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İdrar Örneklerinde L-2-Hidroksiglutarik Asitin Kapiler Elektroforez-İndirekt UV Dedeksiyon ile Hızlı Tayini

Yıl 2020, Cilt: 8 Sayı: 2, 234 - 270, 23.08.2020
https://doi.org/10.37696/nkmj.687468

Öz

Amaç: L-2 hidroksiglutarik asidüri (L2HGA) otozomal resesif geçişli, psikomotor gerilik, serebellar ataksi ve değişken makrosefali veya epilepsi ile karakterize edilen nadir görülen nörömetabolik bir hastalıktır. Hastalık biyokimyasal olarak idrarda, beyin omurilik sıvısında ve çok az miktarda plazmada artmış L-2HGA seviyeleri ile tanımlanır. Bu çalışmada, idrarda L-2-hidroksiglutarik asitin tayini için hızlı ve hassas bir kapiler elektroforez yöntemi geliştirilmiştir.
Materyal ve Metot: L-2-hidroksiglutarik asit kromofor gruba sahip olmadığı için, indirekt UV deteksiyon metodu uygulanmıştır. Çalışma tamponu pH 5.6’da 0.1 mmol/L setiltrimetilamonyum bromür (CTAB) içeren 10 mmol/L 2,6-piridindikarboksilik asitten (PDC) oluşmaktadır. PDC iyi bir kromoforik özellik göstermesinden dolayı indirekt deteksiyon için uygundur. CTAB elektro-osmotik akışı (EOF) ters çevirmek için kullanılmıştır.
Bulgular: L-2-hidroksiglutarik asit için tayin limiti 1.16 μmol/L’dir. Geliş zamanı ise 2.0 dakikadan daha az bir süredir. Geliştirilen metod 16 L-2-hidroksiglutarik asidüri hastasından ve 4 sağlıklı gönüllüden toplanan idrar örneklerine uygulanmıştır.
Sonuç: Hasta bireylerin idrarındaki L-2-hidroksiglutarik asitin konsantrasyonu 48.9 – 1,15 µg/mL arasında bulunmuştur.

Kaynakça

  • 1. Duran M, Kamerling JP, Bakker HD, van Gennip AH, Wadman SK. L-2-Hydroxyglutaric aciduria: an inborn error of metabolism?. J. Inherit. Metab. Dis. 1980; 3:109-112. 2. Hoffmann GF, Kölker S. Cerebral organic acid disorders and other disorders of lysine catabolism. In: Saudubray JM, Baumgartner MR, Walter J (eds) Inborn Metabolic Diseases: Diagnosis and Treatment, 6th edn. Springer-Verlag, Heidelberg, 2016 pp. 333-348. 3. Steenweg ME, Salomons GS, Yapici Z, Uziel G, Scalais E, Zafeiriou DI, et al. L-2-Hydroxyglutaric aciduria: pattern of MR imaging abnormalities in 56 patients. Radiology 2009; 251:856-865. 4. Samuraki M, Komai K, Hasegawa Y, Kimura M, Yamaguchi S, Terada N, Yamada M. A successfully treated adult patient with L-2-hydroxyglutaric aciduria. Neurology 2008; 70:1051–1052. 5. Yilmaz K. Riboflavin treatment in a case with l-2-hydroxyglutaric aciduria. Eur J Paediatr Neurol 2009; 13:57–60. 6. Rashed MS, AlAmoudi M, Aboul-Enein HY. Chiral liquid chromatography tandem mass spectrometry in the determination of the configuration of 2-hydroxyglutaric acid in urine. Biomed Chromatogr 2000; 14:317-320. 7. Gibson KM, Brink HJT, Schor DS, Kok RM, Bootsma AH, Hoffmann GF, Jakobs C. Stable-isotope dilution analysis of D- and L-2-hydroxyglutaric acid: Application to the detection and prenatal diagnosis of D- and L-2-hydroxyglutaric acidemias. Pediatr Res 1993; 34:277-280. 8. Kaunzinger A, Rechner A, Beck T, Mosandl A, Sewell AC, Bohles H. Chiral compounds as indicators of inherited metabolic disease: Simultaneous stereodifferentiation of lactic-, 2-hydroxyglutaric- and glyceric acid by enantioselective cGC. Enantiomer 1996; 1:177-182. 9. Bal D, Gryff-Keller A. 1H and 13C NMR study of 2 hydroxyglutaric acid and its lactone. Magn Reson Chem 2002; 40:533-536. 10. Barbot C, Fineza I, Diogo L, Maia M, Melo J, Guimaraes A, Pires MM, Cardoso ML, Vilarinho L. L-2-Hydroxyglutaric aciduria: Clinical, biochemical and magnetic resonance imaging in six Portuguese pediatric patients. Brain Dev 1997; 19:268-273. 11. Soga T, Ross GA. Capillary electrophoretic determination of inorganic and organic anions using 2,6-pyridinedicarboxylic acid: effect of electrolyte's complexing ability, J Chromatogr A, 1997; 767:223-230. 12. Tezcan F, Kolaylı S, Şahin H, Ulusoy E, Erim FB. Evaluation of organic acid, saccharide composition and antioxidant properties of some authentic Turkish honeys. J Food Nutr Res 2011; 50:33-40. 13. Kalaycıoğlu Z, Kaygusuz H, Döker S, Kolaylı S, Erim FB. Characterization of Turkish honeybee pollens by principal componentanalysis based on their individual organic acids, sugars, minerals, andantioxidant activities. LWT-Food Sci Techno 2017; 84:402-408. 14. Nutku MS, Erim FB. Polyethyleneimine-coated capillary electrophoresis capillaries for the analysis of organic acids with an application to beverages. J Microcolumn Sep 1999; 11:541-543. 15. Tezcan F, Gültekin-Özgüven M, Diken T, Özçelik B, Erim FB. Antioxidant activity and total phenolic, organic acid and sugar content in commercial pomegranate juices. Food Chem. 2009; 115:873-877. 16. EURACHEM Guide 1998. The fitness for purpose of analytical methods: A laboratory guide to method validation and related topics. Available at http://eurachem.org/.

