For the assessment of the saccharin contents, aliquots of each sample were analyzed via the HPLC method that is described by Weinert et al. [18 (link)]. The HPLC system used consists of the Jasco liquid chromatography (Jasco Co., Easton, MD, USA) equipped with the Jasco PU-980 pump, an UV-Vis detector set to 254 nm, and the Rheodyne 20-µL injector. The Phenomenex LC-18 analytical column with the parameters of 250 mm × 4.6 mm (i.d.) (Phenomenex Co., Torrance, CA, USA) was used with packing material of a 5-µm particle size. The mobile-phase consisted of a mixture of 20% (v/v) reagent-grade glacial acetic acid in water, which was buffered to a pH 3.0 using a saturated sodium acetate solution. Before the injection, the samples were filtered through the Millex-HV 0.45-µm membrane filter (Millipore Co., Temecula, CA, USA) and this was followed by an ultra-sonication. A calibration curve was also prepared using a standard concentration between 50 and 600 mg/L.
Pu 980 pump
Manufactured by Jasco
Sourced in Japan, United States
The PU-980 pump is a laboratory equipment designed for fluid handling applications. It is a positive displacement pump that can precisely control the flow rate of liquids. The pump features a compact design and is suitable for a variety of laboratory settings.
Automatically generated - may contain errors
Lab products found in correlation
Hplc system, by Jasco (6 mentions)
Uv 975, by Jasco (4 mentions)
Jms 700 instrument, by JEOL (2 mentions)
Uv 970, by Jasco (2 mentions)
Ftir 8400s instrument, by Shimadzu (2 mentions)
600 or 500 hz nmr spectrometer, by Bruker (2 mentions)
P 2300 polarimeter, by Jasco (2 mentions)
Kieselgel 60 f254, by Merck Group (2 mentions)
Ri 2031 plus refractive index detector, by Jasco (2 mentions)
Au 950 autosampler, by Jasco (2 mentions)
Dg 2080 53 degasser, by Jasco (2 mentions)
Co 965 column oven, by Jasco (2 mentions)
Spectrum 100 ftir spectrometer, by PerkinElmer (2 mentions)
Uv 975 detector, by Jasco (2 mentions)
Integrated hplc system, by Jasco (2 mentions)
As 4150 rhplc autosampler, by Jasco (2 mentions)
Md 2015 plus, by Jasco (2 mentions)
Dg 980 50 degasser, by Jasco (1 mentions)
Jnm ecz400 spectrometer, by JEOL (1 mentions)
Uv 2075 plus detector, by Jasco (1 mentions)
30 protocols using pu 980 pump
1
HPLC Analysis of Saccharin Content
2
NMR and GPC Analysis of Polymers
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1 H NMR spectra were recorded on a JNM-ECZ400 spectrometer (JEOL, Tokyo, Japan) using CDCl 3 , d 6 -DMSO, or D 2 O as a solvent. Gel permeation chromatography (GPC) with organic solvent was performed on a HLC-8320 GPC Eco-SEC system equipped with a TSKgel Super AW guard column and TSKgel Super AW (4000, 3000, and 2500) columns (Tosoh, Tokyo, Japan). GPC with water solvent was performed on a JASCO DG-980-50 degasser equipped with a JASCO PU-980 pump (JASCO Co., Tokyo, Japan), a Shodex OHpak SB-G guard column, a Shodex OHpak LB-806 HQ column (Showa Denko, Tokyo, Japan), a JASCO RI-2031 Plus RI detector. GPC analyses were performed by injecting 20 μL of a polymer solution (1 g L -1 ) in DMF buffer containing LiBr (10 mM) or NaNO 3 aqueous solution (100 mM). The buffer solution was also used as the eluent at a flow rate of 0.5 mL min -1 . The GPC system was calibrated using a poly(methyl methacrylate) standard(Shodex) for organic solvent GPC and pullulan standard (Shodex) for aqueous GPC.
