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Detection of anthocyanins/anthocyanidins in animal tissues.

Aqil F, Vadhanam MV, Jeyabalan J, Cai J, Singh IP, Gupta RC - J. Agric. Food Chem. (2014)

Bottom Line: All five anthocyanidins present in the blueberry were detected in the lung tissue using improved methods.The effect of various solvents on the stability of anthocyanins and their recovery from the biomatrix was analyzed.Although anthocyanins were not detected, cyanidin was detected by UPLC-PDA and other anthocyanidins were detected by LC-MS, following conversion to anthocyanidins and selective extraction in isoamyl alcohol.

View Article: PubMed Central - PubMed

Affiliation: James Graham Brown Cancer Center, ‡Department of Medicine, and #Department of Pharmacology and Toxicology, University of Louisville , Louisville, Kentucky 40202, United States.

ABSTRACT
Dietary polyphenols may contribute to the prevention of several degenerative diseases, including cancer. Anthocyanins have been shown to possess potential anticancer activity. The aim of this study was to determine anthocyanin bioavailability in lung tissue of mice fed a blueberry diet (5% w/w) for 10 days or a bolus dose (10 mg/mouse; po) of a native mixture of bilberry anthocyanidins. All five anthocyanidins present in the blueberry were detected in the lung tissue using improved methods. The effect of various solvents on the stability of anthocyanins and their recovery from the biomatrix was analyzed. Detection of anthocyanins and their metabolites was performed by UPLC and LC-MS. Although anthocyanins were not detected, cyanidin was detected by UPLC-PDA and other anthocyanidins were detected by LC-MS, following conversion to anthocyanidins and selective extraction in isoamyl alcohol. The results show that anthocyanins can be detected in lung tissue of blueberry-fed mice and thus are bioavailable beyond the gastrointestinal tract.

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Related in: MedlinePlus

Multiple reaction monitoring (A) and therespective MS (B) andMS/MS (C) of indicated reference anthocyanidins. Peonidin and malvidinwere eluted at the same retention time, and therefore they appearin the same spectrum. Anthocyanidins were separated with a HypersilGOLD C18 column (50 mm × 2.1 mm) in a binary gradient of 1% formicacid and acetonitrile containing 1% formic acid as described under Materials and Methods.
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fig4: Multiple reaction monitoring (A) and therespective MS (B) andMS/MS (C) of indicated reference anthocyanidins. Peonidin and malvidinwere eluted at the same retention time, and therefore they appearin the same spectrum. Anthocyanidins were separated with a HypersilGOLD C18 column (50 mm × 2.1 mm) in a binary gradient of 1% formicacid and acetonitrile containing 1% formic acid as described under Materials and Methods.

Mentions: Reversed-phase chromatographyof anthocyanidinswas performed on a Thermo Scientific (San Jose, CA, USA) Accela LCsystem. The mobile phases consisted of buffer A, water/formic acid(100:0.1, v/v), and buffer B, acetonitrile/formic acid (100:0.1, v/v).Five microliters of sample was injected onto a Hypersil GOLD C18 column (50 × 2.1 mm, 1.9 μm, 175 Å) fromThermo Scientific. A step gradient at a flow rate of 100 μL/minwas used to elute the compounds. The gradient started at 5% bufferB and increased to 40% buffer B in 10 min, then increased to 90% bufferB in 3 min, and was maintained at 90% buffer B for 7 min. Elute fromthe LC was directed to an LTQ-Orbitrap XL mass spectrometer (ThermoScientific). The compounds were ionized by electrospray ionizationand detected by Orbitrap at 30000 mass resolution (full scan, m/z 220–1000) or by multiple reactionmonitoring (MRM). The spray voltage was 4.0 kV, and the capillarytemperature was 250 °C. The sheath, auxiliary, and sweep gasflows were set to 15, 5, and 0, respectively. In MRM, molecular ionsof anthocyanidins were selected with 3.0 m/z isolation window and fragmented by collision-induced dissociation(CID). For CID, collision energy, activation Q, andactivation time (mS) were 35, 0.25, and 30, respectively. Full scanMS/MS spectra of the compounds (m/z 100–400) were acquired by Orbitrap at 7500 mass resolution.The transitions used for anthocyanidin detection are shown in panelC of Figure 4. Thelimits of detection by LC-MS for Dp and Cy were >0.5 and 0.25 ng,respectively, whereas those for Pt, Pe, and Mv were 2.5 pg.


