Sphingosine 1-phosphate (S1P1 ) modulators provide an emerging therapeutic 
approach for various autoimmune disorders such as multiple sclerosis and 
psoriasis. Fingolimod is the first approved orally active, selective and potent 
drug of this class. Other drugs belonging to this class include siponimod, 
ponesimod, ceralifimod, amiselimod, CS-0777 and GSK2018682. However, owing to 
the high protein binding, polarity and zwitter-ionic nature of the phosphate 
metabolite of parent drugs, it becomes challenging to optimize the extraction 
method for this class of compounds. Although, there are individual published 
bioanalytical methods for the analysis of selected S1P1 modulators to support 
preclinical and clinical drug development, no extensive review compiling all the 
bioanalytical methods for the important drugs in the class is available. Thus, 
we attempted to prepare a comprehensive review on various bioanalytical methods 
for selected S1P1 modulators which will provide all the relevant bioanalytical 
information as required by bioanalytical researchers. This review focuses on the 
various liquid chromatography with tandem mass spectrometry methods that have 
been used to quantify S1P1 modulators in various biological matrices. Extraction 
methods included liquid-liquid extraction, solid-phase extraction and one-step 
protein precipitation for extracting the analytes. This review captures key 
information regarding sample processing options and chromatographic/detection 
conditions.