Zorbax eclipse aaa c18 column

Sample extraction was performed based on the method published by Battal and Tileklioğlu [43 ]. For the analysis, 80% of methanol was adjusted to −40 °C and added to 1 g of sample. The samples were homogenized with an Ultra-Turrax (IKA, Mescit Mah, Fettah Başaran, Turkey) homogenizer for 10 min, and then, it was incubated for 24 h in the dark. These samples were dried at 35 °C with evaporator pumps and dissolved using 0.1-M KH₂PO₄ (pH 8.0). Separation was performed using the Sep-Pak C-18 (Waters, Milford, MA, USA) cartridge, and the adsorbed hormones were transferred to amber vials using 80% methanol. Hormones were analyzed by HPLC using a Zorbax Eclipse-AAA C-18 column (Agilent, Santa Clara, CA, USA). The mobile phase for the analysis was 13% acetonitrile at pH 4.98. The flow rate was set to 1.2 mL min⁻¹ and the column temperature to 25 °C. The determination of the salicylic acid (SA), gibberellic acid (GA), abscisic acid (ABA), indole-3-acetic acid (IAA), cytokinin (CK), jasmonic acid (JA), and zeatin concentrations was performed using a UV detector at 265 nm.

High performance liquid chromatography (HPLC) using a Thermo Scientific UltiMate 3000 system (Thermo Fisher Scientific, Massachusetts, United States) with a Zorbax Eclipse AAA C18 column (Agilent Technologies, Santa Clara, USA) was used to quantify penicillin V and phenoxyacetic acid concentration with a buffer as described elsewhere (Ehgartner, Fricke [19 ]). A flow rate of 1.0 ml/min was applied and the temperature of the column oven was 30 °C. The UV/VIS detector for determining penicillin and phenoxyacetic acid peaks via absorption was set to 210 nm.

After crushing S. rugosoannulata through a 50-mesh sieve (the aperture is 0.3 mm), add 2 ml of concentrated hydrochloric acid, mix 2 to 3 drops of phenol and evacuate heat melt the glass hydrolysis tube while keeping the hydrolysis tube in a high vacuum state, and complete the sealing. Place the hydrolysis tube in an oven at 110 °C. After hydrolyzing for 24 h, take it out and cool it to room temperature. Transfer the hydrolyzed sample to a 25ml volumetric flask. Wash the hydrolysis tube repeatedly with pure water and fill the volume up to the mark. After the constant volume of the sample is passed through a 0.22 µm microporous membrane, put it into the sample vial to be tested. Analysis and identification of amino acids in biological tissues utilizing a high-performance liquid chromatograph (Model 1100; Agilent, USA). The chromatographic conditions are: chromatographic column: Zorbax Eclipse AAA C18 column (75 mm × 4.6 mm, 3.5 µm, Agilent, USA); mobile phase: A is 40 mmol/L NaH₂PO₄ solution, pH 7.8; B is acetonitrile-methanol-water (volume ratio of 4.5:4.5:1). Mobile phase gradient elution program: 0.0–1.0 min (B: 0%), 1.0–9.8 min (B: 0%–57%), 9.8–10.0 min (B: 57% ∼100%), 10.0–12.0 min (B: 100%), 12.0–12.5 min (B: 100% ∼0%), 12.5–14.0 min (B: 0%); flow rate: two mL/min; column temperature: 40 °C; injection volume: 18 µl.