The 3-IS content was estimated in the serum, urine, and kidney tissues using high-performance liquid chromatography (HPLC). The extraction of the samples was performed using 70% acetonitrile, an organic solvent, in a multi-step process. The extracted samples were mixed thoroughly with 2-naphthalene sulfonic acid used as an internal standard, and measured using HPLC (Gilson, LC-321322350) at 280 nm. The retention time of 3-IS was 6.8 min, and the extraction recovery was 84%. The HPLC apparatus used in the present study comprised a Gilson pump (LC-321322350 pump), an autosampler (Gilson-234), and a UV (UV/VIS-151) detector (Gilson, France). A C18 column from Agilent (Torrance, CA, USA) with a pre-column (250 mm 4.6 mm, and 5 m) was used for detection and quantification with a flow rate of 0.9 mL/min at room temperature. A 70:30 v/v combination of acetonitrile and 0.1% trifluoroacetic acid in Milli Q water was employed as the isocratic mobile phase.
Uv vis 151
Manufactured by Gilson
Sourced in United States
The UV-Vis-151 is a spectrophotometer that measures the absorbance of a sample across the ultraviolet and visible light spectrum. It is capable of analyzing the absorption characteristics of liquid samples.
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Lab products found in correlation
Ft ir 4200 spectrometer, by Jasco (5 mentions)
Jms 700 mass spectrometer, by JEOL (4 mentions)
Ri 150, by Gilson (4 mentions)
U 3010 spectrophotometer, by Hitachi (4 mentions)
P 1020 polarimeter, by Jasco (3 mentions)
6130 quadrupole mass spectrometer, by Agilent Technologies (2 mentions)
P 2000 polarimeter, by Jasco (2 mentions)
Avance 600 mhz spectrometer, by Bruker (2 mentions)
Lc 321 322 350 pump, by Gilson (1 mentions)
Lc 321 322 350, by Gilson (1 mentions)
C18 column, by Agilent Technologies (1 mentions)
L 6200, by Hitachi (1 mentions)
Fc204, by Gilson (1 mentions)
Rw 0525g, by Jeio Tech (1 mentions)
Lambda 35 spectrophotometer, by Jasco (1 mentions)
Chirascan v100 cd spectrometer, by Applied Photophysics (1 mentions)
R axis rapid diffractometer, by Rigaku (1 mentions)
1200 series hplc, by Phenomenex (1 mentions)
Mci gel ca08f, by Mitsubishi (1 mentions)
1200 series hplc, by Agilent Technologies (1 mentions)
8 protocols using uv vis 151
1
3-IS Quantification in Biological Samples
2
High-Speed Counter-Current Chromatography
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The mobile phase of solvent system was selected to organic lower phase. The multilayer coil column was filled first with the stationary phase. Then, the mobile upper phase was pumped into the column by a chromatographic pump (L-6200, Hitachi, Japan) at a flow rate of 3 mL/min while preparative HSCCC apparatus (TBE-1000A, Shanghai Tauto Biotech, Co., Ltd., China) was rotated at a revolution speed of 500 rpm. After hydrodynamic equilibrium was established, prepared sample solution was subjected to HSCCC apparatus. The monitoring of HSCCC peak fractions was performed by combining effluent line of the HSCCC apparatus to the UV detector (UV/VIS-151, Gilson Inc., Middleton, WI, USA) at 330 nm. The eluent from the UV detector was collected by a fraction collector (FC-204, Gilson Inc.) in 3 min per each test tube. HSCCC systems were kept the internal column temperature at 25°C by circulatory temperature regulator (RW-0525G, Jeio Tech., Korea).
