1H NMR spectra were recorded on a Bruker AVANCE 300 MHz or AVANCE II 600 MHz and the spectra were recorded at 300 K. Mass spectra were taken on a Bruker microTOF or amazon SL mass spectrometer. IR spectra were recorded on a JASCO FT/IR-4600 spectrometer. Gel permeation chromatography (GPC) measurements were conducted using a system consisting of a JASCO PU-2089 pump, a CO-2065 column oven, an RI-2031 refractive index detector, and a Shodex KD-804 (8.0 mm × 300 mm) column. DMF containing 10 mM LiBr was used as the eluent at a flow rate of 0.5 mL min−1 at 50 °C. Poly(methyl methacrylate) samples were used as standards. Transmittance was recorded using a JASCO V-550 UV-vis spectrometer. A 0.5 wt % polymer aqueous solution was filtered through a membrane filter (0.45 µm), then the transmittance of the sample solution was measured in a quartz cell (cell length: 10 mm) at 500 nm while heating at a rate of 0.5 °C min−1.
Pu 2089 pump
Manufactured by Jasco
Sourced in Japan
The PU-2089 pump is a laboratory-grade peristaltic pump designed for fluid transfer applications. It features a compact and durable construction, allowing for precise and reliable operation. The pump's core function is to precisely control the flow rate of liquids through the use of a rotating rotor and removable tubing.
Automatically generated - may contain errors
Lab products found in correlation
Hplc system, by Jasco (3 mentions)
Co 2065 column oven, by Jasco (3 mentions)
Fp 2020 fluorescence detector, by Jasco (2 mentions)
As 2057 automated injector, by Jasco (2 mentions)
Md 2018 multiwavelength diode array detector dad, by Jasco (2 mentions)
V 550 uv vis spectrometer, by Jasco (1 mentions)
Avance 2 600 mhz, by Bruker (1 mentions)
Ft ir 4600 spectrometer, by Jasco (1 mentions)
Avance 300 mhz, by Bruker (1 mentions)
Biospin av 300 spectrometer, by Bruker (1 mentions)
Fp 6500 spectrometer, by Jasco (1 mentions)
U 2000 spectrometer, by Hitachi (1 mentions)
Microtof q 3 spectrometer, by Bruker (1 mentions)
Ri 2031 refractive index detector, by Jasco (1 mentions)
Fp 6500 fluorometer, by Jasco (1 mentions)
Av 600 spectrometer, by Bruker (1 mentions)
5c18 ms 2 column, by Nacalai Tesque (1 mentions)
Ohpak sb 804 hq column, by Resonac (1 mentions)
Hplc grade, by Merck Group (1 mentions)
Supelcosil lc si column, by Merck Group (1 mentions)
12 protocols using pu 2089 pump
1
Characterization of Polymer Materials
2
Microbial Growth and Galactose Quantification
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Cell growth was measured at 600 nm by spectrophotometer (Eppendorf, Germany). Cell culture samples were centrifuged at 13,000 rpm for 10 min, and the supernatant was then conserved for additional research. The concentration of galactose was determined using an HPLC system (Jasco, France) consisting a PU-2089 pump, AS-2057 auto-injector, and RI-2031 differential refractive index (RI) detector (JASCO, France). Aminex HPX 87-H column (Bio-Rad, Richmond, USA) was equipped and 5 mM H2SO4 (0.5 mL/min) was used as the mobile phase at 60 °C.
3
Physicochemical Characterization of Polymer Materials
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NMR spectra were recorded using a Bruker (MA, USA) BioSpin AV-300 spectrometer. Gel permeation chromatography (GPC) measurements were conducted using a system consisting of a JASCO (Tokyo, Japan) PU-2089 pump, a JASCO CO-2065 column oven, a JASCO RI-2031 refractive index detector, and a Shodex OHpak SB-804 HQ column (8.0 × 300 mm, SHOWA DENKO, Tokyo, Japan). A phosphate buffer (20 mM, pH 7.0) containing 20% N,N-dimethylformamide was used as the eluent at a flow rate of 0.5 mL/min at 30 °C. Pullulan samples were used as standards. Transmittance was recorded using a Hitachi (Tokyo, Japan) U-2000 spectrometer. Dynamic light scattering (DLS) analyses were conducted using an Otsuka Electronics (Osaka, Japan) ELSZ-1000 at 667 nm. The samples for transmittance and DLS analyses were prepared using 0.5 wt % in aqueous media and filtered through a membrane filter (0.45 μm). The fluorescence intensity was recorded using a JASCO FP-6500 spectrometer.
