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Bashmil, Y. Phenolic Compounds from Australian Grown Bananas. Encyclopedia. Available online: (accessed on 07 December 2023).
Bashmil Y. Phenolic Compounds from Australian Grown Bananas. Encyclopedia. Available at: Accessed December 07, 2023.
Bashmil, Yasmeen. "Phenolic Compounds from Australian Grown Bananas" Encyclopedia, (accessed December 07, 2023).
Bashmil, Y.(2021, October 22). Phenolic Compounds from Australian Grown Bananas. In Encyclopedia.
Bashmil, Yasmeen. "Phenolic Compounds from Australian Grown Bananas." Encyclopedia. Web. 22 October, 2021.
Phenolic Compounds from Australian Grown Bananas

Bananas are an essential source of staple food and fruit worldwide and are widely regarded as the world’s largest fruit crop, with more than 100 million tons total annual production. Banana peel, a by-product that represents about 40% of the entire banana’s weight, and pulp are rich in bioactive compounds and have a high antioxidant capacity.

banana polyphenols antioxidant activities characterization identification quantification HPLC-PDA LC-MS/MS

1. Introduction

Banana is a tropical seasonal fruit that encompasses a diverse range of species belonging to the genus Musa of the Musaceae family. It is among the world’s most popular fruits and the 4th most cultivated crop on a worldwide scale [1]. Almost all recognized cultivars are descended from two diploid species; Musa balbisiana and Musa acuminata, the most widespread of which are the Cavendish type [2]. Asia is the world’s largest banana producer, accounting for 54.4 percent of global banana production, as per the recent Food and Agriculture Organization (FAO) statistics. With an average usage of 12 kg per person per annum [3], bananas are one of the world’s primary food crops behind rice, wheat, and maize. In Australia, particularly in Queensland, banana production is one of the largest agricultural industries, with over 390,000 tonnes in 2015 [3] and around 600 merchant banana suppliers. The region of Australian banana cultivation varied between 10,000 and 16,000 ha during the period 1997–2015 [4]. Overall, 95% of banana genome groups in Australia are Cavendish, the remaining are Lady Fingers (AAB), Goldfingers (AAAB), Ducasse (ABB), Red Dacca (AAA), Sucrier (AA), and the Pacific Plantain (AAB) sub-group, and 95% of production is in North Queensland (QLD) [5].
The banana fruit is divided into two parts: the peel and the pulp. The peel, which is the fruit’s primary secondary product, accounts for around 40% of the fruit’s overall mass. Banana peels served no purpose and were discarded as waste, resulting in vast quantities of organic materials going into landfill. The central part, the pulp, the consumable portion of the fruit, is highly nutritious. Various research on banana pulp has examined various features, from its usage as a food fortification component to the extraction and recovery of numerous healthy constituents, including multiple kinds of starch, cellulose, and bioactive phytochemicals [6]. It is well established that banana flesh and skin comprise several secondary metabolites, such as catecholamines [7], phenolics [8], and carotenoids [9]. These bioactive substances promote probiotic development and help to reduce the risk of cancer and cardiovascular disease [10]. Pulp and peel parts include phenolic compounds, carotenoids, flavonoids, biogenic amines, phytosterols, and other phytochemicals [11]. Bananas have a greater antioxidant potential than various berries, herbs, and vegetables, attributed to the prevalence of these components [12]. Amongst the existing carotenoids in banana fruit, α-carotene, β-carotene, and β-cryptoxanthin have provitamin A activity, whereas others such as lycopene and lutein have a significant antioxidant capability [13]. Later, 171 various cultivars of Musa spp. were analysed for provitamin A carotenoids and 47 cultivars for two minerals, iron and zinc. It has been found that there is a great variability in provitamin A amongst the various genotypes, but a low variability in iron and zinc, regardless of the soil type and growing environmental conditions [14]. Moreover, banana pulp includes numerous antioxidants, such as phenolic compounds and vitamins, for example, catechin, epicatechin, lignin and tannin, and anthocyanin [15]. The catechins (epicatechin and gallocatechin) are most prevalent in the pulp when analysed by high-performance liquid chromatography [16]. Russel et al. have detected many phenolics in banana, such as ferulic, sinapic, salicylic, gallic, p-hydroxybenzoic, vanillic, syringic, gentisic, and p-coumaric acids as major components. Nevertheless, ferulic acid concentration was the highest among other phenolics [17]. According to Tsamo et al., hydroxycinnamic derivatives, such as ferulic acid-hexoside, were the major ones (4.4–85.1 µg/g DW) in plantain pulp. They discovered large differences in the phenolic contents among the tested varieties. In the plantain peels, rutin was the most abundant flavonol glycoside (242.2–618.7 µg/g DW). Therefore, the banana peel and pulp both are good sources of health-promoting phenolic compounds [18]. The main flavonoids detected in bananas are as follows: quercetin, myricetin, kaempferol, and cyanidin, which provide health benefits mostly due to their action as free radical scavengers [19]. Most of these phenolics, as natural antioxidants, have several biological effects, containing antiviral, antibacterial, antiallergenic, anti-inflammatory, vasodilatory and antithrombotic functions [20][21].
