Agilent 1220 infinity hplc

A total of 0.05 g of ISGs were injected into 10 mL of PBS (pH 7.4) at 37 °C with an agitation speed of 50 rpm. At different time points, 2 mL of the dissolution medium was withdrawn and replaced with the same amount of fresh PBS (pH 7.4). The released IM was quantified using Agilent 1220 Infinity HPLC (C8 column, Dr. Maisch, 4.6 × 150 mm, 5 µm) at a flow rate of 1 mL/min with a mixture of acetonitrile and 0.1% orthophosphoric acid (17:83). The experiment was performed in triplicate. To study the release mechanisms, the release profile was fitted to various release mechanism models (zero-order, first-order, Higuchi square root of time, and power law) using DD-solver software.

The analysis of the enzymatic product (i.e., acetophenone) was conducted using an Agilent1220 Infinity HPLC coupled with a C12 reverse-phase column (150 × 2.0 mm). The UV detection was conducted at 245 nm. A mobile phase comprised acetonitrile (100%) and 0.1% (v/v) formic acid in water (3:7) with an isocratic elution flow rate of 0.3 mL/min at 30 °C. The injection volume was 10 µL.

Berries were crushed and the must sieved for soluble solids and tritatable acidity analysis. Soluble solids were measured by refractometer (Atago PR32) at 20 °C. Titratable acidity (expressed as g L⁻¹ of tartaric acid equivalents) was determined using a Micro TT 2022 automatic titrator (Crison, Barcelona, Spain) by titration with 0.1N NaOH. Anthocyanins were extracted from the skin of 30 berries and analysed^{76 (link)}. For each replicate, an aliquot of 0.1000 g (±0.0005 g) of frozen ground berry skin was extracted in 1.0 mL of 50% (v/v) methanol in water for 20 min with sonication. The extracts were then centrifuged (10 min at 13,000 × g) and 200 µL of the supernatant transferred to HPLC auto-sampler vials. Samples were analysed on an Agilent 1220 infinity HPLC (Agilent Technologies), equipped with a diode array detector (DAD) and a C-18 SS Wakosil (150 mm × 4.6 mm, 3 µM; SGE, Ringwood, Australia). Anthocyanins were quantified at 520 nm using an external calibration curve with malvidin-3-glucoside chloride as the standard (Sigma-Aldrich).