Development of a method for the detection of zinc in Brassica oleracea using solid phase extraction and size-exclusion chromatography inductively coupled plasma mass spectrometry (SEC-ICP-MS).

The aim of this work is the development of a suitable method to extract and detect zinc-bound compounds from cabbage, broccoli and kale (family Brassicaceae, species oleracea) using solid phase extraction (SPE) and size-exclusion chromatography inductively coupled plasma mass spectrometry (SEC-ICP-MS). Tris [2-Amino-2-(hydroxymethyl)-1,3-propanediol]/hydrochloric acid (Tris/HCl) or ammonium nitrate were used as extractants added to the freeze-dried samples, which were then sonicated and centrifuged. An enzymatic mixture was added to the extracts and then incubated for 5- and 18 h prior to analysis by SEC-ICP-MS. Results showed a good coefficient of variation (CV) of the elution time (0.06-0.9%), concentration (4.7-16.9%) and molecular size (0.4-5.4%). The limit of detection (LOD) and the limit of quantitation (LOQ) were 0.9 µg L-1 and 2.8 µg L-1, respectively. The proposed method is reliable and robust and can be applied to samples with difficult matrices like vegetables and soil.•Good precision, stability and reproducibility.•Easy to execute and suitable for analysis of vegetables and other samples with complex matrices, eg. soil.•This method contributed to good maintenance of the instrument and to minimal cleaning time.

Qualitative in vitro study on the degradation of mineral complexes in vegetables.

Mechanisms of degradation and absorption of mineral complexes by the human digestive system are complex and still under investigation. The elaborate matrix of vegetables, and the presence of phytates and other inhibitors make study of these mechanisms difficult. In this qualitative study, extracts from freeze-dried savoy cabbage, broccoli, kale and spinach were subjected to digestion in vitro at pH 2.0 and pH 7.5 and analysed using SEC-ICP-MS. The results suggest that low molecular weight species (peak 6), related to the iron and zinc fractions, which appeared after acidic digestion in all vegetables, except in kale, were considerably reduced after digestion at pH 7.5. Low molecular weight species (peak 9), related to the phosphorus fraction, were present in all vegetables, except in kale, after alkaline digestion. While cabbage, broccoli and spinach showed similar degradation patterns, kale showed a different degradation behaviour.

Development of a reproducible method of analysis of iron, zinc and phosphorus in vegetables digests by SEC-ICP-MS.

Vegetables contain iron, zinc and phosphorus as complexes with phytates limiting their availability from a vegetarian diet, meaning non-haem iron deficiency anaemia and zinc deficiency immune malfunction are a risk. Although these elements have been analysed previously in biological fluids and cereal using LC-ICP-MS, there is no method suitable for analysing iron, zinc and phosphorus simultaneously in vegetables because of their complex matrix. In this study, we analysed iron, zinc and phosphorus in cabbage, broccoli, pepper, spinach, kale and rocket after a simulated gastrointestinal digestion using a newly optimised SEC-ICP-MS method. Ammonium nitrate, as the mobile phase, and a suitable rinsing regime, allowed good reproducibility and maintenance of the equipment. The method showed good reproducibility and can be easily adapted to other vegetables, as required.

Phytoestrogen content of cereals and cereal-based foods consumed in the UK.

Dietary phytoestrogens may be involved in the occurrence of chronic diseases. Reliable information on the phytoestrogen content in foods is required to assess dietary exposure and disease risk in epidemiological studies. However, there is little information on isoflavone, lignan, and coumestrol content of cereals and cereal-based foods, leading to an underestimation of intake. This is the first study of phytoestrogens (isoflavones: biochanin A, daidzein, formononetin, genistein, glycitein; lignans: matairesinol, secoisplariciresinol; coumestrol) in a comprehensive selection of 101 cereals and cereal-based foods-including breads, breakfast cereals, biscuits, pasta and rice-consumed in the UK using a sensitive LCMS technique with 13C-labelled internal standards. Phytoestrogens were detected in all foods analyzed; bread contained the highest amount of phytoestrogens-many as isoflavones-with an average content of 375 +/- 67 microg/100 g wet weight (excluding soya-linseed bread with 12,000 microg/100 g). Most other foods contained less than 100 microg/100 g, many as lignans. Our study shows that all foods analyzed contained phytoestrogens, with the highest amount found in breads, making them one of the main sources of dietary phytoestrogens in the UK. These results will allow a more accurate estimation of exposure to dietary phytoestrogens.

Phytoestrogen content of foods of animal origin: dairy products, eggs, meat, fish, and seafood.