Although the concentration of phenolics in different banana cultivars has previously been investigated, a thorough profile of these compounds, especially in cultivars grown in Australia, remains inadequate due to their complicated composition and nature. Chemical analysis of these phenolics is typically performed by liquid chromatography-mass spectrometry (LC-MS) following sample extraction and filtering. Mass spectrometry is a well-established analytical technique used to infer unknown chemicals from complicated samples of various plant components, including banana [22].
To accomplish the study’s purpose, the polyphenols in six Australian grown banana cultivars, Cavendish, Ladyfinger, Red Dacca, Plantain, Monkey, and Ducasse were defined by TPC, TFC and TTC. In addition, the antioxidant capacity of all varieties was evaluated by ferric reducing antioxidant power (FRAP), 2,2′-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) assay (ABTS), ferrous ion chelating assay (FICA), reducing power assay (RPA), hydroxyl radical scavenging assay (OH-RSA) and total antioxidant capacity assay (TAC). Furthermore, the characterization and classification of phenolics were conducted using LC-ESI-QTOF-MS/MS, whereas the quantification of selected phenolics from various banana cultivars conducted using HPLC-PDA. As banana peel and pulp have a high antioxidant capacity, this study reinforces their usage as an origin of polyphenols in various industries, such as food, nutraceuticals, pharmaceuticals, cosmetics, and animal feed.

2. Polyphenols Assessment of Banana

The assessment of polyphenols in various banana cultivars was attained via TPC, TFC, and TTC. Diets high in fruits and vegetables have been associated with a decreased risk of heart diseases and cancer. The antioxidant components of fruit and vegetables, for example, phenolic acids, flavonoids, tannins, lignans and stilbenes with pharmacological and biological attributes, have been credited with their protecting impact [23]. Multiple factors, such as plants’ varieties, degree of ripening, growing environment, and initial treatments, can affect the composition and concentrations of these components [24].
The highest phenolic contents were found in the ripe Ducasse peel and pulp, unripe Ladyfinger peel, ripe Plantain peel, unripe Monkey peel and unripe Ladyfinger pulp, which contained 1.32, 1.28, 1.15, 0.87, 0.79 and 0.76 mg GAE/g, respectively, whereas the lowest value of TPC was found in ripe Red Dacca pulp (0.40 mg GAE/g) and ripe Plantain pulp (0.38 mg GAE/g) (Table 1).
Table 1. Polyphenol contents of six Australian grown bananas.
Banana Samples TPC (mg GAE/g) TFC (mg QE/g) TTC (mg CE/g)
Ripe Cavendish 0.54 ± 0.03 g–i 0.01 ± 0.001 h–j 0.19 ± 0.02 gh
Unripe Cavendish 0.71 ± 0.04 d–f 0.01 ± 0.0004 h–j 0.5 ± 0.05 d–f
Ripe Ladyfinger 0.56 ± 0.05 gh 0.01 ± 0.00 g–i 0.12 ± 0.01 hi
Unripe Ladyfinger 1.15 ± 0.04 b 0.007 ± 0.001 ij 0.12 ± 0.01 hi
Ripe Red Dacca 0.64 ± 0.05 e–g 0.01 ± 0.0005 d–f 0.38 ± 0.02 ef
Ripe Plantain 0.87 ± 0.01 c 0.03 ± 0.001 a 0.53 ± 0.03 c–e
Ripe Monkey 0.56 ± 0.03 gh 0.006 ± 0.0004 j 0.56 ± 0.01 cd
Unripe Monkey 0.79 ± 0.05 cd 0.013 ± 0.001 de 0.44 ± 0.03 d–f
Ripe Ducasse 1.32 ± 0.10 a 0.02 ± 0.0004 b 3.34 ± 0.2 a
Ripe Cavendish 0.43 ± 0.01 h–j 0.01 ± 0.0003 e–g 0.02 ± 0.01 i
Unripe Cavendish 0.55 ± 0.03 g–i 0.01 ± 0.001 f–h 0.66 ± 0.08 c
Ripe Ladyfinger 0.42 ± 0.01 ij 0.001 ± 0.0001 k 0.42 ± 0.012 d–f
Unripe Ladyfinger 0.76 ± 0.02 c–e 0.01 ± 0.001 g–i 1.52 ± 0.09 b
Ripe Red Dacca 0.40 ± 0.01 j 0.01 ± 0.0001 f–i -
Ripe Plantain 0.38 ± 0.01 j 0.01 ± 0.001 d -
Ripe Monkey 0.43 ± 0.03 h–j 0.02 ± 0.001 c 0.001 ± 0.0 i
Unripe Monkey 0.58 ± 0.03 fg 0.02 ± 0.001 c 0.02 ± 0.01 i
Ripe Ducasse 1.28 ± 0.03 a 0.03 ± 0.001 a 0.34 ± 0.04 fg
