Alkylresorcinols Research Articles

Application of hydrolases and probiotic Pediococcus acidilactici BaltBio01 strain for cereal by-products conversion to bioproduct for food/feed

The aim of this study was to apply the enzymatic treatment and fermentation by Pediococcus acidilactici BaltBio01 strain for industrial cereal by-products conversion to food/feed bioproducts with high amount of probiotic lactic acid bacteria (LAB). LAB propagated in potato media and spray-dried remained viable during 12 months (7.0 log10 cfu/g) of storage and was used as a starter for cereal by-products fermentation. The changes of microbial profile, biogenic amines (BAs), mycotoxins, lactic acid (L+/D−), lignans and alkylresorcinols (ARs) contents in fermented cereal by-product were analysed. Cereal by-products enzymatic hydrolysis before fermentation allows to obtain a higher count of LAB during fermentation. Fermentation with P. acidilactici reduce mycotoxins content in fermented cereal by-products. According to our results, P. acidilactici multiplied in potato juice could be used for cereal by-products fermentation, as a potential source to produce safer food/feed bioproduct with high amount of probiotic LAB for industrial production.

Novel urinary alkylresorcinol metabolites as biomarkers of whole grain intake in free-living Swedish adults.

Most studies on the role of whole grain for health rely on self-reported intake data, which are prone to measurement errors. There is a need for dietary biomarkers that can provide an objective measure of intake. Alkylresorcinols (AR) and their main metabolites 3,5-dihydroxybenzoic acid (DHBA) and 3-(3,5-dihydroxyphenyl)-propanoic acid (DHPPA) have been proposed as biomarkers for whole grain (WG) wheat and rye intake. The medium-term reproducibility and relative validity of four putative urinary AR metabolites (3,5-dihydroxycinnamic acid (DHCA), 5-(3,5-dihydroxyphenyl) pentanoic acid (DHPPTA), 2-(3,5-dihydroxybenzamido)acetic acid (DHBA-glycine) and 3,5-dihydroxycinnamic acid amide (DHCA-amide)) as biomarkers for WG intake were investigated. Three-day weighed food records and 24-h urine samples from two occasions 2-3 months apart were obtained from 69 Swedish adults. WG intake was calculated and urinary AR metabolites were analyzed. The medium-term reproducibility determined for DHCA, DHPPTA, and DHBA-glycine varied from moderate-to-excellent (intra-class correlation coefficient = 0.63-0.85). Moreover, DHCA and DHPPTA excretion correlated well with self-reported total WG intake (r=0.55, p<0.001 and r=0.42, p<0.001, respectively). DHCA or DHPPTA excretion in 24-h urine might be a suitable medium- to long-term biomarker of WG wheat and rye intake. These findings need to be confirmed in populations with low and infrequent WG intake.

Growth suppression of Fusarium culmorum, Fusarium poae and Fusarium graminearum by 5-n-alk(en)ylresorcinols from wheat and rye bran

Alk(en)ylresorcinols (AR), a class of phenolic lipids, are regarded as antifungal compounds showing high potential for the use in plant protection, especially against Fusarium head blight (FHB). In view of the very limited knowledge of the activity of single AR against Fusarium species, the antifungal effect of crude extracts, fractions and isolated homologues from wheat and rye bran was determined. It was shown that the saturated AR are the active compounds in the extracts, whereas the presence of unsaturated molecules leads to an antagonistic effect. The activity of single saturated AR is dependent on the chain length, but for highest antifungal efficiency a mixture of saturated homologues is required. Affecting the stage of germination, these molecules reduce, and may even completely prevent, the growth of the tested Fusarium species.

Separation of alk(en)ylresorcinols from rye bran with saturated, monoenoic, dienoic, trienoic and hydroxylated monoenoic side chains using an octyl phase in ultra-high performance liquid chromatography and their differentiation by tandem mass spectrometrie

