Vitamin D, a secosteroid, has recently been implicated in the stimulation of erythroid precursors and ultimately the rate of erythropoiesis. However, there are a paucity of randomised controlled trials (RCT), investigating the effect of vitamin D supplementation iron status, especially in populations at risk of iron deficiency. An eight-week, double-blind RCT was carried out in 50 female (mean age (± SD): 27 ± 9 years), iron-deficient (plasma ferritin concentration < 20 μg/L) participants, randomised to consume an iron-fortified cereal containing 9 mg of iron, with either a vitamin D supplement (1,500 international units (IU)/day, 38 μg/day) or placebo. The effect of dietary vitamin D supplementation on haematological indicators was investigated. Blood samples were collected at baseline, 4-weeks and 8-week timepoints for measurement of iron and vitamin D status biomarkers. The effect of intervention was analysed with a mixed-model repeated measures ANOVA using IBM SPSS statistical software (Version 21, IBM Corporation, New York, USA). Significant increases were observed in two haematological parameters: haemoglobin concentration and haematocrit level from baseline to post-intervention in the vitamin D group, but not in the placebo group. The increase from baseline to post-intervention in haemoglobin concentration in the vitamin D group (135 ± 11 to 138 ± 10 g/L) was significantly higher than in the placebo group (131 ± 15 to 128 ± 13 g/L) (P ≤ 0.05). The increase in haematocrit level from baseline to post-intervention was also significantly higher in the vitamin D group (42.0 ± 3.0 to 43.8 49 ± 3.4%) compared to the placebo group (41.2 ± 4.3 to 40.7 ± 3.6%) (P ≤ 0.05). Despite non-significant changes in plasma ferritin concentration, this study demonstrates that dietary supplementation with 1,500IU vitamin D, consumed daily with an iron-fortified cereal led to improvement in haemoglobin concentration and haematocrit levels in women with low iron stores. Further long-term studies are required, however, these findings suggest a potential role for improvement of vitamin D status as an adjunct therapy for recovery of iron status in iron-deficient populations.

Worldwide, cardiovascular disease (CVD) is the number 1 cause of mortality and is associated with insulin resistance (IR). Emerging biomarkers such as FGF21 and adiponectin are associated with cardiometabolic risk. Low carbohydrate, high fat (LCHF) diets have been reported to reduce cardiometabolic risk markers; however, few studies have compared a LCHF diet vs. a high carbohydrate (HC), lower fat diet under ad libitum conditions on adiponectin and FGF21. The purpose of this study was to investigate the effects of an ad libitum LCHF vs. HC diet on IR, FGF21 and adiponectin in 16 healthy adults. Ethical approval: Liverpool John Moores University Research Ethics Committee (16/ELS/029); registered with ClinicalTrials.gov (Ref. NCT03257085). Participants were randomly assigned to a HC diet (n = 8, the UK Eatwell guidelines; ≥ 50% of energy from carbohydrates) or a LCHF diet (n = 8, consume < 50 g/day of carbohydrates). All provided plasma samples at 0, 4 and 8 weeks. FGF21 (R&D Systems) was analysed via ELISA and adiponectin, insulin and glucose were analysed via immunoassay technology (Randox Evidence Investigator™ Metabolic Syndrome Arrays I & II). Mann Whitney, Friedmans, Wilcoxon tests and 2×3 ANOVA (IBM SPSS 25®) were undertaken to investigate significant differences between and within groups. The homeostatic model assessment (HOMA) was used to calculate IR. FGF21 significantly (P = 0.04) decreased (Mdn, IQR:148.16, 78.51–282.02 to 99.4, 39.87–132.29 pg/ml) after 4 weeks and significantly (P = 0.02) increased (Mdn, IQR:167.38, 80.82–232.89 pg/ml) by 8 weeks vs. baseline with LCHF. No significant differences (P > 0.05) were observed between groups. Adiponectin was significantly (P = 0.03) different at week 4 only between groups. Adiponectin increased after 4 weeks (Mdn, IQR:13.44, 9.12–25.47 to 16.64, 11.96–21.51 ng/ml) but was only significantly (P = 0.03) different by 8 weeks vs. baseline in the HC group (Mdn, IQR:16, 10.8–27.43 ng/ml). Adiponectin remained unchanged (P = 0.96) in the LCHF group. HOMA significantly decreased with both diets after 8 weeks only (mean ± SD, LCHF: 2.9 ± 1.3 to 1.8 ± 0.8, HC: 2.5 ± 0.6 to 1.9 ± 0.6, P = 0.008) but was not significantly (P = 0.60) different between groups. These preliminary data reveal that while both diets improved insulin sensitivity, they may do so by different mechanisms. Future studies are warranted to investigate further, how a LCHF vs. HC diet affects FGF21 and adiponectin, and the subsequent regulation of IR. Furthermore, studies that extend these findings by determining the impact of LCHF vs. HC on peripheral metabolism to determine potential nutrition-mediated mechanisms of metabolic adaptation are warranted.

