Acquity pda detector

All solvents and reagents were used as received from commercial suppliers, unless noted otherwise. The compounds were named using the Biovia Draw 2016 package (IUPAC).
NMR spectra were recorded on a Bruker Avance III HD 500 MHz or 250 MHz spectrometer.
The chemical-shifts (δ) reported are given in parts per million (ppm) and the coupling constants (J) are in Hertz (Hz). The spin multiplicities are reported as s = singlet, d = doublet, t = triplet, q = quartet, dd = doublet of doublet, ddd = doublet of doublet of doublet, dt = doublet of triplet, td = triplet of doublet, and m = multiplet.
uPLC-MS was performed on a Waters Acquity UPLC system coupled to a Waters Acquity PDA detector, an ELS detector and an MSD (Scan Positive: 150–850). Method (pH 3): Phenomenex Kinetix-XB C18 (2.1 x 100 mm, 1.7 μm) column. Elution with a linear gradient of Water + 0.1% Formic acid and Acetonitrile + 0.1% Formic acid at a flow rate of 0.6 mL/min. Chiral SFC analysis: Waters Thar 3100 SFC system connected to Waters 2998 PDA detector, Chiralcel OD-H 25 cm. Chiral SFC separation: Water Thar SFC system with a Waters Thar FDM pump, Waters Thar Alias autoinjector, Waters Thar fraction collector and a Waters 2998 PDA detector.

The ABTS+ cation radical method was used to determine the antioxidant activity of the extracts. A 2 mm ABTS+ stock solution containing 3.5 mM potassium sulfate (VI) was prepared by diluting the stock solution eight times in methanol and incubating it overnight at room temperature in the dark to allow radical stabilization. An Acquity UPLC liquid chromatography system (Waters, Milford, MA, USA) with a Waters Acquity PDA detector (Waters, Milford, MA, USA) and an Acquity UPLC^® BEH C18 column (150 mm × 2.1 mm, particle size 1.9 μm) (Waters, Dublin, Ireland) were used. The gradient started with A—0.1% aqueous formic acid solution in acetonitrile and B—0.1% aqueous formic acid (10:90, %, v/v to 40:60, %, v/v) for 15 min. The chromatogram was recorded at a 280 nm wavelength. The injection volume of the sample was 10 µL, and the flow rate was 0.4 mL/min. After running through the column, the UPLC elution returned to baseline. The sample was then re-injected with ABTS+. This time, the chromatogram was recorded at a 734 nm wavelength. The antioxidant activity was calculated from the difference in peak areas of both chromatograms and compared with the standard curve for TROLOX. Each commercial compound standard was obtained from Sigma (St. Louis, MO, USA). The obtained results were converted into 1 g of extract [27 (link),28 (link),29 (link)].

Avastin^® (AVT/AVT08, Genentech, San Francisco, CA, USA, Lot: 33808339) and SIMAB054 (produced by ILKO ARGEM Biotechnology R&D Center, Istanbul, Turkey) samples were diluted to 9.4 mg/mL with ultrapure distilled water and injected into the HPLC directly. CEX separation was performed on BioPro SPF Non-porous Column (5 µm, 100 × 4.6 mm, YMC). Mobile phase A:100 mM NaH₂PO₄·2H₂O (Merck), mobile phase B:100 mM Na₂HPO₄·2H₂O (Merck, Kenilworth, NJ, USA), and mobile phase C:1 M NaCl (Merck) were prepared with mass spectrometry (MS) grade water. Auto Blend method was used to perform a salt gradient from 0 mM to 200 mM sodium phosphate at constant pH of 5.7. The flow rate was 0.5 μL/min, and the column temperature was 25 °C. Detection was achieved using a PDA detector (Waters, ACQUITY PDA Detector) at 280 and 214 nm wavelengths. Raw data were processed by EMPOWER Software. The acidic and basic variants of AVT and SIMAB054 in CEX analysis were collected by fraction manager equipment in the HPLC system. Samples were loaded to HPLC at high concentration to get high yield fractions. The method was adapted from Ref. [36 (link)].