HPLC method with fluorescence detection for the quantitative determination of flaxseed lignans

Mukker, Jatinder Kaur; Kotlyarova, Valeriya; Singh, Ravi Shankar Prasad; Alcorn, Jane

doi:10.1016/j.jchromb.2010.09.012

Cited by 20 publications

(16 citation statements)

References 35 publications

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“…Serum samples were transferred to microcentrifuge tubes and stored at − 80°C until analysis. Lignans were quantified using a HPLC fluorescence method ( 27 ) . The bioanalytical assay was tested for within- and between-assay accuracy and precision, and quality-control (QC) samples were used as acceptance criteria for running an individual analysis.…”

Section: Methodsmentioning

confidence: 99%

“…After equilibration, the serum samples were transferred onto micropartition kit membranes and centrifuged using a fixed angle (25°) rotor (TA-14-50) in a Beckman Coulter Allegra 25R centrifuge (Beckman Coulter) for 8 min at 1000 g . About 70 μl of the filtrate were collected and analysed using the HPLC fluorescence method described below ( 27 ) .…”

Section: Methodsmentioning

confidence: 99%

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Comparative pharmacokinetics of purified flaxseed and associated mammalian lignans in male Wistar rats

et al. 2015

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Consumption of flaxseed lignans is associated with various health benefits; however, little is known about the bioavailability of purified lignans in flaxseed. Data on their bioavailability and hence pharmacokinetics (PK) are necessary to better understand their role in putative health benefits. In the present study, we conducted a comparative PK analysis of the principal lignan of flaxseed, secoisolariciresinol diglucoside (SDG), and its primary metabolites, secoisolariciresinol (SECO), enterodiol (ED) and enterolactone (EL) in rats. Purified lignans were intravenously or orally administered to each male Wistar rat. SDG and its primary metabolites SECO, ED and EL were administered orally at doses of 40, 40, 10 and 10 mg/kg, respectively, and intravenously at doses of 20, 20, 5 and 1 mg/kg, respectively. Blood samples were collected at 0 (pre-dose), 5, 10, 15, 20, 30 and 45 min, and at 1, 2, 4, 6, 8, 12 and 24 h post-dosing, and serum samples were analysed. PK parameters and oral bioavailability of purified lignans were determined by non-compartmental methods. In general, administration of the flaxseed lignans SDG, SECO and ED demonstrated a high systemic clearance, a large volume of distribution and short half-lives, whereas administration of EL at the doses of 1 mg/kg (intravenously) and 10 mg/kg (orally administered) killed the rats within a few hours of dosing, precluding a PK analysis of this lignan. PK parameters of flaxseed lignans exhibited the following order: systemic clearance, SDG , SECO , ED; volume of distribution, SDG , SECO , ED; half-life, SDG , ED , SECO. The percentage of oral bioavailability was 0, 25 and , 1 % for SDG, SECO and ED, respectively.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Comparative pharmacokinetics of purified flaxseed and associated mammalian lignans in male Wistar rats

et al. 2015

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“…Meanwhile, it is reported that the structural analog of anwuligan, was given to rat i.v. at a dose of 20 mg/kg for pharmacokinetic study [27,28], was given to rat i.g. at a dose of 40 mg/kg for pharmacokinetic study [28].…”

Section: Dose Selectionmentioning

confidence: 99%

Pharmacokinetics and Tissue Distribution of Anwuligan in Rats after Intravenous and Intragastric Administration by Liquid Chromatography-Mass Spectrometry