The influence on health of the human gut microbiota is increasingly recognised, however wheat fibre, consumed frequently in Western diets has traditionally been considered inert with regard to gut microbiota composition and metabolic activity. We undertook a systematic review (PRISMA methodology) of human intervention studies examining the effects of intact cereal fibres on gut microbiota composition among healthy adults.(1) Studies published in the past 20 years were identified on PubMed and Cochrane electronic databases. Inclusion criteria were: healthy adult participants, at least one intact cereal fibre (or its sub-fraction) and measurement of faecal microbiota related outcomes. Out of forty studies meeting inclusion criteria, seventeen manipulated wheat fibre/bran or its key constituent arabinoxylans (AXOS), and ten used a whole diet approach with predominantly wheat fibre. Results from these twenty seven wheat fibre papers are presented here. Eight studies provided wheat bran/fibre (ranging from 5.7g-21g/day wheat fibre or 13g-28g/day wheat bran). Three reported significant effects on gut microbiota abundance and/or diversity (both at phyla and species level) and one showed no effect. Six reported significant increases in fermentation metabolites and one reported no significant change. Ten studies manipulated whole day fibre intake (predominantly wheat but also permitting some oats, rye and rice). Wholegrain intake ranged from 80g-150 g per day and fibre from 13.7g–40 g per day. Six found significant increases in bacterial diversity and/or abundance and five showed significant increases in fermentation metabolites. Two identified that response to high fibre intervention is dependent on baseline gut microbiota richness - those with limited richness exhibiting greater microbiota change over time in response to fibre increase. Two reported no significant effects. Nine studies utilised manipulation of AXOS (2.2g–18.8 g per day) with five demonstrating significant increases in target bacterial species and six significant increases in fermentation metabolites. One reported no significant effect to faecal metabolites. This review supports a role for the wheat fibre found in everyday foods (such as bran breakfast cereal of high fibre breads) promoting both microbiota diversity and abundance. While the healthy microbiome is yet to be defined, consumption of a single daily serving of wheat bran fibre appears sufficient to effect gut microbiota fermentation (with demonstrable effects arising from as low as 6g/day), and promote species diversity, with potential benefit to health.However exploration of stability over longer time frames (> 12 weeks) is now required.

Metabolic syndrome (MetS), which refers to co-occurrence of cardiovascular risks primarily obesity, hypertension, insulin resistance and atherogenic dyslipidemia was previously uncommon in sub-Sahara African. But it is increasingly becoming a public health concern heightened by the epidemiological transition associated with excess energy intake, sedentary lifestyles and urbanization. The aim of this study was to assess the dietary, lifestyle and physiologic risk factors associated with MetS and the effect on perceived quality of life. The design was one-to-one age and sex-matched case-control study. MetS was defined as concomitant adiposity, hypertension and diabetes. Cases (n = 76) were recruited in the diabetic clinic of a municipal hospital in Ghana while controls (n = 76) were recruited in the communities in which the cases lived. Lifestyle habits and physical activity were assessed using a validated lifestyle and habits questionnaire while habitual dietary intakes were assessed using a food frequency questionnaire. Anthropometry, blood pressure and fasting capillary glucose were measured. Differences between cases and controls were tested using McNemar and paired t-test tests. Crude odds ratio (COR) for MetS was estimated using conditional logistic regression.