Values are mean ± standard deviation per gram fresh weight; n = 3 samples per sample. Values within the same column with different superscript letters (a–k) are significantly different from each other (p < 0.05). TPC (total phenolic contents); TFC (total flavonoid contents); TTC (total tannin contents); GAE (gallic acid equivalents); QE (quercetin equivalents); and CE (catechin equivalents).
TPC values ranging from 0.15 to 55.5 mg GAE/g had already been reported in twenty-seven different banana cultivars in which the Brazilian Cavendish has the second highest value (29.2 mg GAE/g) [24]. However, TPC differences can be ascribed to extraction circumstances (solvent kind, concentration, solvent/sample ratio, and time/temperature combination for extracting), cultivar variations, and the geographical location of where the bananas were grown [25]. Flavonoids are the most abundant class of phenolic compounds present in practically all plants, as shown in this study using the aluminium chloride colorimetric method.
Aluminium chloride interacts with the flavonoids’ carbonyl group, generating a stable compound [26]. The TFC values in the banana samples had a similar pattern of TPC results, in which the most prevalent flavonoid compounds were identified in the ripe Ducasse pulp (0.03 mg QE/g), and peel (0.02 mg QE/g), whereas the lowest values were found in ripe Monkey peel (0.006 mg QE/g). TFC values ranged from 8.56 ± 0.22 to 16.15 ± 0.28 mg QE/g in Egyptian banana cultivars, according to Aboul-Enein, et al. [27], which was higher than our Australian samples. This may be due to varieties divergence or type of solvent used for the extraction of phenolic compounds. Comparing to other tropical fruits, it has been found that the mango peel showed the maximum concentration of flavonoid (1.75 ± 0.08 mg QE/g), then pineapple and banana peels (1.47 ± 0.07 and 1.32 ± 0.12 mg QE/g), respectively [28].
Tannins are also a substantial family of phenolic substances that could be partitioned into hydrolysable and non-hydrolysable tannins. In our banana samples, the ripe Ducasse peel had the highest amount (3.34 ± 0.2 mg CE/g), followed by the unripe Ladyfinger pulp (1.52 ± 0.09 mg CE/g) and unripe Cavendish pulp (0.66 ± 0.08 mg CE/g). Most fruits show a reduction in tannins content when they ripen, which is believed to be due to tannin polymerisation [29][30][31]. According to Von Loesecke [32], several forms of tannin and soluble tannin become insoluble throughout ripening. Barnell and Barnell [29] explained that the Latex vessels in both the pulp and peel and small dispersed cells in the peel are two kinds of tannin-containing elements. Furthermore, they suggested that the tannin structure in latex vessels varies, whereas the tannin content of the peel’s tiny dispersed cells tends to have little or no alteration. On the other hand, Kyamuhangire et al. [33] found that Mbidde bananas had a greater tannin content than Matooke bananas at both the green and mature phases. The high tannin content is due to the unusual presence of laticifer cells in the fruit of Mbidde varieties, which is similar to our findings in ripe Ducasse peel. Moreover, their results showed that the amount of extractable tannins improved with ripening in all four banana cultivars, and after ripening, the amount of water-extractable tannins increased significantly in Mbidde bananas, but a very small amount was found in Matooke bananas. The increased amount of extractable tannins appears to indicate that tannins become more accessible as the banana ripens. This contrasts with the observation that bananas lose their astringent flavour during maturation, a phenomenon explained by increasing tannin polymerisation [29] and inactivity [34].

