The reaction between fullerenol (F-ol) and cytochrome C (Cyt C) could be carried out to form a nonphosphorescence compound using tween-80 as photosensitizer which causes the sharp quenching of room temperature phosphorescence (RTP) of F-ol. Bearing this in mind, a novel solid substrate room temperature phosphorimetry (SSRTP) for the determination of trace Cyt C has been proposed in this study. Under the optimum conditions, the linear range of this method is – ？？ , which is directly proportional to of Cyt C-F-ol–tween-80 system, and the detection limit (DL) is ？ . It has been applied to the determination of Cyt C in human serum and forecast of diseases, and the result matches with the enzyme-linked fluorescence immunoassay (ELISA). Meanwhile, the reaction mechanism of SSRTP for the determination of Cyt C and the enhancing effect of tween-80 on RTP of F-ol were also discussed. 1. Introduction In recent years, the studies have been found that the incidence of leukemia and the execution of chemotherapy drugs to leukemia cells are closely related to cell apoptosis [1]. Studies on the clinical examination of Cyt C and the researches about its proapoptotic activity regulating molecular mechanism and the release mechanism have become a focus [2]. There are many methods for the determination of Cyt C, such as synchronous fluorescence spectroscopy (linear range: – ？g？mL？1) [3], CdTe/CdS quantum dots resonance rayleigh scattering spectroscopy (DL: ？g？mL？1) [4], electrochemical probe voltammetric method (DL: ？g？mL？1) [5], cyclic voltammetry [6], adsorption method (DL: ？g mL？1) [7], ED/Au/I？ modified ultramicroelectrode method (DL: ？g？mL？1) [8], capillary zone electrophoretic method (DL: ？g？mL？1) [9], amperometric method (DL: ？g？mL？1) [10], protein blotting method (DL: ？g？mL？1) [11], and ELISA (DL: ？g？mL？1) [12]. All these methods cannot meet the needs of determining the low content of Cyt C in biological samples due to low sensitivity. Therefore, searching for a high sensitive and accurate method for the determination of Cyt C in biological samples and discussing the relativity between the content of Cyt C and cell apoptosis have become research focus with high academic value and great significance. There have been many reports on the synthesis of multihydroxyl C60 derivatives, F-ol derivatives, aminophenol derivatives, dendritic fullerene derivatives [13–15], the fluorescent property of water-soluble F-ol [16], and the determination of alkaline phosphatase [17, 18], glucose [19], As (V) [20], and Mn2+ [21] based on phosphorescent property of F-ol, showing broad
References
[1]
X. M. Fang, M. Z. Chen, R. L. Chen, and Z. L. Ye, “Effect of cytochrome C on HL-60 cell apoptosis and its relationship with the relevant genes bcl-2 and bax,” Experimental Hematology, vol. 13, no. 4, pp. 570–574, 2005.
[2]
D. Wang and Y. F. Liu, “Progress of cytochrome C and apoptosis,” Chinese Children With Blood, vol. 9, pp. 181–184, 2004.
[3]
J. Chou, X. G. Qu, T. Lu, S. J. Dong, and Y. Wu, “Determination of cytochrome C by synchronous fluorescence spectroscopy,” Chinese Journal of Analytical Chemistry, vol. 22, pp. 1238–1240, 1994.
[4]
W. Yan, A. M. Zeng, and H. S. Wang, “Resonance rayleigh scattering spectrometric determination of cytochrome C by its reaction with CdTe/CdS quantum dots,” Phys/Chem Testing, vol. 44, pp. 107–111, 2008.
[5]
L. B. Qu, J. H. Zhao, J. J. Li, and R. Yang, “Determination of cytochrome C by voltammetric method using luteolin as electrochemical probe,” Journal of Chinese Pharmaceutical, vol. 38, pp. 656–658, 2007.
[6]
H. L. Li, X. Q. Wu, Y. Wei, R. Wang, and X. W. Cao, “The electrochemical behaviors of cytochrome C at gold electrode modified with 2-aminoethanethiol self assembly monolayer,” Journal of Shanghai Normal University, vol. 35, pp. 52–55, 2006.
[7]
Y. J. Yin, Y. F. Lv, P. Wu, P. Du, Y. M. Shi, and C. X. Cai, “Immobilization of cytochrome C on the surface of single-wall carbon nanotube and its direct electron transfer and electrocatalysis,” Electrochem, vol. 12, pp. 299–303, 2006.
[8]
M. Zhu, G. Y. Shi, M. Liu, and L. T. Jin, “Study of gold colloid monolayer modified carbon fiber ultramicroelectrode and its application in determination of cytochrome C,” Journal of East China Normal University, vol. 1, pp. 68–73, 2003.
[9]
V. M. Leonardo and R. O. Margarita, “Capillary zone electrophoretic determination of cytochrome c in mitochondrial extracts and cytosolic fractions: application to a digitalis intoxication study,” Talanta, vol. 74, no. 4, pp. 478–488, 2008.
[10]
S. N. Tan and L. Hua, “Amperometric detection of cytochrome C by capillary electrophoresis at a sol-gel carbon composite electrode,” Analytica Chimica Acta, vol. 450, pp. 263–267, 2001.
[11]
J. Huang, X. C. Mo, and H. M. Li, “Determination on cytochrome C in various tissues of mice by western-blotting,” Journal of Guiyang Medical College, vol. 1, pp. 48–50, 2006.
[12]
H. Liu, F. L. Wang, and J. Li, “An experimental study on the cytochrome C changes of brainstem neuron after rat death due to acute brainstem injury,” Chinese Journal of Forensic Medicine, vol. 21, no. 4, pp. 212–214, 2006.
