Arc hplc system

The functional group present in the ethanol extract of mycelial culture was determined by the method followed by Bekiaris et al. [15 (link)] using FTIR (Fourier Transform Infrared) spectroscopy (Bruker FTIR Alpha spectrometer). Briefly, the mycelial sample (4 mg) was kept on the machine lens mirror surface and the lens was adjusted to obtain spectra in the mid-infrared scanning range of 4000–600 cm⁻¹.
The quantification of phenolic compounds, namely gallic acid and rutin, present in A. aegerita extract, was performed by the method followed by Bristy et al. [16 (link)] using high-pressure liquid chromatography (Arc HPLC System, Waters India Pvt. Ltd., Bengaluru, Karnatka). The system consists of a temperature-controlled column chamber, gradient binary pump system, injector (manual), and photodiode array detector (2998) for the quantification and detection of phenol and flavonoids. Empower2 software was used to collect and analyze the obtained data. The C18 column 4.6 × 250 mm, 33 cm (Waters) with a 60 mL/h flow rate was used for rutin and gallic acid analysis. The rutin and gallic acid were quantified at 357 nm and 272 nm, respectively.

100 mg powders of P. chaudocanum stems were dissolved with 10 ml of methanol. A 10 mg ml⁻¹ sample solution was produced by sonicating the prepared sample within 10 min, and then filtering (0.22 µm). The sample solution obtained was subjected to the HPLC experiment.
For determining the crucial compounds presented in P. chaudocanum extract, HPLC analysis was performed. Stock solutions (5 mg ml⁻¹) of piperine were produced by dissolving 250 mg of piperine in 50 ml of methanol. The six different concentrations of 0.125, 0.5, 0.75, 1.0, 1.25 and 1.5 mg ml⁻¹ were prepared by diluting from stock solution. The sample solution above with 10 mg ml⁻¹ concentration was used for HPLC analysis. The method conditions were the mobile phase composition of acetonitrile: water: acetic acid (60:39.5:0.5, v/v), at 1 ml/min of flow rate, and 10 ml of injection volume. By comparing the substances to the reference compounds and utilizing the retention duration, and the absorption spectrum profile, investigated compounds were identified by chromatographic analysis using the HPLC. The chemical substances were analysed by using a Waters Arc HPLC system with a PDA detector (200–800 nm), and a silica gel C₁₈ column (5 µM particle size, 25 cm × 4.6 mm).