Class vp version 6.12 sp1 software

Methanol, Acetonitrile, and ortho-phosphoric acid in ratio of 20:40:40 v/v were used as mobile phase. The flow rate was oscillated from 0.5 to 1.5 mL/min, and the revealing of the mangiferin was made at λ_max 258 nm. The mangiferin solution and selected solvents for mobile phase were passed through HPLC system High Performance Liquid Chromatographic system consisting of HPLC pumps (Shimadzu LC-10AT and LC-10AT VP), and a manually operated 20 mL sample loop and UV–Visible detector (SPD 10A VP) was used. The output signal was integrated by Shimadzu class-VP (version 6.12 SP1 software) for 0.5 h to maintain the equilibrium. The mangiferin solution was injected after passing through 0.22-micron filter. The maintenance of system equilibrium and passage of mobile phase through HPLC is necessary.

For the quantitation of TAN in Sprague-Dawley rat plasma, a Shimadzu (Kyoto, Japan) 2010A Liquid Chromatography was used. This HPLC apparatus is made up of an online degasser, a quaternary gradient low-pressure mixing pump, a column oven, an auto-sampler, a dual-wavelength UV-Vis detector and a system controller. The system is controlled by a personal computer using Shimadzu Class-VP Version 6.12 SP1 software (Shimadzu, Kyoto, Japan). For analysis of chromatographic data, the same software was used. Chromatographic separation was achieved using a reversed phase HPLC column (Agilent Zorbax Eclipse Plus C18: 250 × 4.6 mm i.d., 5 µm), protected by a guard column (Agilent Zorbax Eclipse Plus C18: 12.5 × 4.6 mm i.d., 5 µm) through gradient delivery of a mixture of acetonitrile and water for 13.5 min, at a flow rate of 1.5 mL/min at 55 °C. The gradient schedule was: (a) 0–3.5 min, acetonitrile: 35%; (b) 3.5–6.5 min, acetonitrile: 35–90%; (c) 6.5–10 min, acetonitrile: 90%; (d) 10–13.5 min, acetonitrile: 35%. UV absorbance at 322 nm was used to measure TAN while 312 nm was used as a reference.

For the quantification of SG present in LS and in blood, an HPLC method was devised and validated in mobile phase and in plasma for the purpose of selection of suitable mobile phase along with optimization of flow rate. The designed method was further validated as per ICH guidelines in both the mobile phase and plasma where correlation coefficients (R²) for SG were also found in linearity analysis, and calculation of interday and intraday precisions was also performed. The % recovery of drugs and determination of LOD and LOQ were performed with calculation of %RSD for all the HPLC process validating parameters [16 (link)]. In this study, the high-performance liquid chromatographic system consisting of HPLC pumps (LC-10AT and LC-10AT VP Shimadzu, Japan,) having a manually operated 20 μL sample loop and UV-visible detector (SPD 10A VP) was used. The output signal was integrated by Shimadzu CLASS-VP version 6.12 SP1 software. Before running mobile phase through column-C18 (250 mm × 4.6 mm, 5 μm particle size (Sigma-Aldrich, Darmstadt, Germany)), the mobile phase was first filtered through 0.22 μm membrane filter, and mobile phase was consistently run through system for 20 minutes prior to inject the sample solution to equilibrate the system.