Spd 10a uv vis detector

We analyzed urinary MDA as an oxidative stress biomarker by its adducts with 2-thiobarbituric acid (TBA, CAS number: 504-17-6) as a previously described method [24 (link)] with minor modifications. Briefly, 300 μl of 0.5 M of phosphoric acid was mixed with 50 μl of urine or 1,1,3,3,-tetraethoxypropane (CAS number: 122-31-6) standards and 150 μl of TBA reagent. The mixtures were heated at 100°C for 1 h. The tubes were chilled on ice for 5 min. Five hundred μl of methanol was added to the mixtures, and the mixtures were centrifuged at 13,000 × g for 10 min. The supernatant fractions were transferred to glass autosampler vials and 20 μl of each supernatant was analyzed by HPLC. The levels of TBA-MDA adduct were determined at 532 nm on an isocratic HPLC system: The TSK gel column ODS-80TM (5 μm, 4.6 mm × 150 mm, Tosoh, Tokyo, Japan) was eluted with 50 mol/l potassium phosphate buffer (pH 6.8) and methanol (58:42, v/v). The flow rate of mobile phase was 0.6 ml/min.
The HPLC system was composed of dual Younglin SP930D pumps (Younglin, Seoul, Korea), MIDAS COOL autosampler (Spark Holland, Emmen, Netherlands) and SPD-10A UV–VIS detector (Shimadzu, Kyoto, Japan).

Chromatographic analysis was performed using a Shimadzu SCL-10A HPLC system equipped with a Shimadzu SPD-10A UV/Vis detector, an LC-10AT pump, SIL-20AC HT auto-samplers, and a CTO-10ASVP column oven. Chromatographic separation was performed on a Phenomenex Synergi 4u Hydro-RP 80A column (150 × 4.60 mm, 4 µm particle size) connected to a Phenomenex C18 (10 × 4.6 mm, 5 µm) guard column that maintained the temperature at 35 °C. The isocratic mobile phase was methanol and water (40:60 v/v) at a flow rate of 1.0 mL/min. The injection volume was 10 µL, and the eluates were monitored at 275 nm. The total run time was 8 min.

Optical rotations were measured on a WZZ-2A (Shanghai base solid Instrument Co., Ltd., Shanghai, China). The IR spectra were obtained from a Bruker IFS-55 spectrophotometer (Karlsruhe, Germany) with KBr disks. HR-ESI-MS data were measured on a Micro-mass Autospec-UntimaE TOF mass spectrophotometer (Waters, Milford, MA, USA) and a Bruker Solarix 7.0T FT-ICR MS system (Bruker, Germany). NMR spectra were run on a Bruker AVANCE-400/-600 spectrometer (Karlsruhe, Germany). Analytical HPLC was performed on a Shimadzu LC-10AT (Kyoto, Japan) liquid chromatograph and preparative HPLC separation was carried out on a YMC-Pack ODS-A column (10 × 250 mm, 5 μm; YMC-Pack, Kyoto, Japan), equipped with a Shimadzu LC-8A pump (Kyoto, Japan) and a Shimadzu SPD-10A UV–V is detector (Kyoto, Japan). Sugars analytical HPLC was carried out on a Jasco PU-4180 pump (Kyoto, Japan) and an OR-4090 detector (Kyoto, Japan).

Detection and identification of bioconversion products such as 4-methylumbelliferone-7-O-α-D-glucopyranoside (MUG) and α-arbutin in the reaction mixtures were achieved by HPLC analysis. HPLC analysis was performed with a Zorbax Eclipse XDB-C18 column (5 μm, 4.6 × 250 mm; Agilent, USA) connected to a Shimadzu LC10AD_vp system (Shimadzu, Japan), a SPD-10A UV-VIS detector (set at 320 and 280 nm for 4-methylumbelliferone-7-O-α- MUG and α-arbutin, respectively), and an SPD-LC10 pump. Separation of reaction products was achieved with a gradient of 10-90% acetonitrile in 0.1% formic acid/water for 30 min at a flow rate of 1.0 ml/min. All solvents were filtered, degassed, and stored under pressure.

Nanoparticles were suspended in different pH aqueous solutions (pH 7.4, 6.5, 5.0 and 4.0) at 37 °C and were shaken at 40 rpm. The released drugs from nanoparticles were isolated by dialysis bag (MWCO 6–8K). At different time interval, the isolated solution was analyzed using HPLC system. Inspire C18™ column (5 μm, 250 mm × 4.6 mm) was used to separate the analytes. The wavelength for determining IMQ concentration by Shimadzu SPD 10A UV–VIS detector (Kyoto, Japan) as observed at 244 nm. The Shimadzu RF-10 AXL fluorescence detector (Kyoto, Japan) was used to measure DOX concentration at λ_ex = 480 nm/λ_em = 570 nm. The mobile phase was 70% CH₃COONa (pH = 5.5) and 30% ACN at a flow rate of 0.5 mL/min.