HPLC analytical assay was performed using a Shimadzu (Milan, Italy) LC-10ATVP chromatographic pump and a Shimadzu SPD-10AVP UV–vis detector set at 244 nm. Separation was obtained on a Phenomenex (Torrance, CA, USA) SinergyTM 4 µm Hydro-RP 80Å LC column (150 × 4.60 mm) coupled to a Phenomenex Security Guard C18 guard cartridge (4 × 3.0 mm i.d., 5 μm). The mobile phase consisted of a mixture of acetonitrile/PBS pH 7.4 40:60 (v/v) and it was flushed at a rate of 0.5 mL/min. Manual injections were made using a Rheodyne 7125 injector with a 20 μL sample loop and data analysis was carried out through the CromatoPlus software (Shimadzu Italia, Milan, Italy). Because of the different purposes of this work, more than one calibration curve was obtained. The calibration curve of HC in ethanol/PBS pH 5.5 (1:1 v/v) was characterized by a drug concentration range of 2.625–105 μg/mL, a linearity coefficient (R2) equal to 1, and it was employed to determine HC solubility. The calibration curve of HC in PBS pH 7.4/ ethanol (80:20 v/v), obtained with drug concentrations ranging from 0.10 μg/mL to 41.52 μg/mL, showed a good linearity (R2 = 1) and it was used to evaluate the drug permeated during the in vitro diffusion studies. Limits of detection (LOD) and quantification (LOQ) were 0.14 μg/mL and 0.41 μg/mL, respectively.
Lc 10atvp chromatographic pump
Manufactured by Shimadzu
Sourced in Italy
The LC-10ATVP is a chromatographic pump developed by Shimadzu. It is designed to deliver mobile phases in liquid chromatography applications with precision and reliability.
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3 protocols using lc 10atvp chromatographic pump
1
HPLC Analytical Assay for Hydrocortisone
2
HPLC Quantification of AZT
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Quantification of AZT was performed by HPLC. The chromatographic system was composed of a Shimadzu (Milan, Italy) LC-10ATVP chromatographic pump and a Shimadzu SPD-10AVP UV-vis detector set at 215 nm. Separation was obtained on a Phenomenex (Torrance, CA, USA) Kinetex (150 mm × 4.6 mm I.D., 5 mm) coupled to a Phenomenex (Torrance, CA, USA) Security Guard C18 guard cartridge (4 mm x 3.0 mm I.D., 5 mm). A phosphate-buffered saline was prepared by dissolving KH2PO4 in purified water (0.01 M), and the pH was adjusted to 7.5 by adding 10M KOH. The buffer was then filtered through cellulose nitrate membrane filters (0.45 μm pore size; Sartorius AG, Göttingen, Germany). The mobile phase was composed of a mixture of phosphate-buffered saline at pH 7.5, methanol and acetonitrile (10/50/40, v/v). The flow rate was 0.8 mL/min. Solutions of AZT in methanol at drug concentrations ranging from 13 μg/mL to 400 μg/mL were used to construct a standard curve (R2 = 0.9964). For the evaluation of drug diffused through the skin, another calibration curve of AZT in PBS pH 7.4/ ethanol (80:20 v/v) was obtained, with drug concentrations ranging from 0.5 μg/mL to 50 μg/mL (R2 = 0.9943).
3
Propranolol Hydrochloride Film Quantification
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Primary polymeric layers and bilayered films were measured for thickness through a Mitutoyo pocket thickess gauge (Mitutoyo Mfc. Co. Ltd., Tokyo, Japan) . Then each circle (surface area of 1.33 cm 2 ) was dissolved in 40 mL of phosphate buffer at pH 7.4 and the solutions obtained were analyzed by HPLC method in order to determine the amount of propranolol hydrochloride contained in the film.
The results were expressed as mg/cm 2 . The chromatographic system was composed of a Shimadzu (Milan, Italy) LC-10ATVP chromatographic pump and a Shimadzu SPD-10AVP UV-Vis detector set at 290 nm. Separation was obtained on a Phenomenex (Torrance, CA, USA) Synergi Fusion-RP 80A (150 mm × 4.6 mm I.D., 5 μm) coupled to a Phenomenex (Torrance, CA, USA) SecurityGuard C18 guard cartridge (4 mm × 3.0 mm I.D., 5 μm). The mobile phase was a mixture of 0.5 % (w/v) aqueous triethylamine adjusted with orthophosphoric acid to pH 3.0 and acetonitrile (30:70, v/v). The flow rate was 0.4 mL/min and manual injections were made using a Rheodyne 7125 injector with a 20 μL sample loop. Data processing was handled by means of a CromatoPlus computerized integration system (Shimadzu Italia, Milan, Italy). Calibration curve of concentration versus peak area ratio was plotted at concentration range of 0.1-10 μg/mL and a good linearity was found (R 2 = 0.9998).
The results were expressed as mg/cm 2 . The chromatographic system was composed of a Shimadzu (Milan, Italy) LC-10ATVP chromatographic pump and a Shimadzu SPD-10AVP UV-Vis detector set at 290 nm. Separation was obtained on a Phenomenex (Torrance, CA, USA) Synergi Fusion-RP 80A (150 mm × 4.6 mm I.D., 5 μm) coupled to a Phenomenex (Torrance, CA, USA) SecurityGuard C18 guard cartridge (4 mm × 3.0 mm I.D., 5 μm). The mobile phase was a mixture of 0.5 % (w/v) aqueous triethylamine adjusted with orthophosphoric acid to pH 3.0 and acetonitrile (30:70, v/v). The flow rate was 0.4 mL/min and manual injections were made using a Rheodyne 7125 injector with a 20 μL sample loop. Data processing was handled by means of a CromatoPlus computerized integration system (Shimadzu Italia, Milan, Italy). Calibration curve of concentration versus peak area ratio was plotted at concentration range of 0.1-10 μg/mL and a good linearity was found (R 2 = 0.9998).
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