Alkylresorcinols (ARs) occur in bran of cereals and in fruits from the Anacardiaceae family. Their separation by liquid chromatography is challenging, especially in rye (Secale cereale L.) that has a complex AR composition. An octyl phase (C8) with 1.8μm particles was used for the analysis of an acetone extract of rye bran. The ARs were detected by UV at 205 and 275nm and by MS applying selected ion monitoring (SIM) of known and hypothetical m/z values in positive and negative mode. The compounds found were subjected to product ion scans in a triple quadrupole mass spectrometer. The C8 UHPLC column has a suitable selectivity for the analysis of ARs from rye. In combination with the sub–2μm particles, baseline separation of most ARs was achieved. The MS2 spectra in positive mode show diagnostic fragments that allow identifying the ARs subclasses (saturated, monoenoic, dienoic, trienoic and hydroxylated monoenoic) unambiguously. Several minor ARs were detected for the first time: C23:3, C27:1OH, C20:0, C22:0, C24:0 and some minor alkenylresorcinol isomers. The chromatographic resolution on the C8 column is unprecedented in the field of rye ARs. Thus, isolation and quantification using non-mass-selective detectors is now possible for each AR. Since rye bran has the most complex AR composition, this method is expected to facilitate the analysis of ARs also in other samples.

Alkylresorcinols activate SIRT1 and delay ageing in Drosophila melanogaster

Sirtuins are enzymes that catalyze NAD+ dependent protein deacetylation. The natural polyphenolic compound resveratrol received renewed interest when recent findings implicated resveratrol as a potent SIRT1 activator capable of mimicking the effects of calorie restriction. However, resveratrol directly interacts with fluorophore-containing peptide substrates. It was demonstrated that the SIRT1 activation of resveratrol is affected by the amino acid composition of the substrate. Resveratrol did increase the enzyme activity in cases in which hydrophobic amino acids are at the +1 position to the acetylated lysine in the substrate. Alkylresorcinols (ARs) are compounds that belong to the family of phenolic lipids, and they are found in numerous biological species. Here we show that the natural activators ARs increased the Vmax of recombinant SIRT1 for NAD+ and peptide substrate, and that ARs decreased acetylated histone in human monocyte cells by stimulating SIRT1-dependent deacetylation of substrates. ARs also extended the lifespan of Drosophila melanogaster, which was shown to be dependent on functional Sir2. Our results demonstrated that ARs are natural catalytic activators for sirtuin.

Isolation of a new resorcinolic lipid from Mangifera zeylanica Hook.f. bark and its cytotoxic and apoptotic potential.

Mangifera zeylanica is a plant endemic to Sri Lanka and its bark has been used in traditional medicine to treat some cancers. This study was aimed to isolate potentially cytotoxic compound/s from the hexane extract of the bark of M. zeylanica by bio-activity guided fractionation. The structure of the isolated compound (1) was elucidated using 1H, 13C NMR and mass spectrometric techniques. Compound 1 was identified as a new resorcinolic lipid (5-((8Z, 11Z, 14Z)-hexatriaconta-8, 11, 14-trienyl) benzene-1,3-diol). Apoptotic potential of the isolated compound was determined only in MCF-7 (estrogen receptor positive) breast cancer cells to which it was more cytotoxic than to normal mammary epithelial cells. Oxidative stress markers [reactive oxygen species (ROS), glutathione levels (GSH) and glutathione-S-transferase (GSH)] were also determined in MCF-7 cells treated with compound 1. Treatment with compound 1 led to an increase in caspase 7 activity, morphological features of apoptosis and DNA fragmentation in MCF-7 cells. Furthermore, it also led to an increase in ROS and GST levels while depleting GSH levels. Results of this study suggest that isolated new resorcinolic lipid can induce apoptosis in MCF-7 cells, possibly via oxidative stress mechanism.

Use of HPLC- and GC-QTOF to determine hydrophilic and lipophilic phenols in mango fruit (Mangifera indica L.) and its by-products

Mango industry processing generates high quantities of mango by-products such as peels and seeds (35%–60% of the fruit). Indeed, it is known that mango and its by-products contain different families of bioactive compounds that possess several health benefits. Thus, the aim of this study has been the determination of different families of phenolic derivatives (free and bound phenolic compounds and alk(en)ylresorcinols (ARs)) in mango edible part and its by-products (peel, seed and seed husk) from three different cultivars. This is the first study that evaluates the phenolic compounds and ARs in the four fractions of mango of three different cultivars. Special attention has been paid to the determination of anthocyanins and ARs, because these families of compounds had not been studied in depth in mango. In fact, petunidin rutinoside-(p-coumaric acid) gallate was found in mango pulp, peel, seed and seed husk of the three cultivars and, it had never been described in mango before. It is also important to highlight that this is the first time that the identification and quantification of ARs have been performed in mango seed and seed husk; besides, four and five out of eleven alk(en)ylresorcinols detected in peel and pulp, respectively, were identified for the first time in these mango fractions. Furthermore, antioxidant activity was measured by ABTS and FRAP assays. Seed free and bound phenolic extracts showed the highest antioxidant capacity.

