Analysis of Pyrrolizidine Alkaloids in Stingless Bee Honey and Identification of a Botanical Source as Ageratum conyzoides.

Stingless bee honeys (SBHs) from Australian and Malaysian species were analysed using ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) for the presence of pyrrolizidine alkaloids (PAs) and the corresponding N-oxides (PANOs) due to the potential for such hepatotoxic alkaloids to contaminate honey as a result of bees foraging on plants containing these alkaloids. Low levels of alkaloids were found in these SBHs when assessed against certified PA standards in targeted analysis. However, certain isomers were identified using untargeted analysis in a subset of honeys of Heterotrigona itama which resulted in the identification of a PA weed species (Ageratum conyzoides) near the hives. The evaluation of this weed provided a PA profile matching that of the SBH of H. itama produced nearby, and included supinine, supinine N-oxide (or isomers) and acetylated derivatives. These PAs lacking a hydroxyl group at C7 are thought to be less hepatoxic. However, high levels were also observed in SBH (and in A. conyzoides) of a potentially more toxic diester PA corresponding to an echimidine isomer. Intermedine, the C7 hydroxy equivalent of supinine, was also observed. Species differences in nectar collection were evident as the same alkaloids were not identified in SBH of G. thoracica from the same location. This study highlights that not all PAs and PANOs are identified using available standards in targeted analyses and confirms the need for producers of all types of honey to be aware of nearby potential PA sources, particularly weeds.

Physicochemical homogeneity of stingless bee honey (Heterotrigona itama) produced in the west coast, east coast and inland area of Peninsular Malaysia.

BACKGROUND: The nutritional composition of stingless bee honey (SBH) can be affected by different climates and soil composition across different geographical areas. However, the range of attributes set for a honey quality standard should be inclusive. This study analysed the sugar profile's physiochemical properties, including quantifying the rare sugar trehalulose, organic acid and mineral composition of SBH collected from inland, and west and east coasts of Peninsular Malaysia. Forty-three SBH (Heterotrigona itama) samples were collected and labelled as <20 and <40 West Coast (<20WC, <40WC), <20 and <40 East Coast (<20EC, <40EC) and Inland, according to their distance from the coasts. RESULTS: The moisture, pH and sugar composition of all SBH samples adhered to the Malaysian Kelulut Honey Standard (MS2683:2017) but not to the International Codex Standard (CODEX) for honey. Trehalulose presence in all samples, regardless of geographical area, was predominant alongside fructose and glucose. Only hydroxymethylfurfural (HMF) content and electrical conductivity (EC) results complied with both standards. The principal component analysis biplot showed that the discrimination of SBH according to the five different areas was not feasible, indicating sample homogeneity. CONCLUSION: The physicochemical evaluation of SBH from Peninsular Malaysia shows mainly homogeneous attributes of samples across geographical locations. These findings demonstrated that the current MS2683:2017 is relevant and accommodates all SBH of H. itama species produced in Peninsular Malaysia. Furthermore, the trehalulose range calculated in this study can be implemented as a new benchmark for the indicator of SBH honey quality standard by national and international food standard committees. © 2023 Society of Chemical Industry.

Fractionation and physicochemical characterization of dietary fiber of kenaf (Hibiscus cannabinus L.) seed.

BACKGROUND: Kenaf seeds are underutilized kenaf plant by-products, containing essential nutrients including dietary fiber (DF), which can be potentially utilized as food ingredients. The present study aimed to evaluate the physicochemical characteristics of kenaf seed fiber fractions extracted from kenaf seed. RESULTS: Defatted kenaf seed powder yielded four DF fractions: alkali-soluble hemicellulose (146.4 g kg-1 ), calcium-bound pectin (10.3 g kg-1 ) and acid-soluble pectin (25.4 g kg-1 ) made up the soluble fibre fraction, whereas cellulose (202.2 g kg-1 ) comprised the insoluble fraction. All fractions were evaluated for their physicochemical properties. The DF fractions contained glucose, mannose, xylose and arabinose, and a small amount of uronic acid (1.2-2.7 g kg-1 ). The isolated pectin fractions had a low degree of esterification (14-30%). All the isolated DF fractions had high average molecular weights ranging from 0.3 to 4.3 × 106 g mol-1 . X-ray diffractogram analysis revealed that the fractions consisted mainly of an amorphous structure with a relative crystallinity ranging from 31.6% to 44.1%. The Fourier-transform infrared spectroscopy spectrum of kenaf seed and its DF fractions showed typical absorption of polysaccharides, with the presence of hydroxyl, carboxyl, acetyl and methyl groups. Scanning electron microscopy analysis demonstrated that the raw material with the rigid structure resulted in soluble and insoluble DF fractions with more fragile and fibrous appearances, respectively. The soluble DF demonstrated greater flowability and compressibility than the insoluble fractions. CONCLUSION: These findings provide novel information on the DF fractions of kenaf seeds, which could be used as a potential new DF for the food industry. © 2023 Society of Chemical Industry.

