Optimization of determination of reduced and oxidized glutathione in rat striatum by HPLC method with fluorescence detection and pre-column derivatization

Czasopismo : Acta Chromatographica
Tytuł artykułu : Optimization of determination of reduced and oxidized glutathione in rat striatum by HPLC method with fluorescence detection and pre-column derivatization

Autorzy :
Mansoor, S.
Georgia Institute of Technology School of Chemical and Biomolecular Engineering Atlanta GA 30332-0100 USA,
Tas, C.
Georgia Institute of Technology School of Chemical and Biomolecular Engineering Atlanta GA 30332-0100 USA, ctas@gata.edu.tr,
Patel, D. S.
Pramukh Swami Science and H.D. Patel Arts College Chemistry Department Sarva Vidyalaya Campus Kadi 382 715 Gujarat India,
Sharma, N.
BA Research India Ltd. Bio-Analytical Laboratory Bodakdev, Ahmedabad 380054 Gujarat India,
Patel, M. C.
Pramukh Swami Science and H.D. Patel Arts College Chemistry Department Sarva Vidyalaya Campus Kadi 382 715 Gujarat India,
Patel, B. N.
BA Research India Ltd. Bio-Analytical Laboratory Bodakdev, Ahmedabad 380054 Gujarat India, bhavinpatel27@rediffmail.com,
Shrivastav, P. S.
Gujarat University Department of Chemistry, School of Sciences Navrangpura, Ahmedabad 380009 Gujarat India, pranav_shrivastav@yahoo.com,
Sanyal, M.
St. Xavier’s College Department of Chemistry Navrangpura, Ahmedabad 380009 Gujarat India,
Wang, G. Y.
Chinese Academy of Sciences Key Laboratory of Chemistry of Northwestern Plant Resources and Key Laboratory for Natural Medicine of Gansu Province, Lanzhou Institute of Chemical Physics Lanzhou 730000 P. R. China,
Shi, Y. P.
Lanzhou University of Technology School of Petroleum and Chemical Engineering Lanzhou 730050 P. R. China, shiyp@licp.cas.cn,
Akowuah, G. A.
UCSI University Faculty of Applied Sciences No. 1, Jalan Menara Gading, UCSI Heights 56000 Kuala Lumpur Malaysia, wuahmy@yahoo.com,
Okechukwu, P. N.
UCSI University Faculty of Pharmaceutical Sciences No. 1, Jalan Menara Gading, UCSI Heights 56000 Kuala Lumpur Malaysia,
Chiam, N. C.
UCSI University Faculty of Pharmaceutical Sciences No. 1, Jalan Menara Gading, UCSI Heights 56000 Kuala Lumpur Malaysia,
Raszewski, G.
Institute of Agricultural Medicine Department of Pathophysiology Lublin Poland, grasz@galen.imw.lublin.pl,
Juszczak, M.
Institute of Agricultural Medicine Department of Medical Biology Lublin Poland,
Lemieszek, M. K.
Institute of Agricultural Medicine Department of Medical Biology Lublin Poland,
Matysiak, J.
University of Life Sciences Department of Chemistry Lublin Poland,
Niewiadomy, A.
University of Life Sciences Department of Chemistry Lublin Poland,
Rzeski, W.
Maria Curie-Sklodowska University Department of Virology and Immunology, Institute of Microbiology and Biotechnology Lublin Poland,
Reddy, P. S.
IPDO Dr. Reddy’s Laboratories Ltd. Bachupally, Hyderabad 500072 A.P India, sunilpsr@yahoo.com,
Babu, K. S.
S. K. University Department of Chemistry Anantapur 515055 A.P. India,
Kumar, N.
IPDO Dr. Reddy’s Laboratories Ltd. Bachupally, Hyderabad 500072 A.P India,
Pareek, D.
Rajiv Gandhi Technological University School of Pharmaceutical Sciences Bhopal 462033 Madhya Pradesh India,
Jain, S.
Sagar Institute of Research and Technology Ayodhya bypass, Bhopal 462 041 MP India,
Basniwa, P. K.
Rajiv Gandhi Technological University School of Pharmaceutical Sciences Bhopal 462033 Madhya Pradesh India,
Jain, D.
Rajiv Gandhi Technological University School of Pharmaceutical Sciences Bhopal 462033 Madhya Pradesh India, deepti2515@yahoo.com,
Wróblewski, K.
Medical University of Lublin Department of Inorganic Chemistry 4a Chodźki Str. 20-093 Lublin Poland,
Petruczynik, A.
Medical University of Lublin Department of Inorganic Chemistry 4a Chodźki Str. 20-093 Lublin Poland,
Waksmundzka-Hajnos, M.