Unphosphorylated EIIANtr induces ClpAP-mediated degradation of RpoS in Azotobacter vinelandii.

The nitrogen-related phosphotransferase system (PTSNtr ) is composed of the EINtr , NPr and EIIANtr proteins that form a phosphorylation cascade from phosphoenolpyruvate. PTSNtr is a global regulatory system present in most Gram-negative bacteria that controls some pivotal processes such as potassium and phosphate homeostasis, virulence, nitrogen fixation and ABC transport activation. In the soil bacterium Azotobacter vinelandii, unphosphorylated EIIANtr negatively regulates the expression of genes related to the synthesis of the bioplastic polyester poly-β-hydroxybutyrate (PHB) and cyst-specific lipids alkylresorcinols (ARs). The mechanism by which EIIANtr controls gene expression in A. vinelandii is not known. Here, we show that, in presence of unphosphorylated EIIANtr , the stability of the stationary phase sigma factor RpoS, which is necessary for transcriptional activation of PHB and ARs synthesis related genes, is reduced, and that the inactivation of genes coding for ClpAP protease complex in strains that carry unphosphorylated EIIANtr , restored the levels and in vivo stability of RpoS, as well as the synthesis of PHB and ARs. Taken together, our results reveal a novel mechanism, by which EIIANtr globally controls gene expression in A. vinelandii, where the unphosphorylated EIIANtr induces the degradation of RpoS by the proteolytic complex ClpAP.

GacA regulates the PTSNtr-dependent control of cyst formation in Azotobacter vinelandii.

Azotobacter vinelandii forms cysts resistant to desiccation and produces polyhydroxybutyrate (PHB), alginate and alkylresorcinols (ARs) that are components of mature cysts. The expression of genes involved in the synthesis of these compounds is under the control of the GacA-RsmA global regulatory system where the RsmA protein represses the translation of mRNAs involved in the synthesis of these polymers. The synthesis of PHB and ARs is also controlled by the Nitrogen-regulated phosphotransferase system (PTSNtr) global regulatory system. When unphosphorylated, the Enzyme IIANtr (EIIANtr) protein impairs the synthesis of PHB and ARs. Here we show that cells of gacA mutants, as well as mutants that carry the EIIANtr protein in its unphosphorylated state, have similar encysting negative phenotypes. Interestingly, we found that in the gacA mutant strain, the EIIANtr protein was present in its unphosphorylated state. These data indicated that in addition to the GacA-RsmA system, GacA controls polymer synthesis and encystment by controlling the phosphorylation of the EIIANtr, revealing a previously unrecognized link between GacA and PTSNtr.

Laurdan emission study of the cholesterol-like effect of long-chain alkylresorcinols on the structure of dipalmitoylphosphocholine and sphingomyelin membranes

Long-chain alkylresorcinols (ARs) are commonly found in plant and bacteria cells, and they exhibit a wide variety of biological effects, including antifungal, antitumor, and antiphrastic activities. The cholesterol (Chol)-like effect of ARs with hydrocarbon side-chain lengths ranging from C15 to C25 on the structure of pure and Chol-doped dipalmitoylphosphocholine (DPPC) and sphingomyelin (SM) membranes was investigated by Laurdan fluorescence spectroscopy. The Laurdan emission generalized polarization parameter was analyzed as a function of the temperature and excitation wavelength in DPPC (or SM)/Chol, DPPC (or SM)/AR, and DPPC/Chol/AR systems. It was found that AR incorporation into both DPPC and SM bilayers induces an increase in the temperature of the main lipid phase transition, similar to the effect of Chol molecule incorporation. The phase separation, lipid-chain ordering, and membrane hydration are discussed for the AR-mixed membranes and compared with DPPC (or SM)/Chol membranes.

Identification and quantification of even and odd chained 5-n alkylresorcinols, branched chain-alkylresorcinols and methylalkylresorcinols in Quinoa (Chenopodium quinoa).

