Abstract
We report on a simple and sensitive method for the determination of L-cysteine (Cys). It is based on a redox reaction between the non-fluorescent Cu(II)-calcein complex and Cys which results in fluorescence recovery of calcein. When Cys is added to a solution of the Cu(II)-calcein complex, Cu(II) is reduced to Cu(I), and calcein is released to form a strongly fluorescent complex with Zn(II). The effect was used to develop a fluorescence enhancement method for the determination of Cys. Under the optimum conditions, the increase in signal intensity is linear in the range from 3.0 × 10−7 to 1.2 × 10−5 mol L−1, with a correlation coefficient (R) of 0.9978. The limit of detection (3σ) is 4.0 × 10−8 mol L−1. The relative standard deviation (RSD) in the determination of 11 samples containing 5.0 × 10−6 mol L−1 of Cys was 3.5%. There is little interference by common ions and other amino acids. The method, which is simple, rapid, and sensitive, was successfully applied to the determination of Cys in human serum samples.
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References
Bulaj G, Kortemme T, Goldenberg DP (1998) Ionization-Reactivity Relationships for Cysteine Thiols in Polypeptides. Biochemistry 37:8965
Lau CW, Qin XJ, Liang JY, Lu JZ (2004) Determination of cysteine in a pharmaceutical formulation by flow injection analysis with a chemiluminescence detector. Anal Chim Acta 514:45
Wang WH, Rusin O, Xu XY, Kim KK (2005) Detection of Homocysteine and Cysteine. J Am Chem Soc 127:15949
Perez-Ruiz T, Martinez-Lozano C, Tomas V, Lambertos G (1991) Flow-injection successive determination of cysteine and cystine in pharmaceutical preparations. Talanta 38:1235
Perez-Ruiz T, Martinez-Lozano C, Tomas V, Martin J (2002) Flow injection chemiluminescent method for the successive determination of L-cystine using photogenerated tris(2,2ˊ-bipyridyl) ruthenium (III). Talanta 58:987
Shahrokhian S (2001) Lead Phthalocyanine as a Selecitive Carrier for Preparation of a Cysteine-Selective Electrode. Anal Chem 73:5972
Ruiz-Diaz JJJ, Torriero AAJ, Salinas E, Marchevsky EJ, Sanz MI (2006) Enzymatic rotating biosensor for cysteine and glutathione determination in a FIA system. Talanta 68:1343
Wsaeem A, Yaqoob M, Nabi A (2008) Flow-injection determination of cysteine in pharmaceuticals based on luminol-persulphate chemiluminescence detection. Luminescence 23:144
Teshima N, Nobuta T, Sakai T (2001) Simultaneous flow injection determination of ascorbic acid and cysteine using double flow cell. Anal Chim Acta 438:21
Zhao C, Zhang JC, Song JF (2001) Determination of Cysteine in Amino Acid Mixture and Human Urine by Flow-injection Analysis with a Biamperometric Detector. Anal Biochem 297:170
Amarmath K, Amarmath V, Amarmath K, Valentine HL, Valentine WM (2003) Aspecific HPLC-UV method for the determination of cysteine and related aminothiols in biological samples. Talanta 60:1229
Tcherkas YV, Denisenko AD (2001) Simultaneous determination of several amino acids, including homocysteine, cysteine and glutamic acid, in human plasma by isocratic reversed-phase high-performance liquid chromatography with fluorimetric detection. J Chromatogr A 913:309
Fermo I, Arcelloni C, Paroni R (2002) High-performance liquid chromatographic method to quantify total cysteine excretion in urine. Anal Biochem 307:181
Kusmierek K, Blowback R, Bald E (2006) Analysis of urine for cysteine, cysteinylglycine, and homocysteine by high-performance liquid chromatography. Anal Bioanal Chem 385:855
Ye ML, Xu B, Zhang WD (2011) Sputtering deposition of Pt nanoparticles on vertically aligned multiwalled carbon nanotubes for sensing L-cysteine. Microchim Acta 172:429
Mijidi MR, Zeynali AK, Hafezi B (2010) Sensing L-cysteine in urine using a pencil graphite electrode modified with a copper hexacyanoferrate manostructure. Microchim Acta 169:283