3
Quantitative Analysis of Chiang Mai Propolis
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The quantification of the compounds with antibiofilm activity was performed by analytical high-performance liquid chromatography (HPLC). The analytical HPLC was carried out using an instrument equipped with a Jasco PU-980 pump (Jasco Co., Inc.), a UV-2075 Plus detector (Jasco), and a Capcell Pak C18 reversed-phase column (5 µm, φ 4.6 × 250 mm; Osaka Soda). The 70% EtOH extract of the Chiang Mai propolis was analyzed using the following gradient program, with MeCN/H 2 O (in 0.1% trifluoroacetic acid) = 30:70 (0 min)-100:0 (80 min)-100:0 (90 min). The detection wavelength was set at 320 nm to detect the target compounds. Abyssinone V (1) isolated from Chiang Mai propolis was used as the standard to obtain a calibration curve for the compounds. The LOD and LOQ were determined at signal-to-noise ratios of 3 and 10, respectively. The 70% EtOH extract of the Chiang Mai propolis was analyzed 3 times, and the standard deviation was calculated. To correct for the compound loss occurring during the sample preparation, a spike test was conducted using 1. The recovery rate was calculated from the area value of the chromatogram obtained by HPLC (n = 3) and the amount of 1 added.
4
Quantification of Brain Monoamines
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Dissected rat brain tissues (SNc, striatum and PC) were weighed and homogenized in appropriate volumes of homogenizing solution (0.1 M perchloric acid containing 400 mM sodium metabisulphite) and then microcentrifuged. Determination of DA and NA levels was performed with a Jasco PU-980 pump HPLC coupled to electrochemical detection (Coulochem II-ESA model 5011 analytical cell), equipped with a Capital Hypersil column (250 mm x 4.6 mm id 5 μm) using a mobile phase (0.01 M sodium dihydrogen orthophosphate dehydrate, 0.9 mM 1-octanesulfonic acid sodium salt, 0.1% dibutylamine, 12.5% methanol, pH 3.2) at a flow rate of 1 ml min-1. Concentrations of the monoamines were expressed as ng/g ± SEM and were calculated by reference to an internal standard, dihydroxybenzylamine (DHBA – 0.5 ng). Data were captured using Antec’s Scientific Clarity software.
5
Determination of Cellular Lipid Peroxidation
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Lipid peroxide content was determined in the form of malondialdehyde (MDA) as a thiobarbituric acid reactive substance (TBARS). In brief, an aliquot of cell lysates was pipetted into a test tube containing 0.6 mL of 0.44 M phosphoric acid. After mixing, 0.2 mL of a 42 mM thiobarbituric acid solution was placed in a 95°C dry bath for 1 h. The samples were then neutralized with 1 N NaOH in methanol before a high-performance liquid chromatography (HPLC) analysis was conducted. An aliquot of 20 μL of a supernatant was injected into a C18 column (4.6 × 250 mm, with a particle size of 5 μm) by using a Jasco PU-980 pump (Easton, MD, USA) with a solvent system composed of methanol and a 50 mM phosphate buffer (pH 6.8) (4 : 6, v/v) at a flow rate of 1 mL/min. The eluent was monitored by fluorescence detector at 525 nm excitation wavelength and 550 nm emission wavelength.
6
Chromatographic Profiling of Polyphenols
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P90, E90, E80, and E70 chromatographic profiles were obtained with an HPLC system (Jasco, Tokyo, Japan) equipped with a PU‐980 pump, an UV‐1575 detector, and a degasser Populaire DP4003. Data were obtained and processed with PowerChrom 2.5 (eDaq Technologies, NZ, Australia) software and MATLAB R2019b software (MathWorks Inc., USA). Chromatographic separation was performed on a Kromasil C‐18 semi‐preparative column (8 × 250 mm) at 30°C. A gradient elution using water (A) and methanol (B) consisted of 0–3 min (99% A, 1% B), 30 min (5% A, 95% B), and 40 min (99% A, 1% B). Freeze‐dried samples (2 mg/ml) were diluted in EtOH 20% (v/v) and filtered through 0.45 µm syringe filters. Injection volume was 50 µl, detection wavelength 266 nm, and solvent flow rate 0.4 ml/min. Polyphenols content was calculated with standard molecules of phloroglucinol and resorcinol; additionally, glucose and glucuronic acid standards were injected to check for possible interference of UA and sugars (Gonçalves‐Fernández et al., 2019 (link)). To ensure the reproducibility of the assays, a minimum of four injections of each extract were carried out.
