Degradation studies of levosimendan isolation, identification, and structure confirmation of stress degradation products using LCMS, mass mediated Prep-HPLC, NMR, HRMS, SFC and FTIR. Issue 1 (20th January 2020)

• Record Type: Journal Article
• Authors: Reddy, G. Mahesh Kumar; Singamsetti, Jcmknn Murty; Kaliyaperumal, Muralidharan; Doddipalla, Raju; Ivaturi, Ramu; Rumalla, Chidananda Swamy; Korupolu, Raghu Babu; Babu, B. Kishore
• Is Part Of: Journal of liquid chromatography & related technologies. Volume 43:Issue 1/2(2020)
• Page Start: 17
• Page End: 28
• Publication Date: 2020-01-20
• Subjects: Characterization -- forced degradation -- HRMS -- identification -- isolation -- levosimendan -- NMR; Liquid chromatography -- Periodicals; Chromatographic analysis -- Periodicals; Separation (Technology) -- Periodicals; Biotechnology -- Periodicals; Chromatography, Liquid -- Periodicals; 543.8405
• DOI: 10.1080/10826076.2019.1613429
• Languages: English
• ISSNs: 1082-6076

Abstract

Abstract: The present work describes the stress degradation behavior of Levosimendan as per ICH guidelines. Under thermal and photolytic conditions drug had a fair stability and in other stress conditions degradation products (DP) were observed. Initial identification of the DP was performed by UPLC-MS and isolation by MDAP. Various 1 D and 2 D NMR experiments along with supporting FT-IR and HRMS data were performed to characterize the DP. A novel degradant product observed in acidic conditions was isolated and characterized as (R, Z)-2(2-(4-(4-amino-2-methyl-4-oxobutanoyl) phenyl) hydrazono)-2-cyanoacetic acid (DP-1) with molecular formula C14 H14 N4 O4 . Additionally, two more DP were observed in both basic and peroxide conditions which are basically reported impurities. (R, E)-2-amino-N'-(4-(4-methyl-6-oxo-1, 4, -5, 6-tetrahydropyridazin-3-yl)phenyl)-2-oxoaceto hydrazonoyl cyanide (DP-2), a Cyano to amide converted product and (R)-5-methyl-6-phenyl-4, 5-dihydropyridazin-3(2H)-one (DP-3) formed as result of cleavage of aryl nitrogen bond of benzene substituted dicyano hydrazine. GRAPHICAL ABSTRACT: Highlights: Levosimendan was subjected to force degradation under acidic, basic, oxidative, photolytic and thermal conditions as per ICH guidelines. In Acid, base and oxidative conditions three degradant products were observed. DP-1 is identified and is novel. It is not reported in any literature. DP-2 and DP-3 are known and reported earlier, however this known degradation products only identified by LCMS. In present paper all the three degradation products have been identified and characterized by using various high-end analytical techniques such as HRMS, NMR, SFC and FT-IR.