Agilent series 1260 hplc dad instrument

A high performance liquid chromatography analysis was performed in order to recognize and quantify the major phenolic compounds of the olive leaf extract. The phenolic profile was taken following the method of Souilem et al. [19 (link)] by using an Agilent series 1260 HPLC-DAD instrument (Agilent Technologies, Waldbronn, Germany). The mobile phase was made of both phase A (0.1% acetic acid in water) and phase B (100% acetonitrile). The elution conditions were as follows: the flow rate set at 0.5 ml/min, injection volume of 10 ml, and operating temperature of 40°C. The running gradient was as follows: 0–22 min, 10–50% B; 22–32 min, 50–100% B; 32–40 min, 100% B; 40–44 min, 100–10% B.
Detection was conducted by a diode array detector (DAD) while the chromatograms were recorded at λ = 280 nm for oleuropein and hydroxytyrosol and at 330 nm for flavonoids. The act of quantification was carried out by external calibration with particulars standards. The purity level of oleuropein and hydroxytyrosol was determined using the LCMS/MS (Agilent 1100 LC, Germany) equipment according to Bouallagui et al. [14 (link)].

Chromatographic analyses were achieved on an Agilent series 1260 HPLC-DAD instrument (Agilent, Waldbronn, Germany). The instrument includes a quaternary pump, an online degasser, an autosampler, and a thermostatically controlled column compartment. Chromatographic separation was carried out on a ZORBAX Eclipse XDB-C18 column serial number USNH027266 (4.6 mm I.D. × 250 mm × 3.5 μm particle size). The elution conditions were as follows: mobile phase A (0.1% acetic acid in water) and mobile phase B (100% acetonitrile), flow rate of 0.5 mL/min, sample injection volume of 10 μL, and operating temperature 40°C. The running gradient was as follows: 0–22 min, 10%–50% B; 22–32 min, 50%–100% B; 32–40 min, 100% B; 40–44 min, 100–10% B. Reequilibration duration lasted 6 min. The DAD detector scanned from 190 to 400 nm and the samples were detected at 254, 280, and 330 nm.