Xterra ms c18

Concentrations of E1, E2 and Adione were measured using LC-MS/MS at ASKA Pharma Medical Co., Ltd (Kawasaki, Japan). Briefly, weighed brain tissues (40–200 mg) were homogenized, and each internal standard (100 pg of each internal standard, E1-¹³C₄, E2-¹³C₄, and Adione-d7) and ethanol were added, and then shaken at 50 °C for 2 hrs. The obtained extracts were applied to a Bond C18 cartridge column (Varian, Harbor City, CA), and the steroid fraction was eluted with 80% acetonitrile. The steroids fraction was loaded onto a mixed-mode cartridge (Oasis MAX, Waters, Milford, MA) to separate the neutral and phenol fractions containing androgens and estrogens, respectively. Estrogens were measured by LC-MS/MS, an API-5000 triple stage quadrupole mass spectrometer (Appied Biosystems, Foster City, CA) connected to an LC-20AD pump and SI HTC autosampler (Shimadzu, Kyoto, Japan), and electrospray ionization ion source devices, using the column, a Xterra MS C₁₈ (2.1 mm × 100 mm I.D.3.5 μm; Waters) at 40 °C. Adione was measured by an LC-MS/MS instrument, API-4000 (Appied Biosystems) equipped with an ESI ion source and an Agilent 1100 HPLC system (Agilent Technologies, Santa Clara, CA) with an HTC PAL auto-sampler (CTC Analytical, Zwingen, Switzerland), using the column, Cadenza CD-C₁₈ (150 mm × 3 mm I.D., 3 μm; Imtakt, Kyoto, Japan) at 40 °C.

Atrazine was extracted three times from the soil samples with dichloromethane. The mixed extracts were concentrated and dried by evaporation. Blow-dried by N₂, the constant volume was adjusted by mobile phase. HPLC (Agilent 1260) was used to measure the concentration of Atrazine from each sample. The wavelength was set to 222 nm on the UV detector and a reverse-phase column C₁₈ (4.6 × 250 mm, 5 μm) used a flow rate of 1.0 mL min^–1 (methanol/water = 60/40, v/v), a column temperature of 30°C, and an injection volume of 10 μL.
Liquid Chromatography-Mass Spectrometry (LC-MS/MS, UV-8030) was used to detect the Atrazine metabolites. The procedures of the chromatography analysis were performed by following those of previous studies [10 , 23 (link)] with minor modifications. The chromatographic separations were made based on stationary phase with the analytical chromatographic column of XTerra MS C₁₈ (3.9 mm × 100 mm × 3.5μm, Waters, USA). The mobile phase was composed of a mixture of an aqueous solution containing 0.1% of formic acid and the acetonitrile under eluent (1:1, v/v). The flow rate of the mobile phase was 1.0 mL min^–1.

A reversed phase HPLC method was developed and validated using a Shimadzu HPLC, consisting of LC-10AT pump, LC-20AT autosampler and UV–Vis detector (SPD-20AV). A Waters XTerra MS C₁₈ analytical column with 3.5 μm particle size, 150 mm length and 4.6 mm internal diameter was used with an isocratic mobile phase comprising phosphoric acid in water with pH (2.75 ± 0.05) (60%) and acetonitrile (40%). The mobile phase was run at a flow rate of 1 mL/min for 10 min with a volume of injection of 20 μL. The temperature was maintained at 40 °C and a wavelength of 220 nm was used for detection of MUP. The HPLC method was validated according to ICH Topic Q2 (R1) guidelines [25 ].

