Alkylresorcinols in Latvian and Finnish breads.

The alkylresorcinol (AR) content and relative homologue composition were determined in 9 Latvian and 11 Finnish soft breads. ARs were extracted with hot 1-propanol and quantified, using a gas chromatography-mass spectrometry method. The total AR content (µg/g dry matter) varied from 560 to 840 in rye breads, from 500 to 700 in Finnish mixed rye and wheat flour breads, from 200 to 300 in Latvian mixed rye and wheat flour breads and from 25 to 30 in white wheat breads. Rye and white wheat breads in the two countries varied only slightly in AR content, but there were wide variations in AR content in mixed flour breads. The AR contents in soft breads could be indicators of bran or fibre content, but not of whole-grain flour content.

Are there endogenous estrone fatty acyl esters in human plasma or ovarian follicular fluid?

BACKGROUND: Estrone and its sulfated esters are the most abundant estrogens in blood in men and in women after the menopause. However, previous studies on the esterification of estrone with fatty acids have yielded conflicting results, some studies reporting high nanomolar concentrations of estrone fatty acyl esters in plasma. METHODS: We developed an estrone radioimmunoassay (RIA) method to determine endogenous concentrations of estrone and after saponification, applied it to male and female plasma. In addition, the concentration of estrone fatty acyl esters in ovarian follicular fluid was analyzed by gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). RESULTS: By estrone RIA, we did not find measurable amounts of estrone fatty acyl esters in male or female plasma, except for one premenopausal woman who had the highest plasma concentration of nonesterified estrone. The concentration of hydrolyzed estrone fatty acyl esters determined by LC-MS/MS in follicular fluid obtained from women undergoing ovarian stimulation was below the limit of quantification of <10 pmol/l (<2.7 ng/l). CONCLUSIONS: In contrast to previous data by others, our study suggests that estrone fatty acyl esters are in most cases not detectable in plasma of healthy men or healthy nonpregnant women.

Pharmacokinetics of alkylresorcinol metabolites in human urine.

Wholegrain cereals are reported to promote beneficial health effects. Wholegrain wheat and rye are almost exclusive sources of alkylresorcinols, and intact alkylresorcinols together with their plasma and urinary metabolites, 3-(3,5-dihydroxyphenyl)-1-propanoic acid (DHPPA) and 3,5-dihydroxybenzoic acid (DHBA), have been proposed as biomarkers of the intake of these foods in humans. The pharmacokinetics of alkylresorcinols and their metabolites in plasma have been determined but not that of the urinary metabolites. We aimed to characterise the urinary pharmacokinetics of alkylresorcinol metabolites in humans to evaluate their potential as biomarkers of wholegrain wheat and rye. A group of fifteen volunteers followed a low-alkylresorcinol diet for 2 d before ingesting a single dose of rye bread, containing 100 mg alkylresorcinols. Urine was collected between baseline (0 h) and 25 h after administration. Thereafter alkylresorcinol metabolites were quantified by HPLC with coulometric electrode array detection. Maximum excretion rates were observed at 5-6 h for both metabolites, DHPPA being predominant over DHBA and also possessing a greater area under the curve0-25 h. Total urinary recovery between 0 and 25 h yielded 43 % of ingested alkylresorcinols, and at 25 h significant amounts of metabolites were still retained in the body, suggesting that even a spot urine sample may be sufficient to indicate whether or not wholegrain wheat or rye is a daily dietary component. These results support the use of urinary DHPPA and DHBA as biomarkers of wholegrain wheat and rye and enable new potential for studying the association between wholegrain intake and diseases, even in the absence of dietary data.

Quantitative determination of dehydroepiandrosterone fatty acyl esters in human female adipose tissue and serum using mass spectrometric methods.

