Glutation: metody przygotowania próbek dla analiz z wykorzystaniem technik chromatograficznych i elektroforezy kapilarnej

• Autorzy: Błońska-Sikora E. , Oszczudłowski J. , Witkiewicz Z. , Wideł D.

Warianty tytułu

EN

Glutathione: methods of sample preparation for chromatography and capillary electrophoresis

• Języki publikacji: PL

Abstrakty

PL

Zredukowana forma glutationu jest jednym z najważniejszych przeciwutleniaczy występujących w organizmie człowieka, a niedobór tego związku prowadzi często do poważnych zaburzeń. Glutation bierze również udział w modulowaniu odporności, procesach detoksykacji, a także reguluje aktywność innych białek i enzymów. Jest on czułym wskaźnikiem wrażliwości organizmu na stres oksydacyjny a jego stężenie w poszczególnych narządach i komórkach może wskazywać na stopień ich uszkodzenia. Oznaczanie stężenia glutationu z wykorzystaniem technik chromatograficznych stanowi uzupełnienie a niejednokrotnie alternatywę w stosunku do innych metod analitycznych: enzymatycznych, spektrofotometrycznych, spektrofluorymetrycznych, elektromigracyjnych, chemiluminescencyjnych, czy magnetycznego rezonansu jądrowego. Do najważniejszych etapów przygotowania próbki należą: zabezpieczenie aminotioli przed wysoką temperaturą i działaniem substancji utleniających pochodzących z matrycy, odbiałczanie, redukcja disulfidów, a także derywatyzacja (z wyjątkiem technik elektromigracyjnych i tandemowej spektrometrii mas).

EN

Reduced form of glutathione is one of the most important antioxidants, which is found in human body. Deficiency of this compound can often lead to serious disorders. It plays a central role in immunity modulation, detoxification capacities and regulation of activity of another peptides and enzymes. In every live organism it acts as a sensitive rate of susceptibility to oxidative stress and its concentration in each organ and cell shows the level of their damage. Determination of glutathione using chromatographic methods has been a complement and many a time is also an alternative to different widely used methods, such as: enzymatic, spectrophotometric, spectrofluorometric, electromigrative, chemiluminescence, nuclear magnetic resonance. The most important steps in the sample preparation process for analysis are: amino thiols protection from high temperature and oxidation caused by substances from matrix, precipitation, reduction of disulfides and also derivatization (except for in vivo assays and those based on electrochemical and tandem mass spectrometry). Several methods have been presented in order to identify and quantify glutathione forms in human biological samples such as liquid chromatography with electrochemical, UV, fluorometric, mass spectrometric detection capillary electrophoresis and gas chromatography with mass spectrometry. This review presents different techniques for glutathione routine analysis in hospitals and also useful in clinical researches with large groups of patients.

Słowa kluczowe

PL

glutation zredukowany; glutation utleniony; przygotowanie próbki; techniki chromatograficzne; elektroforeza kapilarna

EN

reduced glutathione; oxidized glutathione; sample preparation; chromatographic methods; electrophoretic methods

• Wydawca: Stowarzyszenie Inżynierów i Techników Przemysłu Chemicznego, ZW CHEMPRESS-SITPChem
• Czasopismo: Chemik
• Rocznik: 2012
• Tom: Vol. 66, nr 9
• Strony: 929--942
• Opis fizyczny: Bibliogr. 70 poz.

Twórcy

autor

Błońska-Sikora E.

autor

Oszczudłowski J.

autor

Witkiewicz Z.

autor

Wideł D.