Binder samples were applied on glass panels and dried for 72 h at ambient condition. Approximately 0.5 g of the dried binder sample was taken in a 100 ml beaker. To extract DM, 25 ml of HPLC grade acetonitrile was added and sonicated for 5 min. After filtration the binder sample was extracted again with 25 ml acetonitrile. Filtrates were combined and transferred in to a 50 ml volumetric flask, and volume was made up by acetonitrile. One ml of the solution was filtered through 0.22 micron syringe filter (Millipore Inc., USA) and analyzed by HPLC. HPLC system (Waters, USA) equipped with 1,525 binary pump, 2,487 tunable dual wavelength UV detector, and Rheodyne injector with 10 μl loop was used in the present study. Analysis was performed by isocratic elution (methanol and water 8:2 v/v) on XTerra MS C18 (4.6 × 250 mm, 5 μm) reverse phase HPLC column (Waters, USA). Wavelength and flow rate were set at 280 nm and 1 ml/min, respectively. Standard solution was prepared by dissolving 14.78 mg of DM in 20 ml of acetonitrile. DM content was determined by comparing the corresponding peak area of sample with standard.

Ultrasonic-assisted extraction was performed using an ultrasonic bath (ScientTech, Labotec, Midrand, South Africa). Separation of the compounds was performed using an Agilent HPLC 1260 system (Agilent Technologies, Waldbronn, Germany) which consisted of a binary high-pressure pump, autosampler, a thermostatted column compartment, a diode array detector and a fluorescence detector. Instrument control, data collection and processing were achieved using the ChemStation (version 1.9.0) software. The separation of the mixture was performed on an XTerra^® MS C18 (150 mm × 4.6 mm, 3.5 µm) analytical column (Waters Corporation, Milford, MA, USA). The mobile phase used for the separation was 0.1% formic acid in water (A) and acetonitrile (B). The following gradient elution mode was used to separate the compounds: 0 min 25% (B), 1 min 35% (B), 2 min 45% (B), 3 min 55% (B), 5 min 100% (B). Injection volume was 5 µL; temperature was 25 °C; and flow rate was 1.3 mL min ⁻¹. The compounds were monitored at 254 and 331 nm. A typical chromatogram for the separation of the pure standards in the solvent is shown in Figure 6.

The standard substances were prepared as follows. We weighed Rg1 (5.278 mg), coptisine (0.608 mg), palmatine (0.267 mg), berberine (0.689 mg), Rb1 (5.225 mg), and Rc (5.254 mg), mixed them, and dissolved them in 25 mL methanol in a conical flask. Then, we selected and weighed 10 different batches of Coptidis Rhizoma and Panax ginseng concentrated granules. These samples were dissolved with the same volume of methanol. Finally, we prepared the HRD solution by mixing and dissolving Coptidis Rhizoma and Panax ginseng. The examination condition was as follows: chromatographic column, WATERS XTERRA ®MS C18 (4.6 mm × 250 mm, 5 μm); mobile phase, acetonitrile-0.05mol/L KH2PO4 (pH value adjusted to 4 with phosphoric acid); detection wavelength, 203nm; column temperature, 30°C; flow rate, 0.9mL/min. We added 10μL of the standard substance solution and different sample solutions in the column and obtained the results.

DOTANOC was obtained from ABX GmbH (DOTA-NOC acetate Cat-N° 9712). The peptide was dissolved in MilliQ water to obtain a stock solution of 1 mM. The radiolabeling with ¹⁵²Tb was performed by direct addition of 12 μL DOTANOC stock solution to a ¹⁵²Tb-α-HIBA solution (120 MBq in ~500 μL, pH 4.7). The reaction mixture was incubated at 95 °C for 15 min. Labeling of DOTANOC with n.c.a. ¹⁷⁷Lu (Isotope Technologies Garching ITG GmbH, Germany) was performed under standard conditions in a mixture of HCl (0.05 M) and Na-acetate (0.5 M) at pH 4.5 and elevated temperature (15 min, 95 °C). High-performance liquid chromatography (HPLC) with a C-18 reversed-phase column (Xterra™ MS, C18, 5 μm, 150 × 4.6 mm; Waters) was used for quality control of the radiolabeled peptides. The mobile phase consisted of MilliQ water containing 0.1 % trifluoroacetic acid (A) and acetonitrile (B). A gradient from 95 % A and 5 % B to 20 % A and 80 % B over a period of 15 min was used with a flow rate of 1.0 mL/min. The product peak corresponding to ¹⁵²Tb-DOTANOC and ¹⁷⁷Lu-DOTANOC, respectively, appeared with a retention time (R_t) of 11.1 min.