Degradation studies of levosimendan isolation, identification, and structure confirmation of stress degradation products using LCMS, mass mediated Prep-HPLC, NMR, HRMS, SFC and FTIR
Creators: name: G. Mahesh Kumar Reddy
name: Jcmknn Murty Singamsetti
name: Muralidharan Kaliyaperumal
name: Doddipalla, Raju
name: Ramu Ivaturi
name: Chidananda Swamy Rumalla
name: Raghu Babu Korupolu
name: B. Kishore Babu
Publisher: name: Taylor & Francis
Year: 2019
Subjects: subject: Biochemistry
subject: Biotechnology
subject: 59999 Environmental Sciences not elsewhere classified
subject: FOS: Earth and related environmental sciences
subject: FOS: Earth and related environmental sciences
subject: 39999 Chemical Sciences not elsewhere classified
subject: FOS: Chemical sciences
subject: FOS: Chemical sciences
subject: 69999 Biological Sciences not elsewhere classified
subject: FOS: Biological sciences
subject: FOS: Biological sciences
subject: Science Policy

description: 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 C<sub>14</sub>H<sub>14</sub>N<sub>4</sub>O<sub>4</sub>. 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. HighlightsLevosimendan 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. 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.