A sustainable and efficient preparation process of anthocyanins from blue honeysuckle fruit and comprehensive bioactivity assessment

https://doi.org/10.1016/j.jtice.2020.10.029Get rights and content

Highlights

  • Efficient DES-NPCE method was developed for anthocyanins extraction from blue honeysuckle.

  • Extraction process parameters were determined by rational experimental design methods.

  • Yields of anthocyanins and cyanidin-3-glucoside reached 6.601 mg/g and 4.839 mg/g.

  • Multiple kinetic models were used to analyze the extraction process and thermal stability.

  • Anthocyanins product exhibited powerful chemical and cellular antioxidant activities.

Abstract

In the present study, an integrated process for preparation of anthocyanins components from blue honeysuckle fruits using deep eutectic solvent-based negative pressure cavitation extraction (DES-NPCE) was explored rationally. Single factor optimization combined with Box-Behnken Design (BBD) were applied to confirm the influential parameters. The maximum extraction yields of total anthocyanins and main constituent cyanidin-3-glucoside (C3G) reached 6.601 mg/g and 4.839 mg/g, which were 1.197–1.565 folds of other conventional methods. Moreover, the anthocyanins products exhibited higher stability from DES-NPCE extraction than that of conventional extraction solvents, which was conducive to the preservation and utilization of anthocyanins. After enrichment with macroporous resin D101, the contents of total anthocyanins and C3G reached 25.72% and 19.05%, and the recovery yields were 81.35% and 82.24%, respectively. Furthermore, the anthocyanins products exhibited more powerful chemical and cellular antioxidant activities, which were similar or stronger than that of positive control Vc or BHT. These results revealed that the proposed process was definitely an efficient and sustainable strategy for the stabilization preparation of thermo sensitive biologically active natural products. The developed method has also proved to have sufficient potential for application in food, nutrition and pharmaceutical industries.

Introduction

Lonicera caerulea L. (blue honeysuckle) is a high nutritionally valuable northern temperate zone bush, which is widely planted in Russia, China, and Japan for function fruits. Recent research has supported the biological activities of blue honeysuckle fruit such as hypertension, gastrointestinal disorders, and bacterial infections [1]. Phytochemical researches have shown that anthocyanins are the main bioactive constituents in blue honeysuckle fruit, including cyanidin-3-rutinoside, cyanidin-3, 5-diglucoside and cyanidin-3-glucoside. Cyanidin-3-glucoside (C3G) is the main anthocyanin in blue honeysuckle fruits extract, accounting for 79–88% of total anthocyanins [2]. These anthocyanins compounds possess antioxidant, aging resistance and obesity control, anti-cancer, lower heart diseases and diabetes risk [3], [4], [5]. Due to these promising healthy values, it is quite essential to expore an eco-friendly and efficient process for anthocyanins extraction and enrichment [6].

Extraction, as the first and key procedure for the preparation and recovery of active constituents from plant, has attracted wide attention in recent years [7, 8]. It is necessary to utilize sufficiently green and sustainable solvents to replace toxic extraction solvents and to meet the stringent demands in medicinal and food industries [9]. Deep eutectic solvent (DES), which is common consists of three or two green and cheap constituents, has become a promising alternative to conventional organic solvents in extraction prccedure [10]. DESs with different physico-chemical properties can be prepared to meet specific research demands by adjusting the types and ratios of hydrogen bond acceptor and donor (HBA and HBD). Although considering the obvious advantages of DES, it is more convincing to explore the extraction performances of tailored-DESs in natural products preparation process [11]. Over the last decades, many traditional extraction techniques have been widely used, including heat-refluxing, ultrasound-assisted and microwave-assisted extraction etc. However, these methods usually have many shortcomings can't be ignored [12]. Negative-pressure cavitation extraction (NPCE) device, researched and applied by our own lab, can generate cavitation effect under negative pressure. In this device, enhanced collision, turbulence and mass transfer promote the mixing of plant materials with solvent as well as movement of bioactive components from the plant cell [13, 14]. Compared with traditional methods, NPCE has shown many superiorities including shorter time cost, lower solvent consumption, superior extraction efficiency and easy operation in large scale. Moreover, NPCE process can be conducted in relatively lower temperature, it is beneficial for the thermo sensitive constituents extraction, especially for the anthocyanins production [15]. Until now, there is no reporter to apply DESs based NPCE (DES-NPCE) process for the extraction of anthocyanins compounds from herbs.