Dehydroepiandrosterone-fatty acyl esters (DHEA-FAE) are naturally occurring water-insoluble metabolites of DHEA, which are transported in plasma exclusively by lipoproteins. To find out whether DHEA, like estradiol, might be stored in adipose tissue in FAE form, we set up a mass spectrometric method to quantify DHEA-FAE and free DHEA in human adipose tissue and serum. The method consists of chromatographic purification steps and final determination of hydrolyzed DHEA-FAE and free DHEA, which was carried out by gas chromatography-mass spectrometry (GC-MS) or liquid chromatography-tandem mass spectrometry (LC-MS/MS). Our results showed that no detectable amounts of DHEA-FAE could be found in adipose tissue although 32-178 pmol/g of free DHEA were determined by GC-MS and LC-MS/MS. The DHEA-FAE concentrations in serum quantified by GC-MS were 1.4±0.7 pmol/ml in premenopausal women (n=7), and 0.9±0.4 pmol/ml in postmenopausal women (n=5). Correspondingly, the free DHEA concentrations were 15.2±6.3 pmol/ml and 6.8±3.0 pmol/ml. In addition, the mean proportions of DHEA-FAE of total DHEA (DHEA-FAE+free DHEA) in serum were 8.6% and 11.2% in pre- and postmenopausal women, respectively. Serum DHEA-FAE concentration was below quantification limit for LC-MS/MS (signal-to-noise ratio, S/N=10), while free DHEA concentrations varied between 5.8 and 23.2 pmol/ml. In conclusion, the proportion of DHEA-FAE of total DHEA in serum was approximately 9%. However, in contrast to our previous findings for estradiol fatty acid esters in adipose tissue which constituted about 80% of total estradiol (esterified+free), the proportion of DHEA-FAE of total DHEA was below 5%. Four to ten times higher concentrations of free DHEA were quantified in adipose tissue compared to those in serum.

Plasma and urinary alkylresorcinol metabolites as potential biomarkers of breast cancer risk in Finnish women: a pilot study.

Alkylresorcinols (ARs) are shown to be good biomarkers of consumption of rye and whole-grain wheat products in man. The aim of this pilot study was to investigate AR metabolites as potential biomarkers of breast cancer (BC) risk in Finnish women since intake of cereal fiber and its components has been proposed to reduce this risk through an effect on the enterohepatic circulation of estrogens. This was a cross-sectional and observational pilot study. A total of 20 omnivores, 20 vegetarians, and 16 BC women (6-12 mo after operation) were investigated on 2 occasions 6 mo apart. Dietary intake (5-days record), plasma/urinary AR metabolites [3,5-dihydroxybenzoic acid (DHBA) and 3-(3,5-dihydroxyphenyl)-1-propanoic acid (DHPPA)] and plasma/urinary enterolactone were measured. The groups were compared using nonparametric tests. We observed that plasma DHBA (P = 0.007; P = 0.03), plasma DHPPA (P = 0.02; P = 0.01), urinary DHBA (P = 0.001; P = 0.003), urinary DHPPA (P = 0.001; P = 0.001), and cereal fiber intake (P = 0.007; P = 0.003) were significantly lower in the BC group compared to the vegetarian and omnivore groups, respectively. Based on measurements of AR metabolites in urine and in plasma, whole-grain rye and wheat cereal fiber intake is low in BC subjects. Thus, urinary and plasma AR metabolites may be used as potential biomarkers of BC risk in women. This novel approach will likely also facilitate studies of associations between rye and whole-grain wheat cereal fiber intake and other diseases. Our findings should, however, be confirmed with larger subject populations.

Plasma alkylresorcinol metabolites as potential biomarkers of whole-grain wheat and rye cereal fibre intakes in women.

It has been demonstrated that intact plasma alkylresorcinols (AR) and urinary AR metabolites could be used as biomarkers of whole-grain intake. Thereafter, we developed the method for the plasma AR metabolites, which is more convenient and requires less sample pretreatment than the analysis of intact plasma AR. The aim of the present study is to evaluate whether AR metabolites measured in plasma, in the same population, could also be considered as useful biomarkers of cereal fibre. Fifty-six women were recruited in a cross-sectional and observational study. Dietary intake (5-d record) and plasma AR metabolites (3,5-dihydroxybenzoic acid, DHBA; 3-(3,5-dihydroxyphenyl)-1-propanoic acid, DHPPA) were measured. The relationship between plasma AR metabolites and cereal fibre intake was examined using partial correlation and stepwise regression. Cereal fibre intake correlated significantly with plasma DHBA (r 0.411; P = 0.002) and DHPPA (r 0.463; P = 0.000) even after adjustment for BMI and age. Thus, plasma AR metabolites correlate with cereal fibre intake as noted with plasma intact AR and urinary AR metabolites. We observed that plasma DHPPA was the independent predictor of cereal fibre intake, explaining 18 % of the variance (adjusted r(2) 0.176; P = 0.002). In epidemiological screening, it might be easier to obtain and to collect plasma than urine samples. In addition, the plasma AR metabolites half-life seems longer than those of intact plasma AR, and their measurements are more convenient, and faster. Thus, sum of plasma AR metabolites and more specifically plasma DHPPA seems to be good and specific biomarkers of cereal fibre intake.

