Adrian A. Franke

We report a new method for fast and sensitive analyses of biologically relevant fatty acids (FAs) in red blood cells (RBC) by liquid chromatography mass spectrometry (LC−MS). A new chemical derivatization approach was developed forming picolylamides from FAs in a quantitative reaction. Fourteen derivatized FA standards, including saturated and unsaturated FAs from C14 to C22, were efficiently separated within 15 min. In addition, the use of a recently introduced benchtop orbitrap mass spectrometer under positive electrospray ionization (ESI) full scan mode showed a 2−10-fold improvement in sensitivity compared with a conventional tandem MS method, with a limit of detection in the low femtomole range for saturated and unsaturated FAs. The developed method was applied to determine FA concentrations in RBC with intra- and interday coefficients of variation below 10%.

Background: Isoflavonoids (IFL) may protect against chronic diseases, including cancer. IFL exposure is traditionally measured from plasma (PL), but the reliability of urine is uncertain. We assessed whether IFL excretion in overnight urine (OU) or spot urine (SU) reliably reflects IFLs in PL and the usefulness of the three matrices to determine soy intake compliance. Methods: In a randomized, double-blind, placebo-controlled soy intervention trial with 350 postmenopausal women, IFLs (daidzein, genistein, glycitein, equol, O-desmethylangolensin, dihydrodaidzein, dihydrogenistein) were analyzed by liquid chromatography/mass spectrometry in OU, SU, and PL collected at baseline and every 6 months over 2.5 years. Results: High between-subject intraclass correlations between all three matrices (median, 0.94) and high between-subject Pearson correlations (median r_OU-PL = 0.80; median r_SU-PL = 0.80; median r_OU-SU = 0.92) allowed the development of equations to predict IFL values from any of the three matrices. Equations developed from a randomly selected 87% of all available data were valid because high correlations were found on the residual 13% of data between equation-generated and measured IFL values (median r_OU-PL = 0.86; median r_SU-PL = 0.78; median r_OU-SU = 0.84); median absolute IFL differences for OU-PL, SU-PL, and OU-SU were 8.8 nmol/L, 10.3 nmol/L, and 0.28 nmol/mg, respectively. All three matrices showed highly significant IFL differences between the placebo and soy intervention group at study end (P < 0.0001) and highly significant correlations between IFL values and counted soy doses in the intervention group. Conclusions: OU and SU IFL excretion reflect circulating PL IFL levels in healthy postmenopausal women accurately. Impact: Noninvasively-collected urine can be used to reliably determine systemic IFL exposure and soy intake compliance. Cancer Epidemiol Biomarkers Prev; 19(7); 1775–83. ©2010 AACR.

Sporadic intake and short half-lives of serum or urinary biomarkers may make serum and urinary isoflavones quite unreliable indicators of longer-term dietary soy intake. In 26 participants in the Adventist Health Study-2 (AHS-2) we obtained two measures of fasting morning serum isoflavones, 1–2 years apart. In another 76 subjects we obtained an overnight urine sample and six 24-h dietary recalls over a period encompassing the time of the urine sample. Intraclass correlations (ICC) values for serum isoflavones were 0.11 (log(daidzein)) and 0.28 (log(genistein)). Assuming that the correlation (true dietary intake, true urinary excretion) <0.90, it is shown that this implies an ICC for urinary estimates that exceeds 0.56. As expected, the previous day’s soy intake, and its timing, influenced the next morning’s serum levels. These results suggest that fasting morning serum isoflavone estimates will provide a poor index of long-term soy intake, but that overnight urinary estimates perform much better.

Introduction: Assessments by the handful of prospective studies of the association of serum antioxidants and breast cancer risk have yielded inconsistent results. This multiethnic nested case-control study sought to examine the association of plasma carotenoids, retinol, and tocopherols with postmenopausal breast cancer risk. Methods: From the biospecimen subcohort of the Multiethnic Cohort Study, 286 incident postmenopausal breast cancer cases were matched to 535 controls on age, sex, ethnicity, study location (Hawaii or California), smoking status, date/time of collection and hours of fasting. We measured prediagnostic circulating levels of individual carotenoids, retinol, and tocopherols. Conditional logistic regression was used to compute odds ratios and 95% confidence intervals. Results: Women with breast cancer tended to have lower levels of plasma carotenoids and tocopherols than matched controls, but the differences were not large or statistically significant and the trends were not monotonic. No association was seen with retinol. A sensitivity analysis excluding cases diagnosed within 1 year after blood draw did not alter the findings. Conclusions: The lack of significant associations in this multiethnic population is consistent with previously observed results from less racially-diverse cohorts and serves as further evidence against a causal link between plasma micronutrient concentrations and postmenopausal breast cancer risk.

Isoflavone (IFL) intake may provide numerous health benefits, but IFL bioavailability differences among soya foods remains uncertain. Urinary IFL excretion (UIE) was shown to provide a reliable surrogate for systemic IFL exposure and therefore can be used as a measure of ‘apparent bioavailability’ (AB). We investigated the AB of IFL in fourteen healthy adults, consuming two liquid and two solid soya foods in a crossover designed study. Volunteers consumed the foods with a self-selected breakfast, which was kept identical for all four soya items (soya nuts, soya milk, soya protein bar and soya protein powder drink in water; average 23·7 mg IFL, 88–96 % glycosides, by HPLC analysis) and collected all urine up to 26 h. Liquid foods showed initially higher UIE values than solid foods, but this difference was considerably reduced or disappeared entirely after 24–26 h. Conclusive AB results were obtained only after 24–26 h; earlier collections were not reliable. At 26 h, adjusted UIE values for daidzein (DE) were 20 μmol in the milk and bar and 17 μmol for the nut and powder; urinary genistein excretion was the highest in the milk group (10 μmol) followed by the nut, bar (both 6 μmol) and powder groups (5 μmol); the UIE for glycitein was the highest for bars (4 μmol), followed by powder and nuts (3 μmol), and milk (2 μmol). DE makes the largest contribution to urinary total IFL. The AB of IFL was found to be variable depending on the analyte and soya food consumed.

Isoflavones (IFLs) are natural products to which humans have been traditionally exposed predominantly through soy foods; more recently humans are also exposed to them through soy protein addition to processed foods or through supplements. They are structurally similar to steroidal estrogens and can exert estrogenic or antiestrogenic effects depending on their concentrations and on the tissue considered. These properties qualify IFLs to be classified as phytoestrogens and are believed to account for many of the biological effects observed for soy and/or IFL exposure including benefits for bone and heart health or prevention of menopausal symptoms and certain types of cancer. In order to evaluate the function of IFLs, alone or when exposure happens through soy intake, pharmacokinetics and bioavailability are critical issues to be considered in epidemiologic and clinical research. For this purpose precise, accurate, robust, fast, and affordable techniques for IFL analyses are required. Copyright © 2009 John Wiley & Sons, Ltd.

Epidemiologic evidence supports a role of soy foods in breast cancer etiology. Because chronic inflammation appears to be a critical component in carcinogenesis, we examined the potential anti-inflammatory effects of soy foods.