Response surface method (RSM) uses rational experimental design methods to combine certain data obtained from experiments, and then uses multiple quadratic regression equations to fit the functional relationship between factors and response values [16], [17], [18]. It is a statistical method that solves multivariate problems and determines the optimal response value and corresponding parameters through the analysis of the regression equation [19], [20], [21]. Box-Behnken Design (BBD) is a type of RSM and every factor has only three levels. Normally, it is very economical when the number of factors is 3, but when the number of factors exceeds 4, it is inappropriate due to the excessive number of experiments.

Anthocyanins have been widely recognised as edible functional foods and nutritional supplements in health care and food fields because of its outstanding biological activity [22]. Therefore, the biological activity assessment of anthocyanins product is quite important for further utilization. Many studies have shown the instability of anthocyanins during thermal process. Antioxidant activity, as a representative activity of anthocyanins, is a key index of anthocyanins preparation products evaluation.

In this study, the parameters of DES-NPCE process were confirmed systematically using single factor optimization with Box-Behnken design after Plackett–Burman design. Then the stability of anthocyanins in extraction solution was also evaluated. Subsequently, anthocyanins compounds were recovered from the DES extract through the resin enrichment process. DPPH, ABTS, FRAP and cellular level measures were conducted to assess the antioxidant activity of anthocyanin products. Up to our knowledge, the integrative preparation process of anthocyanins from blue honeysuckle fruit by DES-NPCE has not been reported yet. This efficient and sustainable method for preparing anthocyanin products from blue honeysuckle fruits, with excellent stability and antioxidant activity, could provide new thoughts in food, nutritional and medicinal industries.

Section snippets

Materials and chemicals

Mature blue honeysuckle fruit were picked from Heilongjiang Academy of Agricultural Sciences China. Ethylene glycol, 1,3-butanediol, 1,4-butanediol, Glucose, Sucrose, Sorbitol, Malic acid, Lactic acid, Citric acid were purchased from Tokyo Chemical Industry Co. Ltd. (Tokyo, Japan). Choline chloride was obtained from Aladdin Co., Ltd. (Shanghai, China). Resins were purchased from Bonchem (Hebei, China) and Nankai Hecheng S & T (Tianjin, China). DPPH, ABTS, BHT and VC were obtained from

Selection of DESs system

The extraction yields by the organic acids-based DESs were higher than that sugar-based and polyalcohols-based, which was conformity with the strong polar characteristics of anthocyanin components (Fig. 1A). Furthermore, the pH of organic acids-based DESs is the lowest among these nine DESs. Many studies have proven that acidic conditions can make the structure of anthocyanins compounds more stable and achieve better distribution effect. We can find that the highest extraction yields

Conclusion

This study was the first report to develop a sustainable and high-efficiency preparation process of anthocyanins compounds from blue honeysuckle fruits using DES-NPCE. The extraction yields of total anthocyanins and C3G from blue honeysuckle fruit reached 6.601 mg/g and 4.839 mg/g in DES-ChCl/ lactic acid (1:2, mol/mol) 20% aqueous solution under the optimized conditions: extraction pressure −0.08 MPa, liquid/solid ratio 16 mL/g and extraction temperature 53 °C, which was 1.197–1.565 folds to

Declaration of Competing Interest

The authors declare no conflict of interest.

Acknowledgements

The authors gratefully acknowledge the financial supports by National Key R & D Program of China (2017YFD0600205), Fundamental Research Funds for the Central Universities (2572019AA17) the National Natural Science Foundation of China (31930076) and the 111 Project (B20088), Academy of Changbai Mountain Science (201702), Heilongjiang Touyan Innovation Team Program (Tree Genetics and Breeding Innovation Team).

References (36)

Cited by (10)

  • Modifiers based on deep eutectic mixtures: A case study for the extraction of anthocyanins from black bean hulls using high pressure fluid technology

    2022, Journal of Supercritical Fluids
    Citation Excerpt :

    On the other hand, DES have low cost, simplicity, stability, and adjusted versatility [7]. In recent research, different DES has proven to be excellent solvents for anthocyanin extractions [1,11–13]. However, the wide range of HBA and HBD compounds increases the difficulty of choosing the most promising solvent.

View all citing articles on Scopus
1

These authors contributed equally to this work.

View full text