Research Article: Optimization of a polyphenol extraction method for sweet orange pulp (Citrus sinensis L.) to identify phenolic compounds consumed from sweet oranges

Date Published: January 30, 2019

Publisher: Public Library of Science

Author(s): Lisard Iglesias-Carres, Anna Mas-Capdevila, Francisca I. Bravo, Gerard Aragonès, Begoña Muguerza, Anna Arola-Arnal, Vijai Gupta.


The consumption of sweet oranges has been linked to several health benefits, many of which are attributed to hesperidin, a flavanone that is present in high amounts in these fruits. However, other phenolic compounds can contribute to the bioactivity of sweet orange. To link those effects to their phenolic profile, the complete characterization of the phenolic profile is mandatory. Although many studies have profiled the phenolic composition of orange juices, their pulps, which retain phenolic compounds, are overlooked. This fact is particularly relevant because dietary guidelines recommend the consumption of whole fruits. Therefore, this study aimed to develop a specific method for the optimal extraction of phenolics from orange pulp and to use this method to characterize these fruits grown at different locations by HPLC-ESI-MS/MS. The extraction conditions that reported the highest total polyphenol content (TPC) and hesperidin contents were 20 mL/g, 55 °C, and 90% methanol. The extraction time and number of sequential steps were further evaluated and optimized as 20 min and two extraction steps, respectively. Although lower extraction rates were achieved when using ethanol as the extraction solvent, high TPC and hesperidin yields were obtained, suggesting the potential use of this methodology to produce phenolic-rich extracts for the food industry. By applying the optimized methodology and analyzing the extracts by HPLC-ESI-MS/MS, geographic cultivation regions were demonstrated to affect the phenolic profiles of oranges. In short, we developed a quick, easy-to-perform methodology that can be used to extract orange phenolics from pulp for their identification and quantification and to evaluate the factors that affect the phenolic profile in sweet orange pulps.

Partial Text

Phenolic compounds are secondary plant metabolites that can enter the human diet though the consumption of vegetal products. Importantly, a wide range of health-related activities have been reported for phenolic compounds. In this sense, the phenolic compounds present in citrus fruits have received attention due to their anti-oxidant, anti-inflammatory and cardioprotective activities [1]. Sweet oranges are a rich source of flavanones, and the effects of their consumption are related to cardioprotective effects, among others [2,3]. Remarkably, hesperidin, the most abundant flavanone in sweet oranges [1,4,5], is believed to be the responsible for many of the biological effects linked to sweet orange consumption [1–3]. Other relevant compounds in sweet oranges that can contribute to their health-promoting activities include narirutin, phenolic acids and flavonols [4,6,7].

Citrus polyphenols such as hesperidin are known to exert beneficial health effects when consumed [1–3]. The proper quantification of a food matrix is essential to correlate its consumption and an observed health effect. The diversity of phenolic compounds found among fruits [1], fruit varieties [5] and even fruit parts [31] makes it difficult to develop a universal extraction method. Therefore, specific extraction methods must be developed to accurately profile the phenolic composition of a plant matrix. Moreover, no optimized extraction methods have been developed for orange pulps: they not only contain substantial quantities of phenolic compounds [29,30], but their consumption is also recommended over that of their juices [32,33]. To determine the optimal extraction conditions for orange pulp, RSM was applied to optimize the TPC and as the main compound occurring in sweet oranges, hesperidin [5]. Throughout the optimization process, NOs were used due to their higher phenolic contents.

In this study, we developed an optimized extraction method that is capable of extracting Citrus sinensis phenolics from its pulp. This method consists of two relatively fast sequential extractions (20 min each) using a relatively mild temperature (55 °C) and a high methanol concentration (90%). Importantly, our method is able to extract high quantities of phenolic compounds, which is essential for determining the accurate phenolic profile present in the fruit. Given that no previous extraction methods for orange pulps are reported in the literature, we propose this method as a routine methodology to study the phenolic profile of the edible parts of sweet oranges. Additionally, this method could be useful for producing phenolic-rich extracts with potential bioactive functions. This study also indicates that the geographical region affects the phenolic profile of orange pulps.




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