RAPID DETERMINATION OF L-2-HYDROXYGLUTARIC ACID IN URINE SAMPLES BY CAPILLARY ELECTROPHORESIS WITH INDIRECT UV DETECTION

Yıl 2020, Cilt: 8 Sayı: 2, 234 - 270, 23.08.2020
https://doi.org/10.37696/nkmj.687468

Öz

Aim: L-2-hydroxyglutaric aciduria (L2HGA) which is autosomal recessive and characterized by psychomotor retardation, cerebellar ataxia, variable macrocephaly, and epilepsy is a rarely seen neurometabolic disease. The disease is biochemically identified by slightly increased L-2HGA levels in urine, cerebrospinal fluid, and plasma. A sensitive and rapid capillary electrophoretic technique was used for the determination of L2HGA in urine.
Materials and Method: Since the L2HGA lacks of chromophore, an indirect UV detection method was applied. The running electrolyte consisted of 10 mmol/L 2,6-pyridinedicarboxylic acid (PDC) at pH 5.6 containing 0.1 mmol/L of cetyltrimethylammonium bromide (CTAB). PDC was chosen because of its good chromophoric property for indirect UV detection. CTAB was used as electro-osmotic flow (EOF) reversal.
Results: The method was well-validated in the selected conditions. The detection limit of the method L2HGA was 1.16 μmol/L. The migration time of L2HGA was less than 2.0 min. The developed method was applied to the urine samples collected from 16 patients who suffer from L-2-hydroxyglutaric aciduria and 4 healthy volunteers.
Conclusion: L-2-hydoxyglutaric acid concentration was found between 48.9 – 1,15 µg/mL in patients.

