Shandong Provincial Key Laboratory of FluorineChemistry Chemistry ...
Shandong Provincial Key Laboratory of Fluorine Chemistry and Chemical Materials University of Jinan 106 Ji Wei Road Jinan, Shandong 250022 P.R . China
Qin Wei Professor Phone: 86-531-82765730 Fax: 86-531-82765969 Email: [email protected]
The extended abstract number is 042 Tetra Methoxyl hydroxyphenyl II Fluorimetric Determination of Proteins in Milk Using UsingTetra Tetra- (3 (3-Methoxyl Methoxyl-44-hydroxyphenyl hydroxyphenyl)) porphyrin porphyrin- Zn ((II II)) as a Spectral Probe Yuxue Dai, Yan Li, Yanyan Cai, Ru Li, Yanfang Zhao, Kexia Mao, Qin Wei* Protein is a fundamental element of life and plays important roles in physiological processes in all organisms. The quantitative analysis of serum albumin , which is the most abundant carrier protein in plasma, is of great importance in clinical medicine tests and analytical biochemistry research. Porphyrinis widely used in the analysis and testing of metal
ions and biological
ma cromolecules as chromogenic reagent or indicator. In this paper, T(3-MO-4HP )P-Zn is used for proteins determination as a spectral probe by fluorescence spectra. The fluorescence spectra of T(3-MO-4HP)P-Zn -porphyrinBSA system were shown in Fig. 1. From Fig. 1, it can be seen that T(3-MO-4HP)P-Zn -porphyrin complex had an emission peak at 611 nm and the characteristic fluorescence intensity was weak. The declined fluorescence intensities were proportional to the concentration of BSA. Based on the decline of the intensity, the reaction conditions of the system were studied in a series of experiments. Finally it was found that 1.00 mL pH 4.39 HAcNaAc buffer solution was suitable, 0.60 mL (10 g· L-1) AOT micelle was the optimum medium and 0.40 mL chromogenic reagent was used for the measure ment .
According to experimental method, the addition order of 1.00 mL HAc-NaAc (pH 4.39), 0.40 mL T(3-MO-4HP)P -Znporphyrin, 0.30 mL BSA and 0.60 mL AOT micelle was optimum. And 10 min was recommended. Then it was found that compared with the natural protein systems, ΔI F of heat denaturation of protein reduced slightly, indicating that T(3MO-4HP)P-Zn-porphyrin could enhance the structural stability of BSA. Under the optimum conditions, linear relationships were obtained between the fluorescence intensity difference and the concentration
of
bovine
serum
albumin(BSA),
egg
albumin(Ova) and γ -globulin(γ -G)(concentrations were all 100 μg·mL -1). The fluorescence of this system was proportion to the concentration of BSA in the range 0.10 ~ 3.40 µg· mL -1, 0.05 ~ 2.60 µg·mL -1 for Ova, 0.02 ~ 2.30 µg·mL -1 for γ-G, the detection limits were 0.08 µg·mL -1, 0.03 µg· mL -1, 0.01 µg·mL -1, respectively. Under the conditions given above, the co-existing substances were investigated by adding different substances. This method was used for the determination of proteins in real samples, we can see the recovery was in range of 99.0 % ~ 101.0 %, and the relative standard deviation RSD was less than 5.0 %. The result was satisfactory. Acknowledgements This study was supported by the Natural Science Foundation of
China (20577016), the Natural Science
Foundation of Shandong Province (Y2008B44), and the Key Subject Research Foundation of Shandong Province (XTD0705) Fig.1 The emission spectra of the interaction of Porphyrins with Ova.1.T(3-MO-4HP)P -Zn ; 2.T(3-MO-4HP)P -Zn + Ova; 3.T(3-MO4HP)P-Zn +AOT micelles; 4.T(3-MO-4HP)P -Zn +AOT micelles + Ova; 5. HAc-NaAc+T(3-MO-4HP )P-Zn +AOT micelles; 6. HAc-NaAc+T(3MO-4HP)P-Zn +BSA+AOT m icelles; Conditions: 0.40 mL T(3-MO4HP)P-Zn porphyrin, 5.0×10-5 mol⋅ L-1; 0.30 mL Ova, 100 µg⋅mL -1;
and all the authors express their deep thanks.