4
Analytical Characterization of Compounds
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The NMR spectra were recorded using Bruker BioSpin AV-300 and AV-600 spectrometers. The ESI-MS spectra were recorded using a Bruker Daltonics micrOTOF Q-III spectrometer (Bruker Daltonics, Billerica, MA, USA). The HPLC and GPC measurements were conducted using a system consisting of a JASCO PU-2089 pump and a JASCO CO-2065 column oven (JASCO Corporation, Tokyo, Japan). A JASCO UV-2075 ultraviolet detector and a JASCO RI-2031 refractive index detector were used for the HPLC and GPC analyses, respectively. A 5C18-MS-II column (ɸ4.6 × 250 mm, Nacalai Tesque, INC.) was used for the HPLC analysis. 5, 4, 3, and 10 % MeCN-containing water were used as the eluents at a flow rate of 1.0 mL/min at 30 °C to analyze the enzymatic reaction with 1a – d , respectively. A Shodex OHpak SB-804 HQ column (ɸ8.0 × 300 mm, Showa Denko K.K., Tokyo, Japan) was used for the GPC analysis of 3a – c using a phosphate buffer (20 mM, pH 7.0) as the eluent at a flow rate of 0.5 mL/min at 30 °C. Pullulan samples were used as standards. A Shodex KD-804 column (ɸ8.0 × 300 mm, Showa Denko K.K.) was used for the GPC analysis of 3d using N , N -dimethylformamide (DMF) containing 10 mM lithium bromide as the eluent at a flow rate of 0.5 mL/min at 50 °C. Poly(methylmethacrylate) samples were used as standards. The fluorescence intensity was recorded using a JASCO FP-6500 fluorometer for the lectin binding tests.
5
Vitamin E Analysis in Oils by HPLC
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Analogously, vitamin E values were determined by HPLC analysis in oil samples according to Alves et al. [24 (link)]. Briefly, about 20 mg of oil was diluted in 1 mL of n-hexane (HPLC-grade, Merck, Darmstadt, Germany), where 20 µg/mL of tocol was added as internal standard. Then, 20 µL was injected to perform the separation on a normal-phase SupelcosilTM LC-SI column (3 µm; 75 × 3.0 mm; Supelco, Bellefonte, PA, USA). The used equipment was an HPLC system (Jasco, Tokyo, Japan) equipped with an AS-2057 automated injector, a PU-2089 pump, and an MD-2018 multiwavelength diode array detector (DAD) coupled with an FP-2020 fluorescence detector (Jasco, Japan). They were programmed for excitation at 290 nm and emission at 330 nm. Lastly, the identification of the compounds was accomplished by a comparison with commercial standards. Analyses were performed in duplicate, and results are expressed as mg/kg of oil.
6
Quantitative Analysis of Phenolic Compounds
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The column C18 (5 µm × 0.46 cm × 25 cm) connected to the HPLC (JASCO, Tokyo, Japan) system with a LC-Net II/ADC controller, a PU-2089 pump and an UV-2075 detector was used for quantifying the concentrations of PA and VA. The leaf and root extracts in methanol (1 mg/mL) were filtered, and a volume of 5 µL was injected. A gradient elution of absolute methanol (A) and 0.1% acid acetic (B) was run at 1 mL/min speed with a linear increase of A from 5–10% for 5 min, then 10–90% for 45 min, and 10 min cleaning step with only solvent A. The phenolic standard curves were established from different concentrations from 0 to 100 µg/mL for quantification of identified phenolics in the samples.
