Ods 3

Manufactured by GL Sciences

Sourced in Japan

The ODS-3 is a reversed-phase column used for high-performance liquid chromatography (HPLC) analysis. It features an octadecylsilane (ODS) stationary phase, which provides high selectivity for a wide range of organic compounds. The column is designed for reliable and consistent separation performance.

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Lab products found in correlation

Pu 2080, by Jasco (2 mentions) Acquity uplc h class system, by Waters Corporation (2 mentions) Xevo tqd, by Waters Corporation (2 mentions) Isolera one purification system, by Biotage (2 mentions) Acquity uplc beh c18 1.7 μm, by Waters Corporation (2 mentions) Snap ultra c18 column, by Biotage (1 mentions) Jnm ecz400 spectrometer, by JEOL (1 mentions) Gf254, by Merck Group (1 mentions) Or 2090plus, by Jasco (1 mentions) Ft 710 fourier transform infrared spectrophotometer, by Horiba (1 mentions) Cosmosil 75c18 opn, by Nacalai Tesque (1 mentions) Ltq orbitrap xl, by Thermo Fisher Scientific (1 mentions) P 1030 polarimeter, by Jasco (1 mentions) Diaion hp 20, by Mitsubishi (1 mentions) Avance 500, by Bruker (1 mentions) Lc 2010c, by Shimadzu (1 mentions) Jnm la400 spectrometer, by JEOL (1 mentions) Precoated silica gel plate, by Merck Group (1 mentions) Unity inova 500 ft nmr, by Agilent Technologies (1 mentions) Apex 2 mass spectrometer, by Bruker (1 mentions)

9 protocols using ods 3

Comprehensive Analytical Techniques for Chemical Compounds

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NMR spectra were recorded using a JEOL JNM-ECZ400 spectrometer at 400 MHz for ¹H NMR and 100 MHz for ¹³C NMR. Mass spectra (MS) were obtained using JEOL JMS-T100LP AccuTOF LC-plus 4 G (ESI).
Preparative HPLC was performed on an Inertsil ODS-3 (10.0 × 250 mm) column (GL Sciences Inc.) using an HPLC system composed of a pump (PU-2080, JASCO) and a detector (MD-2015). Preparative MPLC was performed on an Isolera One purification system (Biotage) equipped with a Biotage SNAP Ultra C18 column (for reverse phase separation) or on an MPLC system comprising a pump and detector (EPCLC AI-580S, Yamazen) and equipped with a silica gel column (silica gel 40 μm or Amino 40 μm, Yamazen) (for normal phase separation). LC-MS analysis was performed on an Acquity UPLC H-Class system (Waters) equipped with an Acquity UPLC BEH C18 1.7 μm (2.1 × 50 mm) column (Waters) and an MS detector (QDa or Xevo TQD, Waters).

Yanagi K., Komatsu T., Fujikawa Y., Kojima H., Okabe T., Nagano T., Ueno T., Hanaoka K, & Urano Y. (2023). Development of pathway-oriented screening to identify compounds to control 2-methylglyoxal metabolism in tumor cells. Communications Chemistry, 6, 68.

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Phenolic Profiling of Propolis Extracts

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The phenolic
profile of the propolis extracts was performed using RP-HPLC according
to the method described by Kocabey et al.^{41 (link)} with few modifications. An amount of 20 μL of extract samples
was injected into the column (ODS3, 250 × 4.6 mm, 5 μm;
GL Science, Tokyo, Japan) set at 28 °C with a flow rate of 1
mL/min. Two mobile phases were used with solvent A (0.1% H₃PO₄ in water) and solvent B (0.1% H₃PO₄ in acetonitrile). The flow rate of the mobile phase started
at 8% (solvent B) and increased to 11% within 4 min. Subsequently,
the flow rate of solvent B was adjusted to 35% at 25 min, 60% at 30
min, and 35% at 45 min and returned to 11% at 50 min and 8% at 55
min. Detection was carried out at wavelengths of 280, 320, and 360
nm. Authentic phenolic standards were used for the calculation of
each individual phenolic content.

, & Yurt B. (2023). Effect of Hydrogen-Enriched Solvents on the Extraction of Phytochemicals in Propolis. ACS Omega, 8(15), 14264-14270.

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Spectroscopic Analyses and Chromatographic Techniques