• Instytut Chemii, Uniwersytet Jana Kochanowskiego w Kielcach

Bibliografia

• 1. Serru V., Baudin B., Ziegler F., David J. P., Cals M. J., Vaubourdollee M., Mario N: Quantification of Reduced and Oxidized Glutathione in Whole Blood Samples by Capillary Electrophoresis. Clin. Chem. 2001, 47, 7,1321-1324.
• 2. Pastore A., Federici G., Bertini E., Piemonte F: Analysis of glutathione: implication in redox and detoxification. Clin. Chim. Acta. 2003, 333, 1, 19-39.
• 3. Bilska A., Kryczyk A., Włodek L.: The different aspects of the biological role of glutathione. Post. Hig. Med. Dośw. 2007, 61, I, 438-453.
• 4. Winiarska K.: Glutation: niezwykłe funkcje pospolitego tripeptydu. Post. Bioch. 2000, 46, 318-326.
• 5. Radwańska-Wala B., Buszman E., Drużba D.: Reactive oxygen species in pathogenesis of central nervous system diseases. Wiad. Lek. 2008, 66, 13,67-72.
• 6. Tylec, A., Jarząb A., Stryjecka- Zimmer M., Wójcicka, A.: Stress oxidative in schizophreni. Pol. Merk. Lek. 2007, 23, 133, 74-76.
• 7. Kałużna-Czaplińska J., Grys W., Szymańska A., Rynkowski J.: Role of oxidative stress in autism. Nowa Pediat. 2008, 4, 67-70.
• 8. Miller E., Mrowicka M., Zołyński K., Kedziora J.: Oxidative stress in multiple sclerosis. Pol. Merk. Lek. 2009, 27, 162, 499-502.
• 9. Nolin T. D., McMenamin E., Himmelfarb J.: J. Chromatogr. B. 2007, 852, 1-2,554-561.
• 10. Sakhi A.K., Blomhoff R., Gundersen T. E.: Simultaneous and trace determination of reduced and oxidized glutathione in minute plasma samples using dual mode fluorescence detection and column switching high performance liquid chromatography. J. Chromatogr. A. 2007, I 142, 2, 178-184.
• 11. Iwasaki Y., Saito Y., Nakano Y., Mochizuki K., SakataO., Ito R„ Saito K., Nakazawa H.: Chromatographic and mass spectrometric analysis of glutathione in biological samples. J. Chromatogr. B. 2009, 877, 28, 3309-3317.
• 12. Overbo K., Sorbye H., Svardal A., Grong K., Svanes K.: Glutathione and N-acetylcysteine reduce gastric mucosal blood flow in rats. Dig. Dis. Sci. 1997 42,8, 1765-1774.
• 13. Lenton K., Therriault H., Wagner J. R.: Analysis of glutathione and glutathione disulfide in whole cells and mitochondria by postcolumn derivatization high-performance liquid chromatography with ortho-phthalaldehyde. Anal. Biochem. 1997, 274, I, 125-130.
• 14. Hiraku Y., Murata M., Kawanishi S.: Determination of intracellular glutathione and thiols by high performance liquid chromatography with a gold electrode at the femtomole level: comparison with a spectroscopic assay. Biochim. Biophys. Acta. 2002, 1570, 1,47-52.
• 15. Lakritz J., Plopper C.G., Buckpitt A.R.: Validated high-performance liquid chromatography-electrochemical method for determination of glutathione and glutathione disulfide in small tissue samples. Anal. Biochem. 1997, 247, 63-68.
• 16. Lazzarino G., Amorini A.M., Fazzina G., Vagnozzi R.,Signoretti S., Donzelli S., Di Stasio E., Giardina B., Tavazzi B.: Single-sample preparation for simultaneous cellular redox and energy state determination. Anal. Biochem. 2003, 322(1), 51-59.
• 17. Potesil D., PetrlovaJ., Adam V, Vacek J., Klejdus B., Zehnalek J., Trnkova L., Havel L., Kizek R.: Simultaneous femtomole determination of cysteine, reduced and oxidized glutathione, and phytochelatin in maize (Zea mays L.) kernels using high-performance liquid chromatography with electrochemical detection. J. Chromatogr. A. 2007, 1084(1), 134-144.
• 18. Mendoza J., Soto R, Ahumada I., Garrido T: Determination of oxidized and reduced glutathione, by capillary zone electrophoresis, in Brassica juncea plants treated with copper and cadmium. Electrophoresis. 2004, 25(6), 890-896.
• 19. Monostori P., Wittmann G., Karg E., Turi S.: Determination of glutathione and glutathione disulfide in biological samples: An in-depth review. J. Chromatogr. B. 2009, 877,3331-3346.
