An aqueous containing emodin glucuronide and emodin was extracted 3 times with dichloromethane to get rid of emodin. The extracted aqueous sample was subsequently split into two equal parts, one aspect was incubated with water and then analyzed by UPLC and the other one by hydrolysis with glucuronidase at 37 C for 30 min and then analyzed by UPLC. The Bosutinib clinical trial difference in peak regions of metabolite and emodin received from the samples before and after the hydrolysis, which were represented as Peak areaM and Peak areaE, was calculated to be the ratio K Peak areaM Peak areaE e T. Thus, the concentration of metabolite may be calculated using emodin standard curve. The common SD conversion factor was 1. 0054 0. 023 at a wavelength of 254 nm, established separately at three different concentrations. LC and uplc MS/MS Analysis of Emodin and its Glucuronides The conditions used to evaluate emodin and its metabolites were as follows: process, Waters Acquity UPLC with Empower application and photodiode array detector, line, BEH C18, 1. 85%A, wavelength, 254 nm for emodin and its glucuronide and testosterone, and treatment volume, 10 L. The check linear Retroperitoneal lymph node dissection response range was 0. 625 C100 M for emodin. The mass spectrometer boundaries were established as follows: capillary voltage, 4. 5KV, ion resource temperature, 350 C, desolvation temperature, 108 D, nebulizer gas, nitrogen, 40 psi, turbo gas, argon gas, 20 psi. Identification of Emodin and NMR A combination of reaction and its Glucuronide Metabolite by LC MS/MS products in aqueous solution was extracted with dichloromethane three times. The aqueous fraction was cleaned using pure water and loaded onto an ODS column. The mono glucuronide emodin was eluted using a solvent of H2O/MeOH. The structure of mono glucuronide emodin was recognized by UPLC ESI Q TOF MS and 1H NMR. The mass spectrometer guidelines were set as follows: capillary voltage, 4. 5KV, ion supply temperature, 350 H, desolvation GW0742 temperature, 108 C, nebulizer gas, nitrogen, 40 psi, turbo gas, argon gas, 20 psi. Kinetic Analysis Permeability of emodin was represented by R eff, which was acquired as described previously. Amounts of percentage metabolized values, quantities of glucuronidated emodin excreted to the intestinal lumen, and the percentage absorbed and emodin absorbed were determined as described previously. Fleetingly, Mgut and Mab were expressed as Eqs. 1 and 2: Mab Qt CAin CAout e T e1T Mgut QtCMout e2T where Q is the flow rate of perfusion, may be the period time of testing, CAin and CAout are the inlet and outlet concentrations of emodin, and CMout may be the concentration of emodin 3 O glucuronide. 1% Metabolized and self-absorbed were assessed as: 1% Absorbed in the intestine Mab Mtotal e3T 1% Metabolites excreted in the intestine Mgut Mtotal e4T where Mtotal may be the total quantity of element perfused within the first 30 min period.