Validated LC‐MS/MS method of Sphingosine 1‐phosphate quantification in human serum for evaluation of response to radiotherapy in lung cancer
Xiaohui Tang, Hai‐Sheng Chen, Guanxuan Chen, Cunxian Duan, Qing Fan, Hui Li, Yanhong Wang, Zhijun Li, Wenna Shi, Yuguo Liu
Abstract
BACKGROUND: Sphingosine 1-phosphate (S1P), a bioactive lipid, has been shown to mediate cancer processes. Therefore, accurate qualitative and quantitative determination is essential. The current assay method is still cumbersome to be of practical use worldwide and the aim of this study was therefore to develop a fast, accurate, precise and efficient LC-MS/MS method for targeted analyses of S1P in serum samples. METHODS: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is an established method used for monitoring and analyzing S1P levels in serum. We determined the level of serum S1P in 256 patients with lung cancer and 36 healthy donors, and used Spearman';s rank correlation analysis to evaluate the difference in serum S1P levels between radiotherapy and nonradiotherapy patients. RESULTS: ) greater than 0.9996. The lower limit of quantifications (LLOQs) was 25 ng/mL. The intra- and interbatch precisions and accuracy was less than 10% for S1P. The recoveries of the method were found to be 80%-98%. Serum S1P levels in healthy donors were different from those in patients (P < 0.001). Of 256 lung cancer patients, 124 (48.4%) received radiotherapy and were identified to have concomitant low serum S1P levels (222.13 ± 48.63), whereas 132 (51.6%) who had not received radiotherapy were identified to have high levels (315.16 ± 51.06). The serum S1P levels were therefore associated with radiotherapy (Spearman's Rho = -0.653, P < 0.001). CONCLUSIONS: Our results indicated that this new LC-MS/MS method is rapid, sensitive, specific and reliable for the quantification of S1P levels in serum samples. The level of S1P in serum samples of patients with lung cancer who received radiotherapy was significantly lower than that in patients who did not receive radiotherapy. KEY POINTS: An improved method was established to quantify S1P levels in human serum by LC-MS/MS, which enabled the change in serum S1P levels in lung cancer patients to be monitored, in combination with radiotherapy, and their clinical significance to be analyzed.