Agilent 6224 accurate mass tof lc ms

Emodin was purchased from Fluorochem Ltd. (Glossop, UK). All other reagents and solvents were of reagent-grade quality and were obtained from commercial suppliers Honeywell (Seelze, Germany) and Sigma-Aldrich (Taufkirchen, Germany). TLC was performed on Merck-60-F₂₅₄ plates (Merck, Darmstadt, Germany) using mixtures of EtOAc:EtOH (10:1), CH₂Cl₂:EtOH (100:1) or EtOAc:MeOH (20:1). Crude Emodin preparations were purified by column chromatography on silica gel (63–200 µm, 70–230 mesh ASTM; Honeywell, Seelze, Germany). The isolated compounds were characterized by ¹H, ¹³C NMR spectra, HRMS and IR analysis. ¹H and ¹³C NMR spectra were recorded on Bruker Avance III 500 instruments (Bruker, Billerica, MA, USA). IR spectra were recorded on Bruker Alpha II FTIR Instrument (Bruker, Billerica, MA, USA). HR-MS were recorded on LC MS system Agilent 6224 Accurate Mass TOF LC/MS (Agilent Technologies, Santa Clara, CA, USA).

The drug materials were purchased and identified according to the rigid specifications set by Chinese Pharmacopeia (2010 Edition). The CFDA-approved single-herb granules of Rhizoma Acori Tatarinowii (AT), Poria cum Radix Pini (PRP) and Radix Polygalae (RP) were obtained from Tianjiang Pharmaceutical, Jiangyin, China. The granule-mixed Smart Soup (SS-G) were prepared by mixing 10 g of AT, 10 g of PRP and 10 g of RP granules to a concentration of 1 g/ml in water.
The chemical constituent identification of each batch of SS was performed using HPLC-TOF/MS. In detail, an aliquot of 1 ml of SS-G was centrifuged at 12,000 rpm. The supernatant was filtered and used for analysis. HPLC-TOF/MS was performed on a 1200 Series HPLC instrument (Agilent, Waldbronn, Germany) coupled with an Agilent 6224 Accurate-Mass TOF LC/MS. The chromatographic separations were performed at 25°C on an Apollo-C18 reversed-phase column (4.6×250 mm i.d., 5 µm, Grace) connected to an EasyGuard Kit C18 guard column (4×2 mm, Grace). The separation was conducted with an acetonitrile/water gradient with 0.5% formic acid. The injection volume was 20 µl for MS analyses.