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Analysis of isoflavones and flavonoids in human urine by UHPLC.

Baranowska I, Magiera S - Anal Bioanal Chem (2010)

Bottom Line: Recovery was 70.35-96.58%.It can be routinely used for simultaneous determination of these five isoflavones and seven flavonoids in human urine.The method can also be applied to studies after administration of pharmaceutical preparations containing isoflavones and flavonoids to humans.

View Article: PubMed Central - PubMed

Affiliation: Department of Analytical Chemistry, Silesian University of Technology, 7 M. Strzody Str., 44-100 Gliwice, Poland. irena.baranowska@polsl.pl

ABSTRACT
A rapid, ultra high-performance liquid chromatographic (UHPLC) method has been developed and validated for simultaneous identification and analysis of the isoflavones genistein, daidzein, glycitin, puerarin, and biochanin A, and the flavonoids (±)-catechin, (-)-epicatechin, rutin, hesperidin, neohesperidin, quercitrin, and hesperetin in human urine. Urine samples were incubated with β-glucuronidase/sulfatase. UHPLC was performed with a Hypersil Gold (50 × 2.1 mm, 1.9 μm) analytical column. Elution was with a gradient prepared from aqueous trifluoroacetic acid (0.05%) and acetonitrile. UV detection was performed at 254 and 280 nm. The calibration curves were indicative of good linearity (r(2) ≥ 0.9992) in the range of interest for each analyte. LODs ranged between 15.4 and 107.0 ng mL(-1) and 3.9 and 20.4 ng mL(-1) for flavonoids and isoflavones, respectively. Intra-day and inter-day precision (C.V., %) was less than 3.9% and 3.8%, respectively, and accuracy was between 0.03% and 5.0%. Recovery was 70.35-96.58%. The method is very rapid, simple, and reliable, and suitable for pharmacokinetic analysis. It can be routinely used for simultaneous determination of these five isoflavones and seven flavonoids in human urine. The method can also be applied to studies after administration of pharmaceutical preparations containing isoflavones and flavonoids to humans.

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Mean urine concentration–time profiles (±S.D) of (a) GLY, (b) DA, and (c) RUT in healthy volunteers after oral administration of pharmaceutical preparations
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Fig4: Mean urine concentration–time profiles (±S.D) of (a) GLY, (b) DA, and (c) RUT in healthy volunteers after oral administration of pharmaceutical preparations

Mentions: Analysis of the 24-h urinary excretion of GLY, DA, and GT demonstrated that the cumulative amounts gradually increased during the collection period after oral administration of these isoflavones to the subject. The mean urine concentration–time profiles of GLY, DA, and RUT are shown in Fig. 4. The flavonoid and isoflavone concentrations reached a maximum between 5 and 8 h after dosing with a level of 5482.33 ± 10.78 ng mL−1 for GLY, 228.03 ± 5.34 ng mL−1 for DA, and 780.18 ± 10.39 ng mL−1 for RUT. Assuming the mean 24-h urine volume is 1.4 L [17], the approximate average daily output is 1.73 mg, 0.14 mg, and 0.19 mg of GLY, DA and RUT, respectively. The percentage of GLY, DA, and RUT excreted in urine within 24 h after ingestion of the tablets was 30%, 17%, and 1%, respectively, of the administered dose. The corresponding pharmacokinetic data for GLY, DA, and RUT are presented in Table 3.Fig. 4


Analysis of isoflavones and flavonoids in human urine by UHPLC.

Baranowska I, Magiera S - Anal Bioanal Chem (2010)

Mean urine concentration–time profiles (±S.D) of (a) GLY, (b) DA, and (c) RUT in healthy volunteers after oral administration of pharmaceutical preparations
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3044235&req=5

Fig4: Mean urine concentration–time profiles (±S.D) of (a) GLY, (b) DA, and (c) RUT in healthy volunteers after oral administration of pharmaceutical preparations
Mentions: Analysis of the 24-h urinary excretion of GLY, DA, and GT demonstrated that the cumulative amounts gradually increased during the collection period after oral administration of these isoflavones to the subject. The mean urine concentration–time profiles of GLY, DA, and RUT are shown in Fig. 4. The flavonoid and isoflavone concentrations reached a maximum between 5 and 8 h after dosing with a level of 5482.33 ± 10.78 ng mL−1 for GLY, 228.03 ± 5.34 ng mL−1 for DA, and 780.18 ± 10.39 ng mL−1 for RUT. Assuming the mean 24-h urine volume is 1.4 L [17], the approximate average daily output is 1.73 mg, 0.14 mg, and 0.19 mg of GLY, DA and RUT, respectively. The percentage of GLY, DA, and RUT excreted in urine within 24 h after ingestion of the tablets was 30%, 17%, and 1%, respectively, of the administered dose. The corresponding pharmacokinetic data for GLY, DA, and RUT are presented in Table 3.Fig. 4

Bottom Line: Recovery was 70.35-96.58%.It can be routinely used for simultaneous determination of these five isoflavones and seven flavonoids in human urine.The method can also be applied to studies after administration of pharmaceutical preparations containing isoflavones and flavonoids to humans.

View Article: PubMed Central - PubMed

Affiliation: Department of Analytical Chemistry, Silesian University of Technology, 7 M. Strzody Str., 44-100 Gliwice, Poland. irena.baranowska@polsl.pl

ABSTRACT
A rapid, ultra high-performance liquid chromatographic (UHPLC) method has been developed and validated for simultaneous identification and analysis of the isoflavones genistein, daidzein, glycitin, puerarin, and biochanin A, and the flavonoids (±)-catechin, (-)-epicatechin, rutin, hesperidin, neohesperidin, quercitrin, and hesperetin in human urine. Urine samples were incubated with β-glucuronidase/sulfatase. UHPLC was performed with a Hypersil Gold (50 × 2.1 mm, 1.9 μm) analytical column. Elution was with a gradient prepared from aqueous trifluoroacetic acid (0.05%) and acetonitrile. UV detection was performed at 254 and 280 nm. The calibration curves were indicative of good linearity (r(2) ≥ 0.9992) in the range of interest for each analyte. LODs ranged between 15.4 and 107.0 ng mL(-1) and 3.9 and 20.4 ng mL(-1) for flavonoids and isoflavones, respectively. Intra-day and inter-day precision (C.V., %) was less than 3.9% and 3.8%, respectively, and accuracy was between 0.03% and 5.0%. Recovery was 70.35-96.58%. The method is very rapid, simple, and reliable, and suitable for pharmacokinetic analysis. It can be routinely used for simultaneous determination of these five isoflavones and seven flavonoids in human urine. The method can also be applied to studies after administration of pharmaceutical preparations containing isoflavones and flavonoids to humans.

Show MeSH