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. 2018 Dec;8(6):357-365.
doi: 10.1016/j.jpha.2018.03.001. Epub 2018 Mar 15.

LC and LC-MS/MS studies for the identification and characterization of degradation products of acebutolol

Uday Rakibe  1 , Ravi Tiwari  2 , Anand Mahajan  3 , Vipul Rane  1 , Pravin Wakte  1
Affiliations

LC and LC-MS/MS studies for the identification and characterization of degradation products of acebutolol

Uday Rakibe et al. J Pharm Anal. 2018 Dec.

Abstract

The aim of the present investigation was to demonstrate an approach involving use of liquid chromatography (LC) and liquid chromatography-mass spectrometry (LC-MS) to separate, identify and characterize very small quantities of degradation products (DPs) of acebutolol without their isolation from the reaction mixtures. The drug was subjected to oxidative, hydrolytic, thermal and photolytic stress conditions as per International Conference on Harmonization (ICH) guideline Q1A(R2). Among all the stress conditions the drug was found to be labile in hydrolytic (acidic & basic) and photolytic stress conditions, while it was stable in water-induced hydrolysis, oxidative and thermal stress conditions. A total of four degradation products were formed. A C18 column was employed for the separation of all the DPs on a gradient mode by using high-performance liquid chromatography (HPLC). All the DPs were characterized with the help of their fragmentation pattern and the masses obtained upon LC-MS/MS and MSn analysis. All the hitherto unknown degradation products were identified as 1-(2-(2-hydroxy-3-(isopropylamino)propoxy)-5-(amino)phenyl)ethanone (DP-I), N-(4-(2-hydroxy-3-(isopropylamino)propoxy)-3-acetylphenyl)acrylamide (DP-II), 1-(2-(2-hydroxy-3-(isopropylamino)propoxy)-5-(hydroxymethylamino)phenyl)ethanone (DP-III) and 1-(6-(2-hydroxy-3-(isopropylamino)propoxy)-2,3-dihydro-2-propylbenzo[d]oxazol-5-yl)ethanone (DP-IV). Finally the in-silico carcinogenicity and hepatotoxicity predictions of the drug and all the DPs were performed by using toxicity prediction softwares viz., TOPKAT, LAZAR and Discovery Studio ADMET. The results of in-silico toxicity studies revealed that acebutolol (0.967) and DP-I (0.986) were found to be carcinogenic, while acebutolol (0.490) and DP-IV (0.437) were found to be hepatotoxic.

Keywords: Acebutolol; Degradation pathway; In-silico toxicity; LC; LC–MS/MS; Stress testing.

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Figures

Fig. 1
Fig. 1
Structure of acebutolol.
Fig. 2
Fig. 2
HPLC chromatogram of all the acebutolol products (I-IV) and acebutolol.
Fig. 3
Fig. 3
Line spectrum of acebutolol obtained in MS study.
Fig. 4
Fig. 4
Fragmentation pathway of acebutolol.
Fig. 5
Fig. 5
Line spectra of degradation products: (A) DP-I, (B) DP-II, (C) DP-III and (D) DP-IV obtained in LC–MS/MS studies.
Fig. 6
Fig. 6
Fragmentation pathway of DP-I-IV.
Fig. 7
Fig. 7
Degradation pathway of acebutolol.
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