CAPILLARY ELECTROPHORESIS METHOD FOR THE DETERMINATION OF LACTULOSE AND MANNITOL RATIO FOR ESTIMATING INTESTINAL PERMEABILITY

Bu çalışmada intestinal geçirgenliğin incelenmesi amacıyla idrarda laktuloz ve mannitol tayini için basit, hızlı ve doğru bir kapiler elektroforez yöntemi geliştirilmiştir. İdrar örnekleri katı faz özütleme yöntemi ile analize hazırlanmıştır. Analizler için 6 mmol/L sorbik asit, 1.25 mmol/L hekzadesil trimetil amonyum bromür (HDTMA), 50 mmol/ L LiOH içeren pH sı 12.5 e karşılık gelen destek elektrolit kullanılmıştır. Ayrım 45 cm 50 µm iç çaplı fused silika kolonda 20 °C sıcaklıkta -5 kV ayırma potansiyeli kullanılarak 20 dk içerisinde gerçekleştirilmiştir. Geliştirilen yöntem laktuloz / mannitol derişim oranlarını ölçmek için bir seri idrar numunesine uygulanmıştır.

CAPILLARY ELECTROPHORESIS METHOD FOR THE DETERMINATION OF LACTULOSE AND MANNITOL RATIO FOR ESTIMATING INTESTINAL PERMEABILITY

In this study, a simple, rapid and accurate capillary electrophoresis method has been developed for the determination of lactulose and mannitol in urine for estimating the intestinal permeability. The urine samples were prepared to analyze by using solid phase extraction. For analysis a background electrolyte (BGE) consisted of 6 mmol/L sorbic acid, 1.25mmol/L hexadecyltrimeth ylammoniumbromide (HDTMA), 50 mmol/L LiOH (pH 12.5) was used. Separation was performed at -5 kV over 20 min within the fused capillary column (45 cm, 50 µm I.D.) at 20 °C. The method developed was applied a series of urine sample to measure the lactulose/mannitol concentration rate.

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  • 1. I. Bjarnason, Intestinal permeability, Gut, Sup I, S18-S22,(1994).
  • 2. Renata D’Inca, M.D., Vincenza Di Leo, M.D., Giovanni Corrao, M.D., Diego Martines, M.D., Anna D’Odorico, M.D., Cinzia Mestriner, M.D., Carla Venturi, B.S., Guiseppo Longo, M.D., and Giacomo Carlo Sturniolo, M.D., Intestinal Permeability Test as a Predictor of Clinical Course in Crohn’s Disease, The American Journal of Gastroenterology, Vol. 94, No. 10, 2953, (1999).
  • 3. R. Paroni, I. Fermo, L. Molteni, L. Folini, M.R. Pastore, A. Mosca, E. Bosi, Lactulose and mannitol intestinal permeability detected by capillary electrophoresis, Journal of Chromatography B, 834, 183-187, (2006).
  • 4. M.S. Barboza Jr., T.M.J. Silva, R.L. Guerrant, A.A.M. Lima, Measurement of intestinal permeability using mannitol and lactulose in children with diarrheal diseases, Brazilian Journal of Medical and Biological Research, 32, 1499-1532, (1999).
  • 5. S.C. Fleming, M.S. Kapembwa, M.F. Laker, G.E. Levin, G.E. Griffin, Rapid and Simultaneous determination of lactulose and mannitol in urine by HPLC with pulsed amperometric detection for use in studies of intestinal permeability, Clin. Chem. 36/5, 797 – 799, (1990).
  • 6. T.H. Iqbal, K. O. Lewis, J.C. Gearty, B.T. Cooper, Small intestinal permeability to mannitol and lactulose in the three ethnic groups resident in West Birmingham, Gut, 39, 199- 203, (1996).
  • 7. Cox MA, Lewis KO, Cooper BT., Measurement of small intestinal permeability markers, lactulose, and mannitol in serum: results in celiac disease, Dig Dis Sci. Feb;44(2):402-6, (1999).
  • 8. Marsilio R, D’Antiga L, Zancan L, Dussini N, Zacchello F., Simultaneous HPLC determination with light-scattering detection of lactulose and mannitol in studies of intestinal permeability in pediatrics, Clin Chem. Aug;44(8 Pt 1):1685-91,(1998).
  • 9. M. Celli, P. D’Eufemia, R. Dommarco, R. Finocchiaro, D. Aprigliano, F. Martino, E. Cardi, O. Giardini, Rapid gas – chromtaographic assay of lactulose and mannitol for estimating intestinal permeability, Clin. Chem., 41/5, 752-756, (1995).
  • 10. Menzies IS. Absorption of intact oligosaccharide in health and disease. Biochem Soc Trans., 2: 1042-1047, (1974).
  • 11. Vilela EG, Torres HO, Ferrari ML, Lima AS, Cunha AS. Gut permeability to lactulose and mannitol differs in treated Crohn’s disease and celiac disease patients and healthy subjects. Braz J Med Biol Res. Dec;41(12):1105-9, (2008).
  • 12. A. Hayırlı, H.D. Umucalılar, N. Gulsen, O.B. Citil, Role of lactulose as a modifier in rumen fermentation, Journal of Animal and Veterinary Advances, 9 (19), 2537 – 2545, (2010).
  • 13. G. Chen, L. Zhang, X. Wu, J. Ye, Determination of mannitol and three sugars in Ligustrum lucidum Ait. By capillary electrophoresis with electrochemical detection, Analytica Chimica Acta, 530, 15-21, (2005)