Bradley W. Bolling

TREE NUT PHYTOCHEMICALS: COMPOSITION, ANTIOXIDANT CAPACITY, BIOACTIVITY, IMPACT FACTORS. A SYSTEMATIC REVIEW OF ALMONDS, BRAZILS, CASHEWS, HAZELNUTS, MACADAMIAS, PECANS, PINE NUTS, PISTACHIOS AND WALNUTS

Tree nuts contain an array of phytochemicals including carotenoids, phenolic acids, phytosterols and polyphenolic compounds such as flavonoids, proanthocyanidins (PAC) and stilbenes, all of which are included in nutrient databases, as well as phytates, sphingolipids, alkylphenols and lignans, which are not. The phytochemical content of tree nuts can vary considerably by nut type, genotype, pre- and post-harvest conditions, as well as storage conditions. Genotype affects phenolic acids, flavonoids, stilbenes and phytosterols, but data are lacking for many other phytochemical classes. During the roasting process, tree nut isoflavones, flavanols and flavonols were found to be more resistant to heat than the anthocyanins, PAC and trans-resveratrol. The choice of solvents used for extracting polyphenols and phytosterols significantly affects their quantification, and studies validating these methods for tree nut phytochemicals are lacking. The phytochemicals found in tree nuts have been associated with antioxidant, anti-inflammatory, anti-proliferative, antiviral, chemopreventive and hypocholesterolaemic actions, all of which are known to affect the initiation and progression of several pathogenic processes. While tree nut phytochemicals are bioaccessible and bioavailable in humans, the number of intervention trials conducted to date is limited. The objectives of the present review are to summarise tree nut: (1) phytochemicals; (2) phytochemical content included in nutrient databases and current publications; (3) phytochemicals affected by pre- and post-harvest conditions and analytical methodology; and (4) bioactivity and health benefits in humans.

Review of nut phytochemicals, fat-soluble bioactives, antioxidant components and health effects

The levels of phytochemicals (total phenols, proanthocyanidins, gallic acid + gallotannins, ellagic acid + ellagitannins, flavonoids, phenolic acids, stilbenes and phytates), fat-soluble bioactives (lipid, tocols, phytosterols, sphingolipids, carotenoids, chlorophylls and alkyl phenols) as well as natural antioxidants (nutrient and non-nutrient) present in commonly consumed twelve nuts (almond, Brazil nut, cashew, chestnut, hazelnut, heartnut, macadamia, peanut, pecan, pine nut, pistachio and walnut) are compared and reported. Recent studies adding new evidence for the health benefits of nuts are also discussed. Research findings from over 112 references, many of which have been published within last 10 years, have been compiled and reported.

Aronia melanocarpa (chokeberry) polyphenol–rich extract improves antioxidant function and reduces total plasma cholesterol in apolipoprotein E knockout mice

We hypothesized that a polyphenol-rich chokeberry extract (CBE) would modulate hepatic lipid metabolism and improve antioxidant function in apolipoprotein E knockout (apoE(-/-)) mice. ApoE(-/-) mice were fed diets containing 15% fat with 0.2% cholesterol alone or supplemented with 0.005% or 0.05% CBE for 4 weeks. CBE polyphenol content was determined by the total phenols, 4-dimethylaminocinnamaldehyde, and ultra high-performance liquid chromatography-mass spectrometry methods. The 0.05% CBE diet provided mice with mean daily doses of 1.2 mg gallic acid equivalents of total phenols, 0.19 mg anthocyanins, 0.17 mg phenolic acids, 0.06 mg proanthocyanidins (as catechin-equivalents), and 0.02 mg flavonols. The 0.05% CBE group had 12% less plasma total cholesterol concentrations than the control. Despite the hypocholesterolemic effect of CBE, hepatic mRNA levels of low-density lipoprotein receptor, hydroxyl-3-methylglutaryl coenzyme A reductase and cholesterol 7α-hydroxylase in CBE-fed mice were not significantly different from controls. Dietary CBE did not alter hepatic lipid content or the hepatic expression of genes involved in lipogenesis and fatty acid β-oxidation such as fatty acid synthase, carnitine palmitoyltransferase 1 and acyl-CoA oxidase. Plasma paraoxonase and catalase activities were significantly increased in mice fed 0.05% CBE. Both CBE diets increased hepatic glutathione peroxidase (GPx) activity but the 0.05% CBE group had 24% less proximal intestine GPx activity relative to controls. Thus, dietary CBE lowered total cholesterol and improved plasma and hepatic antioxidant function at nutritionally-relevant doses in apoE(-/-) mice. Furthermore, the CBE cholesterol-lowering mechanism in apoE(-/-) mice was independent of hepatic expression of genes involved in cholesterol metabolism.

Betalains, phase II enzyme-inducing components from red beetroot (Beta vulgaris L.) extracts.