7
Quantification of Tocopherols in Wheat Germ Oil
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Tocopherols (α, β, γ and δ) content of WGO was quantified according to the method of Ng, Lean-Teik [27 (link)] with some modifications. Briefly, 1.0 g of WGO sample was dissolved in 10 mL hexane, filtered (0.45 μm) and injected (20 μL) into a HPLC system (MD-910 multi wavelength detector, PU-980 pump, DG-980-50-3 line degasser, and LG-980-02 ternary gradient unit, Jasco, Japan) with a silica gel column (Nucleosil Si, 5 μm i.d., 200 × 4mm). The mobile phase was n-hexane/ isopropanol (98.5:1.5, v/v) and the flow rate was 1.0 mL/min. Individual tocopherols quantified as mg/kg of oil with corresponding external standards.
8
Biophysical Characterization of Organic Compounds
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A JASCO P-2300 polarimeter (Tokyo, Japan) and shimazu FTIR-8400S instrument (Columbia, MD 21046, USA) was operated for optical rotation and IR spectra, respectively. Then, 1D and 2D NMR spectra were recorded on a Bruker 600 or 500 Hz NMR spectrometer (MA, USA). HR-MS spectra were obtained on a JEOL JMS-700 instrument (Tokyo, Japan). For Chromatography: column chromatography (CC) [silica gel 60 (Merck, 230–400 mesh, Merck, Darmstadt, Germany)]; TLC analysis: [precoated silica gel plates (Merck, Kieselgel 60 F254, 0.25 mm, Merck, Darmstadt, Germany]. High-performance liquid chromatography (HPLC) equipped with a Jasco PU-980 pump, a Jasco UV-970 intelligent UV/VIS detector at 210 nm and a semi preparative reversed-phase column (Cosmosil C18 column 250 × 10 mm, 5 μm).
9
Analytical Characterization of Organic Compounds
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For optical rotation the JASCO P-2300 polarimeter (JASCO, Tokyo, Japan) was used; for IR spectra, the Shimazu FTIR-8400S instrument (Shimazu, Columbia, MD 21046, USA) was used. A Bruker 600 or 500 Hz NMR spectrometer was used to record 1D and 2D NMR spectra (Bruker, MA, USA). A JEOL JMS-700 equipment was used to obtain HR-MS spectra (Tokyo, Japan). TLC analysis was conducted with precoated silica gel plates (Merck, Kieselgel60 F254, 0.25 mm, Merck, Darmstadt, Germany); chromatography (CC) was conducted with silica gel 60 (Merck, 230–400 mesh, Merck, Darmstadt, Germany). HPLC was carried out with a Jasco PU-980 pump, a Jasco UV-970 intelligent UV detector at 210 nm, and a semi-preparative reversed-phase column (Cosmosil C18, column 250 × 10 mm, 5 μm, Nacalai Tesque, Kyoto, Japan).
10
Spectroscopic Characterization of Cellulose
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The 1H-NMR spectra of the products were measured with a Varian Gemini-2000 (400 MHz) or a Bruker (500 MHz) spectrometer (Bruker, Billerica, MA, USA) in pyridine-d5 at 80 °C. IR spectra were obtained with a JASCO FT/IR-620 spectrophotometer (JASCO, Tokyo, Japan). The CD spectra of 3,5-dimethylphenylcarbamates of cellulose were measured on a JASCO J-720 spectrophotometer using a quartz cell with a path length of 0.10 mm. The concentrations of samples in THF were 3.3–4.6 mg/5 mL. Optical rotation was measured with a JASCO P-1030 polarimeter. Thermal analysis of products was performed on a SEIKO SSC/5200TG instrument. The molecular weights (Mn) and its distribution (Mw/Mn) or DPs of the celluloses were estimated as the tris(3,5-dimethylphenylcarbamate)s by SEC on Shodex GPC system-21 at 40 °C using polystyrene standards and tetrahydrofuran (THF) as solvent. Two SEC columns, Shodex KF-806 and KF-803, were used in a series. For the samples with DP 40 and 52, the SEC on Waters 600 GPC system was used with three SEC columns, Styragel HR 0.5, HR 3 and HR 4. The HPLC analysis of racemates was carried out with a JASCO HPLC system equipped with a PU-980 pump and a JASCO OR-990 polarimetric detector or a JASCO CD-2095 detector.
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