İdrarda Orotik Asit’in Temassız İletkenlik Dedektörü Kullanılarak Kapiler Elektroforez Yöntemi İle Hızlı Tayini

Year 2020, Volume: 15 Issue: 2, 194 - 202, 29.11.2020

Mehmet Şerif Cansever , Nevin Öztekin , Veselina Adımcılar , Tanyel Zübarioğlu , Ertuğrul Kıykım , Ayşe Aktuğlu

https://doi.org/10.29233/sdufeffd.695869

Abstract

İdrarda orotik asidin aşırı atılımı üre döngüsü bozukluğu ve pirimidin sentez bozukluğu gibi doğumsal metabolik hastalıklarda görülür. Bu nedenle orotik asidin hızlı bir şekilde tayini hayati öneme sahiptir. Bu çalışmada idrar örneklerinde orotik asidin kantitatif düzeylerinin belirlenmesi için yeni, basit, ucuz ve hızlı bir kapiler elektroforetik analiz yöntemi geliştirildi. Orotik asidin temassız iletkenlik dedektörü (C4D) kullanılarak kapiler elektroforez yöntemi ile ayrımı için pH’ı 6,5 olan 20 mM 4-Morfolinetansülfonik asit (MES), 10 mM Histidin ve 0,1 mM setil trimetilamonyum bromür (CTAB) içeren bir ça-lışma tamponu ile geliştirildi. Orotik asidin ayrılması yaklaşık 3,5 dakika gibi kısa bir sürede tamamlandı. Göç zamanlarının ve düzeltilmiş pik alanlarının tekrarlanabilirliği sırasıyla %RSD olarak 1,25 ve 4,31 olarak bulundu. En düşük dedeksiyon limiti (LOD) 0,733 mg / L ve en düşük tayin limiti de (LOQ) 2,443 mg / L olarak bulundu. Metodun idrarda orotik asit düzeylerinin tayin edilmesinde kolaylıkla ve hassasiyetle kullanılabileceği gösterildi.

Keywords

Kapiler Elektroforez, Orotik Asit, Temassız İletkenlik Dedektörü (C4D), Üre Döngüsü Bozuklukları, Doğumsal Metabolik Hastalık, İdrar