Abstract: Crude aqueous and ethanolic extracts of root tissue of red (Rd) and high-pigment (HP) beet (Beta vulgaris L.) strains exhibited antioxidant and phase II enzyme-inducing activities, and these extracts were fractionated using Sephadex LH-20 chromatography. These bioactivities tended to become co-enriched in early and late eluting fractions, comprising 5-25% of the material recovered from the column. Liquid chromatography-mass spectrometry (MS) was used to resolve and identify multiple betalain components in the most potent quinone reductase (QR)-inducing fractions. Active fractions were found to contain vulgaxanthins I and II, and (iso)betanin, but other components remained unidentified. Two of the isolated active fractions were incorporated into rodent diets at 10-150 ppm over a 2-mo period to assess bioavailability and in vivo efficacy for phase II enzyme induction in various organs. No statistically significant effect of diet was obtained, and wide ranges of tissue enzyme levels among individual animals were observed. This lack of effect and diversity in response to diet may be related to the wide range in absorptive capacity of and/or insufficient level or enrichment of the active agents or to difficulties in assessing such activity in vivo. Subsequent to the animal studies, betanin was isolated in pure form, identified by MS analysis, and confirmed to be QR inducers in the bioassay.

The influence of roasting, pasteurisation, and storage on the polyphenol content and antioxidant capacity of California almond skins☆

Polyphenols and antioxidant activity of skins from California almonds subjected to roasting, pasteurisation, and storage were determined by LC-MS quantification, total phenols (TP), and ferric reducing antioxidant power (FRAP). Pasteurisation did not significantly change TP, FRAP, or flavonoids and phenolic acids (FP). Roasted almonds had 26% less TP and 34% less FRAP than raw, but equivalent FP (n = 12). Storing almonds at 4 and 23 °C for 15 mo resulted in gradual increases in FP, up to 177% and 200%, respectively (n = 13). At 4 °C and 15 mo, polyphenols increased 18-fold for p-hydroxybenzoic acid, whilst others were 45-200% higher compared to baseline values. Isorhamnetin-3-O-rutinoside accounted for 48% of the increase in FP. After 15 mo, FRAP and TP increased to 200% and 190% of initial values. Accelerated ageing of whole almonds increased FP content by 10% after 3 days, but TP and FRAP values were not significantly different from baseline to day 10. Thus, in almond skins, roasting decreases TP and FRAP but not FP, whilst storage for up to 15 mo doubles FP.

Quantification of Almond Skin Polyphenols by Liquid Chromatography‐Mass Spectrometry

Reverse phase HPLC coupled to negative mode electrospray ionization (ESI) mass spectrometry (MS) was used to quantify 16 flavonoids and 2 phenolic acids from almond skin extracts. Calibration curves of standard compounds were run daily and daidzein was used as an internal standard. The inter-day relative standard deviation (RSD) of standard curve slopes ranged from 13% to 25% of the mean. On column (OC) limits of detection (LOD) for polyphenols ranged from 0.013 to 1.4 pmol, and flavonoid glycosides had a 7-fold greater sensitivity than aglycones. Limits of quantification were 0.043 to 2.7 pmol OC, with a mean of 0.58 pmol flavonoid OC. Mean inter-day RSD of polyphenols in almond skin extract was 6.8% with a range of 4% to 11%, and intra-day RSD was 2.4%. Liquid nitrogen (LN(2)) or hot water (HW) blanching was used to facilitate removal of the almond skins prior to extraction using assisted solvent extraction (ASE) or steeping with acidified aqueous methanol. Recovery of polyphenols was greatest in HW blanched almond extracts with a mean value of 2.1 mg/g skin. ASE and steeping extracted equivalent polyphenols, although ASE of LN(2) blanched skins yielded 52% more aglycones and 23% less flavonoid glycosides. However, the extraction methods did not alter flavonoid profile of HW blanched almond skins. The recovery of polyphenolic components that were spiked into almond skins before the steeping extraction was 97% on a mass basis. This LC-MS method presents a reliable means of quantifying almond polyphenols.

Tea and health: preventive and therapeutic usefulness in the elderly?

Purpose of reviewTo update the growing literature suggesting that tea and its constituent flavonoids are inversely related to the risk of chronic diseases common among the elderly. Recent findingsResults are provided from recent observational studies and clinical trials on the relationship of tea and tea catechins to body weight control and energy metabolism, impaired glucose tolerance and diabetes, cardiovascular disease, bone mineral density, cognitive function and neurodegenerative disease, and cancer. The evidence for the efficacy and potency of tea and tea extracts in benefiting these outcomes ranges from compelling for cardiovascular disease to equivocal at best for some forms of cancer. SummaryAlthough randomized clinical trials of tea have generally been of short duration and with small sample sizes, together with experimental and epidemiological studies, the totality of the data suggests a role for tea in health promotion as a beverage absent in calories and rich in phytochemicals. Further research is warranted on the putative benefits of tea and the potential for synergy among its constituent flavonoids, L-theanine, and caffeine.