Detection of anthocyanins/anthocyanidins in animal tissues.

Aqil F, Vadhanam MV, Jeyabalan J, Cai J, Singh IP, Gupta RC - J. Agric. Food Chem. (2014)

Multiple reaction monitoring (A) and therespective MS (B) andMS/MS (C) of indicated reference anthocyanidins. Peonidin and malvidinwere eluted at the same retention time, and therefore they appearin the same spectrum. Anthocyanidins were separated with a HypersilGOLD C18 column (50 mm × 2.1 mm) in a binary gradient of 1% formicacid and acetonitrile containing 1% formic acid as described under Materials and Methods.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4334289&req=5

fig4: Multiple reaction monitoring (A) and therespective MS (B) andMS/MS (C) of indicated reference anthocyanidins. Peonidin and malvidinwere eluted at the same retention time, and therefore they appearin the same spectrum. Anthocyanidins were separated with a HypersilGOLD C18 column (50 mm × 2.1 mm) in a binary gradient of 1% formicacid and acetonitrile containing 1% formic acid as described under Materials and Methods.
Mentions: Reversed-phase chromatographyof anthocyanidinswas performed on a Thermo Scientific (San Jose, CA, USA) Accela LCsystem. The mobile phases consisted of buffer A, water/formic acid(100:0.1, v/v), and buffer B, acetonitrile/formic acid (100:0.1, v/v).Five microliters of sample was injected onto a Hypersil GOLD C18 column (50 × 2.1 mm, 1.9 μm, 175 Å) fromThermo Scientific. A step gradient at a flow rate of 100 μL/minwas used to elute the compounds. The gradient started at 5% bufferB and increased to 40% buffer B in 10 min, then increased to 90% bufferB in 3 min, and was maintained at 90% buffer B for 7 min. Elute fromthe LC was directed to an LTQ-Orbitrap XL mass spectrometer (ThermoScientific). The compounds were ionized by electrospray ionizationand detected by Orbitrap at 30000 mass resolution (full scan, m/z 220–1000) or by multiple reactionmonitoring (MRM). The spray voltage was 4.0 kV, and the capillarytemperature was 250 °C. The sheath, auxiliary, and sweep gasflows were set to 15, 5, and 0, respectively. In MRM, molecular ionsof anthocyanidins were selected with 3.0 m/z isolation window and fragmented by collision-induced dissociation(CID). For CID, collision energy, activation Q, andactivation time (mS) were 35, 0.25, and 30, respectively. Full scanMS/MS spectra of the compounds (m/z 100–400) were acquired by Orbitrap at 7500 mass resolution.The transitions used for anthocyanidin detection are shown in panelC of Figure 4. Thelimits of detection by LC-MS for Dp and Cy were >0.5 and 0.25 ng,respectively, whereas those for Pt, Pe, and Mv were 2.5 pg.

Bottom Line: All five anthocyanidins present in the blueberry were detected in the lung tissue using improved methods.The effect of various solvents on the stability of anthocyanins and their recovery from the biomatrix was analyzed.Although anthocyanins were not detected, cyanidin was detected by UPLC-PDA and other anthocyanidins were detected by LC-MS, following conversion to anthocyanidins and selective extraction in isoamyl alcohol.

View Article: PubMed Central - PubMed

Affiliation: James Graham Brown Cancer Center, ‡Department of Medicine, and #Department of Pharmacology and Toxicology, University of Louisville , Louisville, Kentucky 40202, United States.

ABSTRACT
Dietary polyphenols may contribute to the prevention of several degenerative diseases, including cancer. Anthocyanins have been shown to possess potential anticancer activity. The aim of this study was to determine anthocyanin bioavailability in lung tissue of mice fed a blueberry diet (5% w/w) for 10 days or a bolus dose (10 mg/mouse; po) of a native mixture of bilberry anthocyanidins. All five anthocyanidins present in the blueberry were detected in the lung tissue using improved methods. The effect of various solvents on the stability of anthocyanins and their recovery from the biomatrix was analyzed. Detection of anthocyanins and their metabolites was performed by UPLC and LC-MS. Although anthocyanins were not detected, cyanidin was detected by UPLC-PDA and other anthocyanidins were detected by LC-MS, following conversion to anthocyanidins and selective extraction in isoamyl alcohol. The results show that anthocyanins can be detected in lung tissue of blueberry-fed mice and thus are bioavailable beyond the gastrointestinal tract.

Show MeSH
Related in: MedlinePlus