3. Antioxidant Capacity of Banana

Polyphenols are essential antioxidants found in plants and have a variety of biological actions. They are proven to have several functions because they work in biological systems as reducing chemicals, metal chelators, hydrogen atom donors, scavengers of radical oxygen. The antioxidant capacity of banana was further studied using a variety of methods, including radical scavenging ability and reducing power of the sample. For this aim, DPPH, FRAP, ABTS, RPA, FICA, OH-RSA and TAC experiments were performed, and the results presented in Table 2.
Table 2. Antioxidant activity of six Australian grown banana.
Banana Samples DPPH
(mg AAE/g)
(mg AAE/g)
(mg AAE/g)
(mg AAE/g)
(mg EDTA/g)
(mg AAE/g)
(mg AAE/g)
Ripe Cavendish 0.67 ± 0.02 d 1.28 ± 0.06 de 2.41 ± 0.19 a–c 2.22 ± 0.23 de 0.11 ± 0.01 e 104.32 ± 1.72 ab 0.12 ± 0.006 jk
Unripe Cavendish 0.49 ± 0.01 ef 1.77 ± 0.04 c 2.29 ± 0.22 a–d 2.26 ± 0.15 de 0.04 ± 0.003 fg 105.95 ± 0.72 a 0.17 ± 0.005 ij
Ripe Ladyfinger 0.59 ± 0.01 de 1.04 ± 0.06 e–g 2.69 ± 0.23 a 6.15 ± 0.50 a 0.38 ± 0.03 b 103.60 ± 0.18 ab 0.17 ± 0.01 ij
Unripe Ladyfinger 0.47 ± 0.04 f 1.35 ± 0.12 d 2.49 ± 0.17 ab 4.99 ± 0.04 b 0.32 ± 0.02 c 98.02 ± 1.57 cd 0.13 ± 0.01 jk
Ripe Red Dacca 0.13 ± 0.002 h 0.87 ± 0.02 g–i 1.76 ± 0.08 f–i 1.76 ± 0.09 ef 0.04 ± 0.0005 fg 96.58 ± 1.72 de 0.18 ± 0.01 h–j
Ripe Plantain 0.66 ± 0.01 d 1.20 ± 0.03 d–f 2.17 ± 0.16 b–f 2.37 ± 0.12 d 0.04 ± 0.002 fg 96.58 ± 0.36 de 0.25 ± 0.01 gh
Ripe Monkey 0.66 ± 0.02 d 0.93 ± 0.09 gh 0.59 ± 0.07 k 2.36 ± 0.08 d 0.59 ± 0.05 a 96.04 ± 0.79 de 0.24 ± 0.01 g–i
Unripe Monkey 0.66 ± 0.01 d 1.02 ± 0.10 e–g 1.67 ± 0.09 g–i 1.67 ± 0.17 fg 0.12 ± 0.01 e 95.14 ± 0.95 de 0.12 ± 0.01 jk
Ripe Ducasse 1.06 ± 0.09 b 2.31 ± 0.10 b 1.49 ± 0.13 hi 4.33 ± 0.17 c 0.26 ± 0.02 d 106.67 ± 1.09 a 0.08 ± 0.01 k
Ripe Cavendish 0.13 ± 0.002 h 0.69 ± 0.06 hi 1.82 ± 0.16 e–h 0.20 ± 0.05 i 0.06 ± 0.002 f 94.78 ± 0.48 de 0.39 ± 0.03 e
Unripe Cavendish 0.16 ± 0.01 h 1.004 ± 0.02 fg 2.20 ± 0.09 b–e 1.71 ± 0.21 f 0.05 ± 0.003 f 103.06 ± 0.48 a–c 0.58 ± 0.03 c
Ripe Ladyfinger 0.26 ± 0.01 g 0.86 ± 0.06 g–i 1.77 ± 0.19 f–h - - 99.46 ± 6.04 b–d 0.27 ± 0.003 fg
Unripe Ladyfinger 0.79 ± 0.05 c 1.42 ± 0.05 d 1.98 ± 0.14 c–g 1.33 ± 0.06 f–h 0.03 ± 0.002 fg 97.48 ± 0.36 de 1.03 ± 0.08 a
Ripe Red Dacca 0.09 ± 0.01 h 0.36 ± 0.02 j 1.34 ± 0.07 ij 0.89 ± 0.07 h 0.02 ± 0.001 fg 96.58 ± 1.18 de 0.34 ± 0.004 ef
Ripe Plantain 0.069 ± 0.004 h 0.62 ± 0.01 ij 2.35 ± 0.05 a–c 1.12 ± 0.03 h 0.06 ± 0.005 f 97.12 ± 1.003 de 0.29 ± 0.003 fg
Ripe Monkey 0.26 ± 0.01 g 0.80 ± 0.02 g–i 0.98 ± 0.05 jk 0.84 ± 0.07 h 0.05 ± 0.003 f 94.96 ± 0.65 de 0.40 ± 0.017 de
Unripe Monkey 0.32 ± 0.01 g 0.87 ± 0.05 g–i 1.91 ± 0.06 d–h 1.18 ± 0.10 gh 0.06 ± 0.002 f 92.25 ± 0.65 e 0.47 ± 0.03 d
Ripe Ducasse 1.68 ± 0.06 a 2.85 ± 0.28 a 1.86 ± 0.14 d–h 5.67 ± 0.04 a 0.04 ± 0.003 fg 106.85 ± 1.77 a 0.69 ± 0.04 b
