An aliquot of honey (5 g) was rigorously weighted and mixed with deionised water (50.0 mL). The mixture was filtered through a PTFE membrane (0.22 µm) before HPLC analysis. The integrated HPLC system used (Jasco, Tokyo, Japan) was composed by an AS-950 automated injector, a PU-980 pump, and a MD-2010 Plus multiwavelength diode-array detector (DAD). The chromatographic separation was achieved with a RP-Tracer Excel ODS-A column (5 µm; 250 mm × 4 mm), from Teknokroma (Barcelona, Spain), using an isocratic solvent system of water: acetonitrile (80:20, v/v). Elution was performed at a solvent flow rate of 1 mL/min. Chromatograms were recorded at 285 nm. Chromatographic data was analysed using a Borwin-PDA Controller Software (JMBS, Le Fontanil, France).
Pu 980
Manufactured by Jasco
Sourced in Japan, Italy
The PU-980 is a laboratory equipment designed for general scientific applications. It serves as a precision analytical instrument for various measurement tasks. The core function of the PU-980 is to provide accurate and reliable data for research and analysis purposes.
Automatically generated - may contain errors
Lab products found in correlation
Dg 980 50, by Jasco (6 mentions)
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As 950, by Jasco (3 mentions)
Avance 3 400 system, by Bruker (3 mentions)
Voyager pro sf, by Thermo Fisher Scientific (3 mentions)
Hg 980 31, by Jasco (3 mentions)
Ri 1530, by Jasco (3 mentions)
Md 2010 plus, by Jasco (2 mentions)
Uv 970 detector, by Jasco (2 mentions)
As 2055, by Jasco (2 mentions)
Superose 6 hr 10 300 column, by GE Healthcare (2 mentions)
Uv 975 uv vis detector, by Jasco (2 mentions)
Dg 2080 53, by Jasco (2 mentions)
As 1555, by Jasco (2 mentions)
Ohpak sb 804 hq column, by Resonac (2 mentions)
Uv 2077, by Jasco (2 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Integrated hplc system, by Jasco (1 mentions)
Ecs 400, by JEOL (1 mentions)
34 protocols using pu 980
1
HPLC Analysis of Honey Composition
2
HPLC Analysis of Clomipramine Hydrochloride
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We used an HPLC system (JASCO Corporation, Tokyo, Japan) consisting of a model PU‐980 pump, AS‐950 injector, LCSS‐905 system controller, and UV‐970 detector set at 270 nm. Clomipramine hydrochloride and DMCMI were separated with a COSMOSIL 5C18‐AR‐II Paced column (4.6 × 250 mm; Nacalai Tesque, Kyoto, Japan) using a mobile phase consisting of 50% acetonitrile, 50% water, 0.1% trifluoroacetic acid, and 0.01% triethylamine. The mobile phase flow rate was set at 1.0 mL/min.
3
NMR Spectroscopy Characterization Protocol
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NMR spectra were recorded on a JEOL ecs 400, ecz 400, ecz 600, eca 600 spectrometer. Chemical shifts in CDCl3, acetone-d6, DMSO-d6 or CD3OD were reported downfield from tetramethylsilane (TMS) (= 0 ppm) or solvent signal [acetone-d6 (= 2.04 ppm), DMSO-d6 (= 2.49 ppm) or CD3OD ( = 3.30 ppm)] for 1H NMR. Data are reported as follows: chemical shift, multiplicity (s = singlet, d = doublet, t = triplet, m = multiplet, and br = broad), integration and coupling constants in Hz. For 13C NMR, chemical shifts were reported in the scale relative to the solvent signal [CHCl3 (77.0 ppm), acetone-d6 (29.8 ppm), DMSO-d6 (39.5 ppm) or CD3OD (49.0 ppm)] as an internal reference. ESI mass spectra were measured on JEOL AccuTOF LC-plus JMS-T100LP. Optical rotations were measured on a JASCO P-1020 polarimeter. The enantiomeric excess (ee) was determined by HPLC analysis. HPLC was performed on JASCO HPLC systems consisting of the following: pump, PU-980; detector, UV-970; column DAICEL CHIRALPAK OD-3; mobile phase, n-hexane/i-PrOH. Analytical thin layer chromatography was performed on Kieselgel 60F254, 0.25 mm thickness plates. Column chromatography was performed with silica gel 60 N (spherical, neutral 63-210 mesh). Reactions were conducted in dry solvent. Other reagents were purified by the usual methods.