• 20. Kuśmierek K., Chwatko G., Głowacki R., Kubalczyk R, Bald E.: Ultraviolet derivatization of low-molecular-mass thiols for high performance liquid chromatography and capillary electrophoresis analysis. J. Chromatogr. B. 2011, 879, 1290-1307.
• 21. Jones D. P.: Glutathione measurement in human plasma. Evaluation of sample collection, storage and derivatization conditions for analysis of dansyl derivatives by HPLC. Clin. Chim. Acta. 1998, 275, 175-184.
• 22. Camera E., Picardo M.: Analytical methods to investigate glutathione and related compounds in biological and pathological processes. J. Chromatogr. B. 2002, 781, 181-206.
• 23. McDermott G., Francis P. S., Holt K. J., Scott K. L., Martin S. D., Stupka N., Barnett M. W., Conlan X.: Determination of intracellular glutathione and glutathione disulfide using high performance liquid chromatography with acidic potassium permanganate chemiluminescence detection. Analyst. 2011, 136, 2578-2585.
• 24. Harwood D. T., Kettle A., Brenna J. S., Winterbourn C. C.: Simultaneous determination of reduced glutathione, glutathione disulphide and glutathione sulphonamide in cells and physiological fluids by isotope dilution liquid chromatography-tandem mass spectrometry. J. Chromatogr. B. 2009. 877 (28), 3393-3399.
• 25. Kandar R., Żakovaa P., Lotkovab H., Kucerab O., Červinkova Z: Determination of reduced and oxidized glutathione in biological samples using liquid chromatography with fluorimetric detection. J. Pharm. Biomed. Anal. 2007, 43, 1382-1387.
• 26. Chang C., Tseng W.: Gold nanoparticle extraction followed by capillary electrophoresis to determine the total, free, and protein-bound aminothiols in plasma. Anal. Chem. 2010, 82 (7), 2696-2702.
• 27. Marchand S., Guzek A., Leroy R: HPLC study of the host-guest complexation between fluorescent glutathione derivatives and ß-cyclodextrin. J. Incl.. Phenom. Macro. 2009, 66(3-4), 409-418.
• 28. Hansen R.E., Ostergaard R.,Norgaard R, Winther J.R.: Quantification of protein thiols and dithiols in the picomolar range using sodium borohydride and 4,4’-dithiodipyridine. Anal. Biochem. 2007, 363(1), 77-82.
• 29. Głowacki R., Bald E.: Fully automated method for simultaneous determination of total cysteine, cysteinylglycine, glutathione and homocysteine in plasma by HPLC with UV absorbance detection. J. Chromatogr. B. 2009, 877(28), 3400-3404.
• 30. Hansen R.E., Winther J.R.: An introduction to methods for analyzing thiols and disulfides: Reactions, reagents, and practical considerations. Anal. Biochem. 2009,394(2), 147-158.
• 31. Michaelsen J.T., Dehnert S., Giustarini D., Beckmann B., Tsikas. D.: HPLC analysis of human erythrocytic glutathione forms using OPA and N-acetylcysteine ethyl ester: Evidence for nitrite-induced GSH oxidation to GSSC. J. Chromatogr. B. 2009, 877 (28), 3405-3417.
• 32. Jones D.R, Carlson J.L., Mody V.C., Cai J., Lynn M.J., Sternberg Jr. R: Redox State of Glutathione in Human Plasma. Free Radic. Biol. Med. 2000, 28(4), 625- 635.
• 33. Rosenfeld J.: Enhancement of analysis by analytical derivatization. J. Chromatogr. B. 2011, 879 (17-18), 1157-1158.
• 34. Tietze F: Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: Applications to mammalian blood and other tissues. Anal. Biochem. 1969, 27(3), 502-522.
• 35. Rahman I., Kode A., Biswas S.K.: Assay for quantitative determination of glutathione and glutathione disulfide levels using enzymatic recycling method. Nat. Protocols. 2007, 1, 3159 - 3165.
• 36. Eyer P., Podhradsty D.: Evaluation of the micromethod for determination of glutathione using enzymatic cycling and Ellman's reagent. Anal. Biochem. 1986, 153(l), 5766.
• 37. Giustarini D., Milzani A., Dalle-Donne I., Rossi R.: Red blood cells as a physiological source of glutathione for extracellular fluids. Blood Cells Mol. Dis. 2008,40(2), 174-179.
