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Analysis of urinary sphingolipids using liquid chromatography-tandem mass spectrometry in diabetic nephropathy

J Diabetes Investig. 2020 Mar;11(2):441-449. doi: 10.1111/jdi.13154. Epub 2019 Oct 21.

Abstract

Aims/introduction: Sphingolipids, such as ceramides and sphingosine, are involved in the pathogenesis of diabetes; however, the modulation of urinary sphingolipids in diabetic nephropathy has not been fully elucidated. Therefore, we aimed to develop a simultaneous measurement system for urinary sphingolipids using liquid chromatography-tandem mass spectrometry and to elucidate the modulation of urinary sphingolipids in diabetic nephropathy.

Materials and methods: We established a simultaneous measurement system for the urinary sphingosine, dihydrosphingosine, and six ceramide species (Cer d18:1/16:0, Cer d18:1/18:0, Cer d18:1/18:1, Cer d18:1/20:0, Cer d18:1/22:0 and Cer d18:1/24:0), and we examined the urinary sphingolipids in 64 type 2 diabetes patients and 15 control participants.

Results: The established measurement system for the urinary sphingolipids showed good precision for Cer d18:1/16:0, Cer d18:1/20:0, Cer d18:1/22:0 and Cer d18:1/24:0. We observed that the urinary levels of Cer d18:1/16:0, Cer d18:1/18:0, Cer d18:1/20:0, Cer d18:1/22:0 and Cer d18:1/24:0 were elevated in patients with stage 3 of diabetic nephropathy, and were correlated with urinary biomarkers, such as albumin and N-acetyl-β-d-glucosaminidase, and sediment score.

Conclusions: Our method is useful for the measurement of ceramide in urine specimens, and urinary ceramides might be associated with the pathological condition of diabetic nephropathy, such as renal tubular injury.

Keywords: Ceramide; Tandem mass spectrometry; Urine.

MeSH terms

  • Aged
  • Biomarkers / urine
  • Ceramides / urine
  • Chromatography, Liquid / methods*
  • Diabetes Mellitus, Type 2 / complications
  • Diabetes Mellitus, Type 2 / diagnosis
  • Diabetes Mellitus, Type 2 / urine*
  • Diabetic Nephropathies / complications
  • Diabetic Nephropathies / diagnosis
  • Diabetic Nephropathies / urine*
  • Female
  • Humans
  • Male
  • Middle Aged
  • Sphingolipids / urine*
  • Tandem Mass Spectrometry / methods*

Substances

  • Biomarkers
  • Ceramides
  • Sphingolipids