Stingless Bee (Heterotrigona Itama) Honey and Its Phenolic-Rich Extract Ameliorate Oxidant-Antioxidant Balance via KEAP1-NRF2 Signalling Pathway.

Diabetes is associated with an imbalance between oxidants and antioxidants, leading to oxidative stress. This imbalance contributes to the development and progression of diabetic complications. Similarly, renal and liver diseases are characterised by oxidative stress, where an excess of oxidants overwhelms the antioxidant defense mechanisms, causing tissue damage and dysfunction. Restoring the oxidant-antioxidant balance is essential for mitigating oxidative stress-related damage under these conditions. In this current study, the efficacy of stingless bee honey (SBH) and its phenolic-rich extract (PRE) in controlling the oxidant-antioxidant balance in high-fat diet- and streptozotocin/nicotinamide-induced diabetic rats was investigated. The administration of SBH and PRE improved systemic antioxidant defense and oxidative stress-related measures without compromising liver and renal functioning. Analyses of the liver, skeletal muscle and adipose tissues revealed differences in their capacities to scavenge free radicals and halt lipid peroxidation. Transcriptional alterations hypothesised tissue-specific control of KEAP1-NRF2 signalling by upregulation of Nrf2, Ho1 and Sod1 in a tissue-specific manner. In addition, hepatic translational studies demonstrated the stimulation of downstream antioxidant-related protein with upregulated expression of SOD-1 and HOD-1 protein. Overall, the results indicated that PRE and SBH can be exploited to restore the oxidant-antioxidant imbalance generated by diabetes via regulating the KEAP1-NRF2 signalling pathway.

In Vivo Toxicity Assessment of the Probiotic Bacillus amyloliquefaciens HTI-19 Isolated from Stingless Bee (Heterotrigona itama) Honey.

This study evaluated the acute and sub-acute toxicity of B. amyloliquefaciens HTI-19 (isolated from stingless bee honey) in female Sprague Dawley rats. In an acute toxicity study, the rats received a low dosage (1 × 109 CFU·mL-1), medium dosage (3 × 109 CFU·mL-1), or high dosage (1 × 1010 CFU·mL-1) of B. amyloliquefaciens HTI-19 daily orally by syringe-feeding for 14 days. For the subacute toxicity study, rats received a low dosage (1 × 109 CFU·mL-1) or a high dosage (1 × 1010 CFU·mL-1) for 28 days. The probiotic feeding in acute and sub-acute toxicity studies showed no mortality or significant abnormalities in rats throughout the experimental period. In week 2 of the acute study, the body weight of the rats showed a significant increase (p < 0.05) compared to the control. By gross and microscopic examination of organs, no evidently significant changes were observed in the morphology of organs. Serum biochemical tests and blood hematology tests also revealed no treatment-related changes. Overall, these data indicated that oral administration of B. amyloliquefaciens HTI-19 up to 1 × 109 CFU·mL-1 for 28 days can be considered safe.

Differentiation of High-Fructose Corn Syrup Adulterated Kelulut Honey Using Physicochemical, Rheological, and Antibacterial Parameters.