• 38. Winters R.A., Żukowski J., Ercal N., Matthews R.H., Spitz D.R.: Analysis of glutathione, glutathione disulfide, cysteine, homocysteine, and other biological thiols by high-performance liquid chromatography following derivatization by n-(l-pyrenyl)maleimide. Anal. Biochem. 1995, 227(1), 14-21.
• 39. Benkova B., Lozanov V, Ivanov I.P., Todorova A., Milanov I., Mitev V: Determination of plasma aminothiols by high performance liquid chromatography after precolumn derivatization with N-(2-acridonyl)maleimide. J. Chromatogr. B. 2008, 870(1), 103-108.
• 40. Brehe J.E., Burch H.E.: Enzymatic assay for glutathione. Anal. Biochem. 1976,74(1), 189-97.
• 41. Katrusiak A.E., Paterson P.G., Kamencic H., Shoker A, Lyon A.W.: Precolumn derivatization high-performance liquid chromatographic method for determination of cysteine, cysteinyl-glycine, homocysteine and glutathione in plasma and cell extracts. J. Chromatogr. B: Biomedical Sciences and Applications. 2011, 758(2), 207-2012.
• 42. Nozal M.J., Bernal J.L., Toribio L., Marinero R, Moral O., Manzanas L., Rodriguez E.: Determination of glutathione, cysteine and N-acetylcysteine in rabbit eye tissues using high-performance liquid chromatography and postcolumn derivatization with 5,5'-dithiobis(2-nitrobenzoic acid). J. Chromatogr. A 1997, 778(1-2), 347-353.
• 43. Riener K., Kada G., Gruber H.J.: Quick measurement of protein sulfhydryls with Ellman's reagent and with 4,4'-dithiodipyridine. Anal. Bioanal. Chem. 2002, 373(4-5), 266-276.
• 44. Bald E., Chwatko G., Głowacki R., Kuśmierek K.: Analysis of plasma thiols by high-performance liquid chromatography with ultraviolet detection. J. Chromatogr. A. 2004, 1032(1-2), 109-115.
• 45. Iciek M., Chwatko G., Lorenc-Koci E., Bald E., Włodek L: Plasma levels of total, free and protein bound thiols as wellas sulfane sulfur in different age groups of rats. Acta Biochim. Pol. 2004, 51(3), 815-824.
• 46. Kuśmierek K., Głowacki R., Bald E.: Analysis of urine for cysteine, cysteinylglycine, and homocysteine by high-performance liquid chromatography. Anal. Bioanal. Chem. 2006, 385(5), 855-860 .
• 47. Bald E., Głowacki R.: Analysis of saliva for glutathione and metabolically related thiols by liquid chromatography with ultraviolet detection. Amino Acids. 2005,28(4), 431-433.
• 48. McMenamin M.E., Himmelfarb J., Nolin T.: Analysis of multiple aminothiols in human plasma by high performance liquid chromatography with fluorescence detection. J. Chromatogr. B. 2009, 877(28), 3274-3281.
• 49. Tang D., Wen L., Santschi R: Analysis of biogenic thiols in natural water samples by high-performance liquid chromatographic separation and fluorescence detection with ammonium 7-fluorobenzo-2-oxa-l ,3-diazole-4-sulfonate (SBD-F). Anal. Chim. Acta. 2000, 408(1-2), 299-307.
• 50. Fermo I., Paroni R.: Total plasma homocysteine analysis by HPLC with SBD-F precolumn derivatization. Methods Mol. Biol. 2000,159, 237-244.
• 51. Toyo'oka T: Analysis of thiols. J. Chromatogr. B. 2009, 877 (28), 3318-3330.
• 52. Fujikawa Y., Urano Y., Komatsu T., Hanaoka K., Kojima H., Terai T, Inoue H., Nagano T.: Design and Synthesis of Highly Sensitive Fluorogenic Substrates for Glutathione S-Transferase and Application for Activity Imaging in Living Cells. J. Am. Chem. Soc. 2008, 130, 14533-14543.
• 53. Wintner E.A., Deckwerth T.L., Langston W., Bengtsson A., Leviten D., Hill R, Insko M.A., Dumpit R., VandenEkart E., Toombs C.F., Szabo C.: A monobromobimane-based assay to measure the pharmacokinetic profile of reactive sulphide species in blood. Br. J. Pharm. 2010, 160(4), 941-957.
• 54. Conlan X.A., Stupka N., McDermott G.P., Paul S. Francis P.S., Barnett N.W.: Determination of intracellular glutathione and cysteine using HPLC with a monolithic column after derivatization with monobromobimane. Biomed. Chromatogr. 2010, 24(5), 455-457.
