Simultaneous Assay of ρ-Coumaric Acid and Coumarin Co-encapsulated in Lipid-core Nanocapsules: Validation of an LC Analytical Method

Author(s): Ticiane Krapf Boschetti, Edilene Gadelha de Oliveira, Lucas Almeida Rigo, Karine Coradini, Ruy Carlos Ruver Beck*

Journal Name: Current Pharmaceutical Analysis

Volume 15 , Issue 2 , 2019

Become EABM
Become Reviewer
Call for Editor

Graphical Abstract:


Background: Lipid-Core Nanocapsules (LNC) containing co-encapsulated -coumaric acid and coumarin are under development. However, there is a lack of analytical methods to assay these bioactives in nanoformulations.

Objective: The aim of this study was to validate an LC analytical method for the simultaneous determination of ρ-coumaric acid and coumarin in lipid-core nanocapsules.

Methods: The mobile phase was composed of acetonitrile:water (40:60 v/v) adjusted to pH 4 and a C- 18 reversed-phase column was used. Both bioactives were detected at 275 nm. Specificity, linearity, range, precision and accuracy of the method were assessed, according to the official requirements.

Results: Nanocapsules containing ρ-coumaric and coumarin had monomodal particle size distribution, spherical-shape and Z-average size of 207 ± 2 nm. LC method was specific, linear (5 to 30 µg.mL-1), precise (RSD < 5%) and accurate (97 - 103%). It was applied to assay the content and encapsulation efficiency of the bioactive substances in LNC, which were close to 0.5 mg.mL-1 and 72%, respectively.

Conclusion: The proposed analytical method is reliable for the simultaneous assay of ρ-coumaric acid and coumarin in nanocapsules and can be further used in their development.

Keywords: Bioactives, chromatography, ρ-coumaric acid, coumarin, nanocapsules, validation.