Mean age of participants was 57.31 ± 5.46 years (range 45–65 years). Females constituted the majority (95%, n = 144). Among the cases and controls, lateral obesity (100% vs 60.5%), central obesity (98.7% vs 36.8%), hypertension (90.8% vs 2.6%) and diabetes (72.4% vs 1.3%) differed significantly. Fewer cases (10.5%) consumed diversified and healthy diets compared to the controls (77.6%) (p = 0.001). Significant number of cases felt a high risk for illness based on how they assessed their physical health (73.7% vs 2.6%), exercise and fitness regime (88.2% vs 13.2%), nutrition and weight control (14.5% vs 1.3%) and psychological health (19.7% vs 0%). However, none felt socially unhealthy. Lower risk for MetS was found among participants who had at least secondary education (COR:0.17 95% CI:0.04–0.73) and lived in a peri-urban setting (COR:0.22 95% CI:0.09–0.53) while unemployment (COR:9.00 95% CI:1.26–64.35) and central obesity (COR: 48.0 95% CI: 6.62–147.74) were associated with higher risk. MetS adversely affected general sense of wellbeing of the middle age adults. Obesity was an important predictor. Educational interventions to promote diversification of dietary intakes and enhance weight control are crucial to reduce MetS and could improve the quality of life of those affected. These interventions should be integrated into already existing community-based social support, and health systems.

In recent years, research has been focusing on strategies to counteract inflammatory processes and age-related diseases(1). During ageing, a low-grade systemic inflammation is often associated to an altered intestinal permeability (IP) a condition that has been shown to promote inflammation possibly through the translocation of dietary and bacterial factors into the blood stream that activates the immune system(2).In this regard, dietary pattern and environmental factors could play a fundamental role because of their potential ability to modulate inflammation, IP and the gut microbial ecosystem (GME). Moreover, it has been hypothesized that bioactive compounds such as polyphenols may affect IP and GME(3).The MaPLE project (Microbiome mAnipulation through Polyphenols for managing gut Leakiness in the Elderly) aimed to investigate the hypothesis that a polyphenol-rich diet can improve IP condition in a target population with beneficial changes at intestine and systemic level. To this aim, a randomised, controlled, cross-over dietary intervention study (8-week polyphenol-rich diet versus 8-week control diet, separated by a wash-out period) was carried out in a group of older subjects (> 60 years) living in a well-controlled setting (i.e. nursing home). Markers related with IP, inflammation, oxidative stress, vascular function and intestinal microbial ecosystem were investigated in serum, urine and/or fecal samples. Moreover, blood bacteria DNAemia, and serum/urine metabolomics has been assessed. Moreover, a consistent nutritional evaluation of the standard menu (provided by the nursing home) and of weighed food diaries was performed, providing also data on actual polyphenol intake during the intervention. The results show there were higher levels of IP in the older subjects, and that the polyphenol-enriched diet changed the levels of serum zonulin, a marker of IP. In addition, an association between zonulin and blood bacterial load was demonstrated. Ongoing in vitro and in vivo experiments are exploring the potential effects of different polyphenols on IP and the mechanisms involved. The MaPLE project will generate new data to improve the understanding on the role of polyphenols in the modulation of intestinal microbiome and its interactions with the host.

