600 mhz nmr

The ¹H- and ¹³C-NMR spectra for all compounds were obtained on a Varian 600 MHz NMR at 25 °C. Spectra of all compounds were obtained in methanol-d4 and DMSO-d6. Detailed analysis of resolution-enhanced spectra (peak picking, integration, multiplet analysis) was performed using Varian NMR and ACD/NMR processor Academic Edition software. The NMR spectra and chemical shifts of isolated compounds were compared with published data.

TCO-RGD was synthesized according to our previous study^{43 (link)}. High-resolution mass spectra (HRMS) were performed on a Bruker OTOF-Q II mass spectrometer. ¹H NMR spectra were recorded with a Varian 600 MHz NMR with trimethylchlorosilane as an internal standard. The fluorescence spectra were measured in an Edinburgh Instruments’ FLS1000. The imaging experiments in vitro were recorded on an Olympus FV 10i confocal fluorescent microscope, and the fluorescent signals of DAPI were recorded by blue channel (excitation 405 nm, emission 461 nm) and NIR dyes by red channel (excitation 640 nm, emission 665 nm). In vivo fluorescence imaging analysis was carried out in an IVIS Kinetic imaging system (Living Image Software version 4.5, excitation 660 nm, emission 710 nm). FACS analysis was detected with a BD Accuri C6 flow cytometry (BD CFlow Plus Software version 1.0.264.21), and fluorescent signals were recorded by FL4 channel (excitation 640 nm, emission 675 nm).

Standards were prepared by adding 2.0 mg (7.08 mM) KmTx8 in 200μL MeOD with 10μL CDCl₂ to a 3 mm NMR tube. Helium was used to remove dissolved O₂ and achieve cleaner NMR signals. HSQC and ROESY experiments were completed using a Varian 600 MHz NMR at 25°C. The HSQC data for Region II of the molecule (oxygenated carbons) is shown in blue above. In a separate NMR tube, 2.0 mg (7.08 mM) 8 in 200μL MeOD and 0.2 mg (2.46 mM) cholesterol in 10μL CDCl₂ were combined and data acquired identically to the standard. The HSQC is shown in red above. The two data sets were then overlaid utilizing MestReNova 7.0.3–8830. This process was repeated using 0.3 mg cholesterol dissolved in CDCl₂.

LC–MS grade water and isopropanol were purchased from Fisher Scientific (Bridgewater, NJ, USA). Nanoparticles of anatase titanium dioxide, zinc oxide (<100 nm BET or <50 nm XRD), rutile titanium dioxide single crystal substrate with exposed <100> facet (10 × 10 × 0.5 mm) and juglone were purchased from Sigma-Aldrich (MO, USA). 4, 4′-dichlorodiphenyltrichloroethane was purchased from Dr Ehrenstorfer GmbH (Bgm, Schlosser, Germany). Acetone and ethanol was purchased from Guoyao (Tianjing, China). In the low-mass region of negative ion mode, the mass spectrometer was calibrated with free fatty acids. Conductive sticky alumina tape was purchased from Junke (Shanghai, China). All standard fatty acids (including C4:0, C6:0, C8:0, C10:0, C12:0, C14:0, C16:0, C18:0, C20:0 and C22:0) were purchased from Nu-ChekPrep Inc (MN, USA). In the low-mass region of positive ion mode, the mass spectrometer was calibrated with polyethylene glycol (PEG) standard that was purchased from Waters (Milford, USA). Surfaces of rutile titanium dioxide crystalline were characterized with a Jeol JSM-6700F scanning electron microscope (MA, USA). NMR experiments were performed on a Varian 600 MHz NMR (CA, USA).

Optical rotation was measured on a JASCO P-1020 digital polarimeter (Tokyo, Japan). UV data were recorded on a JASCO V-530 UV/VIS Spectrophotometer (Tokyo, Japan). CD spectrum was acquired on a JASCO J-815 CD spectrometer (Tokyo, Japan). High-resolution ESIMS data were obtained on a Bruker APEX II spectrometer (Billerica, MA, USA). IR spectrum was measured on a Perkin Elmer system 2000 FT-IR spectrophotometer (Waltham, MA, USA). NMR spectra were obtained by Varian 600 MHz NMR (San Carlos, CA, USA). Merck silica gel 60 (Billerica, MA, USA) and Sephadex LH-20 (Stockholm, Sweden) were used for column chromatography. The instrumentation for HPLC was composed of a Shimadzu LC-10AD pump (Kyoto, Japan) and a Shimadzu SPD-M10A PDA detector (Kyoto, Japan).

Sephadex (GE healthcare) LH-20 and silica gel 60 (Merck KGaA, Darmstadt, Germany) were used for open-column chromatography (CC). Luna C₁₈, phenyl-hexyl (5 μm, 250 × 10 mm, Phenomenex, Torrance, CA, USA) semi-preparative columns were used for high-performance liquid chromatography (HPLC). HPLC instrument used a Shimadzu LC-10AT pump with an SPD-20A UV-Vis detector. The UV spectra were obtained by using a Jasco UV-530 ultraviolet spectrophotometer (Jasco, Tokyo, Japan), whereas the IR spectra were obtained on a Jasco FT-IR-4600 spectrophotometer (Jasco, Tokyo, Japan). Optical rotations were measured with a Jasco P-1020 digital polarimeter (Jasco, Tokyo, Japan). NMR spectra were obtained using JEOL JNM ECS 400 MHz (JEOL, Tokyo, Japan), Varian 600 MHz NMR (Varian, Palo Alto, CA, USA), and Bruker AVIIIHD700X 700 MHz spectrometers (Bruker, Bremen, Germany). ESI–MS data were collected on a VG Biotech Quattro 5022 mass spectrometer (VG Biotech, Altrincham, UK). High-resolution ESI–MS data were obtained with a Bruker APEX II spectrometer (Bruker, Bremen, Germany). Circular dichroism spectra were recorded on a JASCO J-810 spectrophotometer (Jasco, Tokyo, Japan).