Values are mean ± standard deviation per gram fresh weight; n = 3 samples per sample. Values within the same column with different superscript letters (a–k) are significantly different from each other (p < 0.05). DPPH (2,2′-diphenyl-1-picrylhydrazyl assay); FRAP (ferric reducing antioxidant power assay); ABTS (2,2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid assay); TAC (total antioxidant capacity); AAE (ascorbic acid equivalents); RPA (reducing power assay); FICA (ferrous ion chelating activity); OH-RSA (hydroxyl-radical scavenging activity); and EDTA (ethylenediaminetetraacetic acid).
The DPPH, FICA, ABTS, and OH-RSA tests have been primarily utilized to assess the scavenging activity of bioactive metabolites, primarily polyphenols [35]. In contrast, the FRAP assay assesses samples’ ability of electrons donation to convert a ferric-TPTZ complex to a blue ferrous-TPTZ complex. DPPH can supply hydrogen ions to the biological system or act as a scavenger of free radicals. When DPPH is combined with banana samples, it accepts hydrogen atoms, and diminishing violet colour occurs. The DPPH, ABTS, and FRAP values of ripe Ducasse pulp and ripe Ladyfinger peel (1.68 mg AAE/g, 2.69 mg AAE/g, and 2.85 mg AAE/g, respectively) were considerably greater than those of other listed cultivars. In contrast, ripe Plantain pulp, ripe Red Dacca pulp, and ripe Monkey peel had the lowest DPPH, FRAP, and ABTS values (0.069 mg AAE/g, 0.36 mg AAE/g and 0.59 mg AAE/g), respectively.
The DPPH values of 23 variant banana cultivars grown in different countries, ranging from 0.68 to 66.9 mg AAE/g, were recorded in a previous review, and our findings were close to some of those varieties [24]. The ability of banana peels to scavenge free radicals (DPPH) decreases as the fruit progresses from green to ripe to overripe [36][37]. Nevertheless, the electron transfer/hydrogen donating capacity of phenolics’ nature is also recognized to be connected to their DPPH radical scavenging activity. Numerous substances of botanical origin have an antioxidant activity proportional to their phenolic content, indicating a causative correlation between total phenolic concentration and antioxidant activity [38]. Furthermore, our banana FRAP values were within the range of other banana cultivars studied (0.32–17.64 mg AAE/g) by Vu, Scarlett and Vuong [24]. The FRAP value of the Brazilian Cavendish peel, which is yellow with a green edge, was higher (14 mg AAE/g) [39]. Guo [33] mentioned that most fruit peels demonstrated 2- to 27-fold greater antioxidant activity than the fruit flesh, and variations between varieties can occur because the antioxidant activity of fruits can be influenced by geographical location, variety, and harvesting or storing period. Furthermore, it is well established that phenolics’ antioxidant capacity depends on their molecular weight, the amount of aromatic rings, and the number and position of hydroxyl groups attached to aromatic ring [40]. Additionally, it has been claimed that parameters such as the stereoselectivity of the radicals or the solubility of phenolics in various testing conditions influence the capacity of extracts to react with and eliminate various radicals [41]. According to Hagerman, et al. [42], tannins, as high molecular weight phenolics, have a greater capacity to scavenge free radicals (ABTS•+).
