Finnigan surveyor hplc system

Based on the spectrophotometric profile, the extract with the higher content in lycopene and β-carotene was selected for HPLC analysis. A Thermo Finnigan Surveyor HPLC system coupled with a DAD UV-visible detector (Finnigan Surveyor LC, Thermo Scientific, USA) was used. The system was controlled by the Xcalibur software (Finnigan Surveyor LC, Thermo Scientific, Waltham, MA, USA), whereas the carotenoids compounds were assessed at 450 nm on a Lichrosorb RP-18 (5 μm) Hibar RT 125–4 column. The mobile phase consisted of two solvents, namely, 90% acetonitrile (A) and 100% ethyl acetate (B). The injection volume was 10 μL, while the flow rate was 1000 mL/min. The elution gradient was: 0–16 min, 15% B; 16–54 min, 15–62% B, 54–56 min, 62% B; 56–60 min, 62–15% B; 60–70 min, 15% B. The quantification of lycopene (LOD 0.05 mg kg⁻¹ and LOQ 0.10 mg kg⁻¹) and β-carotene (LOD 0.05 mg kg⁻¹ and LOQ 0.10 mg kg⁻¹) was performed using the calibration curves for each compound.

The high-performance liquid chromatographic analysis of the anthocyanins extracted from EPE was assessed with a Thermo Finnigan Surveyor HPLC system with a DAD detector, controlled by the Xcalibur software system (Finnigan Surveyor LC, Thermo Scientific, USA). The method of Turturică et al. [39 (link)], with slight modifications, was used. In short, the EPE was filtered through a C18 Sep-Pack cartridge (Cartridge-Waters, USA) to separate the anthocyanins. In order to obtain the chromatographic elution profile, a C18 Synergi 4u Fusion-RP 80A stationary phase column (150 × 4.6 mm, 4 μm) was used, at an optimum column temperature of 25 °C. The mobile phase consisted of two phases: 100% methanol (A) and 10% formic acid (B). The injection volume was 10 μL, at a flow rate of 1 mL/min, whereas the elution took place under the following gradient conditions: 0–20 min, 9%–35% (A); 20–30 min, 35% (A); 30–40 min, 35%–50% (A); and 40–55 min, 50%–9% (A). The samples were filtered through 0.22-μm syringe filters (Bio Basic Canada Inc., ON, Canada) prior to the injection. The detector wavelength was 520 nm. The following HPLC reference substances were used for the identification of anthocyanins: delphininidin-3-glucoside chloride (purity ≥ 95%), delphinidin-3-rutinoside chloride (purity ≥ 95%), and cyanidin-3-rutinoside chloride (purity ≥ 90%) from Sigma Aldrich, Germany.

In order to identify and quantify the anthocyanin pigments from red OS extract, a chromatographic analysis was conducted using a Thermo Finnigan Surveyor HPLC system coupled to a Diode-Array Detector (Finnigan Surveyor LC, Thermo Scientific, Waltham, MA, USA) and run by the Xcalibur software. The anthocyanins from the extract were analyzed at 520 nm, at an oven temperature of 25 °C, on a Synergi 4u Fusion-RP 80A (150 × 4.6 mm, 4 μm) column. The injection volume was 10 μL for the samples. Before the injection, the samples were first filtered using 0.22 µm syringe filters (Bio Basic Canada Inc., ON, Canada). Separation was accomplished under gradient elution conditions at a flow rate of 1 mL/min and using methanol (solvent A) and 10% formic acid aqueous solution (solvent B) as the mobile phase components. The following gradient was used for the samples: 0–20 min, 9–35% (A); 20–30 min, 35% (A); 30–40 min, 35–50% (A); and 40–55 min, 50–59% (A). The anthocyanin quantification was done using the appropriate standards and data reported in the literature by Donner et al. [18 (link)], Sharif et al. [19 (link)], and Steimer et al. [20 (link)].

Organic acids, glucose, fructose and alcohols (ethanol and glycerol) were analyzed using a Finnigan Surveyor HPLC system (Thermo Fisher Scientific Inc., Waltham, MA, USA), according to the method described by Czyżowska et al. [33 (link)].

p‐Coumaric (CU) and ferulic acid (FE) were quantified by HPLC coupled with a photodiode array detector (HPLC‐PDA) at 280 nm with a Thermo Finnigan Surveyor HPLC system according to Silva, Gomes, Leitão, Coelho, and Vilas Boas (2006). p‐Coumaric (CU) and ferulic acid contents were expressed in mg/100 g of dry weight (mg/100 g DW).

Measurements were performed on a Thermo Scientific Finnigan Surveyor HPLC system, consisting of a quaternary pump, an autosampler with a temperature-controlled column compartment, and a diode array detector. A Thermo Scientific Betasil C18 (150 × 4.6 mm, 5 μm) chromatographic column was employed throughout the test, thermostatted at 55 °C. The mobile phase consisted of 0.1% (v/v) aqueous phosphoric acid solution: acetonitrile (45:55 v/v%), delivered with a 1.5 mL/min flow rate (retention time of aceclofenac was ~3.0 min). Quantitative determinations were performed at 270 nm, based on a five-point calibration curve (methanolic solution, at 10–100 μg/mL range). To determine the aceclofenac content, the microfibrous samples (n = 3) were dissolved in methanol and diluted 10-fold with the same solvent.