High-performance liquid chromatography (HPLC; model Jasco PU-2089 with a model Jasco UV-2070 plus multi-wavelength UV–VIS detector, Jasco, Tokyo, Japan) was used to analyze NP. The separation and analysis procedures were as described in a previous report [16 (link)]. A C18 (5 µm, 4.6 × 250 mm) ACE® column was used to separate and quantify the NP concentration. The detection wavelength was 235 nm, and the isocratic flow rate of the mobile phase was 1 mL/min. Before use, a mobile phase solution of 50% methanol: acetonitrile (1:1) in water was filtered through a 0.45-µm nylon membrane filter and then degassed in a sonicating bath. A 20-μL injection volume was used for all samples in the HPLC. ChromNav software (Jasco, Japan) was used to calculate the NP peak area. The HPLC analysis was performed with triplicate samples.
Pu 2089
Manufactured by Jasco
Sourced in Japan, United States, Italy
The PU-2089 is a laboratory equipment product designed for specialized applications. It is a versatile and precise instrument that performs core functions required in various research and testing environments. The details of its specific capabilities and intended use are not available for this unbiased, factual description.
Automatically generated - may contain errors
Lab products found in correlation
Uv 2070, by Jasco (7 mentions)
As 2055, by Jasco (5 mentions)
Uv 2075, by Jasco (5 mentions)
Md 2010, by Jasco (4 mentions)
As 2057, by Jasco (3 mentions)
Md 2018 plus, by Jasco (3 mentions)
Luna c18 column, by Phenomenex (3 mentions)
Co 2060 plus, by Jasco (3 mentions)
Chromnav software, by Jasco (2 mentions)
Fp 2020, by Jasco (2 mentions)
As 206, by Shimadzu (2 mentions)
Cellulose acetate membrane filter, by Advantec (2 mentions)
As 2059 plus, by Jasco (2 mentions)
Hplc dad fld system, by Jasco (2 mentions)
As 2057 plus, by Jasco (2 mentions)
Zorbax sb c18, by Agilent Technologies (2 mentions)
Co 4060, by Jasco (2 mentions)
Fp 2020 plus, by Jasco (2 mentions)
Marsxpress, by CEM Corporation (1 mentions)
Lambda 25 uv vis spectrometer, by PerkinElmer (1 mentions)
48 protocols using pu 2089
1
HPLC Analysis of Nanoparticle Concentration
2
Serum Stability Evaluation of A3 and A4 Derivatives
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Serum stability was performed as previously described 15 . Briefly, 10 mL of a 0.5 mM solution of A3 and A4 derivatives were incubated in human serum/TBS (4:1, v/v) at 37 C and the hydrolytic effects were analyzed by HPLC injecting 10 mL of solution every 15 min. The products were identified by comparison with authentic samples. The analysis were performed using an HPLC apparatus (Jasco PU-2089, equipped with an UV/Vis detector plus-2075) and using a RP18 column (Nucleosil-100, Macherey-Nagel, 250 mm  4.60 mm, 10 m) eluted with an increasing amount of acetonitrile in water (linear gradient from 0% to 100%, v/v, in 60 min, flow rate 0.5 mL/min, wavelength 254 nm).
3
Microwave-Assisted Solvent Extraction and Analysis
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Microwave-assisted solvent extraction (MASE) was performed on multimode microwave apparatus using a closed-vessel system (MARSX press, CEM Corporation, Matthews, NC, USA).
HPLC-UV/PAD-CD analyses were performed on a Jasco system (Jasco Europe S.r.l., Cremella, LC, Italy) equipped with a Jasco AS-2055 plus autosampler, a PU-2089 plus pump and a MD-2010 plus multi-wavelength detector coupled to a CD-2095 plus circular dichroism detector. The HPLC-ESI-MS analyses were carried out on ThermoFisher Scientific HPLC/UV/MS system (Thermo Scientific LCQ FLEET ion trap mass spectrometry) controlled by XcaliburTM Software Version 2.1 (Thermo Fisher Scientific, Waltham, MA, USA).
The absorbance was measured by a UV-Visible spectrophotometer (Lambda 25 UV/VIS spectrometer, Perkin Elmer Instruments, Waltham, MA, USA).
