Highly sensitive rapid determination of orotic acid in urine samples using a field-amplified sample stacking approach in capillary electrophoresis coupled with contactless conductivity detection

Adımcılar V., Saygılı M. T., CANSEVER M. Ş., Öztekin N.

Journal of Pharmaceutical and Biomedical Analysis, vol.238, 2024 (SCI-Expanded) identifier identifier

  • Publication Type: Article / Review
  • Volume: 238
  • Publication Date: 2024
  • Doi Number: 10.1016/j.jpba.2023.115826
  • Journal Name: Journal of Pharmaceutical and Biomedical Analysis
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Academic Search Premier, Analytical Abstracts, Aquatic Science & Fisheries Abstracts (ASFA), BIOSIS, Biotechnology Research Abstracts, CAB Abstracts, Chemical Abstracts Core, Chimica, International Pharmaceutical Abstracts, Veterinary Science Database
  • Keywords: Inborn error, Orotic acid, Orotic aciduria, Sample stacking, Urine
  • Istanbul Technical University Affiliated: Yes


Orotic aciduria is a severe, hereditary, life-threatening condition, particularly in newborns. An increased orotic acid (OA) content in urine may be a strong indicator of this condition. In this study, we developed a rapid, simple, highly sensitive diagnostic method for use in monitoring the OA levels in urine samples, which were successfully determined using capillary electrophoresis combined with capacitively coupled contactless conductivity detection (CE-C4D). A straightforward analysis with an increased sensitivity towards OA and an analysis time of approximately 5 min were realized, and the limit of detection of the developed method was 0.014 mg/L in aqueous solution. The optimized composition of the separation electrolyte was 20 mM 2-(N-morpholino)ethanesulfonic acid/histidine with 0.1 mM cetyltrimethylammonium bromide at a pH of 6.5. The sensitivity of the developed method was significantly increased using a sample stacking approach with a 10% acetonitrile (v/v) plug solution. The method was validated, and satisfactory recoveries of 80.0–92.3% were obtained. The amounts of OA in five urine samples were successfully determined.