A review covering different methods for the analysis of phyto-oestrogens in biological matrices i... more A review covering different methods for the analysis of phyto-oestrogens in biological matrices is presented. Sample pretreatment and analysis of isoflavonoids and lignans by HPLC and GC with various detection methods are discussed. The immunoassay method is also briefly presented.
Soy-derived isoflavone phytoestrogens, such as genistein (4′,5,7-trihydroxyisoflavone), have been... more Soy-derived isoflavone phytoestrogens, such as genistein (4′,5,7-trihydroxyisoflavone), have been shown to protect low-density lipoprotein from oxidation. In addition, human plasma was previously shown to be capable of converting genistein into lipophilic fatty acid esters that accumulate in lipoproteins in vitro. We developed a method for the quantitation of genistein fatty acid esters in plasma. Furthermore, the method was utilized to measure genistein ester concentrations in monkey plasma following administration of genistein or genistein 4′,7-O-dioleate. After extraction from plasma, genistein fatty acid esters were separated from unesterified genistein by Sephadex LH-20 column chromatography. The genistein ester fraction was hydrolyzed by saponification and purified by a second chromatography on Sephadex LH-20. The hydrolyzed genistein esters were measured by time-resolved fluoroimmunoassay. Adult female rhesus monkeys (n = 10) received a subcutaneous injection of genistein (24 mg, n = 2) or genistein 4′,7-O-dioleate (71 mg, n = 3) or an oral dose of genistein (24 mg, n = 2) or genistein 4′,7-O-dioleate (71 mg, n = 3). Plasma was collected at 4, 8, and 24 h post-dosing. Following subcutaneous administration of genistein 4′,7-O-dioleate, the plasma concentrations of genistein esters became elevated in two out of three monkeys with 8-h values exceeding 7.5 nmol/L and 24-h values above 12 nmol/L. Other treatments resulted in lower plasma values ranging between 2.7 and 6.1 nmol/L. The lower limit of detection for the method was 1.44 nmol/L. Subcutaneously administered genistein 4′,7-O-dioleate was also converted to water-soluble conjugates, but oral administration did not elevate plasma genistein fatty acid ester levels. The results suggest that it may be possible to introduce intact genistein ester molecules into plasma by parenteral but not oral administration.
LDL antioxidant mono- and dioleates and -stearates of the isoflavones genistein and daidzein are ... more LDL antioxidant mono- and dioleates and -stearates of the isoflavones genistein and daidzein are synthesised in high yield with excellent regioselectivity.
Journal of Steroid Biochemistry and Molecular Biology, 2003
Epidemiological studies have associated high soy intake with a lowered risk for certain hormone-d... more Epidemiological studies have associated high soy intake with a lowered risk for certain hormone-dependent diseases. Soy and soy foods are rich sources of isoflavones, which have been shown to possess several biological activities. In this study, the metabolism of soy isoflavones daidzein, genistein and glycitein was investigated in human subjects. The aim was to find and identify urinary phase I metabolites of isoflavones, which have an intact isoflavonoid skeleton, and which might possess some bioactivity. Six volunteers included three soy bars per day into their normal western diet for a 2-week period. Daily urine samples were collected before, and after the supplementation period. Urine samples were hydrolyzed with Helix pomatia, extracted with diethyl ether, purified with Sephadex LH-20 chromatography, and analyzed as trimethylsilyl derivatives using gas chromatography–mass spectrometry (GC–MS). The structures of the isoflavone metabolites were identified using authentic reference compounds. The metabolites, for which authentic reference compounds were not available, were identified by the interpretation of mass spectra. Several new isoflavone metabolites were identified, and the presence of previously reported metabolites confirmed. The metabolic pathways of daidzein, genistein and glycitein are presented on the basis of the identification of the metabolites in human urine after soy supplementation.
