Cto 10asvp column oven

For HPLC, CTO-10ASvp column oven, SIL-HTc autosampler (Shimadzu, Japan), and Shimadzu LC-20AD binary pump were used in the study. An Agilent ZORBAX Eclipse XDB-C18 (5 μm, 4.6 × 150 mm) column was used for accomplishing the chromatographic separation with the mobile phase consisting of 70% acetonitrile-water (comprising 10 mmol ammonium acetate and 0.1% formic acid) at a flow rate of 0.5 mL/min. The injection volume was 15 μL, and the column temperature was held at 30°C.
In a positive ion mode, an API 4000 triple quadrupole tandem mass spectrometer with an ESI source was used, and the acquisition and analysis of data were done with Analyst 1.6.2 software (Applied Biosystems Sciex, USA). Multiple reaction monitoring (MRM) parameters for the DIG and lappaconitine hydrobromide (IS) were optimized and are summarized in Table 1. The other parameters for ionization were as follows: curtain gas, 20 psi; collision gas, 6 psi; ion source gas 1, 50 psi; ion source gas 2, 50 psi, respectively, with a temperature of 500°C and an ion spray needle voltage of 5500 V.

An acidic mobile phase was used for detection of insulin. The mobile phase consisted of 0.2 M sodium sulphate anhydrous (adjusted to pH 2.3 with ortho phosphoric acid) and acetonitrile (74:26). The aqueous solution was filtered through 0.45 µm pore size nylon membrane filter (Whatman international, Maidstone, UK) under vacuum and degassed prior to use. The analysis was run at a flow rate of 1.2 mL/min and sample injection volume of 20 µL. The detector was set a wavelength of 214 nm. The chromatographic analysis was performed using a Shimadzu Prominence HPLC system (Kyoto, Japan) consisting of an in-line DGU-20A3 Prominence degasser, LC-20AD Prominence solvent delivery pump, SIL-20A HT prominence auto sampler, CTO-10AS VP column oven, and SPD-M20A Prominence UV-VIS detector. Data acquisition and analysis were performed using Shimadzu LC solution software Version 1.24 SP1 (Kyoto, Japan).

A SCIEX Triple Quad 3200 HPLC-MS/MS system (Sciex, Applied Biosystems Inc, USA) and a Shimadzu LC-20AD binary pump equipped with a SIL-HTc autosampler and CTO-10ASvp column oven (Shimadzu, Japan) were used for the present study. An Agilent ZORBAX Eclipse XDB-C₁₈ (4.6 × 150 mm, 5 μm) column was used for all chromatographic separation. A Pro 200 Hand-Held or Post-Mounted Laboratory Homogenizer (PRO Scientific Inc, USA) was used to prepare tissue homogenates.

Serum was separated by centrifuging the blood sample at 3,000 × g for 10 minutes and analyzed for biochemical parameters using an automated analyzer (Samsung LABGEOPT10, Samsung Electronics Co, Suwon, Korea). Reproducibility and accuracy of LABGEOPT10 were documented according to the IVR-PT06 guideline.
Serum and liver MDA levels were measured using HPLC with an LC-20AD pump, SIL-20A autosampler, SPD-20A ultraviolet-visible spectroscopy detector (at C18-ODS-3V and 5 μm with a 4.6 mm × 250 mm column), and a CTO-10ASVP column oven (Shimadzu) as described previously by Sahin et al. [25 (link)]. An aliquot of 20 μl of the supernatant isolated from serum or liver homogenate was injected into an HPLC column. Samples were eluted with a mobile phase containing 30 mM of KH₂PO₄-methanol (82.5 : 17.5, v/v, pH 3.6) at a flow rate of 1.2 ml/minute. Chromatograms were acquired at 250 nm.
To assess the activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), and liver tissues were homogenized in a mixture of 20 mM of HEPES buffer (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid), 1 mM of ethylene glycol tetraacetic acid, 210 mM of mannitol, and 70 mM of sucrose (pH 7.2) and analyzed by ELISA (Biotek Instruments, Inc., Vermont, USA) using a commercial kit according to the manufacturer's instructions (Cayman Chemical, Ann Arbor, MI, USA).

The phenolic composition was investigated by a Shimadzu HPLC system equipped with a CTO-10AS VP column oven, a DGU-20AS prominence degasser, an LC-20AD prominence pump, an SIL-20A HT prominence autosampler, and an SPD-M20A photodiode array (PDA) detector (Kyoto, Japan), following the methodology validated by Moreira et al. [18 (link)]. The results were expressed as mg of each phenolic compound per 100 g of cookies (mg/100 g cookies).

An HPLC instrument consisted of SCL-10Avp (Shimadzu, Kyoto, Japan) chromatographic control system, two LC-10ATvp pumps (Shimadzu), a CTO-10ASvp column oven (Shimadzu), and a LC Solution chromatographic workstation. The system was equipped with a Zorbax SB-C18 column (4.6 mm × 250 mm, 5 μm, Agilent, Palo Alto, CA, USA). The mobile phase was methanol (A) and 0.6% acetic acid in water (B) with a gradient elution of 40–50% A at 0–8 min, 50–83% A at 8–10 min, 83–85% A at 10–25 min, 85% A at 25–50 min. During the elution program, the flow rate was 1 mL/min, the temperature of column oven was 30 °C, and the injection volume was 20 μL. SofTA Model 400 ELSD (SofTA Corporation, Boulder, CO, USA) was connected to the HPLC instrument in series mode. The drift tube temperature of ELSD was 70 °C, nebulizer temperature was 30 °C, and the nitrogen gas pressure was 40 Psi.