İdrarda Orotik Asit'in Temassız İletkenlik Dedektörü Kullanılarak Kapiler Elektroforez Yöntemi İle Hızlı Tayini
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Dosyalar
Tarih
2020
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Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
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.
Excessive excretion of orotic acid in urine arises as a result of some inborn metabolic diseases such as urea cycle disorders and pyrimidine synthesis disorders. Therefore, rapid determination of orotic acid in the neonatal period is vital. Existing methods for measuring orotic acid levels in urine are both time consuming and expensive. Consequently, rapid definition of the orotic acid level in human urine is important. In this current study a novel, simple, cost effective and rapid (fast) capillary electrophoretic analysis method was developed for quantitative measurement of orotic acid content in urine samples. Determination of orotic acid by employing capillary electrophoresis coupled with contactless conductivity detection was achieved by using, 20 mM 4-Morpholineethanesulfonic acid (MES), 10 mM Histidine, 0.1 mM Cetyltrimethylammonium Bromide (CTAB) at pH of 6.5 as optimum electrolyte solution. Orotic acid was well separated from the sample constituents within a short analysis time less than 3.5 minutes. The precisions for the migration times and corrected peak areas were calculated as 1.25 and 4.31 RSD% respectively. Limit of detection and limit of quantification of the proposed method were also calculated as 0.733 mg / L and 2.443 mg / L respectively. The developed sensitive and facile capillary electrophoresis method was validated and applicability of the method was successfully demonstrated by assessment of orotic acid content in urine.
Excessive excretion of orotic acid in urine arises as a result of some inborn metabolic diseases such as urea cycle disorders and pyrimidine synthesis disorders. Therefore, rapid determination of orotic acid in the neonatal period is vital. Existing methods for measuring orotic acid levels in urine are both time consuming and expensive. Consequently, rapid definition of the orotic acid level in human urine is important. In this current study a novel, simple, cost effective and rapid (fast) capillary electrophoretic analysis method was developed for quantitative measurement of orotic acid content in urine samples. Determination of orotic acid by employing capillary electrophoresis coupled with contactless conductivity detection was achieved by using, 20 mM 4-Morpholineethanesulfonic acid (MES), 10 mM Histidine, 0.1 mM Cetyltrimethylammonium Bromide (CTAB) at pH of 6.5 as optimum electrolyte solution. Orotic acid was well separated from the sample constituents within a short analysis time less than 3.5 minutes. The precisions for the migration times and corrected peak areas were calculated as 1.25 and 4.31 RSD% respectively. Limit of detection and limit of quantification of the proposed method were also calculated as 0.733 mg / L and 2.443 mg / L respectively. The developed sensitive and facile capillary electrophoresis method was validated and applicability of the method was successfully demonstrated by assessment of orotic acid content in urine.
Açıklama
Anahtar Kelimeler
Kaynak
Süleyman Demirel Üniversitesi Fen Edebiyat Fakültesi Fen Dergisi
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Cilt
15
Sayı
2
