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Application of Hydrophilic Interaction Liquid Chromatography for the Quantification of Flavonoids in Genista tinctoria Extract.

Sentkowska A, Biesaga M, Pyrzynska K - J Anal Methods Chem (2016)

Bottom Line: In the case of bare silica column more or less apparent dual retention mechanism was observed, depending on the water component content in the mobile phase.ZIC-HILIC showed better selectivity (in comparison to silica column) with the detection limit of 0.01 mg/L (only for rutin was 0.05 mg/L).Finally, this chromatographic procedure was validated and applied for the determination of some flavonoids in Genista tinctoria L. extract.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, University of Warsaw, Pasteura 1, 02-093 Warsaw, Poland.

ABSTRACT
Hydrophilic interaction chromatography (HILIC) was employed to investigate chromatographic behavior of selected flavonoids from their different subgroups differing in polarity. Chromatographic measurements were performed on two different HILIC columns: unmodified silica (Atlantis-HILIC) and zwitterionic sulfoalkylbetaine (SeQuant ZIC-HILIC). Separation parameters such as content and type of organic modifier were studied. On ZIC column retention factors were observed to be inversely proportional to the buffer content in the mobile phase, which is the typical partitioning mechanism. In the case of bare silica column more or less apparent dual retention mechanism was observed, depending on the water component content in the mobile phase. ZIC-HILIC showed better selectivity (in comparison to silica column) with the detection limit of 0.01 mg/L (only for rutin was 0.05 mg/L). Finally, this chromatographic procedure was validated and applied for the determination of some flavonoids in Genista tinctoria L. extract.

No MeSH data available.


Related in: MedlinePlus

Extracted ion chromatograms of luteolin on HILIC and RP columns under isocratic elution. Eluent for HILIC column-ACN/H2O (95/5%, v/v), for Luna-C18 ACN/water (45/55%, v/v), and for C18: ACN/water (20/80%, v/v).
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fig4: Extracted ion chromatograms of luteolin on HILIC and RP columns under isocratic elution. Eluent for HILIC column-ACN/H2O (95/5%, v/v), for Luna-C18 ACN/water (45/55%, v/v), and for C18: ACN/water (20/80%, v/v).

Mentions: The comparison of HILIC and RP mode was also carried out as reversed-phase mode of the most popular chromatographic mode for flavonoids analysis. Two RP columns were employed—typical fully porous Luna C18 (10 cm × 4.6 mm × 5 Am) and core shell Kinetex C18 (100 × 2.1 mm × 2.6 Am). The composition of the mobile phase in RP mode was ACN/H2O (20/80, v/v) to obtain similar retention factors. Figure 4 presents the MS extracted ion chromatograms of luteolin (SRM 285/133) as a model compound obtained on different HILIC and for comparison RP columns under isocratic elution. The significantly higher sensitivity was observed under HILIC conditions due to much higher content of ACN in the mobile phase (95%, v/v). There were no significant differences in the retention times of luteolin analyzed under HILIC and RP mode. Thus, acetonitrile-rich mobile phase (used in HILIC mode) not only affects the retention factors but also has an impact on the MS detection.


Application of Hydrophilic Interaction Liquid Chromatography for the Quantification of Flavonoids in Genista tinctoria Extract.

Sentkowska A, Biesaga M, Pyrzynska K - J Anal Methods Chem (2016)

Extracted ion chromatograms of luteolin on HILIC and RP columns under isocratic elution. Eluent for HILIC column-ACN/H2O (95/5%, v/v), for Luna-C18 ACN/water (45/55%, v/v), and for C18: ACN/water (20/80%, v/v).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4940558&req=5

fig4: Extracted ion chromatograms of luteolin on HILIC and RP columns under isocratic elution. Eluent for HILIC column-ACN/H2O (95/5%, v/v), for Luna-C18 ACN/water (45/55%, v/v), and for C18: ACN/water (20/80%, v/v).
Mentions: The comparison of HILIC and RP mode was also carried out as reversed-phase mode of the most popular chromatographic mode for flavonoids analysis. Two RP columns were employed—typical fully porous Luna C18 (10 cm × 4.6 mm × 5 Am) and core shell Kinetex C18 (100 × 2.1 mm × 2.6 Am). The composition of the mobile phase in RP mode was ACN/H2O (20/80, v/v) to obtain similar retention factors. Figure 4 presents the MS extracted ion chromatograms of luteolin (SRM 285/133) as a model compound obtained on different HILIC and for comparison RP columns under isocratic elution. The significantly higher sensitivity was observed under HILIC conditions due to much higher content of ACN in the mobile phase (95%, v/v). There were no significant differences in the retention times of luteolin analyzed under HILIC and RP mode. Thus, acetonitrile-rich mobile phase (used in HILIC mode) not only affects the retention factors but also has an impact on the MS detection.

Bottom Line: In the case of bare silica column more or less apparent dual retention mechanism was observed, depending on the water component content in the mobile phase.ZIC-HILIC showed better selectivity (in comparison to silica column) with the detection limit of 0.01 mg/L (only for rutin was 0.05 mg/L).Finally, this chromatographic procedure was validated and applied for the determination of some flavonoids in Genista tinctoria L. extract.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, University of Warsaw, Pasteura 1, 02-093 Warsaw, Poland.

ABSTRACT
Hydrophilic interaction chromatography (HILIC) was employed to investigate chromatographic behavior of selected flavonoids from their different subgroups differing in polarity. Chromatographic measurements were performed on two different HILIC columns: unmodified silica (Atlantis-HILIC) and zwitterionic sulfoalkylbetaine (SeQuant ZIC-HILIC). Separation parameters such as content and type of organic modifier were studied. On ZIC column retention factors were observed to be inversely proportional to the buffer content in the mobile phase, which is the typical partitioning mechanism. In the case of bare silica column more or less apparent dual retention mechanism was observed, depending on the water component content in the mobile phase. ZIC-HILIC showed better selectivity (in comparison to silica column) with the detection limit of 0.01 mg/L (only for rutin was 0.05 mg/L). Finally, this chromatographic procedure was validated and applied for the determination of some flavonoids in Genista tinctoria L. extract.

No MeSH data available.


Related in: MedlinePlus