• 55. Sakhi A.K., Berg T.J.: Reduced glutathione concentrations are not decreased in red blood cells of patients with long term type I -diabetes. Scand. J. Clin. Lab. Invest. 2011,71(2), 108-111.
• 56. Kimura Y., Goto Y., Kimura H.: Hydrogen sulfide increases glutathione production and suppresses oxidative stress in mitochondria. Antioxid. Redox Sign. 2010, 12(1), 1-13.
• 57. Djurhuus R., Nossum V., Lundsett N., Hovin W., Svardal A.M., Havnes M.B., Fismen.L., Hjelde A., Brubak A.O.: Simulated diving after heat stress potentiates the induction of heat shock protein 70 and elevates glutathione in human endothelial cells. Cell Stress Chaperones.2010, 15(4), 405-414.
• 58. Hilgier W., Węgrzynowicz M., Ruszkiewicz J., Oja S.S., Saransaari R, Albrecht J.: Direct Exposure to Ammonia and Hyperammonemia Increase the Extracellular Accumulation and Degradation of Astroglia-Derived Glutathione in the Rat Prefrontal Cortex. Toxicol. Sci. 2010, 117(1), 163-168.
• 59. Zu Y.: Molecular and Nanoparticle Postcolumn Reagents for Assay of Low-Molecular-Mass Biothiols using High-Performance Liquid Chromatography. J. Chromatogr. B. 2009, 877, 3358-3365.
• 60. Senft A.P., Dalton T.P., Shertzer H.G.: Determining Glutathione and Glutathione Disulfide Using the Fluorescence Probe o-Phthalaldehyde. Anal. Biochem. 2000, 280, 80-86.
• 61. Cereser C., Guichard J., Drai J., Bannier E., Garcia I., Boget S., Parvaz R, Revol A.: Quantitation of reduced and total glutathione at the femtomole level by high-performance liquid chromatography with fluorescence detection: application to red blood cells and cultured fibroblasts. J. Chromatogr. B. 2001, 752(1), 123-132.
• 62. Leroy P., Nicolas A., Wellmann M., Michelet F, Oster T., Siest G.: Evaluation o-phtholaldehyde as bifunctional fluorogenic post-column reagent for glutathione in LC. Chromatographia. 1993, 36, 130-134.
• 63. Parmentier C., Leroy P., Wellman M., Nicolas A.: Determination of cellular thiols and glutathione-related enzyme activities: versatility of high-performance liquid chromatography-spectrofluorimetric detection. J. Chromatogr. B 1998,719(1-2), 37-46.
• 64. Böhmer A., Jordan J., Tsikas D.: High-performance liquid chromatography ultraviolet assay for humanerythrocytic catalase activity by measuring glutathione as o phthalaldehyde derivative. Anal. Biochem. 201 1,410 (2), 296-303.
• 65. Musenga A., Mandrioli R., Bonifazi P., Kenndler E., Pompei A., Raggi M.A.: Sensitive and selective determination of glutathione in probiotic bacteria by capillary electrophoresis-laser induced fluorescence. Anal. Bioanal. Chem. 2007, 387(3), 917-924.
• 66. Landino L.M., Moynihan K.L., Todd J.V., Kennett K.L.: Modulation of the redox state of tubulin by the glutathione/glutaredoxin reductase system. Biochem. Biophys. Res. Commun. 2004, 314(2), 555-560.
• 67. Wang Y., Xie Y., Bernier M., Wainer I.W.: Determination of free and protein-bound glutathione in HepG2 cells using capillary electrophoresis with laser-induced fluorescence detection. J. Chromatogr. A. 2009, 1216 (16), 3533-3537.
• 68. Zinellu A., Sotgia S., Posadino A.M., Pasciu V., Zinellu E., Usai M.F., Scanu B., Chessa R., Gaspa L., Tadolini B., Deiana L., Carru C.: Protein-bound glutathione measurement in cultured cells by CZE with LIF detection. Electrophoresis. 2007, 28(18), 3277-3283.
• 69. Caussé E., Malatray P., Calaf R., Charpiot P., Candito M., Bayle C., Valdiguié P., Salvayrel R., Couderc F.: Plasma total homocysteine and other thiols analyzed by capillary electrophoresis/laser-induced fluorescence detection: Comparison with two other methods. Electrophoresis. 2000, 28(18), 3277-3283.
• 70. Araujo A., Saraiva, M., Lima, J.: Determination of total and oxidized glutathione in human whole blood with a sequential injection analysis system. Talanta. 2008,74(5), 1511-1519.