Globally, the adverse effects of increased animal-protein consumption on the environment, the society, and the economy are a cause of growing societal concern. As a result, issues of food system sustainability have received considerable attention within the national and international community. For the European community, changes in food lifestyles of consumers, in particular a reduced share of animal-based foods within the diet, are of great importance for the establishment of sustainable food supply structures. Meanwhile, a worldwide trend for alternative nutrition and lifestyles is obvious, which is reflected by a growing number of plant-based nutrition styles and craft-foods. Often, alternative nutrition styles are ethically or health motivated. Nonetheless, changes in regard to nutrition are often not consistent. Up to now, research mainly focused on the quantitative and qualitative aspects of food lifestyle changes. However, extreme, alternative nutrition lifestyles seem to be more disciplined with regard to nutrition transformation. Nonetheless, especially extreme food consumption patterns have not been assessed with regard to a long-lasting sustainable transformation potential. However, we propose that a comprehensive framework of these issues must contain information on the specific motivational themes that lead people to make substantial changes in their diets. Hence, the objective of the proposed study is to assess the transforming potential by in-depth interviews with extreme alternative nutrition styles. We aim in gathering knowledge on the content of the motivational dynamics influencing consumers in adapting alternative food lifestyles like raw-foodism, Paleolithic diet, or Anthroposophic diet. The theoretical framework applied is grounded in self-determination theory, a meta-theoretical construct used for explaining motivational change dynamics in humans. The research questions that will be addressed are: “What motivates people to adapt extreme alternative dietary styles?”, “How do identity and individual worldview influence the transition process?” and “Which strategies were applied to successfully implement new dietary habits into daily routine? In-depth interviews, following an experience interview will be conducted with 18 participants who had practiced an alternative food lifestyle for a long-term period. The transcripts of these interviews will be structured using content analysis. Since data collection will take place in May 2019 results cannot be described by deadline for the call of abstracts. Results and conclusion that will be addressed in the FENS Conference will include the motivational drivers for long-lasting transformation potential and discuss the applicability for society and behavior change communication.

Perinatal depression is a depressive illness that affects 10–15% of women in the UK with an estimated cost of £1.8 billion/year. Zinc deficiency is associated with the development of mood disorders and zinc supplementation has been shown to help reduce the symptoms of depression. Women who are pregnant and breastfeeding are at risk of lower levels of zinc because of the high demand from the developing and feeding baby. However, studies in the perinatal period are limited. With a long-term aim of designing a randomised controlled trial (RCT) to examine if zinc supplementation reduces depressive symptoms in pregnant and lactating women;the objective of this review was to systematically evaluate previous RCTs assessing zinc supplementation and depressive symptoms, in order to establish a zinc dosing regimen with regards to Galenic formulation, unit dose and frequency. The review was conducted by independent reviewers in accordance with PRISMA guidelines and is registered at Prospero (CRD42017059205). The Allied and Complimentary Medicine, CINAHL, Embase, MEDLINE, PsycINFO, PubMed, and Cochrane databases were searched since records began, with no restrictions, for intervention trials assessing Galenic formulation, unit dose and frequency of zinc supplementation to reduce the symptoms of depression. From a total of 66 identified records, 7 articles met the inclusion and exclusion criteria; all assessed the effect of zinc supplementation on mood. Risk of bias was independently assessed using the standard ‘Cochrane risk of bias tool’. Overall, 5 of the 7 papers were rated as high-quality trials; of the other two, one was rated poor and the other fair but both had a number of learning points. Preliminary findings indicate at the end of supplementing zinc, depression scores were reduced significantly. In one study, the Beck score decreased in the placebo group, but this reduction was not significant compared to the baseline. In two of the studies there was a significant correlation between serum zinc and self-reported mood questionnaires. Results also suggest that 25 mg zinc supplementation combined with antidepressant drugs can be effective in the treatment of major depression in women. This supports other work where researchers supplemented 25 mg of elemental zinc for 12 weeks or longer and found a reduction of symptoms in both pregnant and non-pregnant women. Thus, an early conclusion is that 25 mg of elemental zinc is an effective dose for improving low mood and is achievable in a trial setting.