The redox potential of the cell, as well as the antioxidant status of the tissue, are considered ... more The redox potential of the cell, as well as the antioxidant status of the tissue, are considered to be important regulatory constituents in an adaptive response in plants. Here the involvement of active antioxidants ascorbic acid (AA), reduced glutathione (GSH) and α- and β-tocopherols in reactive oxygen species scavenging, and the effect of anoxic stress on their reduction state were studied in 4 anoxia-tolerant and -intolerant plant species: Iris germanica L., Iris pseudacorus L., wheat (Triticum aestivum L. cv. Leningradka) and rice (Oryza sativa L. cv. VNIIR). The initial antioxidant content (both AA and GSH) was higher in the rhizomes of the more anoxia-tolerant Iris spp., as compared with that of the roots of the cereals. The predominant form of ascorbate was dehydroascorbic acid (DHA) in the cereals and AA in the Iris spp. Imposition of anoxia with subsequent reoxygenation resulted in an overall depletion of the reduced forms of antioxidants. No concurrent increase in oxidised forms (DHA and conjugated glutathione) was observed in anoxic samples. α-tocopherol content in Iris spp. was in the range 1–2 μg g−1 fresh weight, while β-tocopherol content was higher in the anoxia-intolerant I. germanica (7.2 μg g−1 fresh weight) as compared with the tolerant I. pseudacorus (1.5 μg g−1 fresh weight). In I. pseudacorus, a significant decrease in α- and β-tocopherol levels was observed only after long-term (45 days) anoxia. The results suggested exclusion of AA and GSH from the redox cycling under prolonged anoxia, and a concomitant decrease in the redox state, as well as an anoxia-induced depletion of α- and β-tocopherols.
The isoflavonoids, equol, formononetin, daidzein, genistein, biochanin A, and O-demethylangolensi... more The isoflavonoids, equol, formononetin, daidzein, genistein, biochanin A, and O-demethylangolensin (O-DMA), were analyzed from commercial cartons of skimmed Finnish milk by HPLC-diode array detector (DAD)-FL. We found 411 ± 65 ng/mL of equol and traces of formononetin and daidzein in organic skimmed milk whereas conventionally produced milk contained 62 ± 16 ng/mL of equol and no formononetin or daidzein.
A review covering different methods for the analysis of phyto-oestrogens in biological matrices i... more A review covering different methods for the analysis of phyto-oestrogens in biological matrices is presented. Sample pretreatment and analysis of isoflavonoids and lignans by HPLC and GC with various detection methods are discussed. The immunoassay method is also briefly presented.
Soy-derived isoflavone phytoestrogens, such as genistein (4′,5,7-trihydroxyisoflavone), have been... more Soy-derived isoflavone phytoestrogens, such as genistein (4′,5,7-trihydroxyisoflavone), have been shown to protect low-density lipoprotein from oxidation. In addition, human plasma was previously shown to be capable of converting genistein into lipophilic fatty acid esters that accumulate in lipoproteins in vitro. We developed a method for the quantitation of genistein fatty acid esters in plasma. Furthermore, the method was utilized to measure genistein ester concentrations in monkey plasma following administration of genistein or genistein 4′,7-O-dioleate. After extraction from plasma, genistein fatty acid esters were separated from unesterified genistein by Sephadex LH-20 column chromatography. The genistein ester fraction was hydrolyzed by saponification and purified by a second chromatography on Sephadex LH-20. The hydrolyzed genistein esters were measured by time-resolved fluoroimmunoassay. Adult female rhesus monkeys (n = 10) received a subcutaneous injection of genistein (24 mg, n = 2) or genistein 4′,7-O-dioleate (71 mg, n = 3) or an oral dose of genistein (24 mg, n = 2) or genistein 4′,7-O-dioleate (71 mg, n = 3). Plasma was collected at 4, 8, and 24 h post-dosing. Following subcutaneous administration of genistein 4′,7-O-dioleate, the plasma concentrations of genistein esters became elevated in two out of three monkeys with 8-h values exceeding 7.5 nmol/L and 24-h values above 12 nmol/L. Other treatments resulted in lower plasma values ranging between 2.7 and 6.1 nmol/L. The lower limit of detection for the method was 1.44 nmol/L. Subcutaneously administered genistein 4′,7-O-dioleate was also converted to water-soluble conjugates, but oral administration did not elevate plasma genistein fatty acid ester levels. The results suggest that it may be possible to introduce intact genistein ester molecules into plasma by parenteral but not oral administration.
