The melting points were measured on a Thomas-Hoover capillary melting point apparatus (Arthur H. Thomas Company, Philadelphia, PA., U.S.A.). Optical rotations at the sodium D line were measured with a Perkin-Elmer 241 digital polarimeter using quartz cell with a path length of 100 mm at room temperature. Concentrations (c) are given in g/100 mL. IR spectra were measured on a Jasco Fourier Transform IR Spectrometer (FT-IR model 410) loaded with an OMNIC software. UV spectra were collected on a Shimadzu UFLC system with a PDA detector. NMR spectra were recorded on a Bruker AV-400 spectrometer. All chemical shifts were quoted on the δ scale in ppm using residual solvent as the internal standard (methanol-d4: δH 3.30 for 1H NMR, δH 49.90 for 13C NMR). Coupling constants (J) are reported in Hz. For HPLC purification, a C18 semi-preparative HPLC column (Phenomenex C18 column, 250 × 10 mm, 5 µm) and a Shimadzu UFLC system were used. HRESIMS were measured on a Waters SYNAPT hybrid quadrupole/time of flight spectroscopy using positive electrospray ionization. Human melanoma cancer cells MDA-MB-435, human breast cancer cells MDA-MB-231, and human ovarian cancer cells OVCAR3 were purchased from the American Type Culture Collection (Manassas, VA). Molecular models in Figure 2 were generated by Chem3D Pro 14.0 using MM2 force field calculation.
Uflc system
Manufactured by Shimadzu
Sourced in Japan, United States
The UFLC system is an ultra-fast liquid chromatography (UFLC) instrument designed for high-performance liquid chromatography (HPLC) analysis. The UFLC system is capable of rapid separation and detection of various chemical compounds with high resolution and sensitivity.
Automatically generated - may contain errors
Lab products found in correlation
Analyst 1, by AB Sciex (4 mentions)
Sciex 5500 qtrap mass spectrometer system, by AB Sciex (4 mentions)
Ovcar3, by American Type Culture Collection (3 mentions)
Mda mb 231, by American Type Culture Collection (3 mentions)
Mda mb 435, by American Type Culture Collection (3 mentions)
C18 column, by Shimadzu (2 mentions)
Zorbax sb c18 column, by Agilent Technologies (2 mentions)
Triple toftm 5600 system ms ms, by AB Sciex (2 mentions)
4000 qtrap, by AB Sciex (2 mentions)
Cbm 20a, by Shimadzu (2 mentions)
Wyatt 787 ts minidawn treos, by Wyatt Technology (2 mentions)
Api5000 3 triple quadrupole mass spectrometer, by AB Sciex (2 mentions)
Uplc h class system, by Waters Corporation (2 mentions)
Milli q purification system, by Merck Group (2 mentions)
Xevo g2 s qtof mass spectrometer, by Waters Corporation (2 mentions)
Lcms 8050, by Shimadzu (2 mentions)
Capcell pak c18 ug120 column, by Shiseido (2 mentions)
Labsolutions software, by Shimadzu (2 mentions)
Qtrap 5500, by AB Sciex (2 mentions)
Analyst software version 1, by Thermo Fisher Scientific (2 mentions)
79 protocols using uflc system
1
Characterization of Anticancer Compounds
2
Analytical Characterization of Compounds
3
Pharmacokinetics of Hydroxykynurenine in Mice
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Thirty-two mice were randomly divided into two groups and injected intraperitoneally (IP) with HK dissolved in soybean oil or HKE at a dose of 20 mg/kg. At 0.5, 1, 2, and 4 h after injection, the mice were sacrificed to collect plasma and hearts. Hearts were accurately weighed, homogenized in 0.5 ml deionized water with a tissue homogenizer, and extracted with methanol. Tissue homogenate extracts and plasma were analyzed using an ultra-fast liquid chromatography–triple quadrupole mass spectrometer system (LC/MS) (UFLC system, Shimadzu; AB5500 mass spectrometer, Applied Biosystems). The detailed method is provided in the Supplementary Data .
4
UFLC-MS/MS Analysis of Compounds
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A UFLC system (Shimadzu, Kyoto, Japan) was used for sample analysis. The separation was conducted by an Agilent ZORBAX SB-C18 column (4.6 mm × 250 mm, 5 μm) at 30 °C. The mobile phase consisted of methanol (A) and 0.1% formic acid in water (B) with the following gradient elution: 0–2 min (5–30% A), 2–25 min (30–55% A), 25–30 min (55–5% A) and 30–32 min (5–5% A). The column temperature was 30 ℃. The detection wavelength was 360 nm. The injection volume was 10 μL and the flow rate was 0.8 mL/min.
MS data were recorded using an AB Sciex Triple TOFTM 5600 system-MS/MS (AB SCIEX, Framingham, MA, USA), equipped with an electrospray ionization (ESI) source. The optimized MS conditions in negative ion mode were set as follows: ion source temperature (TEM), 600 °C; flow rate of curtain gas (CUR), 40 psi; flow rate of nebulization gas (GS1) and flow rate of auxiliary gas (GS2), 60 psi; ion spray voltage floating (ISVF), 4500 V; collision energy, −10 V; declustering potential, −100 V. Data were acquired for each sample from 50 to 1500 Da. The data acquisition time was 32 min.