Kelulut (stingless bee) honey (KH) possesses a wide range of benefits for human consumption and can exhibit medical effects. Due to its high value, this premium honey is often adulterated with different types of cheaper sugars, causing low nutrients and potential food safety threats in the final product. This study aims to determine the physicochemical, rheological, and antibacterial properties of sugar-based adulterated KH from the stingless bee species Heterotrigona itama. Adulterated samples were prepared using pure honey mixed with different concentrations of high fructose corn syrup (HFCS), i.e., 10%, 20%, 30%, 40%, and 50%. Water activity, colour, total soluble solids, pH, turbidity, viscosity, and antimicrobial activity of KH were determined. In addition, the primary sugar composition (fructose, glucose, and trehalulose) was determined by high-performance liquid chromatography with evaporative light-scattering detection (HPLC-ELSD). This study shows that the increasing percentage of HFCS addition in the KH samples significantly increases (p < 0.05) the total soluble solids, colour, pH, turbidity, viscosity, glucose, and fructose content; meanwhile, the water activity and trehalulose were reduced significantly (p < 0.05). Antimicrobial activity against S. aureus was reduced significantly (p = 0.006) by an increased percentage of HFCS compared to Control. Antimicrobial activity against P. aeruginosa was also found to be reduced significantly but showed non-significant effect from an increased percentage of HFCS in honey (p = 0.413). The bacterium S. aureus was more vulnerable to treatment with honey from both Control and adulterated groups compared to P. aeruginosa. In conclusion, HFCS-adulterated KH and authentic KH can be differentiated using all the parameters investigated. These data are vital for the governing bodies to ensure that KH sold in the markets is free from HFCS adulteration.

Oilseed meals into foods: an approach for the valorization of oilseed by-products.

The excellent health benefits of oil extracted from seeds have increased its application in foods, pharmaceutical and cosmetic industries. This trend leads to a growing research area on their by-products, oilseed meals, to minimize environmental and economic issues. Examples of these by-products are soybean, peanut, kenaf seed, hemp, sesame, and chia seed meals. It is well known that soybean meals have wide applications in food and non-food industries, while other seed meals are not well established. Most oilseed meals are rich in health beneficial compounds and are potential sources of plant protein, dietary fiber, and antioxidants. Many studies have reported on the valorization of these by-products into value-added food products such as bakery and meat products to increase their nutritional and functional properties. These efforts contribute to the sustainability, development of novel functional food and support the zero-waste concept for the environment. This review aims to provide information on the composition of selected oilseed meals from soybean, peanut, hemp, kenaf, sesame and chia seeds, their potential applications in the bakery, meat, beverage, pasta, and other food products, and to highlight the issues and challenges associated with the utilization of oilseed meals into various food products.

A pilot study of toxicological safety evaluation in acute and subacute (28-day) oral assessments of kenaf seed beverage in rats.

Kenaf (Hibiscus cannabinus L.) seed is a non-conventional edible oilseed that can be valorized into various food products. There is a recent discovery of kenaf seed beverage (KSB) potential as a novel plant-based beverage. KSB had less crude protein than soybean (SB)but more carbohydrate, magnesium, and phosphorus contents.Levels of crude fat, phytates, oxalates, total saponins, and lipid peroxidability in KSB were lower than SB. Sugar content between KSB and SB were comparable, while antioxidant properties of KSB were superior. Ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS) analysis detected gluconic acid, citric acid, palmitic acid, oleic acid, and 13-hydroxyoctadecadienoic acid in both KSB and SB. Considering its novelty, acute and subacute oral toxicity assessments in male Sprague Dawley rats were conducted. The acute toxicity assessment was performed at a single dose of 9.2 ml/kg body weight of KSB. In the following subacute toxicity assessment, different groups of rats consumed different doses of KSB (3.1, 6.1, and 9.2 ml/kg body weight) daily for 28 days. Rats presented normal behavioral and physiological states in both toxicity studies. Growth, food and water intakes, organ weight, and hematological parameters were unaffected. No mortality was reported. Several alterations in serum biochemical parameters were within the normal range, and unassociated with histopathological changes. The oral lethal dose (LD50) and the no-observed-adverse-effect-level (NOAEL) of KSB in rats was greater than 9.2 ml/kg (=1533 mg/kg) body weight. Interestingly, KSB exhibited comparable effects with soybean beverage (SB) on high-density lipoprotein cholesterol and triglycerides which worth further research Follow-up toxicity assessments in animals and human trials are also recommended to ascertain its long term safety.

Utilizing Nutritional and Polyphenolic Compounds in Underutilized Plant Seeds for Health Application.

