Enantiospecific UPC2 SFC-MS method for separation and quantification of R & S-Eliglustat tartrate in presence of its stereo isomers and degradation impurities

Abstract

Abstract Eliglustat tartrate is an oral inhibitor of glucosylceramide synthase used in the treatment of Gaucher’s disease. This drug is administered as enantio-specific trans-(R,R) isomer and there is a possibility to have three more potential stereo isomers due to two chiral centers, owing to the toxicological difference between stereoisomers chiral method with high sensitivity is required to detect low level contaminations. A method for enantiomeric separation and quantification of trans-(R,R) Eliglustat tartrate in presence of other isomers trans-(S,S) Eliglustat, cis-(R,S) Eliglustat, cis-(S,R) Eliglustat was developed and validated using ultra-high performance supercritical fluid chromatography with tandem mass spectrometry (UPC2 SFC-MS) which is termed as a green chromatographic technique. Chiral separation was developed using Chiralcel OX-3 (4.6 * 150 mm) 3 µm column which has 4 chloro, 3 methyl phenyl carbamate as chiral selector, with a flow rate of 3 g/min using CO2 and 0.5% diethyl amine in ethanol: methanol (1:1) as a co-solvent. The effect on the enantioseparation by varying chiral additives, gradient conditions and content of organic modifier were investigated. The method has good linearity with concentration ranges from 0.25 to 7.5 µg mL−1. The detection limit was 0.08 µg mL−1 and quantification limit was 0.25 µg mL−1. Forced degradation studies were performed and all the degradation impurities were separated from product peak, the molecular weight of impurities were identified by using UPC2-Mass spectrometer. The method was validated in line to ICH guidelines in terms of linearity, accuracy, precision, LOQ, LOD and robustness.