Development and Validation of HPLC-DAD and UHPLC-DAD Methods for the Simultaneous Determination of Guanylhydrazone Derivatives Employing a Factorial Design.

Wanessa Azevedo De Brito,Cícero Soares Aragão,Fernando Andrade Nogueira,João Xavier De Araújo-Júnior,Monique Gomes Dantas,Thiago Aquino,Lilian Da Silva Solon,Ana Barreto Gomes,Edeildo Ferreira Da Silva-Júnior,Êurica Adélia Nogueira Ribeiro

doi:10.3390/molecules22091394

Wanessa Azevedo De Brito, Cícero Soares Aragão + Show 8 more

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https://doi.org/10.3390/molecules22091394

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Abstract

Guanylhydrazones are molecules with great pharmacological potential in various therapeutic areas, including antitumoral activity. Factorial design is an excellent tool in the optimization of a chromatographic method, because it is possible quickly change factors such as temperature, mobile phase composition, mobile phase pH, column length, among others to establish the optimal conditions of analysis. The aim of the present work was to develop and validate a HPLC and UHPLC methods for the simultaneous determination of guanylhydrazones with anticancer activity employing experimental design. Precise, exact, linear and robust HPLC and UHPLC methods were developed and validated for the simultaneous quantification of the guanylhydrazones LQM10, LQM14, and LQM17. The UHPLC method was more economic, with a four times less solvent consumption, and 20 times less injection volume, what allowed better column performance. Comparing the empirical approach employed in the HPLC method development to the DoE approach employed in the UHPLC method development, we can conclude that the factorial design made the method development faster, more practical and rational. This resulted in methods that can be employed in the analysis, evaluation and quality control of these new synthetic guanylhydrazones.

Highlights

Guanylhydrazones are widely studied molecules of great scientific interest due to their broad biological potential, which includes antihypertensive, antidiabetic, antimalarial, trypanocidal, anti-bacterial and antitumoral activities [1,2,3,4,5,6,7,8,9,10]
1.27% and those for the inter-day precision ranged from 0.43 to 0.68%. These results indicated a better precision of the UHPLC method than the high performance liquid chromatography (HPLC) method (Table 1)
All guanylhydrazones were synthesized as described in the literature [6] and their purity was confirmed by elemental analysis, 1 H-NMR and melting point

Summary

Introduction

Guanylhydrazones are widely studied molecules of great scientific interest due to their broad biological potential, which includes antihypertensive, antidiabetic, antimalarial, trypanocidal, anti-bacterial and antitumoral activities [1,2,3,4,5,6,7,8,9,10]. Andreani et al synthesized and evaluated the biological activity of some new guanylhydrazones against a series of lung, breast and Molecules 2017, 22, 1394; doi:10.3390/molecules22091394 www.mdpi.com/journal/molecules Molecules. They concluded that the 3-nitrophenyl and 4-nitrophenyl moieties are important pharmacophore groups which increase the affinity for specific tumor receptors [11]. Other which increase the affinity for specific tumor receptors [11].

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