Sensitive CE‐MS method for monitoring of riociguat and desmethylriociguat levels in human serum

• Published in: Electrophoresis Vol. 41; no. 18-19; pp. 1564 - 1567
• Main Authors: Hložek, Tomáš, Štícha, Martin, Bursová, Miroslava, Jelínek, Ivan, Tůma, Petr, Čabala, Radomír
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 01.10.2020
• Subjects: ammonium; Analytical chemistry; antihypertensive agents; Antihypertensives; blood serum; calibration; Capillary electrophoresis; chemical species; Desmethylriociguat; Determination; Electrolytes; Electrophoresis; Electrophoresis, Capillary - methods; formates; High performance liquid chromatography; Humans; Hypertension; Limit of Detection; Linear Models; Liquid-Liquid Extraction; Mass spectrometry; Optimization; Pharmacokinetics; Pharmacology; Pretreatment; Pyrazoles - blood; Pyrazoles - chemistry; Pyrazoles - isolation & purification; Pyrazoles - pharmacokinetics; Pyrimidines - blood; Pyrimidines - chemistry; Pyrimidines - isolation & purification; Pyrimidines - pharmacokinetics; Reproducibility of Results; Riociguat; Scientific imaging; Spectrometry, Mass, Electrospray Ionization - methods; Spectroscopy; Desmethylriociguat; Determination; Capillary electrophoresis; Mass spectrometry; Riociguat
• DOI: 10.1002/elps.202000135
• ISSN: 0173-0835; 1522-2683

Riociguat is novel antihypertensive drug for treatment of pulmonary hypertension. As such, it is still being tested in many clinical and pharmacokinetic trials. Existing methods that determine serum riociguat and desmethylriociguat (DMR) are based solely on liquid chromatography with mass spectrometry. Therefore, we present a novel capillary electrophoresis with mass spectrometry method (CE‐MS) for their determination in human serum as alternative method for ongoing trials. Complete resolution of both analytes was achieved by means of pH optimization of ammonium formate background electrolytes that are fully compatible with ESI/MS detection. Simple liquid‐liquid extraction was used as sample pretreatment. The calibration dependence of the method was linear (in the range of 10–1000 ng/mL), with adequate accuracy (90.1–114.9%) and precision (13.4%). LOD and LOQ were arbitrarily set at 10 ng/mL for both analytes. Clinical applicability was validated using serum samples from patients treated with riociguat in pharmacokinetic study and the results corresponded with reference HPLC‐MS/MS values. Capillary electrophoresis proved to be sensitive and selective tool for the analysis of riociguat and DMR.