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Semicarbazide Hydrochloride as Impurity in Drug Substances: a Validated LC-DAD-UV Method for Its Determination in Carbazochrome and Carbazochrome Sodium Sulfonate

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Abstract

Carbazochrome (2-(1,2,3,6-tetrahydro-3-hydroxy-1-methyl-6-oxo-5H-indol-5-ylidene)-hydrazinecarboxamide, CBZ) and carbazochrome sodium sulfonate (5-[(aminocarbonyl)hydrazono]-2,3,5,6-tetrahydro-1-methyl-6-oxo-1H-indole-2-sulfonic acid sodium salt, CSS) are derivatives of adrenochrome currently employed mainly as hemostatic drugs. Since a semicarbazone group is present in both the structures of CBZ and CSS, semicarbazide hydrochloride (SEM) is used, as reagent, in their synthesis. Belonging to the hydrazine family of chemicals, SEM could possess some toxic potential. Accordingly, literature data showed that SEM causes osteolathyrism and induces toxic effects in the cardiovascular and skeletal systems. Moreover, SEM has a weak genotoxic activity in vitro while in vivo available data are not sufficient to state its genotoxicity. Based on these data, the determination of SEM as a synthesis-related impurity in CBZ and CSS drug substances is mandatory. Since there are no publications dealing with the determination of SEM in CBZ and CSS as well as in any other SEM-containing drugs, a new and unprecedented LC-DAD-UV method for its determination as impurity in CBZ and CSS drug substances was developed and validated. Due to the lack of chromophore, semicarbazide was converted into the corresponding UV-absorbing naphthalene-2-carbaldehyde semicarbazone (NCS) before the chromatographic analysis. The complete separation of NCS from the large concentration of the derivatization reagent and the parent drugs CBZ and CSS was accomplished through a liquid–liquid extraction (LLE) step followed by a chromatographic run of 10 min. The method was then successfully validated and its applicability demonstrated by the determination of SEM in real samples of CBZ and CSS.

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References

  1. Moriuchi H, Kashiwada Y, Arai I, Yuizono T (1995) Effects of carbazochrome sodium sulfonate (AC-17) on oleic acid-induced lung injury. Pharmacol Toxicol 77:238–240

    Article  CAS  Google Scholar 

  2. Sendo T, Itoh Y, Aki K, Oka M, Oishi R (2003) Carbazochrome sodium sulfonate (AC-17) reverses endothelial barrier dysfunction through inhibition of phosphatidylinositol hydrolysis in cultured porcine endothelial cells. Naunyn Schmiedebergs Arch Pharmacol 368:175–180

    Article  CAS  Google Scholar 

  3. Basile M, Gidaro S, Pacella M, Biffignandi PM, Gidaro GS (2002) Troxerutin-carbazochrome combination versus placebo in the treatment of posthemorrhoidectomy symptoms: a single-center, randomized, double-blind, placebo-controlled study. Curr Ther Res 63:527–535

    Article  CAS  Google Scholar 

  4. Maranghi F, Tassinari R, Lagatta V, Moracci G, Macrì C, Eusepi A, Di Virgilio A, Scattoni ML, Calamandrei G (2009) Effects of the food contaminant semicarbazide following oral administration in juvenile Sprague-Dawley rats. Food Chem Toxicol 47:472–479

    Article  CAS  Google Scholar 

  5. The EFSA Journal, 219 (2005) Opinion of the Scientific Panel on Semicarbazide in food:1–36

  6. http://www.efsa.europa.eu/sites/default/files/scientific_output/files/main_documents/428.pdf. Accessed 25 Sept 2016

  7. Abramsson-Zetterberg L, Svensson K (2005) Semicarbazide is not genotoxic in the flow cytometry-based micronucleus assay in vivo. Toxicol Lett 155:211–217

    Article  CAS  Google Scholar 

  8. Takahashi M, Yoshida M, Inoue K, Morikawa T, Nishikawa A, Ogawa K (2014) Chronic toxicity and carcinogenicity of semicarbazide hydrochloride in Wistar Hannover GALAS rats. Food Chem Toxicol 73:84–94