Botelho, M.A.; Barros, G.; Queiroz, D.B.; Carvalho, Q.C.F.; Gouvea, J.; Patrus, L.; Bannet, M.; Patrus, D.; Rego, A.; Silva, I.; Campus, G.; Araújo-Filho, I. Nanotechnology in phytotherapy: anti-inflammatory effect of a nanostructured tymol gel from Lippia sidoides in acute periodontitis in rats. Phytother. Res., 2016, 30, 152-159.
Zorzi, G.K.; Carvalho, E.L.S.; von Poser, G.L.; Teixeira, H.F. On the use of nanotechnology-based strategies for association of complex matrices from plant extracts. Rev. Bras. Farmacogn., 2015, 25, 426-436.
Pohlmann, A.R.; Fonseca, F.N.; Paese, K.; Detoni, C.B.; Coradini, K.; Beck, R.C.R.; Guterres, S.S. Poly (Ɛ-caprolactone) microcapsules and nanocapsules in drug delivery. Expert Opin. Drug Deliv., 2013, 10(5), 623-638.
Paese, K.; Jäger, A.; Poletto, F.S.; Pinto, E.F.; Rossi-Bergmann, B.; Pohlmann, A.R.; Guterres, S.S. Semisolid formulation containing a nanoencapsulated sunscreen: Effectiveness, in vitro photostability and immune response. J. Biomed. Nanotechnol., 2009, 5(3), 240-246.
Coradini, K.; Lima, F.O.; Oliveira, C.M.; Chaves, P.S.; Athayde, M.L.; Carvalho, L.M.; Beck, R.C.R. Co-encapsulation of resveratrol and curcumin in lipid-core nanocapsules improves their in vitro antioxidant effects. Eur. J. Pharm. Biopharm., 2014, 88(1), 178-185.
Boechat, A.L.; Oliveira, C.P.; Tarragô, A.M.; Costa, A.G.; Malheiro, A.; Guterres, S.S.; Pohlmann, A.R. Methotrexate-loaded lipid-core nanocapsules are highly effective in the control of inflammation in synovial cells and a chronic arthritis model. Int. J. Nanomedicine, 2015, 22, 6603-6614.
Rodrigues, S.F.; Fiel, L.A.; Shimada, A.L.; Pereira, N.R.; Guterres, S.S.; Pohlmann, A.R.; Farsky, S.H. Lipid-core nanocapsules act as a drug shuttle through the blood brain barrier and reduce glioblastoma after intravenous or oral administration. J. Biomed. Nanotechnol., 2016, 12, 986-1000.
Venturini, C.G.; Jäger, E.; Oliveira, C.P.; Bernardi, A.; Battastini, A.M.O.; Guterres, S.S.; Pohlmann, A.R. Formulation of lipid core nanocapsules. Colloids Surf. A Physicochem. Eng. Asp., 2011, 375, 200-208.
Poletto, F.S.; Oliveira, C.P.; Wender, H.; Regent, D.; Donida, B.; Teixeira, S.R.; Guterres, S.S.; Rossi-Bergmann, B.; Pohlmann, A.R. How sorbitan monostearate can increase drug-loading capacity of lipid-core polymeric nanocapsules. J. Nanosci. Nanotechnol., 2015, 15, 827-837.
Zanotto-Filho, A.; Coradini, K.; Braganhol, E.; Schröder, R.; Oliveira, C.M.; Simões-Pires, A.; Battastini, A.M.O.; Pohlmann, A.R.; Guterres, S.S.; Forcelini, C.M.; Beck, R.C.R.; Moreira, J.C.F. Curcumin-loaded lipid-core nanocapsules as a strategy to improve pharmacological efficacy of curcumin in glioma treatment. Eur. J. Pharm. Biopharm., 2013, 83, 156-167.
Gressler, L.T.; Oliveira, C.B.; Coradini, K.; Rosa, L.D.; Grando, T.H.; Baldissera, M.D.; Zimmermann, C.E.; Da Silva, A.S.; Almeida, T.C.; Hermes, C.L.; Wolkmer, P.; Silva, C.B.; Moreira, K.L.S.; Beck, R.C.R.; Moresco, R.N.; Da Veiga, M.L.; Stefani, L.M.; Monteiro, S.G. Trypanocidal activity of free and nanoencapsulated curcumin on Trypanosoma Evansi. Parasitol., 2015, 142(3), 439-448.
Santos, S.C.; Krueger, C.L.; Steil, A.A.; Kreuger, M.R.; Biavatti, M.W.; Junior, A.W. LC characterisation of Guaco Medicinal Extracts, Mikania laevigata and M. glomerata, and their effects on Allergic Pneumonitis. Planta Med., 2006, 72, 679-684.
Rufatto, L.C.; Gower, A.; Schwambach, J.; Moura, S. Genus Mikania: chemical composition and phytotherapeutical activity. Rev. Bras. Farmacogn., 2012, 22(6), 1384-1403.
Silva, L.S.; Brumano, L.; Stringheta, P.C.; Pinto, M.A.O.; Dias, L.O.M.; Muller, C.S.M.; Scio, E.; Fabri, R.L.; Castro, H.C.; Amaral, M.P.H. Preparation of dry extract of Mikania glomerata Sprengel (Guaco) and determination of its coumarin levels by spectrophotometry and HPLC-UV. Molecules, 2012, 17, 10344-10354.
Stražišar, M.; Andrenšek, S.; Šmidovnik, A. Effect of β-cyclodextrin on antioxidant activity of coumaric acids. Food Chem., 2008, 110, 636-642.
Rękas, A.; Ścibisz, I.; Siger, A.; Wroniak, M. The effect of microwave pretreatment of seeds on the stability and degradation kinetics of phenolic compounds in rapeseed oil during long-term storage. Food Chem., 2017, 222, 43-52.
Trenor, S.R.; Shultz, A.R.; Love, B.J.; Long, T.E. Coumarins in polymers: from light harvesting to photo-cross-linkable tissue scaffolds. Chem. Rev., 2004, 104, 3059-3077.
Jiang, J.; Qi, B.; Lepage, M.; Zhao, Y. Polymer micelles stabilization on demand through reversible photo-cross-linking. Macromolecules, 2007, 40, 790-792.
Chung, J.W.; Lee, K.; Neikirk, C.; Nelson, C.M.; Priestley, R.D. Photoresponsive coumarin-stabilized polymeric nanoparticles as a detectable drug carrier. Small, 2012, 8(11), 1693-1700.
Marturano, V.; Cerruti, P.; Carfagna, C.; Giamberini, M.; Tylkowski, B.; Ambrogi, V. Photo-responsive polymer nanocapsules. Polymer, 2015, 70, 222-230.
Celeghini, R.M.S.; Vilegas, J.H.Y.; Lanças, F.M. Extraction and quantitative HPLC analysis of coumarin in hydroalcoholic extracts of Mikania glomerata Spreng. (“guaco”) leaves. J. Braz. Chem. Soc., 2001, 12(6), 706-709.
Mattila, P.; Kumpulainen, J. Determination of free and total phenolic acids in plant-derived foods by HPLC with diode-array detection. J. Agric. Food Chem., 2002, 50, 3660-3667.
Bertoldi, F.C.; Deschamps, F.C.; Silva-Junior, A.A.; Correa, A.F.; Franco, M.F.; Eberlin, M.N. Validação de um método analítico rápido por CLAE-UV para determinação de cumarina em guaco (Mikania glomerata Sprengel) confirmado com espectrometria de massas. Rev. Bras. Pl. Med, 2016, 18(1), 316-325.
Vianna, D.; Corvello, F.; Ródio, C.; Bruxel, F.; Velho, A.; Carvalho, E.S.; von Poser, G.; Teixeira, H.F. Spectrophotometric determination of coumarins incorporated in nanoemulsions containing Pterocaulon balansae extract. Lat. Am. J. Pharm., 2011, 30(8), 1487-1491.
International Conference on Harmonisation (ICH). Validation of Analytical Procedures: Text and methodology Q2(R1), 2005.
Krai, J.S.; Andrade, D.F.; Beck, R.C.R. Development and validation of a simple LC-UV method to assay doxazosin in polymeric nanocapsules and tablets. Curr. Pharm. Anal., 2017, 13(3), 264-270.
Guimarães, K.L.; Ré, M.I. In: Nanocosmetics and Nanomedicines; Beck, R.; Guterres, S.; Pohlmann, A. Eds.; Springer-Verlag, V: Berlin Heidelberg, 2011; pp. 101-122.
Benbettaïeb, N.; Chambin, O.; Assifaoui, A.; Al-Assaf, S.; Karbowiak, T.; Debeaufort, F. Release of coumarin incorporated into chitosan-gelatin irradiated films. Food Hydrocoll., 2016, 56, 266-276.
Oliveira, C.P.; Venturini, C.G.; Donida, B.; Poletto, F.S.; Guterres, S.S.; Pohlmann, A.R. An algorithm to determine the mechanism of drug distribution in lipid-core nanocapsule formulations. Soft Matter, 2013, 9, 1141-1150.

Rights & PermissionsPrintExport Cite as

Article Details

Year: 2019
Published on: 04 January, 2019
Page: [145 - 151]
Pages: 7
DOI: 10.2174/1573412913666171027163451
Price: $65

Article Metrics

PDF: 23
PRC: 1