Mnova software version 14

The fraction that exhibited the greatest inhibition of AChE activity was further explored for bioactive compounds through LC-MS analysis, performed at the Sophisticated Analytical Instrument Facility, CSIR-Central Drug Research Institute, Lucknow. The LC-MS and MS/MS simulations were carried out using a Waters Alliance E2695/HPLC-TQD mass spectrometer with a mass range of 100–2000 Da. Separation was achieved on an Agilent Poroshell 120 EC C-18 column (150 mm × 4.6 mm, 2.7 μm). Two mobile phases were used: A-0.1% formic acid in water and B-90% of acetonitrile in water, at a flow rate of 1.500 mL/min. The gradient started with A-5% and B-95%, a linear increase in A% and decrease in B% after 6 min at A-30% and B-70%; during 12–16 min, A-60% B-40%; 20 min, A-80% and B-20%; and 26–30, min A-5% and B-95%. The chromatograms were compared to the available literature to identify the peaks, and then, the output file was subjected to analyze the peaks via Mnova software version 14.2.1 offered by Mestrelab Research, chemistry software solutions.

The ¹H NMR spectra were acquired at 25 °C utilizing D₂O as the solvent and trimethyl silane (TMS) as the internal standard on a Bruker AV 500 MHz (Bruker BioSpin, Zurich, Switzerland) instrument. Mnova software version 14.2.1 (Mestrelab Research, Santiago de Compostela, Spain) was used to process the data. Spectrum Two FTIR spectrometer (Perkin Elmer, Buckinghamshire, UK) with diamond attenuated total reflectance (ATR) crystal (with 16 scans) was used to assess drug-formulation interactions. Spectra were collected from 400 to 4000 cm⁻¹, including both loaded and unloaded formulations, as well as pure forms of the ingredients [62 (link)].