1D (1H, 13C, and DEPT) and 2D (HSQC, HMBC, 1H–1H COSY, and NOESY) NMR spectra were obtained on a Bruker AM-500 spectrometer (Bruker, Germany) in CDCl3 or CD3OD (Qingdao Tenglong, China), and TMS was used as an internal reference. IR spectra were scanned on a Nicolet Magna-IR 550 spectrometer (Thermo Nicolet, United States) with KBr pellets. Optical rotations were measured on a Jasco P-1020 digital polarimeter (Jasco, Japan). HR-ESI-MS spectra were obtained on an Agilent 6230 LC/TOF MS spectrometer (Agilent, United States). The prep-HPLC experiment was performed on an Agilent 1260 pump coupled with an analytical preparative ZORBAXSB column (21.2 × 500 m, 5 µm). Silica gel (300–400 mesh, Qingdao Haiyang, China) was used in column chromatography, and Dragendorff’s reagent was used in TLC analysis (GF254 TLC plates, Qingdao Haiyang, China).
Agilent 6230 lc tof ms spectrometer
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 6230 LC/TOF MS spectrometer is a high-performance liquid chromatography-time of flight mass spectrometry system. It is designed to provide accurate, sensitive, and reproducible mass analysis of a wide range of compounds.
Automatically generated - may contain errors
Lab products found in correlation
Varian 600 mhz, by Agilent Technologies (2 mentions)
Combiflash rf 200, by Teledyne (2 mentions)
400 spectrometer, by Bruker (2 mentions)
Am 500 spectrometer, by Bruker (1 mentions)
Cd3od, by Bruker (1 mentions)
P 1020 digital polarimeter, by Jasco (1 mentions)
Mercury plus 400, by Agilent Technologies (1 mentions)
Mabpac rp column, by Thermo Fisher Scientific (1 mentions)
Redisep column, by Teledyne (1 mentions)
Agilent 1260 infinity hplc, by Agilent Technologies (1 mentions)
4 protocols using agilent 6230 lc tof ms spectrometer
1
Characterization of Isolated Compounds
2
Glycoengineered Antibody-Drug Conjugates
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The enzymes of Endo-S, PNGase-F, and AlfC were expressed in E. coli following the reported procedures8 (link), 37 (link), 42 (link), 43 (link). The glycoengineered azido-herceptin (7 ) and DBCO-MMAE (8 ) were prepared following our previous paper8 (link). DM1 (N2′-deacetyl-N2′-(3-mercapto-1-oxopropyl)- maytansine), MMAE (monomethyl auristain E), and SMCC (N-succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate) were purchased from Levena Biopharma (Nanjing, China). Other chemical reagents and solvents were purchased from Sinopharm Chemical Reagent Co. (Shanghai, China) or Sigma-Aldrich (Shanghai, China) and used without further purification. The MAbPac RP column (4 μm, 3.0 × 100mm) was purchased from ThermoFisher. Nuclear magnetic resonance (NMR) spectra were measured on a Varian-MERCURY Plus-400 or 500 instrument. ESI-HRMS spectra were measured on an Agilent 6230 LC-TOF MS spectrometer.
3
Synthesis and Characterization of PF74
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All commercial chemicals were used as supplied unless indicated otherwise. Compounds were purified via flash chromatography using a Combiflash RF-200 (Teledyne ISCO, Lincoln, NE, USA) with RediSep columns (silica) and indicated mobile phase. 1H and 13C NMR spectra were recorded on a Varian 600 MHz (Agilent Technologies, Santa Clara, CA, USA) or Bruker 400 spectrometer (Bruker, Billerica, MA, USA). Diastereomeric ratio was determined by 1H NMR analysis. Mass data were acquired using an Agilent 6230 TOF LC/MS spectrometer (Agilent Technologies). PF74 was synthesized according to reported procedures43 ,44 (link).
4
Synthetic Protocols for Purified Compounds
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All commercial chemicals were used as supplied unless indicated otherwise. Compounds were purified via flash chromatography using a Combiflash RF-200 (Teledyne ISCO, Lincoln, NE, USA) with RediSep columns (Teledyne ISCO, Lincoln, NE, USA) (silica) and indicated mobile phase. 1H and 13C NMR spectra were recorded on a Varian 600 MHz (Agilent Technologies, Santa Clara, CA, USA) or Bruker 400 spectrometer (Bruker, Billerica, MA, USA). Diastereomeric ratio was determined by 1H NMR analysis. Mass data were acquired using an Agilent 6230 TOF LC/MS spectrometer (Agilent Technologies, Santa Clara, CA, USA). Compound purity analysis was performed using Agilent 1260 Infinity HPLC (Agilent Technologies, Santa Clara, CA, USA) with an Eclipse C18 column (3.5 μm, 4.6 × 100 mm). HPLC conditions: flow rate, 1.0 mL/min; solvent A, 0.1% TFA in water; solvent B, 0.1% TFA in acetonitrile; gradient (B, %): 0–3 min (5–100), 3–11 min (100), 11–13 min (100–5). Determined purity was >85% for all final compounds.
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