HPLC Analysis of Phenolic Compounds

The phenolic compound were analyzed using liquid chromatography systems (RP‐HPLC; Waters), equipped with a photodiode detector (M‐2998) and C18 column (4.6 mm 5 µ × 25 cm). To separate phenolic compounds, a mobile phase consisting 0.01% (v/v) acetic acid, acetonitrile (v/v) (solvent 1), and acidified water (0.1% (v/v) acetic acid) (solvent 2) was used. The constant flow rate was 0.5 ml/min. Methanol 0.1% (v/v) (solvent 2) was used to separate flavonoid compounds. The gradient and elution programs were described by Mekky et al. (2019 (link)).

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Esmaeilzadeh Kenari R, & Razavi R. (2022). Phenolic profile and antioxidant activity of free/bound phenolic compounds of sesame and properties of encapsulated nanoparticles in different wall materials. Food Science & Nutrition, 10(2), 525-535.

Publication 2022

RP‐HPLC M‐2998

Acetic acid Acetonitrile Flavonoid Hplc Liquid chromatography Methanol Solvent

Corresponding Organization : Sari Agricultural Sciences and Natural Resources University

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Variable analysis

independent variables

Mobile phase composition (0.01% (v/v) acetic acid, acetonitrile (v/v) (solvent 1), and acidified water (0.1% (v/v) acetic acid) (solvent 2))
Methanol 0.1% (v/v) (solvent 2)

dependent variables

Separation of phenolic compounds
Separation of flavonoid compounds

control variables

Flow rate (0.5 ml/min)
Column (C18 column, 4.6 mm 5 µ × 25 cm)
Liquid chromatography system (RP-HPLC; Waters)
Detector (photodiode detector M-2998)

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