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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

The relationship between log⁡k of flavonoids and log⁡P. Mobile phase: 95% ACN/ammonium formate or 95% MeOH/ammonium formate. (a) Bare silica column; (b) ZIC column.
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fig3: The relationship between log⁡k of flavonoids and log⁡P. Mobile phase: 95% ACN/ammonium formate or 95% MeOH/ammonium formate. (a) Bare silica column; (b) ZIC column.

Mentions: Since hydrophilic interactions are one of the main mechanisms which governs the retention in the HILIC mode, the hydrophilic properties of a compound should determine its behavior. We examined the relationships between log⁡k and calculated partition coefficient P (Table 1), a parameter that relates single solute partitions between polar and nonpolar phase. Formally, the application of log⁡D, as opposed to the traditional approach of log⁡P, should be taken into consideration for ionized species. However, its value depends on compound pKa as well as pH under investigation. It is worth noticing that the pKa for some of the studied flavonols are known to be difficult to predict and their reported values differ from each other about 1-2 pKa units [28–30]. The retention data were acquired with a mobile phase composed of 95% acetonitrile or methanol and the relations are presented in Figure 3. log⁡k is inversely correlated to log⁡P, indicating that the stronger retained analytes are those with lower log⁡P values, namely, the more hydrophilic analytes.


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)

The relationship between log⁡k of flavonoids and log⁡P. Mobile phase: 95% ACN/ammonium formate or 95% MeOH/ammonium formate. (a) Bare silica column; (b) ZIC column.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig3: The relationship between log⁡k of flavonoids and log⁡P. Mobile phase: 95% ACN/ammonium formate or 95% MeOH/ammonium formate. (a) Bare silica column; (b) ZIC column.
Mentions: Since hydrophilic interactions are one of the main mechanisms which governs the retention in the HILIC mode, the hydrophilic properties of a compound should determine its behavior. We examined the relationships between log⁡k and calculated partition coefficient P (Table 1), a parameter that relates single solute partitions between polar and nonpolar phase. Formally, the application of log⁡D, as opposed to the traditional approach of log⁡P, should be taken into consideration for ionized species. However, its value depends on compound pKa as well as pH under investigation. It is worth noticing that the pKa for some of the studied flavonols are known to be difficult to predict and their reported values differ from each other about 1-2 pKa units [28–30]. The retention data were acquired with a mobile phase composed of 95% acetonitrile or methanol and the relations are presented in Figure 3. log⁡k is inversely correlated to log⁡P, indicating that the stronger retained analytes are those with lower log⁡P values, namely, the more hydrophilic analytes.

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