Guduchyadi medhya yoga-An HRLCMS-orbitrap data

Background and rationale. Guduchyadi Yoga (GY) is a classical eight-drug Ayurvedic formulation traditionally indicated for cognition. Constituents relevant to cognitive impairment and stress include: Tinospora cordifolia (anti-stress; memory support), Achyranthes aspera (antidepressant-like effects), Embelia ribes/embelin (anticonvulsant/CNS-depressant), Convolvulus pluricaulis (memory/stress benefits), Acorus calamus (CNS activity), Terminalia chebula (in-vitro neuroprotection via ROS-scavenging phenolics/tannins), and Asparagus racemosus (neuroprotection; memory support). Classical Rasapañcaka and medhya (cognitive) indications, together with small clinical observations (e.g., MMSE and HAM-A/HAM-D improvements), motivate systematic chemical profiling and computational evaluation.
Hydro-alcoholic extract (HAE). Solvent: ethanol:water (70:30, v/v; LC–MS grade). Weighed 25.00 g GY into a Type-I glass bottle; added 250 mL solvent (1:10 w/v). Extraction: magnetic stirring 300 rpm at 20–25 °C for 8 h (covered/amber-wrapped), then 16 h static maceration. Clarification: Whatman No. 42 filtration followed by 0.22 µm PTFE filtration. Solvent removal: rotary evaporation at 40 °C (reduced pressure) to near-dryness. The dried extract was transferred to sterile amber vials, yield (%) = (extract mass/25 g)×100, optionally N₂-flushed, and stored at −20 °C, protected from light. For LC–MS, aliquots were reconstituted/diluted and filtered per SOP.
UHPLC–HRMS conditions. Thermo Scientific Vanquish UHPLC (autosampler 4 °C; column oven 40 °C) with RP-C18 column (2.1 × 100 mm, 1.9–2.6 µm). Injection: 5 µL. Total run: 30.0 min at 0.300 mL min⁻¹. Mobile phases: A = water (0.1% formic acid); D = programmed organic channel (B,C = 0%). Gradient (A complements D): 0–2 min, 5% D; 2–15 min, 5→95% D; 15–20 min, 95% D; 21 min, return to 5% D; 21–30 min, re-equilibrate. MS: Q Exactive™ Plus (HESI), Full MS with data-dependent MS² (Top-5), acquired in separate positive/negative runs. Full MS: 70,000 (m/z 200), AGC 1e6, max IT 100 ms; dd-MS²: 17,500, AGC 1e5, max IT 100 ms, isolation 1.5 m/z, stepped NCE 30, dynamic exclusion 10 s. Tune files: Cal_positive_29082024.mstune; Cal_negative_29082024.mstune. Divert Valve A: Start in 1–2 (True).
Data acquisition and processing. Data were acquired in Xcalibur/SII (profile). Sequence: blank → system suitability → three QCs → study samples (QC every 8–10) → final blanks. Autosampler: puncture offset 0 µm; tray 4 °C. Raw files were processed in ⟨software/version⟩ with centroiding; mass error ≤ 5 ppm; minimum peak width 6–8 s; RT alignment 0.2–0.3 min; gap-filling enabled. MS² matches were queried against ⟨mzCloud/MassBank/in-house⟩ at predefined score thresholds. Features with QC RSD > 20% were flagged/excluded unless justified. Putative annotations were reported at MSI Level 2; Level 1 required |Δm| ≤ 5 ppm, RT within ±0.20 min of an authentic standard, and concordant MS² fragments.