Shahrokhian S, Karimi M (2004) Voltammetric studies of a cobalt(II)-4-methylsalophen modified carbon-paste electrode and its application for the simultaneous determination of cysteine and ascorbic acid. Electrochim Acta 50:77
Amini MK, Khorasani JH, Khaloo SS, Tangestaninejad S (2003) Cobalt(II)-salophen-modified carbon-paste electrode for potentiometric and voltammetric determination of cysteine. Anal Biochem 320:32
Jin WR, Wang Y (1997) Determination of cysteine by capillary zone electrophoresis with end-column amperometric detection at a gold/mercury amalgam microelectrode without deoxygenation. J Chromatogr 769:307
Kuster A, Teal I, Sweeten S, Roze J, Robins RJ, Darmaum D (2008) Simultaneous determination of glutathione and cysteine concentrations and H-2 enrichments in microvolumes of neonatal blood using gas chromatography–mass spectrometry. Anal Bioanal Chem 390:1403
Guo Y, Shao SJ (2004) A specific colorimetric cysteine sensing probe based on dipyrromethane-TCNQ assembly. Tetrahedron Lett 45:6477
Amamath V, Amarnath K (2002) Specific determination of cysteine and penicillamine through cyclization to 2-thioxothiazolidine-4-carboxylic acids. Talanta 56:745
Lunar ML, Rubio S, Perez-Bendito D, Carreto ML, McLeod CW (1997) Hexadecylpyridinium chloride micelles for the simultaneous kinetic determination of cysteine and cystine by their induction of the iodine-azide. Anal Chim Acta 337:341
Liao WS, Wu FY, Wu YM, Wang XJ (2008) Highly sensitive spectrofluorimetric determination of cysteine by Cu2+-morin complex. Microchim Acta 162:147
Wang H, Wang WS, Zhang HS (2001) A spectrofluorimetric method for cysteine and glutathione using the fluorescence system of Zn(II)–8-hydroxyquinoline-5-sulphonic acid complex. Spectrochim Acta A 57:2403
Wang H, Wang WS, Zhang HS (2001) Spectrofluorimetic determination of cysteine based on the fluorescence inhibition of Cd(II)–8-hydroxyquinoline-5-sulphonic acid complex by cysteine. Talanta 53:1015
Huang YM, Zhang ZJ, Lv JG, Cheng H (2000) Flow-injection analysis-fluorescence detection for the in vivo on-line determination of calcium in blood with microdialysis sampling. Anal Chim Acta 419:175
Ali A, Zhang Q, Dai JS, Huang X (2003) Calcein as a fluorescent iron chemosensor for the determination of low molecular weight iron in biological fluids. Biometals 16:285
Berregi I, Durand JS, Casado JA (1999) Simultaneous spectrofluorimetric determination of the rare earths with calcein. Talanta 48:719
Li HB, Chen F (2000) A highly sensitive fluorimetric method for the determination of fluoride in biological material with Al3+-calcein complex. Fresenius J Anal Chem 368:501
Zhu ZC (2001) Determination of Nucleic Acids Using Calcein-Neodymium Complex as a Fluorescence Probe. Anal Sci 17:1375
Fei N, Ru LJ (2007) Determination of ketotifen by using calcein as chemiluminescence reagent. Anal Chim Acta 592:168
Yu FS, Ding YB, Gao YW, Zheng SS, Chen F (2008) Fluorescence enhancement effect for the determination of DNA with calcein-cetyl trimethyl ammonium bromide system. Anal Chim Acta 625:195
Ozkan Y, Ozkan E, Simsek B (2002) Plasma total homocysteine and cysteine levels as cardiovascular risk factors in coronary heart disease. Int J Cardiol 82:269
Sattarahmady N, Heli H (2011) An electrocatalytic transducer for L-cysteine detection based on cobalt hexacyanoferrate nanoparticles with a core-shell structure. Anal Biochem 409:74
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The authors acknowledge the support from National Natural Science Foundation of China(No. 20805016).
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Chang, L., Wu, T. & Chen, F. Determination of L-cysteine base on the reversion of fluorescence quenching of calcein by copper(II) ion. Microchim Acta 177, 295–300 (2012). https://doi.org/10.1007/s00604-011-0759-0
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DOI: https://doi.org/10.1007/s00604-011-0759-0