Kaynakça

  • 1. Duran M, Kamerling JP, Bakker HD, van Gennip AH, Wadman SK. L-2-Hydroxyglutaric aciduria: an inborn error of metabolism?. J. Inherit. Metab. Dis. 1980; 3:109-112. 2. Hoffmann GF, Kölker S. Cerebral organic acid disorders and other disorders of lysine catabolism. In: Saudubray JM, Baumgartner MR, Walter J (eds) Inborn Metabolic Diseases: Diagnosis and Treatment, 6th edn. Springer-Verlag, Heidelberg, 2016 pp. 333-348. 3. Steenweg ME, Salomons GS, Yapici Z, Uziel G, Scalais E, Zafeiriou DI, et al. L-2-Hydroxyglutaric aciduria: pattern of MR imaging abnormalities in 56 patients. Radiology 2009; 251:856-865. 4. Samuraki M, Komai K, Hasegawa Y, Kimura M, Yamaguchi S, Terada N, Yamada M. A successfully treated adult patient with L-2-hydroxyglutaric aciduria. Neurology 2008; 70:1051–1052. 5. Yilmaz K. Riboflavin treatment in a case with l-2-hydroxyglutaric aciduria. Eur J Paediatr Neurol 2009; 13:57–60. 6. Rashed MS, AlAmoudi M, Aboul-Enein HY. Chiral liquid chromatography tandem mass spectrometry in the determination of the configuration of 2-hydroxyglutaric acid in urine. Biomed Chromatogr 2000; 14:317-320. 7. Gibson KM, Brink HJT, Schor DS, Kok RM, Bootsma AH, Hoffmann GF, Jakobs C. Stable-isotope dilution analysis of D- and L-2-hydroxyglutaric acid: Application to the detection and prenatal diagnosis of D- and L-2-hydroxyglutaric acidemias. Pediatr Res 1993; 34:277-280. 8. Kaunzinger A, Rechner A, Beck T, Mosandl A, Sewell AC, Bohles H. Chiral compounds as indicators of inherited metabolic disease: Simultaneous stereodifferentiation of lactic-, 2-hydroxyglutaric- and glyceric acid by enantioselective cGC. Enantiomer 1996; 1:177-182. 9. Bal D, Gryff-Keller A. 1H and 13C NMR study of 2 hydroxyglutaric acid and its lactone. Magn Reson Chem 2002; 40:533-536. 10. Barbot C, Fineza I, Diogo L, Maia M, Melo J, Guimaraes A, Pires MM, Cardoso ML, Vilarinho L. L-2-Hydroxyglutaric aciduria: Clinical, biochemical and magnetic resonance imaging in six Portuguese pediatric patients. Brain Dev 1997; 19:268-273. 11. Soga T, Ross GA. Capillary electrophoretic determination of inorganic and organic anions using 2,6-pyridinedicarboxylic acid: effect of electrolyte's complexing ability, J Chromatogr A, 1997; 767:223-230. 12. Tezcan F, Kolaylı S, Şahin H, Ulusoy E, Erim FB. Evaluation of organic acid, saccharide composition and antioxidant properties of some authentic Turkish honeys. J Food Nutr Res 2011; 50:33-40. 13. Kalaycıoğlu Z, Kaygusuz H, Döker S, Kolaylı S, Erim FB. Characterization of Turkish honeybee pollens by principal componentanalysis based on their individual organic acids, sugars, minerals, andantioxidant activities. LWT-Food Sci Techno 2017; 84:402-408. 14. Nutku MS, Erim FB. Polyethyleneimine-coated capillary electrophoresis capillaries for the analysis of organic acids with an application to beverages. J Microcolumn Sep 1999; 11:541-543. 15. Tezcan F, Gültekin-Özgüven M, Diken T, Özçelik B, Erim FB. Antioxidant activity and total phenolic, organic acid and sugar content in commercial pomegranate juices. Food Chem. 2009; 115:873-877. 16. EURACHEM Guide 1998. The fitness for purpose of analytical methods: A laboratory guide to method validation and related topics. Available at http://eurachem.org/.
Toplam 1 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Konular Klinik Tıp Bilimleri
Bölüm Araştırma Makalesi
Yazarlar

