Sciex analyst software

Pharmacokinetics of wogonoside was conducted in SD rats (200-250g) by intragastric administration at dose of 20 mg/kg. This concentration was equal to the dosage of 40mg/kg in mice in our study according to a body surface area normalized methods [48 (link)]. The rats were allowed food and water ad libitum before pharmacokinetics experiment. About 200 μL of blood was taken from tail vein of rats at 5 min, 15min, 30 min, 1, 2, 4, 6, 8 h, 10 h, 12 h, 16 h and 24 h post-dose. After centrifugation of blood samples at 13,000 rpm at 4°C for 5 min, plasm samples were transferred into Eppendorf tube. 100 μL of each serum was processed by adding 3-folds excess of methanol containing 10 ul internal standard (IS) solution followed by vortexing for 1 min. The tube was then centrifuged at 13,000 rpm at 4°C for 10 min. Then the supernatant was filtrated through 0.22 m membrane and an aliquot were injected into UPLC-MS/MS for analysis. Quantification was performed in MRM mode of the transitions m/z 461.2→285.2 for wogonoside, m/z 461.2→285.2 for IS solution respectively. SCIEX Analyst software (version 1.4.2) was used for data acquisition and analysis. The pharmacokinetic parameters were calculated by the non-compartmental analysis of plasma concentration versus time data using the DAS 3.0 software (Mathematical Pharmacology Professional Committee of China, Shanghai, China).

Pentobarbital and pentobarbital-d5 certified reference material (CRM) in methanol were purchased from Cayman chemicals (Ann Arbor, MI). Pentobarbital (PB) plasma and brain concentrations were determined using a UHPLC-MS/MS analytical method with the same instrumentation as the midazolam method. Pre-weighed brain samples were homogenized using 4 volumes of LC–MS grade water on a DPS-20 dual processing system from Pro scientific Inc. Sample preparation included protein precipitation with acetonitrile in the presence of pentobarbital-d5 internal standard (IS). Chromatographic separation was achieved using a Phenomenex Kinetex XB-C18 analytical column (2.6 µm, 100 Å, 2.1 × 50 mm) at 40 ± 2 °C. Gradient elution was employed where mobile phase A (MP-A) was 0.1% formic acid in water and mobile phase B (MP-B) was 0.1% formic acid in methanol. A flow rate of 0.4 mL/min and a 5 µL injection volume were used. Detection of PB and IS was carried out in the positive ion mode utilizing electrospray ionization (ESI) MRM, by monitoring the precursor to product ion transition pairs of 225.1–182.1 m/z for PB, and 230.1–187.1 for IS. The analytical data obtained were processed using Sciex Analyst software™ (version 1.7.1).

We employed Sciex Analyst Software (version TF1.7.1) for data analysis. The structural analysis and assessment of calculated mass errors was conducted using a combination of ChemDraw (version 19.0, Perkin Elmer, Waltham, MA, USA) and Excel (version 365, Microsoft, Redmond, WA, USA).