Study On Fluorone Reageant-(VI) As Spectral Probe For Protein Determination

In this paper, some major quantitative analysis methods of protein and their applications in the fields of biochemistry, medicine, food and clinical medicine are presented in details. These methods mainly include Turbidity Method and Kjeldahl, which are applied in the earlier period; Spetrophotometric Method and Flurospetrophotometric Method, which are presently studied and applied widely, and can be easy operated; and Esonance Rayleigh Scattering Method, High Performance Liquid Chromatographic Method, and Capilary Electrophoresis Method, which are newly developed. In our experiments, eleven tri-hydroxyl fluorone reagents with different substituents are selected on the basis of Phenyl Fluorone(PF) and salicylfluorone(SAF), such as Phenyfluorone(PF), o-Nitrophenylfluorone(o-NPF), Salicylfluorone and etc. The complex of Tri-hydroxyl Fluorone and Mo(VI) is used as a spectral probe for protein determination in quantity. The Spetrophotometric Analytical Methods is applied for the systematic study of the reaction conditions (such as acidity, media and dosage, dosage of developer, reaction time, ionic strength, interfering ion, and etc.) for Tri-hydroxyl Fluorone-Mo(VI) and protein, and a series of new methods with higher sensitivity for the determination of protein are established. Microemulsion, as the media, is firstly introduced to the determination of protein, which has increased the sensitivity of system with its sensitization and solubilization.The experimental results indicate that the complex of Tri-hydroxyl Fluorone and Mo(VI), as a spectral probe of protein determination, has very high sensitivity. At present, the relevant reference shows that SAF-Mo(VI)-BSA System has the highest sensitivity in the method of Metallic Ion-dye Spetrophotometry, with the molar absorptivity of 2.86 X 106Lmol-1cm-1. Among the eleven systems studied in this paper, 5'-NSF-Mo(VI)-BSA , o-NPF-Mo(VI)-BSA, DBH-PF-Mo(VI)-BSA have the highest sensitivity, with the molar absorptivities of 8.71 X 106mol-1cm-1, 9.06X106Lmol-1cm-1 and 1.53 X 107Lmol-1cm-1 respectively, which are 3.0, 3.2 and 5.4 times higher than the SAF-Mo(VI)-BSA method.The experiments also indicate that the microemulsion media has more solubilization and sensitization than micelle media. That the microemulsion media has higher molar absorptivity than that of micelle media proves that it has higher sensitization than micelle media.Based on the experiments, the 5'-NSF-Mo(VI), o-NPF-Mo(VI), DBH-PF-Mo(VI), as aspectral probe, have been successfully applied to the determination of protein in human urine, human blood sample and milk powder.This paper features in systematic, scientific, and creative methods.1. Systematic feature:This paper systematically studies the reaction conditions (such as acidity, media and dosage, dosage of developer, reaction time, ionic strength, interfering ion, and etc.) for Tri-hydroxyl Fluorone-Mo(VI) and protein and the effects of substituents on the sensitivity of system, and provides a foundation for quantitative determination of protein with the higher sensitivity.2. Scientific feature:The Spetrophotometric Method with Fluorone Reagent-Metallic Ion as spectral probe for the determination of protein becomes a sensitive way in the micro-determination of protein with higher sensitivity and stronger anti-interference. This paper establishes a more sensitive method based on the large amounts of experiments which has indicated 5.4 times higher sensitivity than that of the reference and been successfully applied to the determination of protein in human urine, human blood sample and milk powder.3. Creative feature:Microemulsion, as the media, is firstly introduced to the determination of protein, which has increased the sensitivity of system with its sensitization and solubilization. The most Fluorone reagents described in this paper are all firstly introduced for the determination of protein and formed a series of new methods.