Plants represent a significant part of the human diet. Humans have utilized every part of plants for survival, and seeds are no exception. Seeds offer high protein, unsaturated fats, fibre, essential vitamins, and minerals for various food applications. They are also a promising reservoir of bioactive compounds, where various phytochemicals, such as polyphenolic compounds, capable of maintaining and improving well-being, are present in abundant quantities. Plants from Malvaceae and Cannabaceae families are known for their fibre-rich stems that benefit humankind by serving numerous purposes. For many centuries they have been exploited extensively for various commercial and industrial uses. Their seeds, which are often regarded as a by-product of fibre processing, have been scientifically discovered to have an essential role in combating hypercholesterolemia, diabetes, cancer, and oxidative stress. Maximizing the use of these agricultural wastes can be a promising approach to creating a more sustainable world, in accordance with the concept of Sustainable Development Goals (SDGs).

Unique physicochemical properties and rare reducing sugar trehalulose mandate new international regulation for stingless bee honey.

Stingless bee honey (SBH) of four stingless bee species (Heterotrigona itama, Geniotrigona thoracica, Tetragonula carbonaria, and Tetragonula hockingsi) from two geographic regions (Malaysia and Australia, n = 36) were studied for their physicochemical parameters, including total phenolic and multi-elemental contents. Sugar analysis confirmed the prominent presence of trehalulose in all samples. All SBH failed to meet the CODEX Standard for honey moisture, free acidity, and total fructose plus glucose levels. One-way ANOVA, principal component analysis (PCA) and hierarchical component analysis (HCA) confirm distinctive differences between Australian and Malaysian SBH with Australian SBH having significantly (P < 0.05) higher pH, total phenolic, total mineral, and electrical conductivity attributes. Toxic elements in all SBH were much lower than standard CODEX limit. The species G. thoracica produced honey with significantly (P < 0.05) higher trehalulose than both Australian bee species. This research will add value to SBH and hasten new international standards ensuring commercial viability, safety, quality, and marketability of SBH.

Application of Heating on the Antioxidant and Antibacterial Properties of Malaysian and Australian Stingless Bee Honey.

In the honey industry, heat treatments are usually applied to maintain honey's quality and shelf life. Heat treatment is used to avoid crystallisation and allow the easy use of honey, but treatment with heat might affect the antioxidant and antibacterial activities, which are the immediate health effects of honey. This study will determine the effect of heat treatment on Malaysian and Australian stingless bee honey (SBH) produced by the common bee species in both countries. Eighteen honey samples were subjected to heat at 45 °C, 55 °C and 65 °C for one hour and subsequently analysed for their total phenolic content (TPC), total flavonoid content (TFC), DPPH radical scavenging activity, ferric reducing antioxidant power (FRAP) and minimum inhibitory concentration (MIC). The results show that all samples had high TPC, TFC and antioxidant activities before the treatment. The heat treatments did not affect (p < 0.05) the TPC, TFC and antioxidant activities in most samples, but did inhibit the antibacterial activities consistently in most of the samples, regardless of the bee species and country of origin. This study also confirms a strong correlation between TPC and TFC with FRAP activities for the non-heated and heated honey samples (p < 0.05). Other heat-sensitive bioactive compounds in SBH should be measured to control the antibacterial properties present.

Establishing Relationship between Vitamins, Total Phenolic and Total Flavonoid Content and Antioxidant Activities in Various Honey Types.

Honey is a well-known natural sweetener and is rich in natural antioxidants that prevent the occurrence of oxidative stress, which is responsible for many human diseases. Some of the biochemical compounds in honey that contribute to this property are vitamins and phenolic compounds such as phenolic acids and flavonoids. However, the extent to which these molecules contribute towards the antioxidant capacity in vitro is inconsistently reported, especially with the different analytical methods used, as well as other extrinsic factors that influence these molecules' availability. Therefore, by reviewing recently published works correlating the vitamin, total phenolic, and flavonoid content in honey with its antioxidant activities in vitro, this paper will establish a relationship between these parameters. Based on the literature, vitamins do not contribute to honey's antioxidant capacity; however, the content of phenolic acids and flavonoids has an impact on honey's antioxidant activity.

Antiviral activity of fermented foods and their probiotics bacteria towards respiratory and alimentary tracts viruses.