7
HPLC Analysis of Vitamin D Metabolites
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Analysis of 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 was performed with high-performance liquid chromatography (HPLC) according to a method previously described.34 (link) The HPLC system consisted of a JASCO PU-2089 pump and a JASCO UV-975 detector set on the wavelength 265 nm (both from Japan Spectroscopic Company, Tokyo, Japan). The column was a Grace Smart RP 18 (100 × 2.1 mm, 3 µm), and the mobile phase consisted of methanol:water (80:20, v/v) and the flow rate was 0.4 mL/min. Twenty microliters of the samples were injected, and the concentration was determined based on a standard curve that was prepared by an in-house reference. Standards of 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 were purchased from Sigma-Aldrich Chemie GmbH (Steinheim, Germany). Together with standards with known concentration, a reference was analyzed together with the samples. The inter-assay and intra-assay coefficient of variation was 3.9% and 5.7%, respectively. As a quality control, we used plasma references from Vitamin D External Quality Control Scheme (DEQAS; http://www.deqas.org ). Because sampling was performed on different occasions over the year, the concentrations of vitamin D were adjusted for what time of the year it was drawn against a standard curve, obtained from results of a British study of 7437 individuals.35 (link)
8
HPLC Quantification of Tomatidine
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Tomatidine was purchased from Sigma-Aldrich (St. Louis, MO, USA). Standard stock solution was prepared at a concentration of 2 mg/mL in 80% methanol. The standard solution was serially diluted with 80% methanol to obtain calibration standard solutions at concentrations of 6.25, 12.5, 25, 50, 100, and 200 µg/mL. An HPLC apparatus equipped with a PU-2089 pump, UV-2075 spectrophotometer, and CO-2065 column oven (Jasco, Tokyo, Japan) was used. Chromatographic separation was achieved using an XTerra RP 18 column (4.6 × 150 mm, 5 µm; Waters, Milford, MA, USA) with the column oven temperature maintained at 35 °C. The mobile phase consisted of acetonitrile (Solvent A) and 25 mM triethylammonium phosphate in water (pH 3.0; Solvent B). The mobile phase flow rate was 0.8 mL/min with gradient elution. The initial percentage composition of Solvent A was 20%. It was gradually increased to 45% for 12 min, to 55% for 5 min, and to 57% for 3 min, followed by equilibration to the initial composition for 5 min. All solvents for HPLC analysis were of HPLC grade and were purchased from J.T. Baker (Phillipsburg, NJ, USA).
9
HPLC-MALDI-TOF MS Peptide Analysis
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A Centriprep YM-3 filter device was purchased from Millipore (Billerica, MA, USA). Acetonitrile (ACN), trifluoroacetic acid (TFA), and protease inhibitor cocktail for mammals were purchased from Sigma–Aldrich (St. Louis, MI, USA). Buffer solutions for the RP-HPLC were filtered through a 0.2 μm membrane filter from Millipore prior to use. Peptide calibration standard II (1–3 kDa) used for calibration of the MALDI TOF-MS, MALDI target plate, and α-cyano-4-hydroxycinnamic acid (α-CHCA) were purchased from Bruker Daltonik GmbH (Bremen, Germany). The HPLC system was a JASCO (Tokyo, Japan) and equipped with a PU-2089 pump, UV 2075 detector, MX 2080-32 dynamic mixer, and a CO-8020 column oven. The TSK gel ODS-80Ts (0.46 × 25 cm) column was from Tosoh Co. (Tokyo, Japan). All other chemicals were of analytical grade.
10
Aerobic Lactic Acid Fermentation
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A basal liquid culture medium (10 ml; 10 g/l yeast extract, 10 g/l KH2PO4, 2 g/l (NH4)2SO4, 0.5 g/l MgSO4•7H2O, pH 4.5) (Okamoto et al., 2010) (link) containing 2% glucose with or without 0.2% CaCO3 was used for the fermentation experiments. As described in the previous report (Wang et al., 2016) (link), cultivation was performed under aerobic and semi-aerobic conditions, and the resulting spent media were subjected to HPLC analysis for quantification of LA, ethanol, and glucose concentrations. HPLC was performed using a JASCO PU-2089 pump with a JASCO RI-2031 detector, fitted Shodex SH 1821 column (8.0 mm × 300 mm, 75 ºC). The eluent was 0.5 mM H2SO4, at 0.6 ml/min. Oxygen concentration in the headspace was analyzed every 4 days by gas chromatography (GC, GL Science GC-3200) on a packed column (Molecular Sieve 5A 30/60, 3 m × 1.6 mm outer diameter) with a thermal conductivity detector (TCD). Argon was used as the carrier gas at a flow rate 35 ml/min. The operational temperatures of the injector, detector, and column were 100, 100, and 80 °C, respectively.
After cultivation, CaCO3 was removed in 0.02N HCL, and residual mycelium washed with distilled water. Then, dried fungal weight was measured. For enzymatic quantification of L(+)-LA, D-lactic-/L-lactic acid test kit (Roche Diagnostics) was used according to an instruction manual.
After cultivation, CaCO3 was removed in 0.02N HCL, and residual mycelium washed with distilled water. Then, dried fungal weight was measured. For enzymatic quantification of L(+)-LA, D-lactic-/L-lactic acid test kit (Roche Diagnostics) was used according to an instruction manual.
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