Quinoa is a pseudocereal grown in the Andean region of South America that is of increasing interest worldwide as an alternative staple food. We have detected a complex mixture of both odd- and even-alkyl chain alkylresorcinols (AR), branched-chain alkylresorcinols (bcAR) and methylalkylresorcinols (mAR) in ethyl acetate extracts of quinoa. We quantified the content of AR in 17 commercial samples of quinoa, and found that the mean±SD content of AR was 58±16μg/g, bcAR was 182±52μg/g, and mAR was 136±40μg/g. AR from quinoa could also be detected in plasma after eating quinoa, indicating that some of these unique AR could be used as biomarkers of quinoa intake in humans. Further work is required to understand the role of these ARs in the quinoa plant and whether any of the novel ARs may be of particular interest in human nutrition.

Characterization and Discrimination of Ancient Grains: A Metabolomics Approach.

Hulled, or ancient, wheats were the earliest domesticated wheats by mankind and the ancestors of current wheats. Their cultivation drastically decreased during the 1960s; however, the increasing demand for a healthy and equilibrated diet led to rediscovering these grains. Our aim was to use a non-targeted metabolomic approach to discriminate and characterize similarities and differences between ancient Triticum varieties. For this purpose, 77 hulled wheat samples from three different varieties were collected: Garfagnana T. turgidum var. dicoccum L. (emmer), ID331 T. monococcum L. (einkorn) and Rouquin T. spelta L. (spelt). The ultra high performance liquid chromatography coupled to high resolution tandem mass spectrometry (UHPLC-QTOF) metabolomics approach highlighted a pronounced sample clustering according to the wheat variety, with an excellent predictability (Q2), for all the models built. Fifteen metabolites were tentatively identified based on accurate masses, isotopic pattern, and product ion spectra. Among these, alkylresorcinols (ARs) were found to be significantly higher in spelt and emmer, showing different homologue composition. Furthermore, phosphatidylcholines (PC) and lysophosphatidylcholines (lysoPC) levels were higher in einkorn variety. The results obtained in this study confirmed the importance of ARs as markers to distinguish between Triticum species and revealed their values as cultivar markers, being not affected by the environmental influences.

Wheat alkylresorcinols reduce micellar solubility of cholesterol in vitro and increase cholesterol excretion in mice

Epidemiological studies have shown that the consumption of whole grains can reduce risk for metabolic disorders. We recently showed that chronic supplementation with wheat alkylresorcinols (ARs) prevents glucose intolerance and insulin resistance with hepatic lipid accumulation induced in mice by a high-fat high-sucrose diet (HFHSD). This study examines the effects of ARs on the micellar solubility of cholesterol in vitro, as well as the effects of transient AR supplementation on faecal lipid excretion and plasma lipid levels in mice. We found that ARs formed bile micelles with taurocholate independently of phospholipids, and dose-dependently decreased the micellar solubility of cholesterol in a biliary micelle model. Transient AR supplementation with HFHSD increased faecal cholesterol and triglyceride contents and decreased plasma cholesterol concentrations. These suggest that one underlying mechanism through which ARs suppress diet-induced obesity is by interfering with the micellar cholesterol solubilisation in the digestive tract, which subsequently decreases cholesterol absorption.

Alkylresorcinol content in whole grains and pearled fractions of wheat and barley

The aim of this work was to investigate the content and the composition of alkylresorcinols (ARs) in different wheat and barley cultivars, and in fractions obtained by progressive pearling. Three commercial winter wheats, characterized by different hardness and technological quality, and three barleys, including hulled and hull-less types, were selected. Two different protocols of sequential pearling were applied. Pearling of wheat and hull-less barley cultivars gave five fractions (each corresponding to 5% of grain weight) and 75% of the residue. In the case of hulled barley eight pearled fractions and 60% of inner kernel were obtained. In wheat ARs were prevalently located in the 5–10% intermediate fraction, while for barley results varied depending on the cultivar. In the hull-less cultivar, the AR content progressively decreased from the outermost fraction (0–5%) towards the inner layers, while for hulled barley the highest AR content was observed in the 10–15% fraction, evidencing lower amounts in the coarse hull (included in the 0–5% and 5–10% fractions). Based on the different localization of ARs in the cereal kernel, progressive pearling can be employed to obtain enriched fractions that could be used to enhance ingredients and products rich in these bioactive compounds.