Mehmet Şerif Cansever 0000-0003-3315-5124

Zeynep Kalaycıoğlu 0000-0002-0967-0997

Tanyel Zübarioğlu 0000-0002-7159-4008

Ertuğrul Kıykım Bu kişi benim 0000-0001-5170-5594

Ayşe Aktuğlu Bu kişi benim 0000-0001-7256-0750

Bedia Berker 0000-0001-9406-6681

Yayımlanma Tarihi 23 Ağustos 2020
Yayımlandığı Sayı Yıl 2020 Cilt: 8 Sayı: 2

Kaynak Göster

APA Cansever, M. Ş., Kalaycıoğlu, Z., Zübarioğlu, T., Kıykım, E., vd. (2020). RAPID DETERMINATION OF L-2-HYDROXYGLUTARIC ACID IN URINE SAMPLES BY CAPILLARY ELECTROPHORESIS WITH INDIRECT UV DETECTION. Namık Kemal Tıp Dergisi, 8(2), 234-270. https://doi.org/10.37696/nkmj.687468
AMA Cansever MŞ, Kalaycıoğlu Z, Zübarioğlu T, Kıykım E, Aktuğlu A, Berker B. RAPID DETERMINATION OF L-2-HYDROXYGLUTARIC ACID IN URINE SAMPLES BY CAPILLARY ELECTROPHORESIS WITH INDIRECT UV DETECTION. NKMJ. Ağustos 2020;8(2):234-270. doi:10.37696/nkmj.687468
Chicago Cansever, Mehmet Şerif, Zeynep Kalaycıoğlu, Tanyel Zübarioğlu, Ertuğrul Kıykım, Ayşe Aktuğlu, ve Bedia Berker. “RAPID DETERMINATION OF L-2-HYDROXYGLUTARIC ACID IN URINE SAMPLES BY CAPILLARY ELECTROPHORESIS WITH INDIRECT UV DETECTION”. Namık Kemal Tıp Dergisi 8, sy. 2 (Ağustos 2020): 234-70. https://doi.org/10.37696/nkmj.687468.
EndNote Cansever MŞ, Kalaycıoğlu Z, Zübarioğlu T, Kıykım E, Aktuğlu A, Berker B (01 Ağustos 2020) RAPID DETERMINATION OF L-2-HYDROXYGLUTARIC ACID IN URINE SAMPLES BY CAPILLARY ELECTROPHORESIS WITH INDIRECT UV DETECTION. Namık Kemal Tıp Dergisi 8 2 234–270.
IEEE M. Ş. Cansever, Z. Kalaycıoğlu, T. Zübarioğlu, E. Kıykım, A. Aktuğlu, ve B. Berker, “RAPID DETERMINATION OF L-2-HYDROXYGLUTARIC ACID IN URINE SAMPLES BY CAPILLARY ELECTROPHORESIS WITH INDIRECT UV DETECTION”, NKMJ, c. 8, sy. 2, ss. 234–270, 2020, doi: 10.37696/nkmj.687468.
ISNAD Cansever, Mehmet Şerif vd. “RAPID DETERMINATION OF L-2-HYDROXYGLUTARIC ACID IN URINE SAMPLES BY CAPILLARY ELECTROPHORESIS WITH INDIRECT UV DETECTION”. Namık Kemal Tıp Dergisi 8/2 (Ağustos 2020), 234-270. https://doi.org/10.37696/nkmj.687468.
JAMA Cansever MŞ, Kalaycıoğlu Z, Zübarioğlu T, Kıykım E, Aktuğlu A, Berker B. RAPID DETERMINATION OF L-2-HYDROXYGLUTARIC ACID IN URINE SAMPLES BY CAPILLARY ELECTROPHORESIS WITH INDIRECT UV DETECTION. NKMJ. 2020;8:234–270.
MLA Cansever, Mehmet Şerif vd. “RAPID DETERMINATION OF L-2-HYDROXYGLUTARIC ACID IN URINE SAMPLES BY CAPILLARY ELECTROPHORESIS WITH INDIRECT UV DETECTION”. Namık Kemal Tıp Dergisi, c. 8, sy. 2, 2020, ss. 234-70, doi:10.37696/nkmj.687468.
Vancouver Cansever MŞ, Kalaycıoğlu Z, Zübarioğlu T, Kıykım E, Aktuğlu A, Berker B. RAPID DETERMINATION OF L-2-HYDROXYGLUTARIC ACID IN URINE SAMPLES BY CAPILLARY ELECTROPHORESIS WITH INDIRECT UV DETECTION. NKMJ. 2020;8(2):234-70.