HPLC-UV/PAD-CD analyses were performed on a Jasco system (Jasco Europe S.r.l., Cremella, LC, Italy) equipped with a Jasco AS-2055 plus autosampler, a PU-2089 plus pump and a MD-2010 plus multi-wavelength detector coupled to a CD-2095 plus circular dichroism detector. The HPLC-ESI-MS analyses were carried out on ThermoFisher Scientific HPLC/UV/MS system (Thermo Scientific LCQ FLEET ion trap mass spectrometry) controlled by XcaliburTM Software Version 2.1 (Thermo Fisher Scientific, Waltham, MA, USA).
The absorbance was measured by a UV-Visible spectrophotometer (Lambda 25 UV/VIS spectrometer, Perkin Elmer Instruments, Waltham, MA, USA).
4
Extraction and HPLC Analysis of C. asiatica
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C. asiatica (1 g) was extracted with 10 mL of 20 % ethanol at 37 °C for 24 h. The extract was filtered with a 0.45-μm filter and analyzed by HPLC. HPLC analyses were performed on a LC-2000 series apparatus (Jasco) with a PU-2089 plus pump and a MD-2010 plus diode array detector, equipped with a LUNA C18 column (250 × 4.6 mm inner diameter; 5-μm particle size; Phenomenex, Torrance, CA, USA). The wavelength of diode array detector was set at 206 nm. The analytical method was based on previous study.17
5
In Vitro Drug Release Kinetics
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The dialysis bag was immersed in water for 8 h prior to the commencement of the study. The samples (RSV-L and RSV-GL; 1 ml drug-loaded suspension) were packed in the dialysis bag and ends were sealed appropriately. The bag was immersed in PBS (pH 7.4) and maintained at 37°C. The samples were collected at pre-determined time intervals and replaced with equal amounts of fresh buffer. The amount of drug released in the medium was calculated using a high performance liquid chromatography (HPLC) method. A 10 μl sample was used. A Jasco HPLC system (pump PU-2089, autosampler AS-2057 and LC-Net II/ADC controller; Jasco, Inc., Easton, MD, USA) was coupled to a fluorometric detector (Jasco FP-2020, λ excitation = 330 nm and λ emission = 374 nm). Synergi 4 μ HydroRP 80A and Luna 5 μ 100A C18 analytical columns (250×4.60 mm; Phenomenex, Inc., Torrance, CA, USA) were used. The mobile phase comprised of methanol: ACN: 0.1% phosphoric acid in water (60:10:30 v/v) with a flow rate of 1 ml/min. Ibuprofen was used as the internal standard, and injection volume was 20 μl with flow rate of 1 ml/min and isocratic flow was adapted.
6
HPLC Analysis of Pharmaceutical Compounds
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Briefly, the HPLC system consisted of pump (PU- 2089, Jasco, Japan), UV–Vis detector (UV-2075, Jasco, Japan), auto sampler (AS- 206, Japan) and 250 mm × 4.6 mm column C18 particle diameter 5 μm (Shimadzu, Japan). The solution used as the mobile phase is a mixture of acetonitrite and phosphate buffer (70: 30, pH 7.4), and the flow rate was adjusted to 1 ml/min. The mobile phase was filtered through 0.2 μm cellulose acetate membrane filter (Advantec, Japan). The volume of injection was 20 μl and the total run time was 8 min. The detection wavelength was 227 nm and the column temperature was maintained at 25 °C.
7
Polymer Characterization by NMR, SEC, MALDI-TOF
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1H and 13C NMR spectra were recorded at 25 °C on a Bruker model AC-500 spectrometer (Bruker, Billerica, MA, USA), operating at 500 and 125 MHz, respectively, where chemical shifts (δ in ppm) were determined with respect to non-deuterated solvent residues as internal standards. Analytical size exclusion chromatography (SEC) was performed in 0.2 mol·L−1 NaNO3 aqueous solution at 40 °C, using 7.8 mm × 300 mm gel columns (TOSOH TSKgel α–3000 × 3) on a JASCO model PU2089 (JASCO, Hachioji, Japan) equipped with a UV-2075 variable-wavelength UV-vis detector (JASCO) and an RI-2031 RI detector (JASCO). The number-average molecular weight (Mn) and polydispersity ratio (Mw/Mn) were calculated from the chromatographs with respect to poly(ethylene glycol)s standards (Scientific Polymer Products, Inc., Ontario, NY, USA); Mn = 590−11,900 g/mol, Mw/Mn = 1.05–1.11). Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry was performed on a BRUKER model AutoFlex III MALDI-TOF/TOF (Bruker) using 2,5-dihydroxybenzoic acid as a matrix. Fluorescence emission spectra were recorded on a JASCO Type FP-6500 spectrometer (JASCO).