Regarding RPA, OH-RSA and FICA assays, Ladyfinger, Monkey, and Ducasse peels had higher antioxidant capacity than other varieties. In RPA, ripe Ladyfinger peel had the highest antioxidant potential, followed by ripe Ducasse pulp and unripe Ladyfinger peel. In FICA assay, Ripe Monkey peel had high antioxidant activity compared to other cultivars. On the other hand, ripe Ducasse peel had higher OH-RSA value (106.67 mg AAE/g), followed by unripe Cavendish peel (105.95 mg AAE/g). To our best knowledge, this is the first time that banana’s antioxidant potential has been analysed through the RPA, OH-RSA and FICA assays. Previously, a thorough phenolic analysis of custard apples cultivated in Australia has been performed by Du et al. [43]. They found that African Pride peel possessed the greatest antioxidant capacity for RPA (5.32 ± 0.14 mg AAE/g), OH-RSA (1.23 ± 0.25 mg AAE/g) and FICA (3.17 ± 0.18 mg EDTA/g). Based on the current iron chelating findings, the extracts may be able to protect against oxidative stress by isolating Fe(II) ions that would otherwise accelerate Fenton-type reactions or engage in metal catalysed hydroperoxide breakdown reactions. The samples’ Fe(II) chelating properties may be a result of their endogenous chelating compounds, primarily phenolics [44]. Grinberg et al. [45] claimed that tea polyphenols’ protective action against OH-dependent salicylate hydroxylation was due to iron chelation. The research on polyphenols’ deleterious effects on iron bioavailability has emphasized that iron binding to flavonoid antioxidants can diminish iron’s accessibility to oxygen molecules. In contrast, non-flavonoid polyphenols can reduce Fe3+ ions and then create inert Fe2+ polyphenol complexes [46].
In the TAC assay, which was conducted by reducing molybdenum (VI) to molybdenum (V) in the presence of phenolics, the pulps of the banana varieties had generally higher values than peels. The highest value was observed in unripe Ladyfinger pulp (1.03 ± 0.08 mg AAE/g), followed by ripe Ducasse pulp (10.43 ± 0.20 mg AAE/g) and unripe Cavendish pulp (0.58 ± 0.03 mg AAE/g).
Remarkably, the antioxidant activity of fruits varies depending on the extraction method that was utilized. Fruit extracts contain a diverse array of bioactive chemicals. The amount of phenolic acids and flavonoids in each type is highly variable and is influenced by cultivar, geographic location, and climates. There are several procedures for determining the antioxidant capacity, and each methodology has several advantages and disadvantages. To summarize, our findings indicate that each banana cultivar’s antioxidant activity varies according to its phenolic profile or the method utilized to quantify it. Numerous in vitro experiments may be used to determine the antioxidant capacity of banana, while modern analytical techniques like as LC-ESI-QTOF-MS/MS can be used to identify and confirm these antioxidant components.
Phenolics are a broad collection of molecules that are garnering attention in research due to their possible health benefits. Due to the antioxidant properties of polyphenols, they may be used to prolong the shelf life of lipid-rich products. Additionally, bananas have diverse antibacterial phenolic compounds that act as preservatives for food during storage. Furthermore, because phytochemicals have a complex structure, no one approach accurately evaluates polyphenols’ antioxidant capacity due to the various pathways and reactions occurring in the biological system. Thus, LC-MS characterization is one of the sophisticated research approaches used to profile polyphenols and better understand the banana’s total antioxidant capacity. The polyphenol contents of banana and their antioxidant capacity indicate that additional research should be undertaken to determine the true contribution of phenolics to the antioxidant potential while excluding or limiting non-phenolic substances’ involvement.

4. Correlation between Polyphenols and Antioxidant Potentials

A regression analysis was used to determine the association between the findings of the experiments undertaken (Table 3).
Table 3. Pearson’s linear correlation between banana phenolic content and their antioxidant capacity.
TFC 0.599 **                
TTC 0.575 ** 0.286              
DPPH 0.790 ** 0.584 ** 0.444            
FRAP 0.859 ** 0.564 * 0.539 * 0.896 **          
ABTS 0.107 −0.040 −0.161 −0.010 0.138        
RPA 0.706 ** 0.360 0.227 0.670 0.659 ** 0.338      
FICA 0.182 −0.197 0.160 0.208 0.053 −0.216 0.513 *    
OH-RSA 0.466 * 0.183 0.417 0.563 * 0.736 ** 0.362 0.603 ** 0.069  
TAC −0.016 0.136 0.033 0.187 0.136 −0.050 −0.171 −0.381 −0.094
* Significant correlation at p ≤ 0.05; ** significant correlation at p ≤ 0.01.