Qin Wei Professor Phone: 86-531-82765730 Fax: 86-531-82765969 Email: [email protected]
The extended abstract number is 042 Tetra Methoxyl hydroxyphenyl II Fluorimetric Determination of Proteins in Milk Using UsingTetra Tetra- (3 (3-Methoxyl Methoxyl-44-hydroxyphenyl hydroxyphenyl)) porphyrin porphyrin- Zn ((II II)) as a Spectral Probe Yuxue Dai, Yan Li, Yanyan Cai, Ru Li, Yanfang Zhao, Kexia Mao, Qin Wei* Protein is a fundamental element of life and plays important roles in physiological processes in all organisms. The quantitative analysis of serum albumin , which is the most abundant carrier protein in plasma, is of great importance in clinical medicine tests and analytical biochemistry research. Porphyrinis widely used in the analysis and testing of metal
ions and biological
ma cromolecules as chromogenic reagent or indicator. In this paper, T(3-MO-4HP )P-Zn is used for proteins determination as a spectral probe by fluorescence spectra. The fluorescence spectra of T(3-MO-4HP)P-Zn -porphyrinBSA system were shown in Fig. 1. From Fig. 1, it can be seen that T(3-MO-4HP)P-Zn -porphyrin complex had an emission peak at 611 nm and the characteristic fluorescence intensity was weak. The declined fluorescence intensities were proportional to the concentration of BSA. Based on the decline of the intensity, the reaction conditions of the system were studied in a series of experiments. Finally it was found that 1.00 mL pH 4.39 HAcNaAc buffer solution was suitable, 0.60 mL (10 g· L-1) AOT micelle was the optimum medium and 0.40 mL chromogenic reagent was used for the measure ment .
According to experimental method, the addition order of 1.00 mL HAc-NaAc (pH 4.39), 0.40 mL T(3-MO-4HP)P -Znporphyrin, 0.30 mL BSA and 0.60 mL AOT micelle was optimum. And 10 min was recommended. Then it was found that compared with the natural protein systems, ΔI F of heat denaturation of protein reduced slightly, indicating that T(3MO-4HP)P-Zn-porphyrin could enhance the structural stability of BSA. Under the optimum conditions, linear relationships were obtained between the fluorescence intensity difference and the concentration
of
bovine
serum
albumin(BSA),
egg
albumin(Ova) and γ -globulin(γ -G)(concentrations were all 100 μg·mL -1). The fluorescence of this system was proportion to the concentration of BSA in the range 0.10 ~ 3.40 µg· mL -1, 0.05 ~ 2.60 µg·mL -1 for Ova, 0.02 ~ 2.30 µg·mL -1 for γ-G, the detection limits were 0.08 µg·mL -1, 0.03 µg· mL -1, 0.01 µg·mL -1, respectively. Under the conditions given above, the co-existing substances were investigated by adding different substances. This method was used for the determination of proteins in real samples, we can see the recovery was in range of 99.0 % ~ 101.0 %, and the relative standard deviation RSD was less than 5.0 %. The result was satisfactory. Acknowledgements This study was supported by the Natural Science Foundation of
China (20577016), the Natural Science
Foundation of Shandong Province (Y2008B44), and the Key Subject Research Foundation of Shandong Province (XTD0705) Fig.1 The emission spectra of the interaction of Porphyrins with Ova.1.T(3-MO-4HP)P -Zn ; 2.T(3-MO-4HP)P -Zn + Ova; 3.T(3-MO4HP)P-Zn +AOT micelles; 4.T(3-MO-4HP)P -Zn +AOT micelles + Ova; 5. HAc-NaAc+T(3-MO-4HP )P-Zn +AOT micelles; 6. HAc-NaAc+T(3MO-4HP)P-Zn +BSA+AOT m icelles; Conditions: 0.40 mL T(3-MO4HP)P-Zn porphyrin, 5.0×10-5 mol⋅ L-1; 0.30 mL Ova, 100 µg⋅mL -1;
and all the authors express their deep thanks.
