal complexes has not yet been reported. Orientin and zinc acetate forming complexes being the research object, the synthetic process conditions, were optimized and by modern advanced technology identification method for complex structure characterization, were studied the antioxidation, antitumor activity outside.1The synthesis of orientin-zinc complexesThe product rate as an index, the single factor was used to investigate and reaction pH, reaction temperature and reaction time, using the effect of orthogonal test was applied to optimize synthesis process conditions.2The structure characterization of orientin-zinc complexesBy using elemental analyzer, plasma emission spectrometer, ultraviolet spectrometer, infrared spectrometer, thermogravimetric analyzer and and nuclear magnetic resonance instrument, was researched the structural characterization of orientin zinc complexes.3The antioxidant activity of orientin-zinc complexesSelect superoxide anion radicals, hydroxyl radicals, DPPH free radical generation system for experimental system, examine the antioxidant activity of orientin and orientin-zinc complexes. With blank control group being added corresponding solvent, and sample group added different concentrations of orientin and orientin-zinc complexes, by using ultraviolet spectrometer test the absorbance degree of each group was measured.4The antitumor activity of orientin zinc complexesUsing CCK8method, after cell being cultured, represented, vaccinated, blank control group was added the same amount of culture, and solvent control group added culture containing DMSO and the DMSO concentration being0.2%, experimental group added different concentrations of orientin and orientin-zinc complexes, concentrations are5.00,10.00,20.00,40.00,80.00μmol·L-1, with respectively the reaction time24h,48h,72h, then add CCK8, using automatic fluorescence enzyme standard instrument to detect the absorbance degree respectively.The results showed that:1The synthesis process of orientin-zinc complexes:Single factor analysis showed that orientin-zinc complexes synthesis yield are affected by reaction temperature, reaction time and reaction system pH, Orthogonal test results indicated that the optimized synthesis process was under the condition of pH8.5in the system of65℃and being mixed8h. vacuum drying under60℃, washing precipitation, yellow solid product, the yield was abtained rate is72.64%.2The content of C, H and Zn of orientin-Zinc complexes results:the content of C, H, and Zn in the experimental value were41.65%,3.21%, and14.68%, the theoretical value43.71%,3.30%and16.91%, The experimental results and theoretical calculation values are consistent, the analysis results showed that orientin and zinc ions may be formed in2:3.Ultraviolet absorption spectra revealed that the two absorption peaks of orientin were250nm and350nm, complexes absorption peak270nm and400nm, it meant the redshift occurred, this showed that the orientin-Zinc complexes was formed. By infrared spectrum data, the four u(C=O) of orientin is1656cm "’, after forming complexes, the absorption peak moves to the low wave number to1629cm-1, showing that four u(C=O) may participate in the reaction. The (C-O-C) of orientin and orientin-Zinc complexes are respectively1193cm-1and1198cm-1. And the characteristic peak of glucose ring are respectively812cm-1,848cm-1and808cm-1,845cm-1, There was no obvious change. The (C=C) of orientin is1613cm-1, after forming complexes to1565cm-1, indicating that (C=C) was affected by the coordination, the conjugate effect increased. The hydroxy association characteristic peak of orientin is3385cm-1, after forming complexes increases to3422cm-1, which shows complexes influenced the association of hydroxy. The absorption peak of orientin-zinc complexes is500cm-1～600cm-1, which is υο-zn.Orientin-zinc complexes of thermogravimetric graph showed that the complexes not containing crystal water, have two stages of weightlessness. The first phase of orientin-zinc complexes weightlessness is120-150℃, with the weightlessness rate of5.68%, equivalent to the loss of4molecule complexing water (theory weightlessness rate is6.24%), the second phase of orientin-zinc complexes weightlessness is260-520℃, with the weightlessness rate of73.54%, equivalent to the loss of2molecule orientin (theory weightlessness rate is76.84%), Zinc is oxidized in the air, and the residue is ZnO.The1H NMR map of orientin and orientin-zinc complexes as shown in the experiment, orientin has δ13.16(C5-OH),δ10.81(C7-OH),510.04(C3-OH),δ9.06(C4-OH), when orientin-zinc complexes was formed,810.81(C7-OH) still appearing, which means the3’,4’and5hydroxyl proton coordinated with Zn2+.By the above analysis, the possible chemical structural formula of orientin zinc complexes can be speculated.3Orientin and orientin-zinc complexes have strong ability of·O2- and-OH and DPPH, and the quantity of a certain effect, with orientin-zinc complexes in each dose group removing more·O2-and·OH and·DPPH than equivalent dose orientin at1.0～8.0μg·ml-1, having statistical significance(P<0.05).4Experimental data showed that DMSO will have little impact on cell activity and proliferation, At the same time, the different concentration of orientin and orientin-zinc complexes to inhibition rate of EC-109and MCF-7increased with the concentration increase of them(P<0.05), At different time, the same concentration of orientin and orientin-zinc complexes to inhibition rate of EC-109and MCF-7increased with the increase of time(P<0.05), at different time, the same concentration of orientin to inhibition rate of EC-109and MCF-7is superior to the same concentration of orientin-zinc complexes(P<0.05).Conclusion:The best coordination reaction optimum process of complexes formed by orientin and zinc acetate is:anhydrous ethanol as solvent, with reaction pH is8,reaction time8h, reaction temperature65℃. Under such circumstances, the chemical formula of the orientin-zinc complexes maybe [Zn3(C21H15O11)2]-4H2O and chemical structural formula was obtained (shown in figure1). Orientin-zinc complexes is superior to orientin in the activity of antioxidation and antitumor. |
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