LDL antioxidant mono- and dioleates and -stearates of the isoflavones genistein and daidzein are ... more LDL antioxidant mono- and dioleates and -stearates of the isoflavones genistein and daidzein are synthesised in high yield with excellent regioselectivity.
Journal of Steroid Biochemistry and Molecular Biology, 2003
Epidemiological studies have associated high soy intake with a lowered risk for certain hormone-d... more Epidemiological studies have associated high soy intake with a lowered risk for certain hormone-dependent diseases. Soy and soy foods are rich sources of isoflavones, which have been shown to possess several biological activities. In this study, the metabolism of soy isoflavones daidzein, genistein and glycitein was investigated in human subjects. The aim was to find and identify urinary phase I metabolites of isoflavones, which have an intact isoflavonoid skeleton, and which might possess some bioactivity. Six volunteers included three soy bars per day into their normal western diet for a 2-week period. Daily urine samples were collected before, and after the supplementation period. Urine samples were hydrolyzed with Helix pomatia, extracted with diethyl ether, purified with Sephadex LH-20 chromatography, and analyzed as trimethylsilyl derivatives using gas chromatography–mass spectrometry (GC–MS). The structures of the isoflavone metabolites were identified using authentic reference compounds. The metabolites, for which authentic reference compounds were not available, were identified by the interpretation of mass spectra. Several new isoflavone metabolites were identified, and the presence of previously reported metabolites confirmed. The metabolic pathways of daidzein, genistein and glycitein are presented on the basis of the identification of the metabolites in human urine after soy supplementation.
The redox potential of the cell, as well as the antioxidant status of the tissue, are considered ... more The redox potential of the cell, as well as the antioxidant status of the tissue, are considered to be important regulatory constituents in an adaptive response in plants. Here the involvement of active antioxidants ascorbic acid (AA), reduced glutathione (GSH) and α- and β-tocopherols in reactive oxygen species scavenging, and the effect of anoxic stress on their reduction state were studied in 4 anoxia-tolerant and -intolerant plant species: Iris germanica L., Iris pseudacorus L., wheat (Triticum aestivum L. cv. Leningradka) and rice (Oryza sativa L. cv. VNIIR). The initial antioxidant content (both AA and GSH) was higher in the rhizomes of the more anoxia-tolerant Iris spp., as compared with that of the roots of the cereals. The predominant form of ascorbate was dehydroascorbic acid (DHA) in the cereals and AA in the Iris spp. Imposition of anoxia with subsequent reoxygenation resulted in an overall depletion of the reduced forms of antioxidants. No concurrent increase in oxidised forms (DHA and conjugated glutathione) was observed in anoxic samples. α-tocopherol content in Iris spp. was in the range 1–2 μg g−1 fresh weight, while β-tocopherol content was higher in the anoxia-intolerant I. germanica (7.2 μg g−1 fresh weight) as compared with the tolerant I. pseudacorus (1.5 μg g−1 fresh weight). In I. pseudacorus, a significant decrease in α- and β-tocopherol levels was observed only after long-term (45 days) anoxia. The results suggested exclusion of AA and GSH from the redox cycling under prolonged anoxia, and a concomitant decrease in the redox state, as well as an anoxia-induced depletion of α- and β-tocopherols.
The isoflavonoids, equol, formononetin, daidzein, genistein, biochanin A, and O-demethylangolensi... more The isoflavonoids, equol, formononetin, daidzein, genistein, biochanin A, and O-demethylangolensin (O-DMA), were analyzed from commercial cartons of skimmed Finnish milk by HPLC-diode array detector (DAD)-FL. We found 411 ± 65 ng/mL of equol and traces of formononetin and daidzein in organic skimmed milk whereas conventionally produced milk contained 62 ± 16 ng/mL of equol and no formononetin or daidzein.
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Papers by Antti Hoikkala