Masshunter workstation qualitative software

A matrix of molecular features characterized by mass to charge ratio (m/z) and retention time (RT) was generated using MassHunter Workstation Qualitative software version B07.00, MassHunter Profinder (version B.08.00), Mass Profiler Professional (MPP version 14.5) and Personal Compound Database Library (PCDL) (Agilent Technologies, CA, USA). Molecular features were extracted and binned/aligned using parameters as follows: peak height ≥ 10,000 counts, compound ion count threshold of two or more ions, compound alignment tolerances 0.00% + 0.15 min for RT and 20.00 mg kg⁻¹ ± 2.00 mDa for mass using Profinder. Molecular features which were present only in three samples out of five were included in the analysis. Compounds were tentatively identified by matching molecular entities with PCDL entries having similar accurate mass, RT, and mass spectra (generated from analytical standards) where possible and the METLIN metabolomics database (version B 07.00, Agilent Technologies, CA, USA) otherwise. All descriptive statistical analyses were performed using Statistix (Statistix software, version 4.1; Analytical Software, Tallahassee, FL, USA) and standard deviations were calculated and reported where possible.

Data obtained from UHPLC-ESI-Q-TOF analysis was primarily characterized manually in Mass Hunter Workstation Qualitative software version B06.00 (Agilent Technologies). Data were obtained using scan mode and MS-MS. Based on accurate mass and retention time (available for standards), a database was generated using averaged retention time of identified and annotated compounds and their accurate mass. Personal Compound Database and Library software (version B 04.00, Agilent Technologies) was used.
Targeted analysis of isohexenylnaphthazarins was performed using scan mode to obtain data and later analysed against personal compound database and library. The find by formula algorithm in MassHunter Workstation software (version B06.00 Qualitative Analysis, Agilent Technologies) was used to screen for compounds of interest with a retention time window of ±0.7 min. A set of standards and blanks was analysed with the samples, and the find by formula algorithm was optimized based on the results generated for standard samples. Following the analysis, chromatograms were analysed manually for presence of ions of interest to ensure annotated ions were found in the spectra. Results were later exported to compound exchange file (CEF) format.

A matrix of molecular features characterized by mass to charge ratio (m/z) and retention time (RT) (Table 5) was generated using MassHunter Workstation Qualitative software version B07.00, MassHunter Profinder (version B.08.00), Mass Profiler Professional (MPP version 14.5) and Personal Compound Database Library (PCDL) (version B 07.00, Agilent Technologies, Santa Clara CA, USA). Parameters were as follows: peak height ≥ 5000 counts, compound ion count threshold two or more ions, compound alignment tolerances were 0.00% + 0.15 min for RT and 20.00 mg·kg⁻¹ + 2.00 mDa for mass using MassHunter Profinder. Identification was performed by comparing molecular entities with PCDL having accurate mass, RT and mass spectra generated from analytical standards where possible and references to published literature (Figure S2). All descriptive statistical analyses were performed using Statistix (ver. 10.0, 2013), standard deviation and standard error of means were calculated and reported where applicable.