Novel two-step derivation method for the synchronous analysis of inherited metabolic disorders using urine

  • Authors:
    • Xiao‑Qi Sheng
    • Yi‑Chao Wang
  • View Affiliations

  • Published online on: March 2, 2017     https://doi.org/10.3892/etm.2017.4167
  • Pages: 1961-1968
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Abstract

The aim of the present study was to conduct preliminary clinical screening and monitoring using a novel two‑step derivatization process of urine in five categories of inherited metabolic disease (IMD). Urine samples (100 µl, containing 2.5 mmol/l creatinine) were taken from patients with IMDs. The collected urine was then treated using a two‑step derivatization method (with oximation and silylation at room temperature), where urea and protein were removed. In the first step of the derivatization, α‑ketoacids and α‑aldehyde acids were prepared by oximation using novel oximation reagents. The second‑step of the derivatization was that residues were silylated for analysis. Urine samples were examined using gas chromatography/mass spectrometry (GC/MS) and a retention time‑locking technique. The simultaneous analysis and identification of >400 metabolites in >130 types of IMD was possible from the GC/MS results, where the IMDs included phenylketonuria, ornithine transcarbamylase deficiency, neonatal intrahepatic cholestasis caused by citrin deficiency, β‑ureidopropionase deficiency and mitochondrial metabolic disorders. This method was demonstrated to have good repeatability. Considering α‑ketoglutarate (α‑KG) as an example, the relative standard deviations (RSDs) of the α‑KG retention time and peak area were 0.8 and 3.9%, respectively, the blank spiked recovery rate was between 89.6 and 99.8%, and the RSD was ≤7.5% (n=5). The method facilitates the analysis of thermally non‑stable and semi‑volatile metabolites in urine, and greatly expands the range of materials that can be synchronously screened by GC/MS. Furthermore, it provides a comprehensive, effective and reliable biochemical analysis platform for the pathological research of IMDs.
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May-2017
Volume 13 Issue 5

Print ISSN: 1792-0981
Online ISSN:1792-1015

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Spandidos Publications style
Sheng XQ and Sheng XQ: Novel two-step derivation method for the synchronous analysis of inherited metabolic disorders using urine. Exp Ther Med 13: 1961-1968, 2017
APA
Sheng, X., & Sheng, X. (2017). Novel two-step derivation method for the synchronous analysis of inherited metabolic disorders using urine. Experimental and Therapeutic Medicine, 13, 1961-1968. https://doi.org/10.3892/etm.2017.4167
MLA
Sheng, X., Wang, Y."Novel two-step derivation method for the synchronous analysis of inherited metabolic disorders using urine". Experimental and Therapeutic Medicine 13.5 (2017): 1961-1968.
Chicago
Sheng, X., Wang, Y."Novel two-step derivation method for the synchronous analysis of inherited metabolic disorders using urine". Experimental and Therapeutic Medicine 13, no. 5 (2017): 1961-1968. https://doi.org/10.3892/etm.2017.4167