Abstract
The antioxidant, antithrombotic and anti-inflammatory effects of the amphiphilic compounds extracted from both avocado juice and by-products, were evaluated. All extracts were assessed for their total phenolic content (TPC) and total carotenoid content (TCC), and for their antioxidant activities by DPPH, ABTS and FRAP assays as well as for their anti-inflammatory and antithrombotic potency in human platelets. The extracts rich in TAC (Total Amphiphilic Content) showed much higher content in phenolics and carotenoids from the extracts of total lipophilic content (TLC), which was reflected by the much stronger antioxidant capacities of TAC extracts. ATR-FTIR spectroscopy revealed the presence of not only phenolics and carotenoids, but also of bioactive polar lipids (PLs) in avocado TAC extracts, the LC-MS based structural analysis of which further revealed a fatty acid composition favourable for unsaturated fatty acids (UFAs) versus saturated ones (SFAs), including monounsaturated fatty acids (MUFAs) like the oleic acid (C18:1n9) and omega-3 (n3) polyunsaturated fatty acids (PUFAs) like the alpha linolenic acid (C18:3n3), with the subsequent anti-inflammatory low values of the n6/n3 PUFA ratio. The presence of such bioactive PLs that are rich in UFA within the TAC extracts of avocado juice and its by-products provide an explanation for the observed potent anti-inflammatory and antithrombotic activities of avocado TAC against thrombo-inflammatory mediators like platelet activating factor (PAF) and against standard platelet agonists like ADP, offering promise for such avocado TAC extracts, as ingredients in functional products for health/promoting applications either in cosmetics or in functional foods and nutraceuticals, or even drugs.
1. Introduction
Avocado, particularly valued for its nutritional richness, includes bioactive compounds located in the pulp, seed, and peel, solidifying its position as a functional food with extensive health benefts [1]. Often referred to as “vegetable buter”, due to its high unsaturated fat content, avocado also contains essential vitamins (notably C and D), minerals (such as copper), and phytochemicals like favonoids, which contribute to its antioxidant and anti-infammatory properties [2,3]. These bioactives are known to enhance immune function, mitigate infammation, and reduce the risks associated with chronic diseases [4,5]. Its wealth of secondary metabolites and active molecules, including phenolics, tannins, carotenoids, and anthocyanins, further bolster avocado s antioxidant profle, which highlights its potential role as a protective dietary component [3].
Researching the bioactive constituents of avocado has emerged as a promising area in the prevention and management of infammation-related chronic diseases, including cardiovascular diseases, cancer, diabetes, and respiratory disorders. These conditions contribute signifcantly to global mortality, particularly in developed and developing nations, where they not only afect health, but also burden daily life and economic systems worldwide [6–8]. Risk factors associated with these diseases, both modifable and non-modifable, can foster persistent infammation and oxidative stress, fuelling disease progression [9]. While medical treatments remain the primary approach, concerns over side efects, costs, and accessibility can limit their efectiveness [10]. Bioactive compounds in foods, and especially fruits, not limited to citrus fruits [11] but also in oily fruits like avocado, however, present a natural alternative for supporting health without adverse efects [12]. For those unable to maintain a daily intake of these bioactives through diet, the development of supplements and nutraceuticals derived from avocado provides a practical and accessible option [13–15].
In terms of specifc health effects, research shows that avocado consumption supports cardiovascular health by enhancing blood lipid profles, improving endothelial function, and reducing risk factors for heart disease [4,5]. Additionally, bioactive compounds found in avocado, such as carotenoids and xanthophylls (e.g., lutein and zeaxanthin), have demonstrated anticancer properties, particularly against tumours in the breast, larynx, and oral regions [1]. Avocado´s benefits also extend to diabetes management, with compounds that inhibit enzymes involved in glucose metabolism, potentially reducing rapid increases in blood sugar levels [16]. The evidence thus supports avocado´s multifunctional health efects, ofering a dietary tool for reducing chronic disease risks through bioactive intake [17].
With over three million tons of avocados produced globally each year, where only the pulp is typically consumed, the seeds and peel often become waste, underscoring the need for sustainable approaches to avocado by-products [1,18,19]. A circular economy approach to these by-products aligns with UN and EU sustainability directives, highlighting their utility as cost-efective sources of bioactives that can be repurposed for health-promoting applications [13–15]. Rich in bioactives like monounsaturated faty acids, tocopherols, polyphenols, and carotenoids, avocado by-products exhibit anti-infammatory and antimicrobial efects, positioning them as valuable resources for pharmaceutical, nanotechnological, and tissue engineering applications [19–21]. Moreover, cafeoylquinic acid from avocado could enhance drug bioavailability, while polysaccharides in seeds and peels may be converted into biomaterials, such as polylactic acid (PLA), further supporting green technology and bioproduct development while lipid-soluble polyphenols that are primarily composed of cafeic acid derivatives such as cafeic acid ethylester are potent antioxidants known to inhibit cancer cell growth and lipid oxidation [19,20,22,23].
Consequently, the aim of the present study was to evaluate the physicochemical characteristics of the fruits, peel and pulp of Persea americana Mill. (avocado) of Cretan biological origin, with particular emphasis on the quantifcation of the content of lipophilic, amphiphilic bioactive constituents (i.e., carotenoids and phenolics, respectively) and the determination through biological platelet aggregometry assays of the diferent lipid extracts of avocado by-products, as well as the evaluation of their antioxidant and anti-infammatory capacity, together with their structural elucidation.
2. Materials and Methods
2.1. Materials, Reagents and Instrumentation
Organic cultured Cretan Hass avocado fruits (three in total) of the Fuerte variety grown in Crete were the plant material from which the samples were prepared. Specifcally, the fruits were sampled from orchards in the prefecture of Rethymnon: Koumpes area of the Municipality of Rethymnon. All reagents (Folin-Ciocalteu, Na2CO3, DPPH, ABTS), solvents (chloroform, methanol, petroleum ether, ethanol, n-octane and isopropanol), phenolic and lipid standards (Trolox, gallic acid, quercetin, catechin, soybean polar lipids and beta-carotene, respectively) were purchased from Sigma Aldrich. All UV-Vis spectroscopy analyses were performed on an LLG-uniSPEC 2 spectrophotometer and ATR-FTIR spectroscopy analyses on a Perkin Elmer Frontier ATR/FT-NIR/MIR spectrometer (Perkin Elmer, Waltham, MA, USA).
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In Vitro Antioxidant, Antithrombotic and Anti-Inflammatory Activities of the Amphiphilic Bioactives Extracted from Avocado and Its By-Products
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Marra, A.; Manousakis, V.; Koutis, N.; Zervas, G.P.; Ofrydopoulou, A.; Shiels, K.; Saha, S.K.; Tsoupras, A. In Vitro Antioxidant, Antithrombotic and Anti-Inflammatory Activities of the Amphiphilic Bioactives Extracted from Avocado and Its By-Products. Antioxidants 2025, 14, 146. https://doi.org/10.3390/antiox14020146
