A high throughput flow gradient LC–MS/MS method for simultaneous determination of fingolimod, fampridine and prednisone in rat plasma, application to in vivo perfusion study

Sphingosine 1-phosphate (S1P 1 ) 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 S1P 1 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 S1P 1 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 S1P 1 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.

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Case Studiesmentioning

confidence: 99%

Section: Scopementioning

confidence: 99%

See 3 more Smart Citations

A review of bioanalytical quantitative methods for selected sphingosine 1‐phosphate receptor modulators

Dash

Srinivas

Rais

2017

A sensitive liquid chromatography–tandem mass spectrometry method for quantitative bioanalysis of fingolimod in human blood: Application to pharmacokinetic study

Gopinath

Narenderan

Kumar

et al. 2020

A simple and sensitive liquid chromatography–tandem mass spectrometry (LC–MS/MS) method has been developed and validated for the determination of fingolimod in human blood. The analyte and internal standard fingolimod‐d4 were extracted from 300 μl of human blood using protein precipitation coupled with solid‐phase extraction method. The chromatographic separation was achieved with a Kinetex biphenyl column (100 × 4.6 mm, 2.6 μm) under isocratic conditions at the flow rate of 0.8 ml/min and column temperature was maintained at 45°C. The detection of analyte and internal standard was carried out by tandem mass spectrometry, operated in positive ion and multiple reaction monitoring acquisition mode. The method was fully validated for its selectivity, precision, accuracy, linearity, stability, detection and quantification limit. The extraction recovery of fingolimod in human blood ranged from 98.39 to 99.54%. The developed method was linear over the concentration range of 5–2500 pg/ml with a detection limit of 1 pg/ml. The developed method was validated and successfully applied for pharmacokinetic study after oral administration of fingolimod capsules.

Liquid chromatography–tandem mass spectrometry for determination of fingolimod and its active metabolite fingolimod phosphate in whole blood of patients with multiple sclerosis

Pesakova,

Brozmanova,

Sistik

et al. 2024

Fingolimod is an oral drug for the escalation of treatment of relapsing–remitting multiple sclerosis in patients with persistent disease activity on first‐line drugs or in patients with rapidly progressive severe relapsing–remitting multiple sclerosis. An ultra‐high‐performance liquid chromatography–tandem mass spectrometry method for determining the concentrations of fingolimod and its active metabolite fingolimod phosphate in whole blood has been developed and validated. The advantages of this method are the easy, fast and cheap sample preparation using protein precipitation from blood with a mixture of acetonitrile–methanol (40:60, v/v). Chromatographic separation was performed on a ultra‐high performance liquid chromatography BEH C18 1.7 μm (100 × 2.1 mm) column. Two modes of ionization, electrospray ionization and atmospheric pressure chemical ionization, were tested and compared. For validation, the electrospray ionization mode was chosen. As internal standard, isotopically labeled fingolimod‐D4 was used to quantify the analytes. The method was validated according to the rules of the European Medicines Agency. The coefficients of variation for fingolimod were in the range of 1.13–11.88%, and the recovery was 98.80–106.00%. The coefficients of variation for fingolimod phosphate were in the range of 2.73–9.31%, and the recovery was 90.08–107.00%. The method is quite easy and fast and can be used for routine analysis.