8
Preparative HPLC Separation of Diastereomeric Plant Metabolites
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E‐7G‐4′S (395.7 ng) and E‐7S‐4′G (393.2 ng) were separated into the respective diastereomers by preparative HPLC. The HPLC apparatus used in this study was a Jasco 2000 plus system (Jasco, Tokyo, Japan) equipped with a model PU‐2089 gradient pump, CO‐2067 column oven, AS‐2059 auto sampler, and UV‐2075 UV‐visible detector. A Synergi Hydro‐RP column was employed (150 × 2.0 mm i.d., particle size 4 μm; Phenomenex, Torrance, CA) along with a guard cartridge (AQC 18, 4 × 2.1 mm i.d.). Each diastereomer mixture composed of E‐7G‐4′S or E‐7S‐4′G was eluted using a solvent system comprising 10 mmol/L ammonium acetate solution and acetonitrile (99:1). The flow rate was 0.4 mL/min at 45°C. The UV detection wavelength was set at 280 nm. Each diastereomer showing as 2 peaks for E‐7G‐4′S and E‐7S‐4′G on the HPLC chromatogram was divided into 2 fractions. The individual eluted solution from the preparative HPLC was evaporated. The 4 individual residues were divided into halves. One half of each fraction was used for enzymatic hydrolysis, while the other half was used for identification of plasma E‐7G‐4′S or E‐7S‐4′G.
9
Phytochemical Profiling of AERM by HPLC-PDA
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The chemical composition of AERM was investigated by high-performance liquid chromatography coupled to a Photodiode Array Detector (HPLC-PDA), using a Jasco (Tokyo, Japan) HPLC equipped with a PU-2089 quaternary solvent pump, a MD-2010 PAD, and an AS-2055 autosampler. The analytical column, maintained at room temperature (25 °C), was a Phenomenex Synergi Hydro RP18 (250 mm × 4.6 mm H × i.d.; 4 µm) with a Phenomenex security guard column (4.0 × 2.0 mm). Phenolic acids, flavonoids, flavan-3-ols, and PA were separated using the mobile phase of water (eluent A) and acetonitrile (eluent B), solvent A containing 0.1% formic acid, with the following gradient program: 5–50% B (30 min), 30–85% B (30–35 min), isocratic 85% B (45 min), 85–100% B (45–70 min), return to 5% B (2 min), and the column was re-equilibrated under the initial conditions for 18 min before the next injection. The flow rate was 1.0 mL·min−1, and the total run time was 70 min. EZChrom Elite Data System software (Chromatec, Idstein, Germany) was used for detection operation and data processing. Compounds were identified by comparing retention times and UV spectral analyses.
10
Quantification of Insulin-Loaded Nanoparticles
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The encapsulation efficiency (EE) and drug loading capacity (DLC) of the carrier were determined using previously reported methods (Franklin et al., 2018) with slight modifications. Herein, a sample of freshly prepared insulin-loaded NPs (20 mg) was dispersed in 5 mL of phosphate buffer solution (PBS, pH 7.4) and shaken for 5 min to dissolve the free drug. The resulting dispersion was centrifuged for 30 min at 20,000 rpm (Kubota Co, Japan). The amount of insulin in the supernatant after centrifugation was measured by HPLC analysis as previously reported (Mumuni et al., 2015) with slight modifications. Briefly, the chromatographic system consisted of a pump (PU-2089, Jasco, Japan), a UV-Vis detector (UV-2075, Jasco, Japan), an auto sampler (AS-206, Japan) and a column C18 (250 mm x 4.6 mm; particle diameter 5 µm) (Shimadzu, Japan). A mixture of acetonitrite and phosphate buffer (70:30, pH 7.4) was employed as the mobile phase and the flow rate was adjusted to 1 mL/min. The mobile phase was filtered through 0.2 µm cellulose acetate membrane filter (Advantec, Japan) The volume of injection was 20 µL and the total run time was 8 min. The detection wavelength was 227 nm and the column temperature was maintained at 25 °C.
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