A previous study has demonstrated that when people thought they were eating a low-calorie milkshake (versus a high-calorie labelled equivalent though same product) their physiological satiety, as measured by the gut peptide ghrelin, was consistent with what they believed they were consuming rather than the actual nutritional content (Crum et al., 2011). If replicated and shown for different food types, this finding could have implications for nutrient and health claims labelling and advertising. The aim of the study was to examine whether satiation (self-reported and physiological) varies depending on the mindset in which one approaches food consumption. On two separate visits (1 week between), participants (n = 50) were asked to consume a 380-calorie yoghurt and granola breakfast product under the pretence that it was either a 500-calorie ‘indulgent’ breakfast (high in fat and sugar) or a 250-calorie ‘sensible’ breakfast (low in fat and sugar). At each visit blood samples were collected at three timepoints to measure acylated ghrelin: after a 20-minute rest period (baseline), after 60-minutes (pre-consumption) and after 90-minutes (post-consumption). Self-reported appetite scales were completed 10 minutes prior to each blood sample. During the first interval (between 20 and 60 minutes) participants rated the breakfast label's appearance and perceived healthiness, and during the second interval (between 60 and 90 minutes) participants consumed the breakfast product while rating its sensory appeal. Participants (mean [SD]: 30.1 [10.4] yrs.) rated the ‘indulgent’ breakfast as more appealing than the ‘sensible’ breakfast (mean difference: 5.00 [95% CI: 0.71, 9.30]; P = 0.024), but felt less healthy when consuming the ‘indulgent’ breakfast (mean difference: -13.17 [95% CI: -18.75, -7.60]; P < 0.001). The breakfasts were not rated differently according to their taste, smell, overall palatability and enjoyment. Participants reported a higher mean change in self-reported fullness for the ‘indulgent’ breakfast than the ‘sensible’ breakfast from pre-consumption to post-consumption (mean difference: 7.19 [95% CI: -0.73, 13.6]; P = 0.030). This relationship was not observed between baseline and post-consumption, or for the other self-reported appetite measures (hunger, satiety, quantity and desire to eat). Mean change in acylated ghrelin was not significantly different between the breakfasts at any timepoint. This study demonstrated an increase in self-reported fullness after consuming the ‘indulgent’ breakfast compared to the ‘sensible’ breakfast despite the fact the two breakfast products were identical. A physiological response, however, was not observed.

The multipurpose plant Moringa oleifera (MO) has shown a variety of in vitro activities potentially beneficial for health and evidences are also numerous in animal models. However, the exact composition of the plant is highly variable depending on the area, culture and post harvesting conditions and the effects in humans are scarcely studied. This is a pilot study of the effect of different doses of MO from an Iberian Peninsula cultivar on postprandial glucose level in non-diabetic subjects. Five adult subjects (40 to 70 years, 3 males) with normal renal and hepatic function were included. Two had normal fasting glucose and 3 had pre-diabetic fasting glucose. A standard breakfast plus, respectively, one placebo capsule (PLC), or 1, 3 and 6 capsules containing 400 mg of MO dry leaf powder (MO-1cap, -3cap and -6cap) were consumed by each subject on 4 consecutive visits, one week apart. The basal and postprandial (30, 60, 90, 120, 150 and 180 min) glucose was measured. The mean glucose change from baseline (ΔGlc) at each measure time was calculated and compared between treatments with the Wilcoxon tests and considered significant at P < 0.05 level. The postprandial curves were also studied separately in pre-diabetic subjects. A difference was found 90 min after breakfast in ΔGlc for the MO-6cap compared to the MO-1cap, being lower in MO-6cap. No more significant differences were observed due to the small number of subjects and high variability in glucose values among subjects. However, the graphical representation of separated curves for the three pre-diabetic participants showed that all three MO doses led to values always below those with PLC. In addition, during the first experimental hour all MO doses showed lower ΔGlc than PLC while this was not the case when all subjects were studied. No adverse events occurred during the experiment. The study, thus, showed a trend to lower glycemic postprandial response with MO leaf powder which seems to be more evident in pre-diabetic subjects than in normoglycemic subjects and especially during the first postprandial hour. The MO-6cap seems to be the election daily dose for a long term intervention study on the glycemic control with MO in pre-diabetic subjects.

Rethinking food systems from production to consumption, in order to provide better nutritional inputs at lower environmental cost, is a priority challenge for a sustainable future. Pulses present benefits that may improve the sustainability of our systems and diets, such as their ability to restore soils in nitrogen and their high contents in proteins, fibers and minerals. However, pulses also contain several bioactive compounds such as phytates or tannins that can negatively affect mineral absorption. Additionally, we recently showed in the laboratory that these bioactives, together with fibers and saponins, could negatively impact fat-soluble vitamin bioavailability. The objective of this study was thus to follow up vitamin D (as a model of fat-soluble vitamin) and iron (as a model of mineral) transfer to the aqueous phase of the bolus during digestion of meal containing or not pulses. To this aim, we performed in vitrodigestion using tests meals made of beef (as a model of meat) and/or semolina (as a model of cereals) and/or chickpeas (as a model of pulses). To identify the compounds responsible for the observed effects, we also performed in vitrodigestion using test meals made of potatoes supplemented or not in fibers, phytates, tannins and saponines. Vitamin D bioaccessibility and iron solubility were expressed as the ratio of vitamin D or iron recovered in the aqueous phase of the digestion on the total amount of vitamin D or iron recovered in the whole digesta, at the end of the digestion.