The recent COVID-19, a viral outbreak calls for a high demand for non-conventional antiviral agents that can reduce the risk of infections and promote fast recovery. Fermented foods and their probiotics bacteria have recently received increasing interest due to the reported potential of high antiviral activity. Several probiotics strains demonstrated broad range of antiviral activities and different mechanisms of action. This article will review the diversity, health benefits, interaction with immune system and antiviral activity of fermented foods and their probiotics bacteria. In addition, the mechanisms of action will be reviewed to determine the broad range potential antiviral activity against the respiratory and alimentary tracts viruses. The probiotics bacteria and bioactive compounds in fermented foods demonstrated antiviral activities against respiratory and alimentary tracts viruses. The mechanism of action was reported to be due to the stimulation of the immune system function via enhancing natural killers cell toxicity, enhance the production of pro-inflammatory cytokines, and increasing the cytotoxic of T lymphocytes (CD3+, CD16+, CD56+). However, further studies are highly recommended to determine the potential antiviral activity for traditional fermented foods.

Dietary Fiber: Fractionation, Characterization and Potential Sources from Defatted Oilseeds.

Dietary fiber (DF) has wide applications, especially in the food and pharmaceutical industries due to its health-promoting effects and potential techno-functional properties in developing functional food products. There is a growing interest in studies related to DF; nevertheless, there is less focus on the fractionation and characterization of DF. The characteristics of DF fractions explain their functionality in food products and provide clues to their physiological effects in food and pharmaceutical industrial applications. The review focuses on a brief introduction to DF and methods for its fractionation. It discusses the characterization of DF in terms of structural, physicochemical and rheological properties. The potential sources of DF from selected defatted oilseeds for future studies are highlighted.

Stingless Bee-Collected Pollen (Bee Bread): Chemical and Microbiology Properties and Health Benefits.

Stingless bee-collected pollen (bee bread) is a mixture of bee pollen, bee salivary enzymes, and regurgitated honey, fermented by indigenous microbes during storage in the cerumen pot. Current literature data for bee bread is overshadowed by bee pollen, particularly of honeybee Apis. In regions such as South America, Australia, and Southeast Asia, information on stingless bee bee bread is mainly sought to promote the meliponiculture industry for socioeconomic development. This review aims to highlight the physicochemical properties and health benefits of bee bread from the stingless bee. In addition, it describes the current progress on identification of beneficial microbes associated with bee bread and its relation to the bee gut. This review provides the basis for promoting research on stingless bee bee bread, its nutrients, and microbes for application in the food and pharmaceutical industries.

Stingless bee honey, a novel source of trehalulose: a biologically active disaccharide with health benefits.

Stingless bee (Meliponini) honey has long been considered a high-value functional food, but the perceived therapeutic value has lacked attribution to specific bioactive components. Examination of honey from five different stingless bee species across Neotropical and Indo-Australian regions has enabled for the first time the identification of the unusual disaccharide trehalulose as a major component representing between 13 and 44 g per 100 g of each of these honeys. Trehalulose is an isomer of sucrose with an unusual α-(1 → 1) glucose-fructose glycosidic linkage and known acariogenic and low glycemic index properties. NMR and UPLC-MS/MS analysis unambiguously confirmed the identity of trehalulose isolated from stingless bee honeys sourced across three continents, from Tetragonula carbonaria and Tetragonula hockingsi species in Australia, from Geniotrigona thoracica and Heterotrigona itama in Malaysia and from Tetragonisca angustula in Brazil. The previously unrecognised abundance of trehalulose in stingless bee honeys is concrete evidence that supports some of the reported health attributes of this product. This is the first identification of trehalulose as a major component within a food commodity. This study allows the exploration of the expanded use of stingless bee honey in foods and identifies a bioactive marker for authentication of this honey in associated food standards.

Kenaf (Hibiscus cannabinus L.) Seed and its Potential Food Applications: A Review.

Kenaf belongs to the family Malvaceae noted for their economic and horticultural importance. Kenaf seed is a valuable component of kenaf plant. For several years, it has been primarily used as a cordage crop and secondarily as a livestock feed. The potential for using kenaf seeds as a source of food-based products has not been fully exploited. Consumers are becoming more interested in naturally healthy plant-based food products. Kenaf seed, the future crop with a rich source of essential nutrients and an excellent source of phytocompounds, might serve suitable roles in the production of value-added plant-based foods. At present kenaf seed and its value-added components have not been effectively utilized for both their nutritional and functional properties as either ingredient or major constituent of food products. This review focuses on the possible food applications of kenaf seed and its value-added components based on their nutritional composition and functional properties available in literature, with the purpose of providing an overview on the possible food applications of this underutilized seed. The review focuses on a brief introduction on kenaf plant, nutritional function, lipids and proteins composition and food applications of the seed. The review elaborately discusses the seed in terms of; bioactive components, antioxidants enrichment of wheat bread, antimicrobial agents, as edible flour, as edible oil and a source of protein in food system. The review closes with discussion on other possible food applications of kenaf seed. The need for food scientists and technologists to exploit this natural agricultural product as a value-added food ingredient is of great significance and is emphasized.