Responsiveness of urinary and plasma alkylresorcinol metabolites to rye intake in finnish women.

Alkylresorcinols [ARs] have been proposed for use as biomarkers of whole-grain intake. The aim here was to examine the responsiveness of AR metabolites to rye intake. Sixty women were divided into three groups according to their rye consumption. We observed significant differences between groups in plasma 3-[3,5-dihydroxyphenyl]-1-propanoic acid [DHPPA] and in urinary DHPPA and 3,5-dihydroxybenzoic acid [DHBA]. In addition, these AR metabolites increased proportionally to rye fiber intake. We conclude that these ARs metabolites are accurate and useful biomarkers of rye fiber intake. Further studies are needed to confirm our results in larger and different populations.

Plasma pharmacokinetics of alkylresorcinol metabolites: new candidate biomarkers for whole-grain rye and wheat intake.

BACKGROUND: Alkylresorcinols are phenolic compounds that are present almost exclusively in rye and wheat fiber. Alkylresorcinols are absorbed and thereafter metabolized to 3,5-dihydroxybenzoic acid (DHBA) and 3-(3,5-dihydroxyphenyl)-1-propanoic acid (DHPPA), which have been detected in human urine and plasma. OBJECTIVE: The objective was to determine the plasma pharmacokinetics of DHBA and DHPPA in human subjects to estimate whether they show potential as biomarkers for whole-grain rye and/or wheat intake. DESIGN: Fifteen human volunteers followed a low-alkylresorcinol diet for 2 d before ingesting a single dose of high-fiber rye bread containing 100 mg alkylresorcinols [corrected]. Plasma samples were collected for 25 h, and the alkylresorcinol metabolites were quantified by HPLC with coulometric electrode array detection. RESULTS: Maximum concentrations were reached at approximately 6 h for both metabolites, although interindividual variation was found. The half-life was significantly (P < 0.0002) longer for DHPPA (16.3 h) than for DHBA (10.1 h). No significant differences were discovered between women and men in the half-life of each metabolite, which, from a pharmacokinetic point of view, is the most important parameter. The area under the curve differed significantly between DHBA and DHPPA (P < 0.0001) and between women and men (P = 0.03 for DHBA and P = 0.01 for DHPPA). However, when corrected for body weight, the difference between sexes was no longer significant. CONCLUSIONS: Our results suggest that DHBA and DHPPA are both good candidate biomarkers for whole-grain rye and/or wheat intake; however, DHPPA is the better indicator because of its longer half-life. This could provide a practical tool when investigating the association between diet and diseases.

Urinary 3-(3,5-dihydroxyphenyl)-1-propanoic acid, an alkylresorcinol metabolite, is a potential biomarker of whole-grain intake in a U.S. population.

5-n-Alkylresorcinols (AR) are a major group of phenolic compounds in whole-grain wheat, rye, and barley. As such, they may serve as potential biomarkers of whole-grain intake, because they are quantifiable intact in plasma and as metabolites in urine. We examined relationships between 12-h urinary excretion of AR metabolite 3-(3,5-dihydroxyphenyl)-1-propanoic acid (DHPPA) and self-reported habitual intake of whole-grain foods measured by 3-d food record (3DFR) and FFQ. Urine samples from 100 men and women were analyzed for DHPPA using HPLC with coularray detection. DHPPA excretion ranged from 1.3 to 99.4 (mean +/- SE, 14.0 +/- 1.5) mumol/12 h. Whole-grain food intake, as determined by 3DFR and FFQ and adjusted for BMI and energy and fiber intake, was significantly associated with 12-h urinary DHPPA excretion. Based on 3DFR, whole-grain wheat + rye consumers had a 44% higher DHPPA excretion than nonconsumers [ratio of excretion (95% CI) = 1.44 (1.04, 1.97); P = 0.029]. Using whole-grain intake estimated by FFQ, a serving increase in whole-grain wheat + rye intake increased DHPPA excretion by 94% [ratio of excretion (95% CI) = 1.94 (1.35, 2.78); P = 0.001] and a serving increase in whole grains as defined more broadly in epidemiologic studies of whole-grain intake and disease risk (whole-grain wheat, rye, oats, and corn) increased DHPPA by 67% [ratio of excretion (95% CI) = 1.67 (1.28, 2.17); P < 0.0001]. This study supports the potential utility of urinary DHPPA as a biomarker of whole-grain intake in a U.S. population.

