Lc 10 system

Silica gel 60 powder and silica gel 60 F₂₅₄ aluminum sheets and glass plates for thin-layer chromatography (TLC) were obtained from Merck Supelco (Darmstadt, Hesse, Germany). Sephadex LH-20 (LH20_100) power was obtained from Millipore (Cytiva, Marlborough, MA, USA). High-pressure liquid chromatography (HPLC) was conducted using a Shimadzu LC-10 system (Tokyo, Japan). Breast cancer cell viability was determined using a cell viability assay kit (EZ-Cytox, DoGenBio, Seoul, Korea). The other chemicals and organic solvents were obtained from Sigma (St. Louis, MO, USA).

Preparative RP-HPLC, using a Shimadzu LC10 system equipped with an SPD-M10Avp photodiode array detector, or an Äkta system (Amersham Biosciences, Uppsala, Sweden) collecting data at 215, 254, and 280 nm, was carried out using a Grace Vydac Everest (250 × 4.6 mm i.d., C18, 5 μm, 300 Å) or a Phenomenex column (250 × 4.6 mm i.d., C18, 5 μm, 300 Å), run with linear gradients from 10 to 60% acetonitrile, with 0.1% trifluoroacetic acid, in water. Fractions were analyzed using MS.

Agilent 1100 HPLC system was used for analyses of the cresol contaminants in soil samples. The analysis was carried out by employing a modular Shimadzu LC-10 system comprised of a LC-10 AD pump, a CTO-10A column oven, a SPD-10A UV-DAD detector with wavelength of 274 nm, a FLD detector, a CBM-10A interface, and a LC-10 Workstation. A LC-18 column (250 mm × 4 mm ID × 5 mm) was employed at 26°C and separations were conducted in the isocratic mode, using acetonitrile:acetate buffer (30:70; v/v) at a flow rate of 0.3 mL min − 1, with an injection volume (“loop”) of 20 mL and an accuracy of ±2%. The concentration of acetate buffer was 266 mM (101 mM of acetic acid and 165 mM of Sodium acetate trihydrate) in the HPLC analyses.

Two milligrams from the aqueous and ethanolic extracts were fractionated into 48 fractions (1 ml x 48) in a 96 position deep well plate (VWR, Sweden) using a Shimadzu LC-10 system equipped with an SPD-M10AVP Photodiode-Array (PDA) detector. A Phenomenex Jupiter C18 HPLC column (5 μm, 100 Å, 4.6 mm × 250.0 mm) was used for the microfractionation of the crude extracts by employing a gradient of 5–95% CH₃CN (0.05% TFA) over 48 minutes at a flow rate of 1 ml/min. Similarly, 10 mg of the organic extracts were dissolved in 1 ml of 100% CH₃CN and a volume of 200 μl was fractionated into 48 fractions using two consecutive gradients from 5–60% CH₃CN (0.05% TFA) over 20 minutes and 60–95% CH₃CN (0.05% TFA) over 25 minutes at a flow rate of 1 ml/min [20 (link)]. One hundred microlitres from each well was transferred into 96-well U-bottom polystyrene plates (Greiner Bio-one, USA) and dried in a centrifugal evaporator (Savant Speed Vac plus SC110A).