Probiotic Properties of Bacillus Strains Isolated from Stingless Bee (Heterotrigona itama) Honey Collected across Malaysia.

This study aimed to isolate, identify, and evaluate the probiotic properties of Bacillus species from honey of the stingless bee Heterotrigona itama. Bacillus spp. were isolated from five different H. itama meliponicultures, and the isolates were characterized through Gram-staining and a catalase test. Tolerance to acidic conditions and bile salt (0.3%), hydrophobicity, and autoaggregation tests were performed to assess the probiotic properties of the selected isolates, B. amyloliquefaciens HTI-19 and B. subtilis HTI-23. Both Bacillus isolates exhibited excellent antimicrobial activity against both Gram-positive and Gram-negative bacteria and possessed significantly high survival rates in 0.3% bile solution for 3 h. Their survival rates in acidic conditions were also comparable to a commercial probiotic strain, Lactobacillus rhamnosus GG. Interestingly, the hydrophobicity and autoaggregation percentage showed no significant difference from L. rhamnosus GG, a commercial probiotic strain. The results from this study suggest that B. amyloliquefaciens HTI-19 and B. subtilis HTI-23 isolated from stingless bee honey have considerably good probiotic properties. Therefore, more studies should be done to investigate the effects of these bacteria cultures on gastrointestinal health.

Compositional analyses of white, brown and germinated forms of popular Malaysian rice to offer insight into the growing diet-related diseases.

Diet-related metabolic diseases, and especially obesity, are metabolic disorders with multifactorial aetiologies. Diet has been a cornerstone in both the aetiology and management of this metabolic disorders. Rice, a staple food for over half of the world's population, could be exploited as part of the solution to check this menace which has been skyrocketing in the last decade. The present study investigated nine forms of rice from three widely grown Malaysian rice cultivars for in vitro and in vivo (glycaemic index and load) properties that could translate clinically into a lower predisposition to diet-related diseases. The germinated brown forms of MRQ 74 and MR 84 rice cultivars had high amylose content percentages (25.7% and 25.0%), high relative percentage antioxidant scavenging abilities of 85.0% and 91.7%, relatively low glycaemic indices (67.6 and 64.3) and glycaemic load (32.3 and 30.1) values, and modest glucose uptake capabilities of 33.69% and 31.25%, respectively. The results show that all things being equal, rice cultivars that are germinated and high in amylose content when compared to their white and low amylose counterparts could translate into a lower predisposition to diet-related diseases from the dietary point of view in individuals who consume this cereal as a staple food.

Comparison of partial least squares and random forests for evaluating relationship between phenolics and bioactivities of Neptunia oleracea.

BACKGROUND: Neptunia oleracea is a plant consumed as a vegetable and which has been used as a folk remedy for several diseases. Herein, two regression models (partial least squares, PLS; and random forest, RF) in a metabolomics approach were compared and applied to the evaluation of the relationship between phenolics and bioactivities of N. oleracea. In addition, the effects of different extraction conditions on the phenolic constituents were assessed by pattern recognition analysis. RESULTS: Comparison of the PLS and RF showed that RF exhibited poorer generalization and hence poorer predictive performance. Both the regression coefficient of PLS and the variable importance of RF revealed that quercetin and kaempferol derivatives, caffeic acid and vitexin-2-O-rhamnoside were significant towards the tested bioactivities. Furthermore, principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA) results showed that sonication and absolute ethanol are the preferable extraction method and ethanol ratio, respectively, to produce N. oleracea extracts with high phenolic levels and therefore high DPPH scavenging and α-glucosidase inhibitory activities. CONCLUSION: Both PLS and RF are useful regression models in metabolomics studies. This work provides insight into the performance of different multivariate data analysis tools and the effects of different extraction conditions on the extraction of desired phenolics from plants. © 2017 Society of Chemical Industry.