4
Serum Peptide Separation by RP-HPLC
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Serum peptides were separated by RP-HPLC comprising a UV-970, LG-980-2, PU-980, and DG-980-50 (JASCO, Tokyo, Japan) and a C18 column (Capcell pak C18 UG 80, 3.0 x 250 mm; Shiseido, Tokyo, Japan). A linear gradient of 0%–90% acetonitrile in the presence of 0.1% TFA was applied at a flow rate of 0.5 mL/min with monitoring at 215 nm. The peptides were collected by a FRC10 fraction collector (Shimadzu, Kyoto, Japan).
5
Characterization of Synthetic Oligonucleotides
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1H nuclear magnetic resonance (NMR) spectra were recorded on a AVANCE III 400 system (Bruker, Billerica, MA, USA). Mass spectra were recorded on a Voyager PRO-SF, (Applied Biosystems, Foster City, CA, USA). HPLC was performed on a Chemcosorb 5-ODS-H column with JASCO PU-980, HG-980-31, DG-980-50 system equipped with a JASCO UV 970 detector (JASCO, Tokyo, Japan) at 260 nm. Reagents for DNA synthesis such as A, G, C, T-β-cyanoethyl phosphoramidite and CPG support were purchased form Glen Research (Sterling, VA, USA).
6
Purification and Characterization of SOR
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Middle pressure chromatography or fast protein liquid chromatography FPLC was performed using ÄKTA FPLC system (GE healthcare bioscience, Uppsala, Sweden). HPLC was conducted on a system comprised of high pressure gradient pumps (PU-980) and photodiode array detector 4015 (JASCO, Tokyo, Japan). Matrix-assisted laser desorption (MALDI) time-of-flight (TOF) mass spectrum was obtained on AXIMA-CFR plus (Shimadzu, Kyoto, Japan) using sinapinic acid as a matrix. Amino acid sequences were analyzed by automated Edman degradation using a gas-phase PPSQ-21A sequencer (Shimadzu). Ultrafree-MC (30,000 NMWL Filter unit, Merck KGaA, Darmstadt, German) was used for ultrafiltration of sample. Cellulose tube (MW 15,000 cut) was used for dialysis. Protein concentrations were determined by BCA protein assay kit (Thermo Fisher Scientific, Waltham, MA, USA) with γ-globulin as a standard. To determine the concentration of purified SOR for biological evaluation, known concentrations of bovine serum albumin (BSA) was stained on SDS-PAGE by Coomassie Brilliant Blue. The bands with different concentration were scanned and the image was quantified by using ImageJ software to obtain a standard curve. The concentration of SOR was estimated by using the curve.
7
Quantification of Fucoxanthin in Emulsions
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The concentrations of fucoxanthin in the emulsions
and bulk oil were quantified using HPLC (JASCO International Co.,
Tokyo, Japan) equipped with an AS-2055 autosampler, a PU-980 pump
system, and a UV-970 UV–vis spectrophotometric detector. A
C-18 reversed phase column (as stationary phase, 4.6 × 250 mm;
Shimpack VP-ODS, Japan) was used with the temperature set at 25 °C.
Fucoxanthin was extracted from emulsions and the dispersed phase prior
to the HPLC analysis using a solvent extraction method: 200 μL
of emulsion or a drop of dispersed phase (mass was analyzed) from
the middle of the glass test tube was diluted to 10 mL with methanol
in a volumetric flask to extract fucoxanthin and then ultrasonicated
for 5 min. The samples were filtered using PTFE syringe filters (0.45
μm) and transferred to 2 mL HPLC vials; 20 μL of the filtered
samples from HPLC vials were injected into the HPLC system. The mobile
phase consisted of 10 wt % of Milli-Q water and 90 wt % of methanol.
The mobile phase flow rate was set at 1 mL/min. UV detection of fucoxanthin
was monitored at 450 nm. Fucoxanthin concentration in samples was
calculated using a standard curve (R2 =
0.9995) and all of the analyses were repeated three times.
and bulk oil were quantified using HPLC (JASCO International Co.,
Tokyo, Japan) equipped with an AS-2055 autosampler, a PU-980 pump
system, and a UV-970 UV–vis spectrophotometric detector. A
C-18 reversed phase column (as stationary phase, 4.6 × 250 mm;
Shimpack VP-ODS, Japan) was used with the temperature set at 25 °C.