Polyphenol-rich black chokeberry (Aronia melanocarpa) extract regulates the expression of genes critical for intestinal cholesterol flux in Caco-2 cells☆

Black chokeberry (Aronia melanocarpa) is a rich source of polyphenols. The hypolipidemic effects of polyphenol-rich black chokeberry extract (CBE) have been reported, but underlying mechanisms have not been well characterized. We investigated the effect of CBE on the expression of genes involved in intestinal lipid metabolism. Caco-2 cells were incubated with 50 or 100 μg/ml of CBE for 24 h for quantitative realtime polymerase chain reaction analysis. Expression of genes for cholesterol synthesis (3-hydroxy-3-methylglutaryl coenzyme A reductase and sterol regulatory element binding protein 2), apical cholesterol uptake (Niemann-Pick C1 Like 1 and scavenger receptor class B Type 1) and basolateral cholesterol efflux [ATP-binding cassette transporter A1 (ABCA1)] was significantly decreased by CBE compared with control. Western blot analysis confirmed that CBE inhibited expression of these proteins. In contrast, CBE markedly induced mRNA and/or protein levels of ABCG5 and ABCG8 that mediate apical cholesterol efflux to the intestinal lumen. Furthermore, CBE significantly increased mRNA and protein levels of low-density lipoprotein (LDL) receptor, and cellular LDL uptake. Expression of genes involved in lipid metabolism and lipoprotein assembly, including sterol regulatory element-binding protein 1c, fatty acid synthase and acyl-CoA oxidase 1, was significantly decreased by CBE in a dose-dependent manner. Concomitantly, CBE significantly increased sirtuin 1, 3 and 5 mRNA levels, while it decreased SIRT-2. Our data suggest that hypolipidemic effects of CBE may be attributed, at least in part, to increased apical efflux of LDL-derived cholesterol and to decreased chylomicron formation in the intestine; and specific isoforms of SIRT may play an important role in this process.

Assay Dilution Factors Confound Measures of Total Antioxidant Capacity in Polyphenol-Rich Juices

UNLABELLED The extent to which sample dilution factor (DF) affects total antioxidant capacity (TAC) values is poorly understood. Thus, we examined the impact of DF on the ORAC, FRAP, DPPH, and total phenols (TP) assays using pomegranate juice (PJ), grape juice (GJ), selected flavonoids, ascorbic acid, and ellagic acid. For ORAC, GJ was comparable to PJ at DF 750, but at DF 2000, the ORAC value of GJ was 40% more than PJ. Increasing DF increased GJ and PJ, DPPH, TP, and FRAP values 11% and 14%, respectively. Increased test concentrations of quercetin and catechin resulted in 51% and 126% greater ORAC values, but decreased naringenin by 68%. Flavonoids, but not ellagic acid or ascorbic acid, may contribute to the dilution effect on the variation of final TAC values. Thus, reporting TAC or TP using a single DF may introduce uncertainty about the confidence of TAC assay values, especially when comparing different juices. These results underscore the importance of using compatible test standards for reporting TAC values. PRACTICAL APPLICATION Total antioxidant capacity (TAC) values such as the ORAC assay are increasingly used for comparison of polyphenol-rich foods and beverages. Choice of standards and test concentrations, even within the linear range of standards, may introduce variation probably due to synergy/antagonism between antioxidant and thereby, confound final TAC values. Thus, test concentration or dilution factors of samples should be considered in the design of TAC assays and interpretation of their results.

Bioavailability of anthocyanins and colonic polyphenol metabolites following consumption of aronia berry extract.

A single-dose pharmacokinetic trial was conducted in 6 adults to evaluate the bioavailability of anthocyanins and colonic polyphenol metabolites after consumption of 500mg aronia berry extract. UHPLC-MS methods were developed to quantitate aronia berry polyphenols and their metabolites in plasma and urine. While anthocyanins were bioavailable, microbial phenolic catabolites increased ∼10-fold more than anthocyanins in plasma and urine. Among the anthocyanins, cyanidin-3-O-galactoside was rapidly metabolized to peonidin-3-O-galactoside. Aronia polyphenols were absorbed and extensively metabolized with tmax of anthocyanins and other polyphenol catabolites from 1.0h to 6.33h in plasma and urine. Despite significant inter-individual variation in pharmacokinetic parameters, concentrations of polyphenol metabolites in plasma and urine at 24h were positively correlated with total AUC in plasma and urine (r=0.93, and r=0.98, respectively). This suggests that fasting blood and urine collections could be used to estimate polyphenol bioavailability and metabolism after aronia polyphenol consumption.