Due to the critical antioxidant effects of polyphenols observed in vegetables and fruits, we studied TPC, TFC, and TTC in Australian bananas. TPC was shown to have a range of 0.40–1.32 mg GAE/g, with an average of 0.68 mg GAE/g (Table 1). The highest TPC value was found in Ducasse peel, while the least was reported in Red Dacca pulp. The antioxidant activity of banana extracts was determined using DPPH, FRAP, ABTS, RPA, ·OH-RSA, FICA, and TAC.
Between total phenolic content and the antioxidant activities of DPPH, FRAP, and RPA, ·OH-RSA, and TFC, a high and significant positive correlation was detected, but TFC was only correlated significantly with DPPH and FRAP. Previously, a positive correlation between antioxidant activity and total polyphenols in herbs and spices was discovered [47]. Notably, there was a negative association between TPC, TFC, and TTC and TAC, ABTS, and FICA.
TAC is also negatively correlated with ABTS, ·OH-RSA, RPA, and FICA. Total phenolics have been shown to be responsible for the antioxidant capacity of plant diets [48]. Numerous variables affect the correlation, including the number of samples analysed, the quantities measured, and the antioxidant assays used. DPPH and FRAP were highly correlated in our research. Kim, Yang, Lee and Kang discovered that TPC had a higher relationship with antioxidant activity than TFC [49].
Principal component analysis (PCA, Figure 2) demonstrated that TPC and TTC were positively correlated with ˙OH-RSA, RPA, and FICA with higher scores. Furthermore, FRAP, TFC, DPPH, and ABTS were positively correlated, while FICA showed negative correlation with TAC, TFC, ABTS, FRAP and DPPH, which revealed the divergence of bioactive components in banana cultivars.
Figure 2. Principal component analysis (PCA) of the phenolic contents (TPC, TFC, TTC, phenolic acids, and flavonoids) and their antioxidant capacities (DPPH, FRAP, ABTS, RPA, OH-RSA, FICA, and TAC) of banana cultivars.
After evaluating the data, it is indicated that the antioxidant activity of banana extracts may also be related to non-phenolic elements. Even though simple phenols have relatively little contribution to antioxidant potentials, they can interact with Folin–Ciocalteu reagent [22]. The current study’s findings reveal that distinct phenolic compounds exhibit varying levels of antioxidant activity based on their structure, synergistic effect, quantity, and hostile attitude towards other components appearing in each banana extract.

5. LC-ESI-QTOF-MS/MS Identification of the Phenolic Compounds

To identify and characterize bioactive substances, such as phenolics from various fruits, vegetables, and herbal medicines, LC-MS/MS has been broadly used. A LC-ESI-QTOF-MS/MS analysis in negative and positive ionization modes was used to obtain a non-targeted qualitative analysis of phenolics from Australian banana cultivars pulp and peel samples (Table 4) utilizing Agilent LC-MS Qualitative Software and Personal Compound Database and Library, phenolics in banana samples were identified and based on their m/z value and MS spectra in both negative and positive modes of ionization ([M–H]/[M+H]+). For further MS/MS detection, (m/z) characterization, and verification, compounds with a mass error of less than ±10 ppm were used. LC-MS/MS was used to identify 24 phenolic compounds in banana samples, which include phenolic acids (6), flavonoids (13), and other polyphenols (5) mentioned in Table 4.
Table 4. Characterization of phenolic compounds from different banana cultivars by LC-ESI-QTOF-MS/MS.