Medical University of Lublin Department of Inorganic Chemistry 4a Chodźki Str. 20-093 Lublin Poland, monika.hajnos@umlub.pl,
Jain, P. S.
R. C. Patel Institute of Pharmaceutical Education and Research Karwand Naka, Shirpur, Dist. Dhule 425 405 MS India, pritash79@yahoo.com,
Chaudhari, A. J.
R. C. Patel Institute of Pharmaceutical Education and Research Karwand Naka, Shirpur, Dist. Dhule 425 405 MS India,
Surana, S. J.
R. C. Patel Institute of Pharmaceutical Education and Research Karwand Naka, Shirpur, Dist. Dhule 425 405 MS India,
Elgorashe, R. E. E.
Sudan University of Science and Technology Department of Chemistry, College of Science Khartoum Sudan,
Idris, A. M.
King Khalid University Department of Chemistry, College of Science Abha Saudi Arabia, abubakridris@hotmail.com,
Abdelrahman, M. A.
Sudan University of Science and Technology Department of Chemistry, College of Science Khartoum Sudan,
Saeed, A. E. M.
Sudan University of Science and Technology Department of Chemistry, College of Science Khartoum Sudan,
Gawlik, M.
Jagiellonian University Medical College Department of Toxicology, Faculty of Pharmacy Medyczna str. 9 30-688 Kraków Poland,
Krzyżanowska, W.
Jagiellonian University Medical College Department of Toxicology, Faculty of Pharmacy Medyczna str. 9 30-688 Kraków Poland,
Gawlik, M. B.
Jagiellonian University Medical College Department of Toxicology, Faculty of Pharmacy Medyczna str. 9 30-688 Kraków Poland, mfgawlik@cyf-kr.edu.pl,
Filip, M.
Jagiellonian University Medical College Department of Toxicology, Faculty of Pharmacy Medyczna str. 9 30-688 Kraków Poland,
Abstrakty : A high-performance liquid chromatography (HPLC) method was used to assess the concentration of reduced and oxidized glutathione (GSH and GSSG) in rat striatum. Following decapitation, striatum was isolated from the male Wistar rat brain and immediately homogenized with double distilled water. GSH level was determined after pre-column derivatization with o-phthalaldehyde (OPA). The optimal incubation time with OPA was tested. The concentration of GSSG was determined after blockage the thiol groups of GSH by N-ethylmaleimide (NEM). The useful time for incubation with NEM was optimized. Next, disulfide bounds of GSSG were reduced by dithiothreitol (DTT), and released GSH is derivatized with OPA. The total glutathione, tGSH (sum of free and bound GSH, GSSG, and other low-molecular-mass aminothiols), was determined after reduction with DTT and then derivatization with OPA. The level of GSSG was calculated of the difference in concentrations of tGSH and GSH, but we showed that the calculated concentration of GSSG was within the range of standard deviation of the mean concentration of tGSH or GSH. Finally, the concentration of GSH was determined after 5-min incubation with OPA and the concentration of GSSG after 30-s incubation with NEM and 5-min incubation with DTT and OPA. The relative standard deviation (RSD) values obtained for the assay of GSH and GSSG were lower than 10%. The values obtained for accuracy for GSH (50–500 nM) and GSSG (0.5–5 nM) were within limits regarded as acceptable for analysis of biological samples (percent of recovery: 95–105%). Mean absolute recovery of GSH and GSSG was ranged from 97.1% to 99%. Limit of detection for GSH was 2.7 nM, and limit of quantification was 8.2 nM. Limit of detection (LOD) for GSSG is twice the value for GSH. Described method allows to determine GSH and GSSG levels in isolated rat brain structures with high level of reliability.

