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KMID : 0370220210650010031
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Yakhak Hoeji
2021 Volume.65 No. 1 p.31 ~ p.40
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Rapid and Sensitive Quantification Method of Sphingosine and Sphingosine-1-phosphate in Human Serum by Ultra High-Performance Liquid Chromatography-Tandem Mass Spectrometry (UHPLC-MS/MS)
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Na Seong-Hoon
Yoon Hye-Ran
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Abstract
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Sphingosine (SPH) and sphingosine-1-phosphate (S1P) are emerging as key players in asthma metabolism and in numerous cellular inflammation processes. To identify potential biomarkers of asthma and inflammatory therapeutics, it is essential to determine their levels. Herein, we developed a rapid and sensitive UHPLC-MS/MS method to simultaneously quantify SPH and S1P in human serum using C17-SPH and C17-S1P as internal standards. After methanol precipitation of serum proteins, the supernatants were analyzed by MS/MS performed in the positive ion mode by multiple reaction monitoring. UHPLC analysis (C18 column) was performed using two mobile phase systems (water containing 0.1% formic acid, and 85% acetonitrile containing 0.1% formic acid) within 5 min of the short run. The calibration curves were linear in the range of 0.002-1.5 μg/mL for S1P and SPH with an R2 greater than 0.9999. The LOD and LOQ were 0.0002 and 0.0004 μg/mL for S1P, and 0.0005 and 0.001 μg/mL for SPH, respectively. The accuracy and precision of the method were in the range of 89.8-100.7% (RSD, 1.5-2.8%) for both SPH and S1P species. We were able to quantify both molecules in serum from healthy and asthmatic patients. These results suggest that SPH and S1P are promising potential biomarkers, and also contribute to the basic data for the construction of an omics-based platform for preventive index prior to asthma diagnosis.
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KEYWORD
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asthma, serum, sphingolipids, sphingosine, sphingosine-1-phosphate, UHPLC-MS/MS
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