Dynamic maceration of Matricaria chamomilla inflorescences: optimal conditions for flavonoids and antioxidant activity

Revista Brasileira de Farmacognosia 28 (2018) 111–117 www.elsevier.com/locate/bjp Original Article Dynamic maceration of Matricaria chamomilla inflorescences: optimal conditions for flavonoids and antioxidant activity Simone Vieira Pereira a , Rayssa A.S.P. Reis a , Danielle Cristina Garbuio b,c , Luis Alexandre Pedro de Freitas a,∗ a Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil Escola de Enfermagem de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil c Faculdades Anhanguera, Valinhos, SP, Brazil b a r t i c l e i n f o Article history: Received 13 August 2017 Accepted 9 November 2017 Available online 20 December 2017 Keywords: Flavonoids DPPH Apigenin Apigenin-7-glycoside Desirability a b s t r a c t The aim of this paper was to study and optimize the dynamic maceration process to obtain Matricaria chamomilla L., Asteraceae, inflorescences extracts with optimum flavonoid content and antioxidant activity using a multivariate approach. Hydroalcoholic extracts were obtained by dynamic maceration in lab scale and the influence of extraction temperature, ratio of plant to solvent, ethanol strength; extraction time and stirring speed on the flavonoid content and antioxidant activity were unveiled using a fractional factorial design. The ethanol strength, ratio of plant to solvent and temperature were the three factors that influenced most the extract properties and were studied by a central composite design. Total flavonoid content and antioxidant activity were affected by the ethanol strength and ranged from 1.49 to 3.95% and 13.3 to 36.2 ␮g/ml, respectively. The desirability functions resulted in an optimal dynamic maceration condition using 1 h extraction at stirring speed of 900 rpm, ethanol 74.7%, temperature of 69 ◦ C and using 36.8% of plant in solvent (w/v). Under this set of conditions, the extract had total flavonoid content of 4.11 ± 0.07%, in vitro antioxidant activity with IC50 of 18.19 ␮g/ml and apigenin and apigenin7-glycoside contents of 2.0 ± 0.1 mg/g and 20.1 ± 0.9 mg/g, respectively. The results showed a low solvent consumption compared to previous works. The model was able to predict extract properties with maximum deviation of 12% and the extraction process developed herein showed to be reliable, efficient and scalable for M. chamomilla inflorescences, enriched with flavonoids, apigenin and apigenin-7-glycoside and high antioxidant activity. © 2017 Published by Elsevier Editora Ltda. on behalf of Sociedade Brasileira de Farmacognosia. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/ 4.0/). Introduction Plant extracts usually present several therapeutic applications because of their richness in bioactive and have been used to treat man’s health problems since ancient times. Recognizing this fact, the World Health Organization (WHO) has also been working to promote the use of these in health systems and recently published guidelines for the quality, safety and efficacy of herbal medicines. The sustainable production and efficacy of medicinal plants have made their extracts a focus of world attention (Ansel et al., 2000; Srivastava et al., 2010; Cortés-Rojas et al., 2015). Among the worldwide known medicinal plants there is the Matricaria chamomilla L., Asteraceae, which is one of the oldest and ∗ Corresponding author. E-mail: (L.A. Freitas). best documented plants in the world (Srivastava et al., 2010) and its inflorescences are widely used to obtain infusions, extracts and essential oils. Due to M. chamomilla diverse therapeutic properties, it stands out as the most cultivated and consumed medicinal plant in the world today, with estimated more than one million cups of its tea consumed daily (Borsato et al., 2005; Srivastava et al., 2010). The extract of M. chamomilla has a wide variety of constituents, which can provide antioxidant, anti-inflammatory, moisturizing and emollient properties that could be used in formulations for topical application. Also because of the numerous benefits to the skin the use of extracts and essential oils in pharmaceutical and cosmetic formulations is increasing. Despite this, there are still few studies on topical formulations containing the extracts of chamomile (Dal’Belo et al., 2006; Nóbrega et al. (2013)). Chamomile extraction can be performed by different processes and conditions that can influence extract characteristics. Harbourne et al. (2009) studied chamomile extraction by water infusion at 90 ◦ C during https://doi.org/10.1016/j.bjp.2017.11.006 0102-695X/© 2017 Published by Elsevier Editora Ltda. on behalf of Sociedade Brasileira de Farmacognosia. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). 112 S.V. Pereira et al. / Revista Brasileira de Farmacognosia 28 (2018) 111–117 20 min comparing fresh and dried flowers and found total phenolic content of 19.7 ± 0.5 mg/g and 13.0 ± 1.0 mg/g for fresh and oven-dried inflorescences, respectively. The M. chamomilla soxhlet, microwave-assisted (MAE), ultrasound-assisted (UAE) and subcritical water (SCW) extractions were compared (Cvetanovic et al., 2014) using the same solvent, drug-to-solvent proportion, time and temperature. SCW at 200 ◦ C and 1.6 bar for 40 min gave the best results with 49.70% extraction yield, phenolic content from 117.31 to 151.45 mg/g and flavonoid content from 51.6 to 64.3 mg/g. Dynamic maceration of M. chamomilla (Srivastava and Gupta, 2009a,b) using methanol, ethanol, water and methanol–water mixtures as solvent at 200 rpm, 37 ◦ C, drug/solvent ratio of 1:20 for 4 h demonstrated the water extract had no apigenin but high contents of apigenin-7-glycoside. Solid liquid extraction using aqueous 0.5 N HCl, 0.5 N NaOH and water at 80 ◦ C and drug/solvent proportion of 1:20 for 3 h resulted in extracts with higher phenolic content in alkaline conditions, but flavonoid content and antioxidation activity was higher in acidic solvent (Osman et al., 2016). Recently, the International Conference on Harmonization (Freitas et al., 2017) had established the guidelines for the quality of pharmaceutical products. The guideline Q8 is devoted to the development of pharmaceutical products and indicates the need for a deep understanding of the process and formulation influences on final product quality. Among the considerations, ICH-Q8 recommends the application of multivariate analysis to fully understand the processes involved. Therefore, the design of experiments, DOE, is an increasingly important tool that allows obtaining a greater number of information using a smaller number of experiments (Peralta-Zamora et al., 2005; Ferreira et al., 2007; Rodrigues and Iemma, 2009). The DOE became a very powerful tool to generate, interpret and apply scientific experiments in most efficient way (Ferreira et al., 2007) with optimization of product and process. The aim of this w (...truncated)