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Optical rotations were measured on a JASCO P-1030 polarimeter (Jasco, Tokyo, Japan). UV and IR spectra were obtained on a Jasco V-520 UV/Vis spectrophotometer and a Horiba FT-710 Fourier transform infrared spectrophotometer (Horiba, Kyoto, Japan), respectively. NMR experiments were measured on a Bruker Avance 500, 600 and 700 MHz spectrometers (Bruker, Billerica, MA, USA), with tetramethylsilane (TMS) as an internal standard. Positive ion HR-ESI-MS spectra were recorded using a LTQ Orbitrap XL mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA). Column chromatography (CC) was performed on Diaion HP-20 (Mitsubishi Chemical Corp., Tokyo, Japan), silica gel 60 (230–400 mesh, Merck, Darmstadt, Germany), and octadecyl silica (ODS) gel (Cosmosil 75C₁₈-OPN (Nacalai Tesque, Kyoto, Japan; Φ = 35 mm, L = 350 mm), and TLC was performed on precoated silica gel plates 60 GF₂₅₄ (0.25 mm in thickness, Merck). HPLC was performed on ODS gel (Inertsil ODS-3, GL-science, 10 mm × 250 mm, flow rate 2.5 mL/min) with a mixture of H₂O, acetone and MeOH, and the eluate was monitored by refractive index and/or a UV detector. An amino column (Shodex Asahipak NH2P-50 4E (4.6 mm × 250 mm), CH₃CN-H₂O (3:1) 1 mL/min) together with a chiral detector (Jasco OR-2090plus) was used for HPLC analysis of sugars obtained after hydrolysis [15 (link)].

Abdulaziz Al-Hamoud G., Saud Orfali R., Sugimoto S., Yamano Y., Alothyqi N., Mohammed Alzahrani A, & Matsunami K. (2019). Four New Flavonoids Isolated from the Aerial Parts of Cadaba rotundifolia Forssk. (Qadab). Molecules, 24(11), 2167.

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Plasma Malondialdehyde Quantification by HPLC

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Malondialdehyde was extracted and analyzed according to a previously described method with slight modifications [49 (link)]. Briefly 40 μL plasma was diluted with 100 μL of H₂O and mixed with 20 μL of 2.8 mmol/L BHT in 95 % ethanol, 40 μL of 81 g/L sodium dodecyl sulfate, and 600 μL of thiobarbituric acid (TBA) reagent consisting of 8 g/L TBA diluted 1:1 with 200 mL/L acetic acid adjusted to pH 3.5 with NaOH. The mixture was immediately incubated in a 90 °C water bath for 60 min and cooled on ice; 200 μL of H₂O and 1 mL of butanol-pyridine (15:1 by vol.) were then added. After vigorous mixing, the organic layer was separated by centrifugation (10 min at 10000 rpm). An aliquot (10 μL) was directly injected onto the high-performance liquid chromatography (HPLC). Calibration curves were constructed using 1,1,3,3-tetraethoxypropane (0.75 μmol/L – 40 μmol/L). The separation of the extracts was performed on an automated Shimadzu HPLC system (VP Series, Kyoto, Japan). The analytical column was a reverse phase silica based C18 column (GL Sciences/Inertsil ODS-3), with column dimensions of 150×4.6 mm, 5 μm. The mobile phase consisted of 70 % 10 mM KH₂PO₄, pH 7.0 and 30 % MeOH. The sample run was 5 min, with a flow rate of 0.8 mL/min, and fluorescence detection at 515 nm (excitation) and 553 nm (emission).

Ceylan D., Tuna G., Kirkali G., Tunca Z., Can G., Arat H.E., Kant M., Dizdaroglu M, & Özerdem A. (2018). Oxidatively-induced DNA damage and base excision repair in euthymic patients with bipolar disorder. DNA repair, 65, 64-72.

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HPLC Analysis of API in Pellets

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An assay of SD-granules and SD-pellets was performed on a Shimadzu LC-2030 3D Plus HPLC system (Shimadzu Corp, Kyoto, Japan) equipped with a C18 reverse-phase column (Inertsil ODS-3, GL Sciences, Tokyo, Japan; 250 mm length, 4.6 mm inner diameter, and 5 µm particle size). For both API, the eluent consisted of acetonitrile and 10 mM phosphate buffer, pH 7.4 (70/30, %v/v) at a flow rate of 1.0 mL/min. The column temperature was set to 40 °C, the autosampler was kept at 25 °C, and injection volume was 10 µL. The detector was set to 220 nm for both KCZ and LRD. Linearity was confirmed from 5 to 50 µg/mL. Prior to analysis, the coatings of 100 mg of pellets were dissolved in 100 mL of methanol and filtered through PTFE syringe filters (pore size 20 µm).

Neuwirth M., Kappes S.K., Hartig M.U, & Wagner K.G. (2023). Amorphous Solid Dispersions Layered onto Pellets—An Alternative to Spray Drying?. Pharmaceutics, 15(3), 764.

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Plasma Lutein Quantification by HPLC

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Blood samples were obtained at baseline and 3 and 6 months. The plasma lutein concentration was measured using a high-performance liquid chromatography (HPLC) system (LC-2010C; Shimadzu Corp., Kyoto, Japan) in Sumika Chemical Analysis Service, LTD (Osaka, Japan). Plasma samples were extracted with a mixture of methyl tertiary butyl ether (4:1) and centrifuged at 3,000 rotations/minute for 5 min at 4 °C. After evaporation of the organic layer under a nitrogen gas stream, the residue was dissolved in a 0.2-mL mobile phase solution for HPLC injection. The column was a GL Science Inertsil ODS-3 (5-µm particle size, 4.6-mm inside diameter × 250 mm). The column temperature was 40 °C. A mobile phase containing acetonitrile, 0.1 M ammonium acetate and methanol, and dichloromethane (71:22:7, v/v/v) was used. The flow rate was 1.0 mL/min. The 450-nm wavelength was used.