Effects of Labisia pumila var alata extracts on the lipid profile, serum antioxidant status and abdominal aorta of high-cholesterol diet rats

BackgroundPrevious studies on Labisia pumila var. alata (LPva) have showed that it could inhibit low-density lipoprotein (LDL) oxidation and provide protection on myocardial infarction in rats. Hypothesis/PurposeWe hypothesized that LPva extracts can modulate the lipid profiles and serum antioxidant status of hypercholesterolemic rats. In the present study, we investigated the effects of aqueous and 80% ethanol extracts of LPva on atherogenic and serum antioxidant parameters as well as changes in abdominal aorta of high-cholesterol diet rats. MethodsThe major components of the extracts, gallic acid, flavonoids and alkyl resorcinols were analyzed by using a validated reversed phase HPLC method. The rats were induced to hypercholesterolemic status with daily intake of 2% cholesterol for a duration of 8 weeks. Three different doses (100, 200 and 400mg/kg) of the extracts were administered daily on the 4th week onwards. The rats were then sacrificed and the blood was collected via abdominal aorta and serum was separated by centrifugation for biochemical analysis. Part of the aorta tissues were excised immediately for histopathological examination. ResultsThe serum of LPva treated rats showed significant reduction in serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) levels and the abdominal aorta showed a significant decrease of atheroma lesions in treated rats. Serum lipid profiles of treated rats showed a decrease in total cholesterol, total triglycerides and low-density lipoprotein (LDL) levels as compared to control group. The atherogenic indices in treated rats were significantly improved along with an increasing level of serum high-density lipoprotein (HDL). The extracts also exhibited significant increase of antioxidant enzymes and decrease of MDA as a product of lipid peroxidation. ConclusionLPva extracts can reduce the risk of dyslipidemia by improving the serum lipid profiles and modulating serum antioxidants.

Plasma Alkylresorcinol Concentrations as a Novel Measure of Compliance to a Gluten‐Free Diet

BackgroundMany celiac disease (CD) patients experience difficulties in adherence to a gluten‐free diet. Methods for testing compliance to a gluten‐free diet are costly and cumbersome.ObjectiveA simple biomarker of gluten intake is needed in a clinical setting and will be useful for epidemiological studies investigating wider effects of gluten intake. The aim was to evaluate plasma total alkylresorcinols (AR) concentrations as a measure of gluten intake.DesignThis randomized, controlled, cross‐over intervention study in 52 Danish adults with features of the metabolic syndrome compared eight weeks gluten‐rich and gluten‐poor diet separated by a wash‐out period of at least six weeks. We measured fasting plasma concentrations of AR to determine if they reflected differences in gluten intake. Additionally, we investigated in 118 Danish adults the cross‐sectional association between self‐reported gluten intake and plasma AR in this and a similar study at baseline.ResultsTotal plasma AR decreased more during the gluten‐poor period compared to the gluten‐rich period −124.8 (−156.5; −93.0) vs. −31.8 (−63.1; −0.4) nmol/L, respectively, p&lt;0.001. Based on the plasma AR‐profile we built a classification tree to objectively determine compliance and found an overall misclassification error of participants of 3.9%. The cross‐sectional study showed a 5.6% (2.4; 8.9) increase in total plasma AR per 1 g‐increase in reported gluten intake (p&lt;0.001).ConclusionWe propose plasma AR as a new tool to monitor compliance to a gluten‐free diet to support clinical work with CD, as well as to help investigations on the possible effects of gluten in the wider population.Support or Funding InformationFunded by the Innovation Fund Denmark, grant no. 0603‐00487B (11‐116163). Some intervention products were sponsored by Kohberg, Lantmännen, AXA, Wasa, Urtekram, Finax, and Doves Farm. MVL is part sponsored by Cereal Partners Worldwide with an unrestricted grant. The study sponsors played no role in the design, methods, data‐management and analysis or in the decision to publish.

Spectroscopic determination of alkyl resorcinol concentration in hydroxyapatite composite

Recently, alkyl resorcinol compounds showed remarkable improvements on dental implant restoration as well as anaesthetic, antiseptic, and anthelmintic applications. In this study, we prepared biologically functional composition of 4-hexylresorcinol (4HR)-loaded hydroxyapatite (HA). Attenuated total reflectance (ATR) Fourier-transform infrared (FT-IR) spectroscopy fully assigned vibrational absorptions of 4-hexylresorcinol as well as the HA. The absorption coefficient of 4HR aqueous solution is estimated to be 1393 ± 61 cm−1 M−1 at highly diluted concentration. The 4HR was loaded with 0.018 % (wt/wt) in the 4HR-HA composite. We quantitatively determined the micromolar concentration of 4HR loaded in the composite based on ultraviolet-visible (UV-Vis) absorption spectroscopy.