Fucoxanthin was extracted from emulsions and the dispersed phase prior
to the HPLC analysis using a solvent extraction method: 200 μL
of emulsion or a drop of dispersed phase (mass was analyzed) from
the middle of the glass test tube was diluted to 10 mL with methanol
in a volumetric flask to extract fucoxanthin and then ultrasonicated
for 5 min. The samples were filtered using PTFE syringe filters (0.45
μm) and transferred to 2 mL HPLC vials; 20 μL of the filtered
samples from HPLC vials were injected into the HPLC system. The mobile
phase consisted of 10 wt % of Milli-Q water and 90 wt % of methanol.
The mobile phase flow rate was set at 1 mL/min. UV detection of fucoxanthin
was monitored at 450 nm. Fucoxanthin concentration in samples was
calculated using a standard curve (R2 =
0.9995) and all of the analyses were repeated three times.
8
Quantifying Serum Biomarkers by LC-MS and HPLC
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To measure IS and TMAO, serum samples were deproteinized with an ethanol solution containing an internal standard and analyzed using liquid chromatography–mass spectrometry (LC–MS) [11 (link)]. The internal standards were IS-d4, IS, and TMAO-d9, TMAO. A high-performance LC (HPLC) system (Acquity UPLC®, Waters Corp., Milford, MA, USA) and a mass spectrometer (LTQ-Orbitrap, Thermo Fisher Scientific GmbH, Bremen, Germany) were used.
To measure PCS, serum samples were deproteinized with methanol and analyzed using HPLC [12 (link)]. An HPLC system equipped with an autosampler (AS-950, JASCO Corp., Tokyo, Japan), a pump (PU-980, JASCO Corp., Tokyo, Japan), and a fluorescence detector (RF-10A, Shimadzu Corp., Kyoto, Japan) was used.
To measure PCS, serum samples were deproteinized with methanol and analyzed using HPLC [12 (link)]. An HPLC system equipped with an autosampler (AS-950, JASCO Corp., Tokyo, Japan), a pump (PU-980, JASCO Corp., Tokyo, Japan), and a fluorescence detector (RF-10A, Shimadzu Corp., Kyoto, Japan) was used.
9
Phenolic Compounds Analysis in Ginseng Seed Oil
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The phenolic compounds in ginseng seed oil were analyzed with high-performance liquid chromatography (PU-980; Jasco, Tokyo, Japan) under the following analytical conditions: a Waters C-18 column (5.0 μm, 4.6 mm × 250 mm; Waters, Milford, MA, USA) was used, with a mobile phase of water with 2% acetic acid (solvent A) and 50% acetonitrile with 0.5% acetic acid (solvent B) or (solvent A) with 2% acetic acid and 50% acetonitrile (solvent B) with 0.5% acetic acid; samples were developed from an initial 100% of A solvent to a 45% gradient after 70 min with a speed of 0.8 mL/min for 80 min. The sample was detected at 280 nm with a UV detector (MD-2010; Jasco) using a 20-μL sample for injection. Each 2 g sample was dissolved in 10 mL n-hexane, and 20 mL of 80% methanol was added to extract the phenolic compounds. Finally, 10 mL of n-hexane was added to the extract to eliminate the remaining lipid constituents, and the solvent in the 80% methanol layer was evaporated completely using a vacuum evaporator. The concentrated extracts were diluted with methanol to 10 mg/mL, and filtered through a 0.45-μm syringe filter (Whatman) for further analysis.
10
Lipoprotein Fractionation by FPLC
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Rat plasma samples were fractionated by fast protein liquid chromatography (FPLC) as previously described (Gerdes et al., 1992 (link)) with minor modifications in order to investigate the changes in lipoprotein particles (VLDL, LDL, and HDL). In brief, the system contained a PU-980 ternary pump with an LG-980-02 linear degasser and a UV-975 UV/VIS detector (Jasco). EDTA plasma was diluted 1:1 with tris-buffered saline, and 300 μl sample/buffer mixture was loaded onto a Superose 6 HR 10/300 column (GE Healthcare, Life Sciences Division) for lipoprotein separation at a flow rate of 0.5 ml/min. Quantitative analysis of the chromatograms was performed with ChromNav chromatographic software, version 1.0 (Jasco). The plots for individual fast-performance liquid chromatography profiles were generated using GraphPad version 8.0.1.
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