Sawa M., Shunto T., Nishiyama I., Yokoyama A., Shigeta R., Miura S, & Kawasaki R. (2020). Effects of Lutein Supplementation in Japanese Patients with Unilateral Age-Related Macular Degeneration: The Sakai Lutein Study. Scientific Reports, 10, 5958.

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NMR, MS, and HPLC/MPLC Characterization

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NMR spectra were recorded using a JEOL JNM-LA400 spectrometer at 400 MHz for ¹H NMR and 100 MHz for ¹³C NMR. Mass spectra (MS) were obtained using JEOL JMS-T100LP AccuTOF LC-plus 4G (ESI).
Preparative HPLC was performed on an Inertsil ODS-3 (10.0 × 250 mm) column (GL Sciences Inc.) using an HPLC system composed of a pump (PU-2080, JASCO) and a detector (MD-2015). Preparative MPLC was performed on an Isolera One purification system (Biotage) equipped with a Biotage SNAP Ultra C18 column (for reverse phase separation) or on an MPLC system comprising a pump and detector (EPCLC AI-580S, Yamazen) and equipped with a silica gel column (silica gel 40 μm or Amino 40 μm, Yamazen) (for normal phase separation). LC-MS analysis was performed on an Acquity UPLC H-Class system (Waters) equipped with an Acquity UPLC BEH C18 1.7 μm (2.1 × 50 mm) column (Waters) and an MS detector (QDa or Xevo TQD, Waters).

Sakamoto S., Hiraide H., Minoda M., Iwakura N., Suzuki M., Ando J., Takahashi C., Takahashi I., Murai K., Kagami Y., Mizuno T., Koike T., Nara S., Morizane C., Hijioka S., Kashiro A., Honda K., Watanabe R., Urano Y, & Komatsu T. (2024). Identification of activity-based biomarkers for early-stage pancreatic tumors in blood using single-molecule enzyme activity screening. Cell Reports Methods, 4(1), 100688.

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Analytical Techniques for Compound Characterization

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Optical rotations were measured on a JASCO P-1020 polarimeter (JASCO, Tokyo, Japan). IR spectra were recorded on a JASCO FT/IR-4100 spectrophotometer (JASCO). Ultraviolet spectra were recorded on a JASCO V-650 spectrophotometer. ESIMS and HRESIMS spectral data were recorded on a BRUKER APEX II mass spectrometer (Bruker, Bremen, Germany). The NMR spectra were recorded on a Varian Unity INOVA 500 FT-NMR at 500 MHz for ¹H and 125 MHz for ¹³C or on a Varian 400 FT-NMR at 400 MHz for ¹H and 100 MHz for ¹³C or on a Bruker AMX-300 FT-NMR at 300 MHz for ¹H and 75 MHz for ¹³C, in CDCl₃ or C₆D₆ using TMS as internal standard (δ in ppm, J in Hz). Silica gel 60 (Merck, 230–400 mesh), precoated silica gel plates (Merck, Darmstadt, Germany, Kieselgel 60 F254, 0.2 mm) were used for open CC and analytical TLC analysis, respectively. Isolation by HPLC was performed by a Hitachi L-2455 instrument equipped with a reversed-phase (RP-18) column (GL Sciences Inc., Tokyo, Japan ODS-3, 5 μm, 250 × 20 mm).

Ahmed A.F., Tsai C.R., Huang C.Y., Wang S.Y, & Sheu J.H. (2017). Klyflaccicembranols A–I, New Cembranoids from the Soft Coral Klyxum flaccidum. Marine Drugs, 15(1), 23.

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Analytical Characterization of Compounds

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All instruments used in this study were properly calibrated and validated by Mechanical Division of the institute. All kind of standard glassware used in this study was properly sterilized using Autoclave (Make-Indfos) at 121 °C for 20 min. To scan the chromatographic strips, 2π-Scanner (Berthold, Germany) was used. Characterization studies were performed using Deluxe Electrophoresis Chamber (Gelman, Germany) system, D-200 Elite HPLC system interconnected with C-18 column 4.6 × 150 mm (Inertsil® ODS-3, GL Sciences) along with NaI(Tl) detector, NMR spectra was recorded using ECS 400 MHz instruments (JEOL Resonance, Japan). ¹H NMR chemical shifts (δ) were recorded relative to DMSO-D₆ (δ = 2.50 ppm). High-resolution mass spectroscopy was performed on ORBITRAP ELITE (Thermo Scientific). Scintigraphy study was performed using Dual-headed Siemens Integrated Gamma Camera (interfaced with high-resolution parallel hole collimator) while blood cell images were captured using LED microscope (Zeiss Primo Star).

Rizvi S.F., Zhang H., Mehmood S, & Sanad M. (2020). Synthesis of 99mTc-labeled 2-Mercaptobenzimidazole as a novel radiotracer to diagnose tumor hypoxia. Translational Oncology, 13(12), 100854.

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