Separation and isolation of saturated and unsaturated 5-n-alk(en)ylresorcinols from rye bran

Alkylresorcinols (AR) are a class of phenolic lipids which are found in a number of plants and microorganisms and which are associated with various biological activities. In view of the very limited availability of reference compounds, we developed a process for the isolation of AR from rye bran. For this purpose, a crude extract was subjected to deep bed filtration at −80°C, whereby the crystallized saturated AR were separated from the unsaturated homologues. Highly purified, individual compounds were subsequently isolated by semi-preparative HPLC. In contrast to previously published approaches, the method reported here is relatively easy to perform and allows the recovery of saturated and unsaturated AR of high purity. Furthermore, it is superior to argentation chromatography because it does not necessitate the removal of silver ions for subsequent bioassays.

Validation of Reported Whole-Grain Intake from a Web-Based Dietary Record against Plasma Alkylresorcinol Concentrations in 8- to 11-Year-Olds Participating in a Randomized Controlled Trial

Background: Whole-grain (WG) intake is important for human health, but accurate intake estimation is challenging. Use of a biomarker for WG intake provides a possible way to validate dietary assessment methods.Objective: Our aim was to validate WG intake from 2 diets reported by children, using plasma alkylresorcinol (AR) concentrations, and to investigate the 3-mo reproducibility of AR concentrations and reported WG intake.Methods: AR concentrations were analyzed in fasting blood plasma samples, and WG intake was estimated in a 7-d web-based diary by 750 participants aged 8–11 y in a 2 school meal × 3 mo crossover trial. Reported WG intake and plasma AR concentrations were compared when children ate their usual bread-based lunch (UBL) and when served a hot lunch meal (HLM). Correlations and cross-classification were used to rank subjects according to intake. The intraclass correlation coefficients (ICCs) between subjects’ measurements at baseline and after the UBL were used to assess reproducibility.Results: Correlations between reported WG wheat + rye intake and plasma AR were 0.40 and 0.37 (P < 0.001) for the UBL and the HLM diets, and 78% and 77% were classified in the same or adjacent quartiles for the UBL and HLM diets, respectively. The ICC over 3 mo was 0.47 (95% CI: 0.38, 0.55) for plasma total ARs and 0.64 (95% CI: 0.58, 0.70) for reported WG intake. Correlations were higher when using the AR C17:0 homolog as a biomarker, reflecting rye intake instead of plasma total ARs [UBL: r = 0.47; HLM: r = 0.43, P < 0.001; ICC = 0.51 (95% CI: 0.43, 0.59)].Conclusions: Self-reported WG wheat + rye intake among children showed moderate correlations with plasma AR concentrations. Substantial intraindividual variation was found in WG intake and plasma AR concentrations. The AR homolog C17:0 may be used as a biomarker for WG intake when the WG intake primarily comes from rye as in the present study. This trial was registered at clinicaltrials.gov as NCT01457794.

Differentiation of bread made with whole grain and refined wheat (T. aestivum) flour using LC/MS-based chromatographic fingerprinting and chemometric approaches

A fuzzy chromatography mass spectrometric (FCMS) fingerprinting method combined with chemometric analysis was established to differentiate between breads made from whole wheat (WW) flour and refined wheat (RW) flour. The chemical compositions of the bread samples were profiled using ultrahigh-performance liquid chromatography with high-resolution accurate-mass multistage mass spectrometry (UHPLC-HRAM-MSn) with atmospheric pressure chemical ionisation (APCI) in positive ionisation mode. Principal component analysis (PCA) and soft independent modelling of class analogy (SIMCA) of the FCMS fingerprints revealed the components responsible for the chemical differences between WW and RW flour/bread samples. Alk(en)ylresorcinols (ARs) were demonstrated to be the most important markers for differentiation between WW and RW flour/breads. Diglycerides (DGs), and phosphatidylethanolamine (PE) also contributed significantly to the classification. Bread crumb and crust showed no difference in chemical profiles in this study. SIMCA, using WW modelling, could be a potent and robust tool for authentication of WW breads.