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Isolation and characterization of degradation products of moxidectin using LC, LTQ FT-MS, H/D exchange and NMR

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Abstract

This study aimed to evaluate the degradation profile and pathways, and identify unknown impurities of moxidectin under stress conditions. During the experiments, moxidectin samples were stressed using acid, alkali, heat and oxidation, and chromatographic profiles were compared with known impurities given in European Pharmacopeia (EP) monograph. Moxidectin has shown good stability under heat, while reaction with alkali produced 2-epi and ∆2,3 isomers (impurities D and E in EP) by characteristic reactions of the oxahydrindene (hexahydrobenzofuran) portion of the macrocyclic lactone. Two new, previously unreported, unknown degradation products, i.e. impurity 1 and impurity 2, detected after acid hydrolysis of moxidectin (impurity 2 was also observed to a lesser extent after oxidation), were isolated from sample matrices and identified using liquid chromatography, NMR, high-resolution FT-ICR MS, and hydrogen/deuterium exchange studies. FTMS analysis showed accurate mass of molecular ion peaks for moxidectin at m/z 640.38412, impurity 1 at m/z 656.37952 and impurity 2 at m/z 611.35684, giving rise to daughter ions traceable up to the seventh levels of MSn experiments and supporting the proposed structures. Both unknown impurities along with moxidectin were fully characterized by 1H, 13C, 1D HMBC and 2D (NOESY, COSY and HSQC) NMR experiments. The interpretation of experimental data positively identified impurity 1 as 3,4-epoxy-moxidectin and impurity 2 as 23-keto-nemadectin. The identification of new impurities and correlation of their chromatographic profiles with the EP method is very useful to establish the stability profile of moxidectin and its preparations, as well as add value to the forthcoming moxidectin finished product European Pharmacopeia monographs.

Acid catalyzed degradation of moxidectin into 23-keto-moxidectin and 3,4-epoxy-moxidectin

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Acknowledgments

This research work was supported by Technology New Zealand (TechNZ). The authors would like to take this opportunity to thank Mr. Robert Holmes and Ancare Scientific Ltd, New Zealand for their continuous support. Also, special thanks to Prof. Saranjit Singh (NIPER, India) for his guidance on characterization protocols.

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Authors and Affiliations

  1. School of Pharmacy, The University of Auckland, Private Bag 92019, Auckland, 1142, New Zealand

    Atul Awasthi, Raida Al-Kassas & Sanjay Garg

  2. Ancare Scientific Ltd, Auckland, PO Box 36240, Northcote, Auckland, 0748, New Zealand

    Majid Razzak

  3. School of Biological Sciences, The University of Auckland, Private Bag 92019, Auckland, 1142, New Zealand

    David R. Greenwood

  4. New Zealand Institute for Plant & Food Research Limited, Private Bag 92169, Auckland, 1142, New Zealand

    David R. Greenwood

  5. School of Chemical and Physical Sciences, Victoria University of Wellington, PO Box 600, Wellington, New Zealand

    Joanne Harvey

  6. School of Pharmacy and Medical Sciences, University of South Australia, PO Box 2471, Adelaide, 5001, Australia

    Sanjay Garg

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  1. Atul Awasthi
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  2. Majid Razzak
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  3. Raida Al-Kassas
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  4. David R. Greenwood
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Correspondence to Sanjay Garg.

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Awasthi, A., Razzak, M., Al-Kassas, R. et al. Isolation and characterization of degradation products of moxidectin using LC, LTQ FT-MS, H/D exchange and NMR. Anal Bioanal Chem 404, 2203–2222 (2012). https://doi.org/10.1007/s00216-012-6393-9

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