Simultaneous diagnostic method for phenylketonuria and galactosemia from dried blood spots using high-performance liquid chromatography-pulsed amperometric detection

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 878; no. 21; pp. 1860 - 1864
• Main Authors: Kim, Nam-Hee, Jeong, Ji-Seon, Kwon, Ha-Jeong, Lee, Yong-Moon, Yoon, Hye-Ran, Lee, Kyoung Ryul, Hong, Seon-Pyo
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.07.2010; Elsevier
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Blood; Chromatography; Chromatography, High Pressure Liquid - methods; Cut-off; Diagnostic systems; Dried blood spot; Electrical measurements; False Positive Reactions; Fundamental and applied biological sciences. Psychology; Galactosemia; Galactosemias - blood; General pharmacology; HPLC; Humans; Infant, Newborn; Linear Models; Liquids; Medical sciences; Neonatal Screening - methods; Newborn screening; Pharmacology. Drug treatments; Phenylalanine; Phenylketonuria; Phenylketonurias - blood; Reproducibility of Results; Sensitivity and Specificity; Simultaneous diagnosis; Specimen Handling - methods; Spots; IMD; PAD; Phenylketonuria; GALT; Dried blood spot; Newborn screening; PKU; ECM; HPLC; Simultaneous diagnosis; Galactosemia; Human; Biological fluid; Nervous system diseases; HPLC chromatography; Metabolic diseases; Medical screening; Enzymopathy; Blood; Genetic disease; Newborn; Amperometry; Carbohydrate; Diagnosis; Aminoacid disorder
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2010.04.038

We developed a simultaneous diagnostic method for phenylketonuria (PKU) and galactosemia through simultaneous determination of phenylalanine (Phe) and galactose (Gal) by high-performance liquid chromatography (HPLC) with pulsed amperometric detection (PAD). The intra- and inter-day precisions were <5.8%, with satisfactory mean recoveries (98.2–105%). For all PKU-positive samples, Phe levels were above the cut-off value (>30.0 mg/L), but Gal levels were nearly zero. For 77% of galactosemia-positive samples, Phe levels were above the cut-off value, but Gal levels were above the cut-off value (>80.0 mg/L) for all samples. Our HPLC-PAD method can reduce the false-positive rate of misdiagnosis for PKU and galactosemia.