MS data were recorded using an AB Sciex Triple TOFTM 5600 system-MS/MS (AB SCIEX, Framingham, MA, USA), equipped with an electrospray ionization (ESI) source. The optimized MS conditions in negative ion mode were set as follows: ion source temperature (TEM), 600 °C; flow rate of curtain gas (CUR), 40 psi; flow rate of nebulization gas (GS1) and flow rate of auxiliary gas (GS2), 60 psi; ion spray voltage floating (ISVF), 4500 V; collision energy, −10 V; declustering potential, −100 V. Data were acquired for each sample from 50 to 1500 Da. The data acquisition time was 32 min.
5
LC-MS/MS Analysis of Panobinostat
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The LC-MS/MS system consists of a QTRAP 4000 mass spectrometer (AB SCIEX) coupled to a Shimadzu UFLC system. LC separation was carried out on an ACE C18 column (50 mm × 2.1 mm, 5 µm) with isocratic elution using a mobile phase composed of 65% water and 35% methanol. 0.1% of formic acid was added to both aqueous and organic phases. The flow rate was set at 0.25 ml/min. Column temperature was 25°C. The analysis time was 2.5 min. The injection volume was 10 µl. The mass spectrometer was operated in the positive mode with multiple-reaction monitoring (MRM). The m/z 350.2→158.1 and 358.3→164.2 transitions were used for Panobinostat and Panobinostat-d8 (IS). Data acquisition and analysis were performed using the Analyst 1.6.1 software (AB SCIEX).
6
Size-Exclusion HPLC Analysis of LtEc HBL
7
HPLC-MS Analysis of Harmine in Mouse Serum
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HPLC-MS was done on
a 3200 QTRAP (Applied Biosystems) equipped with a Shimadzu (Kyoto,
Japan) UFLC System. Mouse serum and harmine were purchased from Sigma-Aldrich.
Pooled CD1 mouse liver (female) microsomes were purchased from XenoTech,
LCC (Lenexa, KS) with protein concentrations of 0.5 mg/mL. HCT116
cell lysates were prepared with PhosphoSafe lysis buffer (Novagen,
San Diego, CA). Protein concentration was determined by using the
BCA Assay. Analysis was carried out similarly as previously described.21 (link)
a 3200 QTRAP (Applied Biosystems) equipped with a Shimadzu (Kyoto,
Japan) UFLC System. Mouse serum and harmine were purchased from Sigma-Aldrich.
Pooled CD1 mouse liver (female) microsomes were purchased from XenoTech,
LCC (Lenexa, KS) with protein concentrations of 0.5 mg/mL. HCT116
cell lysates were prepared with PhosphoSafe lysis buffer (Novagen,
San Diego, CA). Protein concentration was determined by using the
BCA Assay. Analysis was carried out similarly as previously described.21 (link)
8
Carotenoid Analysis in Physalis peruviana
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Carotenoids in Physalis peruviana L. were already identified by HPLC-DAD-APCI-MSn (Etzbach et al., 2018 (link)). They were quantified as described previously (Etzbach et al., 2019 ) using a Prominence UFLC system (Shimadzu, Kyoto, Japan) equipped with two Nexera X2 LC-30AD high-pressure gradient pumps, a Prominence DGU-20A5R degasser, a Nexera SIL-30AC Prominence autosampler (15 °C, injection volume 5 μL and 15 μL for total carotenoids and surface carotenoids, respectively), a CTO-20AC Prominence column oven at 25 °C, and a SPD-M20A Prominence diode array detector. The chromatograms were processed at 450 nm and carotenoids were quantified as β-carotene equivalents using an external calibration curve.
9
HPLC-MS Analysis of Biomolecular Compounds
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An HPLC-MS analysis was performed in a Shimadzu UFLC system coupled to a quadrupole time-of-flight mass spectrometer (micrOTOF QII, Bruker Daltonics, Billerica, MA, USA) using a C18 column (5 µm, XB-C18 Kinetex, 100Å, 150 × 3 mm, Phenomenex). The mobile phase was composed of water (A) and MeOH (B), both with 0.1% formic acid, at a flow rate of 0.75 mL.min−1. The following gradient was employed: 0–23 min, 10–100% B; 23–26 min, 100% B; 26–27 min, 100–10% B; and 27–30 min, 10% B. The column oven was set to 45 °C, and an injection volume of 10 µL was selected. Chromatograms were acquired in both positive and negative ionization modes, and the following parameters were employed for the mass spectrometer: end plate offset, 500 V; capillary voltage, 3200 V for negative mode and 3500 V for positive mode; nebulizer pressure, 4.5 bar; dry gas (N2) flow, 9 L.min−1; dry temperature, 200 °C; mass range, m/z 150 to 1200; MS/MS scan mode; number of precursors, 3; exclusion activation, 1 spectrum; and exclusion release, 36 s.
10
UFLC Analysis of NHPN and NHPNG
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Both NHPN and NHPNG were analyzed by a UFLC system (Shimadzu, Kyoto, Japan), equipped with a CBM-20A communications bus module, two LC-20AD pumps, a DGU-20A5R vacuum degasser, a SIL-20AC autosampler, a CTO-20AC column oven, and an RF-20A fluorescence detector. A Shim-pack VP-ODS C18 column (4.6 mm × 150 mm, 5 μm, Shimadzu) was used in this study, and the column temperature was maintained at 40 °C. The fluorescence signals of NHPN and NHPNG were recorded with the excitation wavelength at 370 nm, while the emission wavelength was set at 520 nm [13 (link)]. The mobile phase was a mixture of acetonitrile (A) and 0.2% formic acid (B). The following gradient elution program was used: 0–0.5 min, 40%–20% B; 0.5–1.8 min, 20%–5% B; 1.8–3.8 min, 5% B; 3.8–5 min, 20%–40% B; and 5–6 min, balance to 40% B.
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