3
Analytical Techniques for Structural Characterization
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Optical rotation values were measured at 25 °C using a JASCO P-2000 polarimeter (Tokyo, Japan) with a 1 cm cell. UV and IR spectra were obtained using a Perkin-Elmer Lambda 35 spectrophotometer (Waltham, MA, USA) and a JASCO FT/IR-4200 spectrometer (Tokyo, Japan), respectively. Circular dichroism (CD) spectra were acquired on an Applied Photophysics Chirascan V100 CD spectrometer (Leatherhead, UK) using a 1 mm CD cell. NMR spectra were collected using Bruker Avance 600 and 850 MHz NMR spectrometers (Billerica, MA, USA) located at the National Center for Inter-University Research Facilities (NCIRF), Seoul National University. Low-resolution electrospray ionization (ESI) data were acquired using an Agilent Technologies 6130 quadrupole mass spectrometer (Santa Clara, CA, USA) coupled with an Agilent Technologies 1200-series HPLC with a reversed-phase C18 column (Phenomenex Luna, 100 × 4.6 mm). High-resolution FAB mass spectrometric data were obtained using a JEOL JMS 700 mass spectrometer (Tokyo, Japan) at the National Center for Inter-University Research Facilities (NCIRF). HPLC isolation was performed on a Gilson 321 (Middleton, WI, USA) equipped with a UV detector Gilson UV-Vis-151 (Middleton, WI, USA). X-ray crystallographics were collected on a Rigaku R-AXIS RAPID diffractometer (Tokyo, Japan) using graphite-monochromated Mo Kα radiation (λ = 0.71075 Å).
4
Characterization of Organic Compounds
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Optical rotations were measured using a JASCO P-1020 polarimeter (Jasco, Tokyo, Japan) with a 1-cm cell. CD spectra were obtained using an Applied Photophysics Ltd. Chirascan Plus spectrometer (Leatherhead, Surrey, UK). UV spectra were acquired using a Hitachi U-3010 spectrophotometer (Hitachi High-Technologies, Tokyo, Japan). IR spectra were recorded on a JASCO 4200 FT-IR spectrometer (Jasco, Tokyo, Japan) using a ZnSe cell. NMR spectra were recorded in MeOH-d3, MeOH-d4 and CDCl3 with the solvent peaks (δH 3.30/δc 47.5 and δH 7.26/δc 77.0) as internal standards on a Bruker Avance 600 MHz spectrometer (Billerica, MA, USA). Proton and carbon NMR spectra were measured at 600 and 150 MHz, respectively. High-resolution FABMS data were obtained at the National Center for Inter-university Research Facilities (NCIRF), Seoul National University, and acquired using a JEOL JMS 700 mass spectrometer (Jeol, Tokyo, Japan) with 6 keV-energy, emission current 5.0 mA, xenon as the inert gas, and meta-nitrobenzyl alcohol (NBA) as the matrix. HPLC separations were performed on a SpectraSYSTEM p2000 equipped with a refractive index detector (SpectraSYSTEM RI-150 (Waltham, MA, USA)) and a UV-Vis detector (Gilson UV-Vis-151 (Middleton, WI, USA)). All solvents used were of spectroscopic grade or were distilled prior to use.
5
Quantifying Urinary 8-OHdG Levels
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The urinary 8-OHdG levels were determined according to our previous study.(23 (link)) Briefly, each urine sample was defrosted to room temperature and centrifuged at 8,500 × g for 5 min. A 50 µl portion of the urine supernatant was mixed with the same volume of a dilution solution, containing the ribonucleoside marker 8-hydroxyguanosine. Afterwards, a 20 µl portion of the prepared solution was fractionated and injected into the first HPLC column (MCI GEL CA08F, 1.5 × 150 mm; Mitsubishi Chemical, Tokyo, Japan). The chromatograph was recorded with a UV detector (Gilson UV/VIS-151, 235 nm). The fraction containing 8-OHdG was then automatically injected into the second HPLC column (InsertsilTM ODS-3, 3 µm, 4.6 × 250 mm; GL Sciences Inc., Tokyo, Japan). The chromatograph was recorded with an electrochemical (EC) detector (Coulochem II; ESA, Chelmsford, MA). The 8-OHdG levels were expressed as the ratios to the urinary creatinine contents (UV detector at 235 nm).
