As 4050 autosampler

A HPLC apparatus equipped with a PU-4180 RHPLC quaternary pump, a UV-4070 UV–Vis, an AS-4050 autosampler and an LC-NetII/ADC interface box (Jasco Europe, Lecco, Italy) was used to assess the analytical quantification of Acyclovir, The samples were eluted under isocratic conditions using an RPC18 column (Mediterranea SEA18 column, size 250 mm × 4.6 mm, 5 µm) operating at room temperature (25 °C) under a flow of 1 mL min⁻¹, and the drug was detected at 251 nm with a retention time of 11,6 min. A mixture of water and acetonitrile (97:3 v/v) containing 0.05% (v/v) of trifluoroacetic acid was filtered through a 0.2 μm polycarbonate filter and used as the mobile phase^{50 (link)}. Data were acquired and processed with ChromNAV v.2.04.01 software (Jasco Inc., Tokyo, Japan), using the linear equation y = 4521.7x + 129,384 with a correlation coefficient (R²) of 0.9999 obtained from the analysis of standard solutions (concentration range of 0.25–25 µg/mL, n = 6). Under the experimental conditions, the limit of detection (LoD) and limit of quantitation (LoQ) were 0.08 µg/mL and 0.25 µg/mL, respectively.
The detection of Methylene Blue was carried out by using a PerkinElmer Lambda 35 ultraviolet–visible (UV–Vis) spectrophotometer at a λmax of 660 nm^{51 (link)}.

Carboxymethoxylanmine hemihydrochloride (CMO), 1,1′-carbonyldiimidazole (CDI), 1-ethyl-3-[3-dimethylaminopropyl]-carbodiimide (EDC), N-hydroxysuccinimide (NHS), sodium bicarbonate, 1,4-butanediol diglycidyl ether, surfactant were purchased from Sigma (St Louis, MO, USA). Formic acid was obtained from Merck (Darmstadt, Germany). Both 50 nm and 70 nm gold nanoparticle were purchased from Cytodiagnostics Inc. (Burlington, Canada). The HPLC equipment consisted of a Jasco PU-4180 HPLC pump, a FP-1520 intelligent fluorescence detector and an AS-4050 autosampler was from Jasco (Tokyo, Japan). The 250 × 4.6 mm i.d., 5 μm, Atlantis T3 C18 reverse phase column was obtained from Waters (Milford, USA). A 40 × 4.0 mm i.d., 5 μm, Lichrospher C18 guard column was obtained from Merck (Darmstadt, Germany). CTN standard (Fig. S1A†) and other reagents were the same as the previous report.¹⁸ All reagents for pilot production of ELISA kit and immunostrip are prepared in our laboratory and sent to the biotechnology company for final immunostrip assembling and vacuum packaging techniques. All of the organic solvents and other chemicals used were reagent grade or better. An immunostrip scan reader CHR-110R was purchased from Kaiwood technology (Tainan, Taiwan). The anti-CTN polyclonal antibody was produced from our previous report.¹⁸

In accordance with the procedures outlined by Schieber et al. [34 (link)] and Fredericks et al. [35 (link)], the analysis of phenolic acids was carried out. A Jasco PU-2089 plus controller, a Jasco UV-2070 plus ultraviolet (UV) detector, and a Jasco AS-4050 autosampler made up the HPLC system used for the analysis. Jasco is based in Easton, Maryland. The column used was an Aqua Luna C18 250 4.6 mm (Phenomenex, Torrance, CA, USA) protected by a Phenomenex 4.0 3.0 mm C18 ODS guard column (Phenomenex, Torrance, CA, USA). The HPLC UV detector was set at a wavelength of 320 nm. Two mobile phases made up the gradient program: A (2% acetic acid) and B (1:50:49 acetic acid, acetonitrile, and water). The gradient program began with 55% A and 45% B for 50 min, then 100% B for 10 min, and finally a decline to 10% B until the completion of the run. Calibration curves were built using external standards of chlorogenic acid, caffeic acid, and ellagic acid for the quantification of phenolic acid concentration. The results were reported in milligrams (mg) of phenolic acids per 100 g of fresh-weight strawberries (mg/100 g FW).