Agilent 1260 infinity hplc series

To analyze and detect the phenol and flavonoid compounds in the Salsola extract, a specific protocol was followed (30 (link)), with little modification, using the Agilent 1260 Infinity HPLC Series (Agilent, Santa Clara, CA 95051, USA), equipped with a quaternary pump. Kinetex® 5 μm EVO C18 100 × 4.6 mm (Phenomenex, Torrance, CA 90501-1430, USA) was used as the column and was operated at 30°C (31 (link)). The separation method was carried out using a ternary linear elution gradient with (A) 2% HPLC-grade water and H₃PO₄ (v/v; 1:1), (B) methanol, and (C) acetonitrile. Then, a final volume of 20 μL was injected. For the detection of phenols and flavonoids, an AVWD detector set at 284 nm was used.

Using specific phenolic standards, the BVJ phenolics were identified and quantified at 284 nm using Agilent 1260 Infinity HPLC series (Agilent Technologies, Palo Alto, CA, USA) [18 ]. The BVJ (20 μL) was separated by a ternary linear elution gradient on a 100 mm × 4.6 mm Kinetex EVO C18 column and eluted using 0.2% H₃PO₄, methanol, and acetonitrile.

Whole plants (WP) of H. tuberculatum collected from the northwest of Egypt in April 2018 were air-dried at room temperature for one week. The dried WP was ground to a fine powder using a small laboratory mill. Approximately 100 g of the powdered WP of H. tuberculatum was extracted by the soaking method [44 (link)] with 200 mL of ethanol solvent for three days. After the extraction process was finished, the extract was filtered through a cotton plug and then with Whatman No. 1 filter paper. The filtered extract was concentrated by evaporating the ethanol solvent to obtain the H. tuberculatum whole plant extract (WPE). To prepare the concentration of the extract, the H. tuberculatum WPE was dissolved in dimethyl sulfoxide (10% DMSO = 10 mL DMSO (99.999%) + 90 mL distilled water), and the concentrations levels of 1%, 2%, and 3% were obtained. Then, 1, 2, and 3 g from the extract were dissolved in 100 mL of 10% DMSO to obtain the extract concentrations of 1%, 2%, and 3%, respectively. The extract was analyzed for its polyphenolic compounds using Agilent 1260 Infinity HPLC Series (Agilent, Santa Clara, CA, USA), equipped with a Quaternary pump and a Zorbax Eclipse Plus C18 column (100 × 4.6 mm i.d.) in H. tuberculatum WPE [15 (link),44 (link),45 (link),46 (link),47 (link),48 ]. The analysis was carried out based on the 23 standard phenolic compounds [47 (link)].

Agilent1260 infinity HPLC Series (Agilent, USA), equipped with Quaternary pump, a Zorbax Eclipse plusC18 column 100 mm × 4.6 mm i.d. (Agilent technologies, Santa Clara, CA, USA), was operated at 30 °C. The separation is achieved using a ternary linear elution gradient with (A) HPLC grade water 0.2% H₃PO₄ (v/v), (B) methanol, and (C) acetonitrile. The injected volume was 20 μL. Detection: Variable Wavelength Detector (VWD) set at 284 nm.

The detection of alkaloids (Vincamine, Catharanthine, Vincracine (Vincristine) and vinblastine) were performed using HPLC according to Liu et al. [132 (link)]. About 10 g sample transferred into 10 mL centrifuge tube and then soaked with 10 extracting solution (2% formic acid: methanol (50:50 v/v) for 2 h. The mixture was sonicated for 30 min and centrifuged for 10 min at 4000 rpm. The supernatant was transferred into another tube filtered with 0.2 um PTFE syringe filter and injected in HPLC. Agilent 1260 infinity HPLC Series (Agilent, Santa Clara, CA 95051, USA), equipped with Quaternary pump, the column used: zorbax Eclipse plus C18 150 × 4.6 mm was operated at 30 °C. The separation was achieved using a ternary linear isocratic elution with (A) HPLC-grade water 0.2% formic (v/v), (B) methanol as mobile phase at a flow rate 1 mL/min. A VWD detector was used and the wavelength 254 nm was selected for the analysis.

The phenolic components and flavonoids in the DBRP extract solution were identified using high-performance liquid chromatography (HPLC). HPLC analysis was performed using an Agilent 1260 Infinity HPLC series (Agilent Technologies, Santa Clara, CA, USA) equipped with a quaternary pump and a Zorbax Eclipse plus C180 column (Agilent Technologies, Santa Clara, CA, USA) with a 100 4.6 mm i.d. A triple linear elution gradient was used to separate the samples: (A) HPLC grade water 0.2 percent H₃PO₄ (v/v), (B) methanol, and (C) acetonitrile. The volume injected was 20 μL. For estimated phenolic acids, all chromatograms were plotted at 284 nm, and for flavonoids, at 330 nm. By comparing peak areas to external standards, all components were recognized and quantified.

The phenolic compounds of the extracted rosemary at 60 °C were determined using Agilent 1260 Infinity HPLC series (Agilent, Santa Clara, CA, USA), equipped with a quaternary pump, a Zorbax Eclipse Plus C18 column 100 mm × 4.6 mm i.d., (Agilent Technologies, Santa Clara, CA, USA) operated at 25 °C was used for phenolic compound analysis. The injected volume was 20 µ. VWD detector set at 284 nm. The separation was achieved using a ternary linear elution gradient with (a) HPLC-grade water 0.2% H₃PO₄ (v/v), (b) methanol, and (c) acetonitrile [25 (link)].