Lc 20ab hplc system

MPTs are UV-active chemicals because they contain benzene rings and conjugated groups in their structures. Maximum absorbance was measured at a wavelength of 200 nm. The reference standard of MPT was run with a concentration of 10 µg/mL to detect the wavelength. A Shimadzu LC 20AB HPLC system (Tokyo, Japan) with a 1000 pump and UV-VIS detector was used in this analysis. The mobile phase consisted of a mixture of acetonitrile and sodium acetate buffer with a pH of 3.2. The stationary phase consisted of an Agela C18 column that was 250 millimeters by 4.6 millimeters and had a particle size of 5 micrometers. The injection volume for HPLC analysis was kept at 20 µL, along with the 1.0 mL/min flow rate, and the UV detector was set to a wavelength of 200 nm. The acquired chromatograms were compared with the reference standard of MPT, and the drug loading efficiency was calculated in triplicate. The proposed technique was effectively used to investigate drugs in plasma samples [37 (link)]. We carried out this assay in rabbit plasma under identical conditions, and the outcomes demonstrated its validity as a technique. According to USP and ICH guidelines, the analytical process was carried out, and numerous parameters, including precision, accuracy, specificity, and linearity, as well as the limit of quantification (LOQ) and limit of detection (LOD), were determined.

The proteins were extracted from EA.hy926 cells using the Lysis buffer (7 mol/L Urea, 2 mol/L Thiourea, 4% CHAPS, 40 mmol/L Tris-HCl, pH 8.5) containing protease inhibitors PMSF (1 mmol/L) and EDTA (2 mmol/L). The protein concentrations were determined using BCA kits (Beyotime Institute of Biotechnology). After trypsin digestion and peptide measurement, iTRAQ labeling was performed according to the manufacturer’s protocol for 8-plex iTRAQ reagent (Applied Biosystems). The labeled samples were mixed and divided into 20 fractions, via strong cation exchange chromatography, using an LC-20AB HPLC system (Shimadzu, Kyoto, Japan). After desalting in a Strata X C18 column (Phenomenex, Torrance, CA, USA), each fraction’s supernatant was loaded on an LC-20AD nanoHPLC system (Shimadzu). Mass spectrometry (MS) using a TripleTOF 5600 System (AB SCIEX, Concord, ON, Canada) was performed after HPLC.
The Mascot search engine (Matrix Science, London, UK; version 2.3.02) was applied to analyze the MS data for protein identification. The proteins that possessed at least two unique spectra were considered for further analysis. p < 0.05 and fold change >1.2 denoted significance.

Each 100 mg of MSN-FP was measured with an analytical balance. MSN-FP was separately combined with 100 mL of methanol while being gently stirred for an hour. A validated HPLC method was used to quantify the filtrate after the combination had been filtered. The Shimadzu LC 20AB HPLC system had a UV-VIS detector and a 1000-pressure pump (Shimadzu Scientific Instruments, Kyoto, Japan). Ammonium acetate buffer (pH = 7.6), methanol, and acetonitrile were employed as the mobile phase in a ratio of 10:50:40 (%v/v) on an Agela C18 column (250 × 4.6 mm, 5 µ) as the stationary phase. The UV detector was set at a wavelength of 254 nm, the injection volume was 20 µL, and the mobile phase flow rate was 1.0 mL/min. Drug loading efficiencies were computed in triplicate and compared with the resulting chromatograms to standards. The nasal spray stability and FP leakage from MSN within the nasal spray were both determined using the same process.
$$\mathbf{Entrapment}\mathbf{efficiency}\mathbf{(}\mathbf{\%}\mathbf{)}\mathbf{=}\frac{Drug added-Free drug}{Drug added}\mathbf{\times }\mathbf{100}$$