HPLC Analysis of Flavonoids in Lime Juice on Ascentis® Express C18
Materials
analytical column
mobile phase component
CONDITIONS
sample preparation
SPE (Solid Phase Extraction)
sample/matrix
lime juice hand-squeezed in laboratory, centrifuged and filtered through a 0.45 μm nylon filter disc (Acrodisc)
column
Ascentis Express C18, 5 cm x 4.6 mm I.D., 2.7 μm particles (53826-U)
mobile phase
[A] water/formic acid (99.9:0.1); [B] water/acetonitrile/isopropanol/formic acid (39.9:20:40:0.1)
gradient
10% B for 3 min; 10 to 34% B in 30 min; 34 to 100% B in 1 min; held at 100% B for 6 min; 100 to10% B in 1 min
flow rate
0.8 mL/min
column temp.
ambient
detector
PDA: 190–370 nm; chromatogram extracted at 283 nm
detector
ESI-MS: mass spectral range, 250-700 m/z; interval, 0.5 s; scan speed, 938 amu/s; nebulizing gas (N2) flow, 1.5 L/min; (interface temperature, 350 °C; heat block, 300 °C; desolvation line (DL) temperature, 300 °C; DL voltage, 34 V; probe voltage, +4.5 kV; Qarray voltage, 1.0 V; RF voltage, 90 V; detection gain, 1.05 kV)
injection
2 μL
Description
Analysis Note
Flavonoids are polyphenolic compounds ubiquitous in nature and classified, according to their chemical structure, into flavonols, flavones, flavanones, isoflavones, catechins, anthocyanidins and chalcones. They are widely present in fruits, vegetables, leaves, nuts, seeds, bark and beverages (tea, coffee, beer, wine and fruit drinks).
Legal Information
Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany