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Simultaneous determination of allantoin and adenosine in human urine using liquid chromatography - UV detection.
Andries, Asmin; De Rechter, Stéphanie; Janssens, Peter; Mekahli, Djalila; Van Schepdael, Ann.
Afiliação
  • Andries A; KU Leuven - University of Leuven, Department of Pharmaceutical and Pharmacological Sciences, Pharmaceutical Analysis, B-3000 Leuven, Belgium. Electronic address: asminandries@gmail.com.
  • De Rechter S; KU Leuven - University of Leuven, Department of Development and Regeneration, Laboratory of Pediatrics, PKD Research Group, B-3000 Leuven, Belgium; University Hospitals Leuven, Department of Pediatric Nephrology, B-3000 Leuven, Belgium.
  • Janssens P; KU Leuven - University of Leuven, Department of Development and Regeneration, Laboratory of Pediatrics, PKD Research Group, B-3000 Leuven, Belgium; University Hospitals of Brussels, Department of Nephrology, B-1000 Brussels, Belgium.
  • Mekahli D; KU Leuven - University of Leuven, Department of Development and Regeneration, Laboratory of Pediatrics, PKD Research Group, B-3000 Leuven, Belgium; University Hospitals Leuven, Department of Pediatric Nephrology, B-3000 Leuven, Belgium.
  • Van Schepdael A; KU Leuven - University of Leuven, Department of Pharmaceutical and Pharmacological Sciences, Pharmaceutical Analysis, B-3000 Leuven, Belgium.
Article em En | MEDLINE | ID: mdl-30176509
ABSTRACT
We report a HPLC-UV method for the quantitative determination of allantoin and adenosine in human urine, validated according to the acceptance criteria of both the USA Food and Drug Administration (FDA) guideline for bioanalytical method validation and the European Medicines Agency (EMA) validation guidelines. Both allantoins and adenosine are compounds of the purine catabolic pathway. Adenosine is situated at the top as a uric acid (UA) precursor, while allantoin is the best-known degradation product of UA. These two compounds are endogenously present in human urine. Chromatographic separation was achieved with a gradient elution at 0.6 mL/min using a Zorbax SB-Aq column coupled to a Zorbax SB-Aq guard column. Three different mobile phases were used mobile phase A consisted of 10 mM KH2PO4 (pH 4.7) in milli-Q water, mobile phase B was 12.5 mM KH2PO4 (pH 4.7) - ACN (8020) and mobile phase C consisted of ACN - H2O (5050). The linear response range in human urine was 14-800 µM for allantoin and 1.25-50 µM for adenosine. The recoveries of allantoin, adenosine and the internal standard were greater than 93.8%. The intra-day accuracy ranged between 99.5 and 104.9% for allantoin and between 96.6 and 107.3% for adenosine, while the inter-day accuracy ranged respectively from 91.2 to 103.0% and from 94.5 to 107.8%. The intra-day precision range was from 0.8 to 6.2% RSD for allantoin and from 0.6 to 15.0% for adenosine. The inter-day precision ranged from 2.1-17.5% for allantoin and from 2.9-17.9% for adenosine. This method was successfully applied to analyze both compounds in urine samples of healthy volunteers. In conclusion, an accurate, precise and stable HPLC-UV method was developed and validated to quantify the endogenously present compounds allantoin and adenosine in human urine samples.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Espectrofotometria Ultravioleta / Adenosina / Cromatografia Líquida de Alta Pressão / Alantoína Tipo de estudo: Diagnostic_studies / Prognostic_studies Limite: Adult / Humans Idioma: En Revista: J Chromatogr B Analyt Technol Biomed Life Sci Assunto da revista: ENGENHARIA BIOMEDICA Ano de publicação: 2018 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Espectrofotometria Ultravioleta / Adenosina / Cromatografia Líquida de Alta Pressão / Alantoína Tipo de estudo: Diagnostic_studies / Prognostic_studies Limite: Adult / Humans Idioma: En Revista: J Chromatogr B Analyt Technol Biomed Life Sci Assunto da revista: ENGENHARIA BIOMEDICA Ano de publicação: 2018 Tipo de documento: Article