Słowa kluczowe : reduced glutathione, oxidized glutathione, HPLC method, rat striatum,
Wydawnictwo : University of Silesia in Katowice
Rocznik : 2014
Numer : Vol. 26, no. 2
Strony : 335 – 345
Bibliografia : 1 I. Dalle-Donne, R. Rossi, G. Colombo, D. Giustarini, and A. Milzani, Trends Biochem. Sci., 34, 85 (2009)
2 G. Wu, Y.Z. Fang, S. Yang, J.R. Lupton, and N.D. Turner, J. Nutr., 134, 489 (2004)
3 M.E. Anderson, Chem. Biol. Interact., 11, 1 (1998)
4 K. Aoyama, M. Watable, and T. Nakaki, J. Pharmacol. Sci., 108, 227 (2008)
5 D.S. Backos, C.C. Franklin, and P. Reigan, Biol. Pharm., 83, 1005 (2012)
6 M. Raffa, F. Atig, A. Mhalla, A. Kerken, and A. Mechri, Psychiatry, 11, 12 (2011)
7 J.W. Gawryluk, J.F. Wang, A.C. Andreazza, L. Shao, and L.T. Young, Int. J. Neuropsychopharmacol., 14, 123 (2011)
8 P.K. Mandal, M. Tripathi, and S. Sugunan, Biochem. Biophys. Res. Commun. (2011) Nov 19. Epub ahead of print
9 K. Aquilano, S. Baldelli, and M.R. Ciriolo, Commun. Integr. Biol., 4, 477 (2011)
10 Y. Ma, Z. Zhang, C. Ren, G. Liu, and X. Chen, Analyst, 137, 485 (2012)
11 S. Robin, N. Leveque, C. Courderot-Masuye, and P. Humbert, J. Chromatogr., B: Anal. Technol. Biomed. Life Sci., 879, 3599 (2011)
12 A.A. Dadkhah-Tehrani and H. Karimi-Maleh, Drug Test Anal. (2011). doi: 10.1002/dta.347
13 K.J. Huang, Q.S. Jing, C.Y. Wei, and Y.Y. Wu, Spectrochim. Acta A. Mol. Biomol. Spectrosc., 79, 1860 (2011)
14 P. Monostori, G. Wittmann, E. Karg, and S. Túri, J. Chromatogr. B, 877, 3331 (2009)
15 C. Cereser, J. Guichard, J. Drai, E. Bannier, I. Garcia, S. Boget, P. Parvaz, and A. Revol, J. Chromatogr. B, 752, 123 (2001)
16 K.J. Lenton, H. Therriault, and J.R. Wagner, Anal. Biochem., 274, 125 (1999)
17 L. Wang, I. Wang, T. Xia, G. Bian, L. Dong, Z. Tang, and F. Wang, Spectrochim. Acta A. Mol. Biomol. Spectrosc., 61, 2533 (2005)
18 A.K. Sakhi, R. Blomhoff, and T.E. Gundersen, J. Chromatogr. A, 1142, 178 (2007)
19 D. Giustarini, I. Dalle-Donne, R. Colombo, A. Milizani, and R. Rossi, Free Radic. Biol. Med., 35, 1365 (2003)
20 R. Paroni, E. De Vecchi, G. Cighetti, C. Arcelloni, I. Fermo, A. Grosii, and P. Bonini, Clin. Chem., 41/3, 448 (1995)
21 R. Rossi, I. Dalle-Donne, A. Milzani, and D. Giustarini, Clin. Chem., 52, 1406 (2006)
22 B. Ates, B.C. Ercal, K. Manda, L. Abraham, and N. Ercal, Biomed. Chromatogr., 23, 119 (2009)
23 B.H. Harvey, C. Joubert, J.L. du Preez, and M. Berk, Neurochem. Res., 33, 508 (2008)
24 P.J. Hissin and R. Hilf, Anal. Biochem., 74, 214 (1976)
25 V.H. Cohn and J. Lyle, Anal. Biochem., 14, 434 (1966)
26 O.H. Lowry, N.J. Rosebrough, A.L. Farr, and R.J. Randall, J. Biol. Chem., 193, 265 (1951)
27 Y. Iwasaki, Y. Saito, Y. Nakano, K. Mochizuki, O. Sakata, R. Ito, K. Saito, and H. Nakazawa, J. Chromatogr. B., 877, 3309 (2009)
DOI :
Cytuj : Mansoor, S. ,Tas, C. ,Patel, D. S. ,Sharma, N. ,Patel, M. C. ,Patel, B. N. ,Shrivastav, P. S. ,Sanyal, M. ,Wang, G. Y. ,Shi, Y. P. ,Akowuah, G. A. ,Okechukwu, P. N. ,Chiam, N. C. ,Raszewski, G. ,Juszczak, M. ,Lemieszek, M. K. ,Matysiak, J. ,Niewiadomy, A. ,Rzeski, W. ,Reddy, P. S. ,Babu, K. S. ,Kumar, N. ,Pareek, D. ,Jain, S. ,Basniwa, P. K. ,Jain, D. ,Wróblewski, K. ,Petruczynik, A. ,Waksmundzka-Hajnos, M. ,Jain, P. S. ,Chaudhari, A. J. ,Surana, S. J. ,Elgorashe, R. E. E. ,Idris, A. M. ,Abdelrahman, M. A. ,Saeed, A. E. M. ,Gawlik, M. ,Krzyżanowska, W. ,Gawlik, M. B. ,Filip, M. , Optimization of determination of reduced and oxidized glutathione in rat striatum by HPLC method with fluorescence detection and pre-column derivatization. Acta Chromatographica Vol. 26, no. 2/2014
facebook