Our results showed that the presence of chickpeas within a meal induced a significant decrease of both vitamin D bioaccessibility (up to -56%, p < 0.05) and iron solubility (up to -28%, p < 0.05) compared to meals containing only meat and/or semolina. However, this effect was largely compensated for vitamin D by the fact that this vitamin was less stable (loss > 50%, p < 0.05) during the digestion of meal containing meat compared to meals containing only plant-based foods (i.e. semolina and chickpeas). Among the different bioactives, tannins appear to be the most deleterious regarding iron solubility, while both phytates and tannins were responsible for a decreased in vitamin D bioaccessibility.

Our results confirm that in some conditions, the presence of pulses within a meal can be deleterious regarding vitamin D and iron bioavailability. These data thus encourage research to propose dietary and technological solutions to tackle pulse negative effects on micronutrient bioavailability.

Over the last years, the consumption of unpasteurised milk has increased in popularity in the Western countries, despite the known risks associated with food-borne pathogens. Some people appear to experience milk-related gastrointestinal symptoms even when tested negative for lactose intolerance and milk allergy. In such cases, processing of milk, most often homogenisation or heat treatment, has been suggested to be involved in the induction of the gastrointestinal symptoms. Commercial milk is generally homogenised for the purpose of physical stability. Homogenisation reduces the size of fat droplets. Breaking of the fat globules creates new interface, which is covered with denaturated proteins. Further denaturation level of the proteins depends on the severity of the heat treatment. In this study entity, the effect of processing of bovine milk on the gastrointestinal symptoms in self-reported milk sensitive individuals was tested in two separate double blind crossover trials. Tested milks included raw milk (unprocessed), pasteurized milk, pasteurized and homogenized milk, and UHT treated and homogenized milk thus including milks at both end of commercially approved liquid milks in Finland. All milks came from the same herd of cows, which nulled the effect of genetic background and feeding of the cows. Ingestible pH-, pressure and temperature measuring probe was used to investigate gastrointestinal pressure. Self-reported diaries were used to measure perceived gastrointestinal symptoms. Additionally inflammation markers were studied from the plasma. In the first trial no significant difference was found in the amount of gastrointestinal symptoms or in the intestinal pressure after the consumption of native and processed milk. However, the obtained results on pressure in the large intestine (P = 0.068) as well as reported symptoms (P = 0.103) suggested that further studies in this area were needed. However, in the second trial where pasteurised, pasteurised and homogenised, and UHT -treated and homogenized milks were compared, the amount, severity or duration of the reported symptoms did not differ. This study demonstrates that the ingestible pressure-, pH and temperature measuring probe provided a novel possibility to study intestinal pressure in nutrition trials. The results of the present studies do not support the hypothesis that cows’ milk processing would induce gastrointestinal symptoms in milk sensitive but lactose tolerant subjects.