[13]
E. Nakamura and H. Isobe, “Functionalized fullerenes in water. The first 10 years of their chemistry, biology, and nanoscience,” Accounts of Chemical Research, vol. 36, no. 11, pp. 807–815, 2003.
[14]
Y. Takaguchi, T. Tajima, and K. Ohta, “Reversible binding of C60 to an anthracene bearing a dendritic poly.amidoamine. substituent to give a water-soluble fullerodendrimer,” Angewandte Chemie, vol. 41, pp. 817–819, 2002.
[15]
Y. Liu, H. Wang, P. Liang, and H. Y. Zhang, “Water-soluble supramolecular fullerene assembly mediated by metallobridged β-cyclodextrins,” Angewandte Chemie, vol. 43, no. 20, pp. 2690–2694, 2004.
[16]
X. Q. Yan, J. L. Qiao, L. Lu, Y. H. Wei, W. J. Jin, and B. S. Xu, “Fluorescence properties of water-soluble fullerols and interaction with various metallic ions,” Spectroscopy and Spectral Analysis, vol. 22, no. 2, pp. 289–291, 2002.
[17]
J. M. Lin, F. Gao, H. H. Huang et al., “Determination of trace alkaline phosphatase by solid-substrate room-temperature phosphorimetry based on triticum vulgare lectin labeled with Fullerenol,” Chemistry and Biodiversity, vol. 5, no. 4, pp. 606–616, 2008.
[18]
J. M. Liu, X. M. Huang, Z. B. Liu et al., “Exploitation of phosphorescent labelling reagent of fullerol-fluorescein isothiocyanate and new method for the determination of trace alkaline phosphatase as well as forecast of human diseases,” Analytica Chimica Acta, vol. 648, no. 2, pp. 226–234, 2009.
[19]
J. M. Liu, H. X. Wang, L. H. Zhang et al., “Fullerol-fluorescein isothiocyanate phosphorescent labeling reagent for the determination of glucose and alkaline phosphatase,” Analytical Biochemistry, vol. 404, no. 2, pp. 223–231, 2010.
[20]
J. M. Liu, F. Gao, T. L. Yang, J. H. Lai, and Z. M. Li, “Catalytic solid substrate-room temperature phosphorimetry for the determination of trace As(V) based on oxidising reaction between hydrogen peroxide and fullerenol using tween-80 as sensitizer,” International Journal of Environmental Analytical Chemistry, vol. 88, no. 9, pp. 613–624, 2008.
[21]
J. M. Liu, X. J. Cui, F. Gao et al., “Solid substrate-room temperature phosphorescence method for the determination of trace Mn(II) based on oxidizing reaction of hydrogen peroxide using α,α′-bipyridine as sensitizer,” Journal of Fluorescence, vol. 17, no. 1, pp. 49–55, 2007.
[22]
J. M. Liu, L. P. Lin, X. X. Wang et al., “Highly sensitive detection of residual chlorpromazine hydrochloride with solid substrate Room temperature phosphorimetry,” Journal of Fluorescence, vol. 22, pp. 1087–1094, 2012.
[23]
J. M. Liu, L. P. Lin, X. X. Wang, W. L. Cai, L. H. Zhang, and S. Q. Lin, “A highly sensitive coupling technique for the determination of trace quercetin based on solid substrate room temperature phosphorimetry and poly (vinyl alcohol) complex imprinting,” Analytica Chimica Acta, vol. 723, pp. 76–82, 2012.
[24]
J. M. Liu, L. P. Lin, H. X. Wang et al., “Catalytic solid substrate room temperature phosphorimetry for the determination of trace rhamnose based on its condensation reaction with calcein,” Spectrochimica Acta A, vol. 84, pp. 221–226, 2011.
[25]
J. M. Liu, S. Q. Lin, X. Lin, and L. Q. Zeng, “Determination of trace carvedilol by solid substrate-room temperature phosphorimetry, based on its activating effect on hypochlorite-oxidizing amaranth using sodium dodecyl benzene sulphonate as sensitizer,” Luminescence, vol. 26, pp. 734–740, 2011.
[26]
D. Y. Sun, Z. Y. Liu, X. H. Guo, Y. M. Yu, Y. Zhou, and S. Y. Liu, “Convenient preparation and properties of C60.OH.x,” Chemical Journal of Chinese Universities, vol. 17, pp. 19–20, 1999.
[27]
J. Luo, L. L. Wu, J. T. Wu, S. H. Huang, and Z. H. Lin, “Effects of pH, ligand CN-, metallic Ions Hg2+, Cd2+ and Pb2+ on electroactivity of cytochrome C,” Electrochem, vol. 2, pp. 61–65, 1996.
[28]
N. Kavathia, A. Jain, J. Walston, B. A. Beamer, and N. S. Fedarko, “Serum markers of apoptosis decrease with age and cancer stage,” Aging, vol. 1, no. 7, pp. 652–663, 2009.
[29]
T. L. Yang, Z. B. Liu, J. M. Liu et al., “Solid substrate-room temperature phosphorimetry for the determination of trace lead using p-nitro-phenyl-fluorone-multi-wall carbon nanotubes-Tween-80 micellae compound and diagnosis about human diseases,” Spectrochimica Acta A, vol. 72, no. 1, pp. 156–164, 2009.
[30]
C. L. Xu and H. Y. Yao, “Study on the interaction mechanism between tween-80 and liposome membrane,” Journal of Xi'an Shiyou University, vol. 20, no. 6, pp. 45–49, 2005.
