Antioxidant, Antimicrobial, and Metabolomic Characterization of Blanched Pomegranate Peel Extracts: Effect of Cultivar.

Hot water blanching at 80 °C for 3 min can be used as a novel pre-treatment step in pomegranate peel to preserve the integrity of the phytochemical content within the peel extracts by lowering or inactivating enzymes such as polyphenol (PPO) oxidase and peroxidase (POD) that are responsible for the break-down of phytochemicals within the peel. The aim of this study was to investigate the effect of hot water blanching pre-treatment on yield, bioactive compounds, antioxidants, enzyme inactivation, and antibacterial activity of 'Wonderful', 'Acco', and 'Herskawitz' pomegranate peel extracts. We used a variety of spectrophotometric-based assays and liquid chromatography mass spectrometry (LC-MS)-based approach to characterize and quantify metabolites within the peel extracts. Blanching significantly (p < 0.05) reduced PPO activity in all peel extracts, with the highest PPO reduction in 'Herskawitz' peel extracts at 0.25 U/mL. Furthermore, higher antioxidant activity in 'Herskawitz' blanched peel extracts using 2,2-diphenyl-1-picryl hydrazyl (DPPH) antioxidant activity, ferric ion reducing antioxidant power (FRAP), and 2,2-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical scavenging activity at 567.78 ± 9.47 µmol Trolox/g DM, 800.05 ± 1.60 µmol Trolox/g DM, and 915.27 ± 0.61 µmol Trolox/g DM, respectively, was noted. 'Herskawitz' blanched peel extracts were recorded with the lowest minimum inhibitory concentration (MIC) value of 80 µg/mL for Gram-positive Bacillus subtilis and Gram-negative Klebsiella pneumoniae bacteria strains. A total of 30 metabolites were present in 'Acco' and 'Herskawitz' peel extracts and were tentatively identified after LC-MS profiling. This study demonstrates that blanched peel extracts from 'Herskawitz' cultivar have great potential for commercial use in value-added products in the nutraceutical, cosmeceutical, and pharmacological industries.

Blanching Pre-Treatment Promotes High Yields, Bioactive Compounds, Antioxidants, Enzyme Inactivation and Antibacterial Activity of 'Wonderful' Pomegranate Peel Extracts at Three Different Harvest Maturities.

'Wonderful' pomegranate (Punica granatum L.) peel contains a wide range of phytochemicals including vitamins, dietary fibre, phenolic compounds, and antioxidant properties. Yet, it is often used as animal feed or discarded in landfills, which is not the best eco-friendly way to utilize this phenolic-rich bioresource. Finding novel ways of utilizing pomegranate peel waste could prove a more profitable and eco-friendlier alternative that is far more beneficial to the economy. Adding a blanching pre-treatment step at optimal conditions prior to processing of pomegranate peel aids in the inactivation of quality changing enzymes such as polyphenol oxidase (PPO) and peroxidase (POD), which are accountable for the degradation reactions that cause breakdown of nutrients and phytochemicals. This study aimed to determine the effect of blanching at 80 °C for 3 min on the yield, polyphenol content, antioxidant properties, enzyme inactivation, and antibacterial activity of 'Wonderful' pomegranate peel ethanolic extracts from three different harvest maturities (unripe, ripe, and over ripe), including a comprehensive characterization and quantification using liquid chromatography-mass spectrometry (LC-MS). The blanched unripe peel extracts exhibited the highest total phenolic content, total tannin content, 2,2-diphenyl-1-picryl hydrazyl (DPPH) antioxidant activity, 2,2-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical scavenging activity and ferric ion reducing antioxidant power (FRAP) at 14.0 mg gallic acid equivalent (GAE)/g dry mass (DM), 1.0 mg GAE/g DM, 359.1 µmol Trolox/g DM, 912.2 µmol Trolox/g DM and 802.5 µmol Trolox/g DM, respectively. There was significant (p < 0.05) decrease in PPO and POD activity of all blanched pomegranate peel extracts. The blanched unripe peel extracts had the lowest PPO activity at 0.2 U/g fresh weight (FW), with a 70% PPO inactivation compared to ripe and over ripe harvest, whereas the highest POD inactivation was recorded at 67% in over ripe peel extracts. All blanched peel extracts, irrespective of harvest maturity, had minimum inhibitory concentration (MIC) values at 160 µg/mL against all four bacteria strains tested, which included two Gram-positive bacterial strains (Bacillus subtilis ATCC 6051 and Staphylococcus aureus ATCC 12600) and two Gram-negative bacteria (Escherichia coli 11775 and Klebsiella pneumonia ATCC 13883). A total of 25 metabolites including phenolic acids (4), organic acids (1), flavonoids (4), ellagitannins (13), and other polyphenols (3) in all three pomegranate peel samples were tentatively identified after LC-MS profiling. The blanched unripe peel extracts showed significantly higher punicalin α and ß, ß punicalagin, catechin, epicatechin content at 414 mg/g, and 678 mg/g, 151 mg/g, 229 mg/g, respectively, compared to peel extracts from other harvest maturities. This study provides supportive information for the commercial utilization of pomegranate fruit peel as source of value-added ingredients for the development of novel food, cosmetics, and pharmacological products.

Processing Factors Affecting the Phytochemical and Nutritional Properties of Pomegranate (Punica granatum L.) Peel Waste: A Review.

Pomegranate peel has substantial amounts of phenolic compounds, such as hydrolysable tannins (punicalin, punicalagin, ellagic acid, and gallic acid), flavonoids (anthocyanins and catechins), and nutrients, which are responsible for its biological activity. However, during processing, the level of peel compounds can be significantly altered depending on the peel processing technique used, for example, ranging from 38.6 to 50.3 mg/g for punicalagins. This review focuses on the influence of postharvest processing factors on the pharmacological, phytochemical, and nutritional properties of pomegranate (Punica granatum L.) peel. Various peel drying strategies (sun drying, microwave drying, vacuum drying, and oven drying) and different extraction protocols (solvent, super-critical fluid, ultrasound-assisted, microwave-assisted, and pressurized liquid extractions) that are used to recover phytochemical compounds of the pomegranate peel are described. A total phenolic content of 40.8 mg gallic acid equivalent (GAE)/g DM was recorded when sun drying was used, but the recovery of the total phenolic content was higher at 264.3 mg TAE/g when pressurised liquid extraction was performed. However, pressurised liquid extraction is costly due to the high initial investment costs and the limited possibility of carrying out selective extractions of organic compounds from complex peel samples. The effects of these methods on the phytochemical profiles of pomegranate peel extracts are also influenced by the cultivar and conditions used, making it difficult to determine best practice. For example, oven drying at 60 °C resulted in higher levels of punicalin of 888.04 mg CE/kg DM compared to those obtained 40 °C of 768.11 mg CE/kg DM for the Wonderful cultivar. Processes that are easy to set up, cost-effective, and do not compromise the quality and safety aspects of the peel are, thus, more desirable. From the literature survey, we identified a lack of studies testing pretreatment protocols that may result in a lower loss of the valuable biological compounds of pomegranate peels to allow for full exploitation of their health-promoting properties in potentially new value-added products.