Fennel (Foeniculum vulgare Mill.) is a well-known aromatic plant, widely used as spice and remedy herb. It provides many beneficial effects and is used as diuretic and expectorant, as well as for various dyspeptic disorders. Beside essential oils, fennel seeds are abundant with various phenols, which are known as strong antioxidants and thereby contribute to human health. Among different extraction methods for phenols isolation, pressurized liquid extraction (PLE) represents a novel technique with numerous advantages over conventional extraction approaches. Its efficiency manifests through combination of high temperature and pressure, enabling extraction time shortening, as well as decreased solvent consumption, thus being environment-friendly technique. In order to achieve maximum extraction yield, it is of great importance to select optimal PLE conditions, e.g., extraction temperature and time. Additionally, solvent selection also affects on content of targeted compounds, where phenols amount depends on polarity of solvent. Hence, the aim of this study was to examine the effect of temperature (75 and 100 °C), static time (5 and 10 min) and cycle number (1, 2 and 3) on total phenols (TP) and total flavonoids (TF) content, as well as the antioxidant capacity (AC) in fennel seeds using three-step exhaustive PLE. Firstly, grinded dry fennel seeds were subjected to PLE using non-polar solvent (hexane) under the mentioned conditions in order to remove seeds' lipid fraction. Afterwards, defatted samples were extracted with polar solvents by the increasing polarity: aqueous acetone solution (30 %, v/v) followed by aqueous methanol solution (30 %, v/v). Obtained acetone (AE) and methanol (ME) extracts (n = 24) were analyzed for TP (Folin-Ciocalteu method), TF (aluminium chloride colorimetric assay) and AC [ferric reducing antioxidant power (FRAP) method]. Multivariate analysis of variance (MANOVA) and Tukey's HSD test (p ≤ 0.05) were applied for statistical analysis of collected data. Expectedly, AE were described with higher values of analyzed parameters (grand means: TP = 416.18 mg/100 g, TF = 41.87 mg/100 g, AC = 359.57 mg AAE/100 g) in comparison with ME (grand means: TP = 80.25 mg/100 g, TF = 16.41 mg/100 g, AC = 96.13 mg AAE/100 g. Furthermore, all examined influences significantly affected TP, TF and AC in all samples, except temperature on TP in ME. Thus, conditions of 100 °C/10 min/3 cycles showed the greatest yield of tested parameters in AE, while 100 °C/5 min/1 cycle were sufficient for maximum TP and AC levels in ME. Regarding TF in ME, slightly longer extraction time was required to achieve the highest efficiency (100 °C/10 min/1 cycle).

Due to nowadays endeavoured green approach, the utilization of environmentally-friendly extraction techniques is encouraged, where pressurized liquid extraction (PLE) provides such possibility. The PLE application allows shorter extraction time and simultaneously less solvent consumption through combined temperature and pressure effect, thus improving energetic and economic efficiency. Accordingly, in order to accomplish satisfactory extraction yield, it is necessary to optimize PLE conditions, depending on sample type and aimed compounds. As fennel (Foeniculum vulgare Mill.) seeds are rich source of structurally different bioactive compounds, this study aimed to examine the influence of PLE temperature (75 and 100 °C), static time (5 and 10 min) and cycle number (1, 2 and 3) on the content of fennel seeds’ polyphenols fractions: total hydroxycinnamic acids (THA), total flavonols (TF) and total flavan-3-ols (TFO). For this purpose, samples of grinded dry fennel seeds were extracted according to the mentioned PLE conditions in serial exhaustive extraction using solvents ordered by its polarity: hexane, aqueous acetone solution (30 %, v/v) and aqueous methanol solution (30 %, v/v). After defatting with hexane, acetone (AE) and methanol (ME) extracts of residues were collected (n = 24) and used for spectrofotometrically determination of THA, TF and TFO. In order to access data about effect of examined PLE conditions on analyzed parameters, results were statistically analyzed using multivariate analysis of variance (MANOVA) and Tukey's HSD test at significance level p ≤ 0.05. Obtained results showed that observed fractions of polyphenols were present in 4–5 folds higher amounts in AE compared to ME, where established grand means in AE were as follows: THA = 312.82 mg/100 g, TF = 113.23 mg/100 g and TFO = 29.01 mg/100 g, while in ME they were THA = 56.90 mg/100 g, TF = 28.78 mg/100 g and TFO were determined in traces. As for influence of extraction temperature and time, content of all tested compounds was significantly affected by examined sources of variation in extracts of both solvent types, except static time on ME’ THA content. Generally, 100 °C/10 min/3 cycles were conditions which yielded the highest THA, TF and TFO in AE, while ME results showed diversity upon examined conditions. Accordingly, the highest THA content was established at lower temperature (75 °C/10 min/2 cycles), while 100 °C/10 min/3 cycles conditions documented the greatest TF content.