Dietary phytoestrogens may be involved in the occurrence of chronic diseases. Reliable information on the phytoestrogen content in foods is required to assess dietary exposure and disease risk in epidemiological studies. However, existing analyses have focused on only one class of these compounds in plant-based foods, and there is only little information on foods of animal origin, leading to an underestimation of intake. This is the first comprehensive study of phytoestrogen content in animal food. We have determined the phytoestrogen content (isoflavones: biochanin A, daidzein, formononetin, genistein, and glycitein; lignans: secoisolariciresinol and matairesinol; coumestrol; equol; enterolactone; and enterodiol) in 115 foods of animal origin (including milk and milk-products, eggs, meat, fish, and seafood) and vegetarian substitutes using liquid chromatography-mass spectrometry (LC-MS) with (13)C-labeled internal standards. Phytoestrogens were detected in all foods analyzed; the average content was 20 microg/100 g of wet weight (isoflavones, 6 microg/100 g; lignans, 6 microg/100 g; equol, 3 microg/100 g; and enterolignans, 6 microg/100 g). In infant soy formula, 19 221 microg/100 g phytoestrogens were detected (compared to 59 microg/100 g in non-soy formula). Our study shows that all foods analyzed contained phytoestrogens and most foods (except for fish, seafood, and butter) contained mammalian phytoestrogens (enterolignans and equol). This is the first comprehensive study of phytoestrogen content of foods of animal origin and will allow for a more accurate estimation of exposure to dietary phytoestrogens.

Phytoestrogen content of beverages, nuts, seeds, and oils.

Phytoestrogens are secondary plant metabolites that have received increasing attention for their bioactivity, in particular due to their structural and functional similarity to 17beta-estradiol. Although urinary and plasma phytoestrogens can be used as biomarkers for dietary intake, this is often not possible in large epidemiological studies or in the assessment of general exposure in free-living individuals. Accurate information about dietary phytoestrogens is therefore important, but there are very limited data concerning food contents. In this study was analyzed a comprehensive selection of tea, coffee, alcoholic beverages, nuts, seeds, and oils for their phytoestrogen content using a newly developed sensitive method based on LC-MS incorporating (13)C 3-labeled standards. Phytoestrogens were detected in all foods analyzed, although the contents in gin and bitter (beer) were below the limit of quantification (1.5 microg/100 g). Lignans were the main type of phytoestrogens detected. Tea and coffee contained up to 20 microg/100 g phytoestrogens and beer (except bitter) contained up to 71 microg/100 g, mainly lignans. As these beverages are commonly consumed, they are a main source of dietary lignans. The results published here will contribute to databases of dietary phytoestrogen content and allow a more accurate determination of phytoestrogen exposure in free-living individuals.

Extraction and quantification of phytoestrogens in foods using automated solid-phase extraction and LC/MS/MS.

Phytoestrogens are a group of polyphenolic plant metabolites that can induce biological responses. Their bioactivity is based on their similarity to 17beta-estradiol and their ability to bind to the beta-estrogen receptor. Although epidemiological data are inconclusive, phytoestrogens are considered to be beneficial for a variety of conditions, for example, hormone-related cancers like breast and prostate cancer. To investigate the biological effects of these compounds and to assess the exposure of larger cohorts or the general public, reliable data on the phytoestrogen content of food is necessary. Previously, food analysis for phytoestrogens was performed using either HPLC-UV or GC/MS. Here, we describe the development of the first generic method for the analysis of phytoestrogens in food, using automated solid-phase extraction and liquid chromatography-tandem mass spectrometry. The presented method shows a good reproducibility and can be easily adapted to other phytoestrogens if required.

Separation of tricyclic antidepressants by capillary zone electrophoresis with N,N,N',N'-tetramethyl-1,3-butanediamine (TMBD) as an effective electrolyte additive.

Five tricyclic antidepressants (TADs), desipramyne, nortriptyline, imipramine, doxepin and amitriptyline, were separated by using the N,N,N',N'-tetramethyl-1,3-butanediamine (TMBD) as additive in the background electrolyte solution. Because the tricyclic antidepressants are similar in structure, mass and pka values, their separation, by capillary zone electrophoresis, requires the careful manipulation of parameters, such as the pH and the composition of the electrolyte solution. As basic drugs, the TADs interact with the silanol groups on the capillary wall giving rise to peak broadening and asymmetry, non reproducible migration times and failing in selectivity. Different concentrations of TMBD (40, 60, 100 and 150 mM) were used at pH 9.5, but only a 100 mM TMBD allowed a good separation and a high efficiency for all the TADs. At this pH the separation was not possible without additive. This result is due to the reduced electroosmotic flow whose mobility is at a value of 10(-9) m(2) V(-1) s(-1).