Plasma alkylresorcinols and urinary alkylresorcinol metabolites as biomarkers of cereal fiber intake in Finnish women.

Alkylresorcinols (AR) could be good biomarkers of consumption of fiber-rich cereal products. The aim of this study was to examine the relationship between plasma ARs or urinary AR metabolites and cereal fiber intake in women consuming their habitual diet. Twenty-five postmenopausal and 31 premenopausal women were recruited. The subjects included also vegetarians (n = 20) to obtain a broad range of cereal intake. Dietary intake, plasma ARs, and urinary AR metabolites [3,5-dihydroxybenzoic acid and 3-(3,5-dihydroxyphenyl)-1-propanoic acid] were measured. Pearson's and Partial correlation tests were done between dietary fiber intake and plasma ARs or urinary AR metabolites. Cereal fiber intake correlated significantly with plasma AR C17:0 (r = 0.387), AR C19:0 (r = 0.350), AR C21:0 (r = 0.428), AR C23:0 (r = 0.409), AR C25:0 (r = 0.283), and total AR (r = 0.406) and with urinary AR metabolites DHBA (r = 0.359) and DHPPA (r = 0.402) even after adjustment for body mass index and age, which could be confounding variables. This is the first study to show a significant correlation between plasma ARs or urinary AR metabolites and cereal fiber intake during consumption of a habitual diet. These results indicate that assay of plasma ARs or urinary AR metabolites may be used as biomarkers in epidemiologic studies in free-living populations to evaluate the role of cereal fiber intake in various diseases.

Quantification of alkylresorcinol metabolites in plasma by high-performance liquid chromatography with coulometric electrode array detection.

This study presents the optimization and validation of a rapid protocol for quantifying alkyresorcinol (AR) metabolites 3,5-dihydroxybenzoic acid (DHBA) and 3-(3,5-dihydroxyphenyl)-1-propanoic acid (DHPPA) in plasma, using high-performance liquid chromatography (HPLC) coupled with a coulometric electrode array detector. Syringic acid (SyrA) serves as the internal standard. The new method is simple and could be used in large epidemiological studies. The summed AR metabolite concentrations measured in plasma correlate significantly with the summed urinary AR metabolite concentrations (R = 0.613; p < 0.001) and with the summed intact AR (C17:0-C25:0) concentrations in plasma (R = 0.686; p < 0.001). Additional investigation is needed to clarify whether the two plasma AR metabolites are useful as biomarkers of whole-grain intake and helpful in the exploration of the association between whole-grain cereal intake and human diseases.

Quantification of alkylresorcinol metabolites in urine by HPLC with coulometric electrode array detection.

BACKGROUND: Whole-grain rye and wheat cereals contain high amounts of alkylresorcinols (ARs), phenolic lipids. ARs can be quantified in plasma. Two recently identified urinary AR metabolites, 3,5-dihydroxyphenylbenzoic acid (DHBA) and 3-(3,5-dihydroxyphenyl)-1-propanoic acid (DHPPA), may be useful as biomarkers of intake of whole-grain rye and wheat. METHODS: We evaluated 4 pretreatment protocols for quantifying urinary DHBA and DHPPA using HPLC coupled with a coulometric electrode array detector. Syringic acid was used as the internal calibrator. RESULTS: Measured urinary concentrations of DHBA and DHPPA were 0.8-115 micromol/L. The mean recoveries of all added concentrations were 85%-104% for DHBA and 86%-99% for DHPPA, depending on the degree of the purification. The protocol versions with less purification correlated well with the protocol including highest purification. The correlation coefficients (r(2)) were 0.9699-0.8153 for DHBA and 0.9854-0.8371 for DHPPA. CONCLUSION: Although the protocol with the most purification steps was most specific, all protocols were suitable for measuring DHBA and DHPPA in urine. The rapid protocol with simple hydrolysis could be used in large-scale clinical studies. Additional investigation is needed to clarify whether these metabolites are useful biomarkers of whole-grain intake and helpful in the exploration of its association with human diseases.