No. Proposed Compounds Molecular Formula RT
Ionization (ESI+/ESI−) Molecular Weight Theoretical (m/z) Observed (m/z) Error
MS2 Product ions Sample Code
  Phenolic acid                  
  Hydroxyphenylpropanoic acids                  
1 3-Hydroxyphenylpropionic acid C9H10O3 30.544 [M–H] 166.0630 165.0557 165.0556 −0.6 165, 121, 119 RCPl
  Hydroxycinnamic acids                  
2 Ferulic acid C10H10O4 9.105 [M–H] 194.0579 193.0506 193.0506 0.0 178, 149, 134 RCP
3 Caffeic acid C9H8O4 41.086 [M–H] 180.0423 179.035 179.035 0.0 143, 133 UCP
4 p-Coumaroyl glycolic acid C11H10O5 62.765 [M+H]+ 222.0528 223.0601 223.0599 −0.9 163 * RCPl, ULPl, RRPl, RPPl, RMP, RDPl
  Hydroxyphenylacetic acids                  
5 3,4-Dihydroxyphenylacetic acid C8H8O4 20.831 [M–H] 168.0423 167.035 167.0341 −5.4 149, 123 * RRPl, UCPl
  Hydroxybenzoic acids                  
6 3,4-O-Dimethylgallic acid C9H10O5 5.557 [M+H]+ 198.0528 199.0601 199.0595 −3.0 153, 139, 125, 111 RLPl
7 Delphinidin 3-O-(6’’-acetyl-galactoside) C23H23O13 82.037 [M–H] 507.1139 506.1066 506.1066 0.0 303, 507 * RPPl, UCPl, UCP, RLP, ULP, UMPl, UMP, RMPl, RMP
8 Pelargonidin 3-O-(6’’-succinyl-glucoside) C25H25O13 40.051 [M–H] 533.1295 532.1222 532.1181 −7.7 533, 473, 443, 383, 353 UCP
9 Cyanidin 3,5-O-diglucoside C27H31O16 89.507 [M+H]+ 611.1612 612.1685 612.1674 −1.8 449, 287 * RCP, RRPl
10 Malvidin 3-O-(6’’-acetyl-glucoside) C25H27O13 6.241 [M+H]+ 535.1452 536.1525 536.1525 0.0 535, 481, 463, 445 RRPl
11 2’-Hydroxyformononetin C16H12O5 24.296 [M+H]+ 284.0685 285.0758 285.0755 −1.1 270, 229 RMP
12 Isorhamnetin 3-O-glucoside 7-O-rhamnoside C28H32O16 29.971 [M–H] 624.169 623.1617 623.1622 0.8 433, 315, 300, 271 UCPl
13 Myricetin 3-O-rutinoside C27H30O17 24.472 [M–H] 626.1483 625.141 625.1397 −2.1 301 UCPl
14 Patuletin 3-O-glucosyl-(1->6)-[apiosyl(1->2)]-glucoside C33H40O22 37.955 [M–H] 788.2011 787.1938 787.1918 −2.5 625, 463, 301, 271 RRPl
15 Quercetin 3-O-xylosyl-glucuronide C26H26O17 38.624 [M+H]+ 610.117 611.1243 611.1238 −0.8 479, 303, 285, 239 UCPl
16 (+)-Gallocatechin 3-O-gallate C22H18O11 3.075 [M–H] 458.0849 457.0776 457.0773 −0.7 305, 169 RRPl
17 Apigenin 7-O-apiosyl-glucoside C26H28O14 88.42 [M–H] 564.1479 563.1406 563.1402 −0.7 503, 443, 383, 353, 325, 297 UCP
18 Chrysoeriol 7-O-glucoside C22H22O11 89.788 [M+H]+ 462.1162 463.1235 463.1224 −2.4 445, 427, 409, 381 RCPl
19 Neoeriocitrin C27H32O15 27.305 [M–H] 596.1741 595.1668 595.1696 4.7 431, 287 RRPl
  Other polyphenols                  
20 Scopoletin C10H8O4 7.392 [M–H] 192.0423 191.035 191.0349 −0.5 176 * RPPl, RCP, UCP
21 Urolithin A C13H8O4 4.563 [M–H] 228.0423 227.035 227.035 0.0 198, 182 UCPl
22 Umbelliferone C9H6O3 35.668 ** [M+H]+ 162.0317 163.039 163.0387 −1.8 145, 135, 119 * RMPl, UCP
23 2’-Hydroxyformononetin C16H12O5 24.296 [M+H]+ 284.0685 285.0758 285.0755 −1.1 270, 229 RMP
24 Isopimpinellin C13H10O5 5.61 [M+H]+ 246.0528 247.0601 247.0583 −7.3 232, 217, 205, 203 ULP
* = compound detected in more than one sample, with data presented only asterisk, ** = compound found in both positive [M+H]+ and negative modes [M–H]. RT stands for “retention time”. Bananas were presented with abbreviations. Ripe Cavendish peel (RCPl), Ripe Cavendish pulp (RCP), Unripe Cavendish peel (UCPl), Unripe Cavendish pulp (UCP), Unripe Ladyfinger peel (ULPl), Ripe red dacca peel (RRPl), Ripe Plantain peel (RPPl), Ripe Monkey pulp (RMP), Ripe Ducasse peel (RDPl), Ripe Ladyfinger peel (RLPl), Ripe Ladyfinger pulp (RLP), Unripe Ladyfinger pulp (ULP), Unripe Monkey peel (UMPl), Unripe Monkey pulp (UMP), and Ripe Monkey peel (RMPl).


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