et al. 2019

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Anwuligan, a natural 2,3-dibenzylbutane lignan from the nutmeg mace of Myristica fragans, has been proved to possess a broad range of pharmacological effects. A rapid, simple, and sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method has been established and successfully applied to the study of pharmacokinetics and tissue distribution of anwuligan after intravenous or intragastric administration. Sample preparation was carried out through a liquid-liquid extraction method with ethyl acetate as the extraction reagent. Arctigenin was used as the internal standard (IS). A gradient program was employed with a mobile phase consisting of 0.1% formic acid aqueous solution and acetonitrile. The mass spectrometer was operated in a positive ionization mode with multiple reaction monitoring. The transitions for quantification were m/z 329.0→205.0 for anwuligan and m/z 373.0→137.0 for IS, respectively. Calibration curves were linear over the ranges of 0.5-2000 ng/mL for both plasma samples and tissue samples (r > 0.996). The absolute bioavailability is 16.2%, which represented the existing of the obvious first-pass effect. An enterohepatic circulation was found after the intragastric administration. Anwuligan could be distributed rapidly and widely in different tissues and maintained a high concentration in the liver. The developed and validated LC-MS/MS method and the pharmacokinetic study of anwuligan would provide reference for the future investigation of the preclinical safety of anwuligan as a candidate drug.

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“…A reliable and simple HPLC method for the quantitative determination of flaxseed lignans was established in our lab 102 . Hence, a partial HPLC method validation was conducted to allow for the quantitative determination of EL in this study.…”

Section: Experiments Two: Hplc Methodsmentioning

confidence: 99%

The Comparison of Rat and Human Intestinal and Hepatic Glucuronidation of Enterolactone Derived from Flaxseed Lignans

Lin¹,

Krol²,

Alcorn

2013

NPJ

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The mammalian lignan, enterolactone (EL), is a gut microbe metabolite of plant lignan secoisolariciresinol diglucoside (SDG), which is most abundant in flaxseed. Numerous epidemiological, experimental and clinical studies suggest the protective effects of EL against various chronic diseases such as cancer, cardiovascular disease, and inflammation. However, EL's oral bioavailability is low and highly variable due to extensive first-pass metabolism, especially glucuronidation, which results in the large amount of glucuronide metabolites but low levels of free EL in human plasma. Hepatocytes and enterocytes express UDP-glucuronosyltransferases (UGT), the enzymes responsible for the conjugation of glucuronic acid to EL. To better understand the contribution of liver and intestine to the first-pass glucuronidation, I conducted an in vitro enzyme kinetic analysis of EL glucuronidation using hepatic and intestinal microsomal fractions from both human and rat.An intrinsic clearance (CL int ) value was calculated using the substrate depletion approach. In addition to monitoring substrate depletion, high-pressure liquid chromatography (HPLC) analysis allowed detection of EL glucuronides, which were further substantiated by LC-MS.EL monoglucuronide was identified in rat and human intestinal and liver microsomes.Enzyme kinetic studies indicated the extent of hepatic microsomal glucuronidation exceeded intestinal glucuronidation in both human and rat, while the human liver CL int value was slightly higher than that of rat liver. The CL int value generated in human intestinal microsomes was only one third of the value of human liver, whereas, the CL int of rat jejunum or colon was one-twentieth of rat liver, suggesting the human intestine makes a greater contribution to EL glucuronidation than rat intestine. These results suggest that both liver and intestine contribute to EL glucuronidation and the human intestine may exert a greater influence on the first-pass glucuronidation of EL than rat intestine, thereby significantly decreasing EL's oral bioavailability. The rat might underestimate the extent of intestinal metabolism of EL relative to human.iii

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HPLC method with fluorescence detection for the quantitative determination of flaxseed lignans

Cited by 20 publications

References 35 publications

Comparative pharmacokinetics of purified flaxseed and associated mammalian lignans in male Wistar rats

Comparative pharmacokinetics of purified flaxseed and associated mammalian lignans in male Wistar rats

Pharmacokinetics and Tissue Distribution of Anwuligan in Rats after Intravenous and Intragastric Administration by Liquid Chromatography-Mass Spectrometry

The Comparison of Rat and Human Intestinal and Hepatic Glucuronidation of Enterolactone Derived from Flaxseed Lignans

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