Related Experiment Video
Updated: Jun 11, 2026
08:53
Quantitative and Qualitative Method for Sphingomyelin by LC-MS Using Two Stable Isotopically Labeled Sphingomyelin Species
Published on: May 7, 2018
UPLC-MS/MS method for analysis of sphingosine 1-phosphate in biological samples
Adele Cutignano1, Ugo Chiuminatto, Filomena Petruzziello
1CNR - Institute of Biomolecular Chemistry, via Campi Flegrei 34, 80078 Pozzuoli (NA), Italy. acutignano@icb.cnr.it
Prostaglandins & Other Lipid Mediators
|July 6, 2010
Summary
A new liquid chromatography-tandem mass spectrometry method accurately quantifies sphingosine 1-phosphate (S1P) and sphinganine 1-phosphate (DH-S1P) in biological samples. This validated technique offers high sensitivity for sphingolipid analysis in plasma.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Lipidomics
Background:
- Sphingosine 1-phosphate (S1P) and its precursor sphinganine 1-phosphate (DH-S1P) are critical bioactive lipids involved in numerous physiological processes.
- Accurate quantification of these lipids in biological matrices is essential for understanding their roles in health and disease.
- Existing analytical methods may lack the sensitivity or simplicity required for routine biological sample analysis.
Purpose of the Study:
- To develop and validate a simple, sensitive, and reliable liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for determining S1P and DH-S1P levels.
- To establish a robust analytical workflow applicable to various biological samples, including human and mouse plasma.
- To provide a sensitive tool for sphingolipid research and clinical diagnostics.
Main Methods:
- Utilized negative ion-electrospray ionization liquid chromatography-tandem mass spectrometry (LC-MS/MS) for analysis.
- Employed a UPLC BEH Hilic column with a binary mobile phase for chromatographic separation.
- Validated the method using synthetic C(17)-sphingosine 1-phosphate (C17-S1P) as an internal standard over a linearity range of 2-100 ng/ml.
Main Results:
- Achieved a lower limit of quantification of 5.0 ng/ml for S1P, with a detection limit below 5 pg (12 fmol).
- Demonstrated high linearity (r>0.999) for the validated method.
- Quantified S1P and DH-S1P in human plasma (176.7±54.0 ng/ml S1P, 81.2±23.3 ng/ml DH-S1P) and mouse plasma (201.0±72.0 ng/ml S1P, 96.5±20.1 ng/ml DH-S1P).
Conclusions:
- The developed LC-MS/MS methodology is simple, sensitive, and suitable for quantifying S1P and DH-S1P in biological samples.
- This validated method provides a reliable tool for sphingolipid research, enabling accurate measurements in plasma.
- The high sensitivity and validated performance support its application in various research settings and potentially in clinical diagnostics.