6
Spectroscopic Analysis of Organic Compounds
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Optical rotations were measured using a JASCO P-1020 polarimeter (Easton, MD, USA) with a 1 cm cell. CD spectra were obtained using an Applied Photophysics Chirascan Plus spectrometer (Applied Photophysics Ltd., Leatherhead, Surrey, UK). UV spectra were acquired using a Hitachi U-3010 spectrophotometer (Tokyo, Japan). IR spectra were recorded on a JASCO 4200 FT-IR spectrometer (Easton, MD, USA) using a ZnSe cell. NMR spectra were recorded in DMSO-d6, with the solvent peaks (δH 2.50/δc 39.50) as internal standards, on a Bruker Avance 600 MHz spectrometer (Billerica, MA, USA). High-resolution FABMS spectrometric data were obtained at the National Center for Inter-university Research Facilities (NCIRF), Seoul National University and acquired using a JEOL JMS 700 mass spectrometer with 6 keV-energy, emission current 5.0 mA, xenon as inert gas, and meta-nitrobenzyl alcohol (NBA) as the matrix. HPLC separations were performed on a SpectraSYSTEM p2000 equipped with a refractive index detector (SpectraSYSTEM RI-150 (Waltham, MA, USA)) and a UV-Vis detector (Gilson UV-Vis-151 (Middleton, WI, USA)). All solvents used were of spectroscopic grade or were distilled prior to use.
7
Spectroscopic Characterization of Compounds
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Optical rotations were measured on a JASCO P-1020 polarimeter (Easton, MD, USA) using a cell with a 1-cm path length. UV spectra were acquired using a Hitachi U-3010 spectrophotometer (Tokyo, Japan). CD spectra were recorded on an Applied Photophysics Ltd. Chirascan plus CD spectrometer (Applied Photophysics Ltd., Leatherhead, Surrey, UK). IR spectra were recorded on a JASCO 4200 FT-IR spectrometer (Easton, MD, USA) using a ZnSe cell. NMR spectra were recorded in DMSO-d6 or CDCl3 solutions on Bruker Avance-400, -500, or -600 instruments (Billerica, MA, USA). High-resolution FABMS data were acquired using a JEOL JMS 700 mass spectrometer (Tokyo, Japan) with 6 keV-energy, emission current 5.0 mA, xenon as inert gas, and meta-nitrobenzyl alcohol (NBA) as the matrix at the Korea Basic Science Institute (Daegu, Korea). Low-resolution ESIMS data were recorded on an Agilent Technologies 6130 quadrupole mass spectrometer (Santa Clara, CA, USA) with an Agilent Technologies 1200 series HPLC (Santa Clara, CA, USA). HPLC separations were performed on a SpectraSYSTEM p2000 equipped with a refractive index detector (SpectraSYSTEM RI-150 (Waltham, MA, USA)) and a UV-Vis detector (Gilson UV-Vis-151 (Middleton, WI, USA)). All solvents used were of spectroscopic grade or were distilled prior to use.
8
Comprehensive Analytical Characterization
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Optical rotation was measured with a JASCO P-2000 polarimeter (Easton, MD) with a 1-cm cell. Ultraviolet (UV) spectra were acquired using a Hitachi U-3010 spectrophotometer (Tokyo, Japan). Infrared (IR) spectra were obtained with a JASCO 4200 FT-IR spectrometer (Easton, MD) using a ZnSe cell. High-resolution electrospray ionization mass spectrometry (HR-ESI-MS) data were acquired at the National Instrumentation Center for Environmental Management (Seoul, Korea) using an AB Sciex 5600 QTOF HR-MS instrument (Sciex, MA). Proton and carbon nuclear magnetic resonance spectroscopy (NMR) and two-dimensional NMR spectra were recorded with a Varian Gemini 2000 300 MHz spectrometer (Palo Alto, CA) or Bruker Avance 500 and 600 MHz spectrometers (Berlin, Germany) using MeOH-d4 with a solvent peak at δH 3.31/δC 49.0 or dimethyl sulfoxide (DMSO)-d6 with a solvent peak at δH 2.50/δC 39.50 as internal standards. High-performance liquid chromatography (HPLC) separation was conducted on a SpectraSYSTEM p2000 instrument equipped with a refractive index detector (SpectraSYSTEM RI-150) and a UV-Vis detector (UV-Vis-151, Gilson, Middleton, WI). All solvents used were of spectroscopic grade or were distilled prior to use.
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