References

• [1] M. Duran, L. Dorland, E. E. Meuleman, P. Allers, R. Berger, ‘’Inherited defects of purine and pyrimidine metabolism: laboratory methods for diagnosis,’’ J Inherit Metab Dis 20:227–236, 1997.
• [2] M. Loeffler, E. Zameitat, ‘’Pyrimidine biosynthesis’’, Encycl Biol Chem, 3:600–605, 2004
• [3] S. Balasubramaniam, J.A. Duley, J. Christodoulou, ‘’Inborn errors of pyrimidine metabolism: clinical update and therapy,’’ J. Inherit. Metab. Dis, 37(5):687-98, 2014.
• [4] R.E. Kelley, H.C. Andersson, ‘’Disorders of purines and pyrimidines,’’. Handb Clin Neurol., 120:827- 38, 2014.
• [5] J. Häberle, N. Boddaert, A. Burlina, A. Chakrapani, M. Dixon, M. Huemer, D. Karall, D. Martinelli , P.S. Crespo, R: Santer, A. Servais, V. Valayannopoulos, M. Lindner , V. Rubio disorders,’’ Orphanet J Rare Dis, 29;7-32, 2012.
• [6] C. Bachmann, J.P. Colombo. ‘’Diagnostic value of orotic acid excretion in heritable disorders of the urea cycle and in hyperammonemia due to organic acidurias,’’ Eur J Pediatr., 134(2):109-13,1980.
• [7] S.W. Brusilow, E. Hauser. ‘’Simple method of measurement of orotic acid and orotidine in urine,’’ J. Chromatogr., 1;493(2):388-91, 1989.
• [8] M. Löffler , E.A. Carrey , E. Zameitat. ‘’Orotic Acid, More Than Just an Intermediate of Pyrimidine de novo Synthesis’’. J Genet Genomics., 20;42(5):207-19, 2015.
• [9] C. Salerno, C. Crifò, ‘’Diagnostic value of urinary orotic acid levels: applicable separation methods,’ J. Chromatogr. B. Analyt. Technol. Biomed. Life Sci., 781, 57–71, 2002
• [10] S. Yin, S. Dragusha, V. Ejupi, T. Shibata, T. Kabashima, M. Kai, ‘’Sensitive and selectivedetermina-tion of orotic acid in biological specimens using a novel fluorogenicreaction,’’ J. Fluoresc., 25; 1005–1011, 2015
• [11] A. Fioravanti, M. Flaviani, C. Gambelunghe, A. Micheletti, M. Sposito, S. Rufini, ‘’High-performance Liquid Chromatographic Determination of Orotic Acid as its Methyl Derivative in Human Urine’’ Journal of chromatography. B, Biomedical sciences and applications, vol. 703, 263–266, 1997
• [12] B. Tavazzi, G. Lazzarino, P. Leone, A.M. Amorini, F. Bellia, C.G. Janson, et al.,’’Simultaneous high performance liquid chromatographic separation of purines, pyrimidines, N-acetylated amino acids,and dicarboxylic acids for the chemical diagnosis of inborn errors of metabolism,’’ Clin. Biochem. 38, 997–1008, 2005
• [13] M.S. Rashed, M. Jacob, M. Al-Amoudi, Z. Rahbeeni, M.A.D. Al-Sayed, L. Al-Ahaidib, et al., ‘’Rapid determination of orotic acid in urine by liquid chromatography-electrospray tandem mass spectro-metry,’’ Clin. Chem., 49, 499–501, 2003.
• [14] G. la Marca, B. Casetta, E. ‘’Zammarchi, Rapid determination of orotic acid in urineby a fast liquid chromatography/tandem mass spectrometric method’’, Rapid Commun. Mass Spectrom., 17, 788-793, 2003.
• [15] O. D'Apolito, D. Garofalo, G. Paglia, A. Zuppaldi, G. Corso, ‘’Orotic acid quantification in dried blood spots and biological fluids by hydrophilic interaction liquid chromatography tandem mass spectrometry’’, J. Separ. Sci., 33, 966–973,2010.
• [16] R. Hušková, P. Barták, L. Čáp, D. Friedecký, T. Adam, ‘’Analytical derivatization-a tool for deter-mination of orotic acid’’, J. Chromatogr. B Anal. Technol. Biomed. Life Sci. 799, 303–309,2004.
• [17] O. Giardini, C. Salerno, P. D. Eufemia, M. Celli, R. Finocchiaro, C. Crifo, ‘’Determination of urinary orotic acid and uracil by capillary zone electrophoresis’’, J. Chromatogr. B., 734,175–178,1999
• [18] J. Sevcík, T. Adam, V. Sázel, ‘’A rapid and simple screening method for detection of orotic aciduria by capillary zone electrophoresis,’’ Clin. Chim. Acta ,259,73–81, 1997.
• [19] H. Khajehsharifi, H. Tavallali, M. Shekoohi, M. Sadeghi, ‘’Spectrophotometric simultaneous deter-mination of orotic acid, creatinine and uric acid by orthogonal signal correction-partial least squares in spiked real samples’’, Drug Test. Anal., 5, 353–360, 2013.
• [20] J. W. Jorgenson, K. D. Lukacs,. “Zone electrophoresis in open-tubular glass capillaries,” Analytical Chemistry, 53, 1298– 1302,1981
• [21] J. W. Jorgenson, K. D. Lukacs. “Free-zone electrophoresis in glass capillaries,” Clinical Chemistry, 27, 1551-1553,1981.
• [22] J. W. Jorgenson, K. D. Lukacs,. “High-resolution separations based on electrophoresis and electroosmosis,” Journal of Chromatography A, 218, 209-216, 1981.
• [23] S. F. Y., Capillary Electrophoresis. Amsterdam: Elsevier, 1992.
• [24] S. Özden, R. Ertan, E. Akı-Şener, G. Yalçın, Farmasötik Kimya Pratikleri. Ankara. Ankara Üniversi-tesi, Eczacılık Fakültesi, 1-2, 2004.
• [25] P. Kubáň, P. C. Hauser, “Contactless conductivity detection in capillary electrophoresis: A review,” Electroanalysis, 16(24), 2009-2021, 2004.
• [26] A. J. Zemann, E. Schnell, D. Volgger, G. K. Bonn, ‘’Conductivity detection for capillary electrophoresis,” Analytical Chemistry, 70(3), 563-567, 1998.
• [27] H.E. Yelkenci, N. Öztekin, ‘’Separation and Sensitive Detection of Lanthanides by Capillary Elect-rophoresis and Contactless Conductivity Detection,’’ Journal of Chromatographic Science, 55 (4), 465– 470, 2017.
• [28] P. Kubáň, P. C. Hauser, “Review: Capacitively coupled contactless conductivity detection for microseparation techniques-recent developments, ” Electrophoresis, 32, 30-42, 2011.
• [29] P.Kubáň, P. C. Hauser, “Ten years of axial capacitively coupled contactless conductivity detection for CZE - a review,” Electrophoresis, 30, 176-188, 2009.
• [30] P. Kubáň, P. C. Hauser, “A review of the recent achievements in capacitively coupled contactless conductivity detection,” Analytica Chimica Acta, 607(1), 15-29, 2008.
• [31] A. A. Elbashir, H. Y. Aboul-Enein, “Recent advances in applications of capillary electrophoresis with capacitively coupled contactless conductivity detection (CE-C4D): An update,” Biomedical Chro-matography, 26, 990-1000, 2012.