RESUMEN
Objective: To analyze the dynamic changes of components in the enzymolysis process of raw products of Raphani Semen. Method: HPLC was employed to analysis of characteristic spectra of Raphani Semen at different enzymolysis time with mobile phase of acetonitrile-0.1% phosphoric acid aqueous solution for gradient elution and detection wavelength at 225 nm.The characteristic peaks were calibrated, meanwhile, the UV spectra of characterstic peaks were extracted, and the difference between UV spectra and the changes of peak areas were compared, and the dynamic changes of characterstic components in Raphani Semen were analyzed. Result: Eleven characteristic peaks were marked from the characteric spectra of raw products of Raphani Semen at different enzymolysis time, and glucoraphenin and sinapine thiocyanate were assigned.Glucoraphenin was enzymatically hydrolyzed fastly by myrosinase, and an intermediate was generated, and then continue to be decomposed into other components.Sinapine thiocyanate did not change significantly during the enzymolysis process, and sinadiosides was also enzymatically degraded. Conclusion: The enzymolysis of Raphani Semen is not only the glucoraphenin, but also the sinadiosides.This paper can provide reference for the property change of Raphani Semen in processing.
RESUMEN
To prepare Tanshinone IIA (TSIIA) molecularly imprinted polymers (MIPs) with affinity and high selectivity to TSIIA and study its mechanism. Taking TSIIA as the template molecular, synthesizing the MIPs at the surface of silica nanoparticles by grafting copolymerization in the toluene solution. A molecular modeling approach was used to elucidate template-monomer interaction. The interaction between template molecule and functional monomer was studied by UV spectrometry. The theoretical molar ratio of template-monomer was 1∶3 and methyl acrylic acid (MAA) was a better functional monomer than acrylamide (AAM) for MIPs synthesis. The MIPs can be used for the extraction and separation of TSIIA.
RESUMEN
Three fragments, viz., BSA-CNBr1 – 183, BSA-CNBr184 – 582, and BSA-T377–582 representing domains I, II + III and III of bovine serum albumin have been isolated and purified. The physicochemical properties have been investigated and compared with their parent albumin molecule. The values of Stokes radii (nm) and intrinsic viscosities (ml/g) have been determined to be 2·36, 3·30; 3·43, 4·36; and 2·40, 3·13 for the fragments BSACNBr1– 183, BSA-CNBr184–582 and BSA-T377–582 respectively. The acid induced unfolding-refolding transitions of intact albumin and the fragment BSA-T 377–582 have been shown to occur in two steps while the fragments BSA-CNBr1 – 183 and BSACNBr184– 582 underwent single step transitions. The formation of the acid denatured states of intact albumin, BSA-CNBr1 – 183 and BSA-CNBr184–582 was accompanied by an increase of about 86, 56 and 44% in the values of intrinsic viscosities respectively. Since all the transitions were reversible, the values of equilibrium constants, KD, were calculated. The analysis of the dependence of KD on pH indicated that the first transition (N–X) of albumin was caused due to the uptake of about 3 protons by the native albumin. ,The intermediate state, X, is converted to acid unfolded state, D, by taking up another two protons. A comparision of the results on intact albumin with that of its fragments revealed that the second transition of the fragment BSA-T377– 582 and the two single step transitions of the fragment BSA-CNBr1–183 and BSA-CNBr184–582 were much closer to the second transition (X-D) of the intact albumin. The first transition of albumin has been attributed to its domain III represented by the fragment BSA-T377–582.