    Article  CAS  Google Scholar 

  9. Vlastos D, Moshou H, Epeoglou K (2010) Evaluation of genotoxic effects of semicarbazide on cultured human lymphocytes and rat bone marrow. Food Chem Toxicol 48:209–214

    Article  CAS  Google Scholar 

  10. Wang Y, Chan HW, Chan W (2016) Facile formation of a DNA adduct of semicarbazide on reaction with apurinic/apyrimidinic sites in DNA. Chem Res Toxicol 29:834–840

    Article  CAS  Google Scholar 

  11. Wang Y, Wong TY, Chan W (2016) Quantitation of the DNA adduct of semicarbazide in organs of semicarbazide-treated rats by isotope-dilution liquid chromatography—tandem mass spectrometry: a comparative study with the RNA adduct. Chem Res Toxicol 29:1560–1564

    Article  CAS  Google Scholar 

  12. De la Calle B, Anklam E (2005) Semicarbazide. Occurrence in food products and state-of-the-art in analytical methods used for its determination. Anal Bioanal Chem 382:968–977

    Article  Google Scholar 

  13. Wang Y, Chan W (2016) Automated in-injector derivatization combined with high-performance liquid chromatography—fluorescence detection for the determination of semicarbazide in fish and bread samples. J Agric Food Chem 64:2802–2808

    Article  CAS  Google Scholar 

  14. Fang Z, Jiang B, Wu W, Xiang Z, Ouyang C, Huang T, Chen J, Zeng L (2013) ELISA detection of semicarbazide based on a fast sample pretreatment method. Chem Commun 49:6164–6166

    Article  CAS  Google Scholar 

  15. Chumanee S, Sutthivaiyakit S, Sutthivaiyakit P (2009) New reagent for trace determination of protein-bound metabolites of nitrofurans in shrimp using liquid chromatography with diode array detector. J Agric Food Chem 57:1752–1759

    Article  CAS  Google Scholar 

  16. Aprile S, Canavesi R, Bianchi M, Grosa G, Del Grosso E (2017) Development and validation of a stability-indicating HPLC-UV method for the determination of thiocolchicoside and its degradation products. J Pharm Biomed Anal 132:66–71

    Article  CAS  Google Scholar 

  17. ICH, 3A (R2) (2006) Impurities in new drug substances. In: International Conference on Harmonisation, Geneva, Oct 2006

  18. ICH, Q2A (R1) (2005) Validation of analytical procedures: text and methodology. In: International Conference on Harmonisation, Geneva, Nov 2005

  19. Beaudet C, Debot F, Lambot H, Toussaint J (1951) The monosemicarbazone of adrenochrome. Experientia 7:293–294

    Article  CAS  Google Scholar 

  20. Center for Drug Evaluation and Research, US Food and Drug Administration (1994) Reviewer guidance, validation of chromatographic methods; FDA, Rockville, MD, Nov 1994

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Acknowledgements

Financial support from Università del Piemonte Orientale, Italy, (MedChem-2015) is gratefully acknowledged.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences and Drug and Food Biotechnology Center, Università del Piemonte Orientale, Largo Donegani 2, 28100, Novara, Italy

    Rossana Canavesi, Silvio Aprile, Erika Del Grosso & Giorgio Grosa

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  1. Rossana Canavesi
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  2. Silvio Aprile
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  3. Erika Del Grosso
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  4. Giorgio Grosa
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Correspondence to Rossana Canavesi.

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Canavesi, R., Aprile, S., Del Grosso, E. et al. Semicarbazide Hydrochloride as Impurity in Drug Substances: a Validated LC-DAD-UV Method for Its Determination in Carbazochrome and Carbazochrome Sodium Sulfonate. Chromatographia 80, 1535–1544 (2017). https://doi.org/10.1007/s10337-017-3368-1

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  • DOI: https://doi.org/10.1007/s10337-017-3368-1

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