Optical rotations were measured using the 241 polarimeter (PerkinElmer, Waltham, MA, USA). Measurements of the UV spectra were carried out using the UV-Vis spectrophotometer UV-2450 (Shimadzu, Kyōto, Japan) and measurements of the ECD spectra were carried out using a J-815 spectropolarimeter (Jasco, Pfungstadt, Germany). NMR spectra were obtained using the Avance III 500 MHz spectrometer equipped with a BBFO (plus) SmartProbe (1H 500 MHz, 13C 125 MHz) and the Avance III 700 MHz spectrometer equipped with a 5 mm TCI cryoprobe (1H 700 MHz, 13C 175 MH) (both Bruker, Billerica, MA, USA). NMR data were referenced to selected chemical shifts of acetonitrile-d3 (1H: 1.94 ppm, 13C: 1.4 ppm), methanol-d4 (1H: 3.31 ppm, 13C: 49.2 ppm), and pyridine-d5 (1H: 7.22 ppm), respectively. HRESIMS mass spectra were recorded with a 1200 series HPLC-UV system (Agilent, Santa Clara, CA, USA) in combination with an ESI-TOF-MS (Maxis, Bruker). Performance of the measurements was conducted with a 2.1 × 50 mm, 1.7 µm, C18 Acquity UPLC BEH (Waters, Milford, MA, USA) column, using MilliQ H2O + 0.1% formic acid as solvent A and MeCN + 0.1% formic acid as solvent B (gradient: 5% B for 0.5 min increasing to 100% B in 19.5 min and maintaining 100% B for 5 min, flow rate: 0.6 mL/min, UV detection: 200–600 nm).
Esi tof ms
Manufactured by Bruker
Sourced in United States, United Kingdom
The ESI-TOF-MS is a mass spectrometry instrument that utilizes electrospray ionization (ESI) to introduce samples into a time-of-flight (TOF) mass analyzer. The core function of the ESI-TOF-MS is to accurately measure the mass-to-charge ratios of ionized molecules in a sample.
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Lab products found in correlation
Avance 3 700 mhz spectrometer, by Bruker (3 mentions)
Acquity uplc beh c18, by Waters Corporation (3 mentions)
Agilent 1200 series hplc uv system, by Bruker (3 mentions)
241 polarimeter, by PerkinElmer (2 mentions)
Bbfo plus smartprobe, by Bruker (2 mentions)
Avance 3 500 mhz spectrometer, by Bruker (2 mentions)
J 815 spectropolarimeter, by Jasco (2 mentions)
Uv 2450, by Shimadzu (2 mentions)
5 mm tci cryoprobe, by Bruker (2 mentions)
Uv 2450 uv vis spectrophotometer, by Shimadzu (2 mentions)
Ez link nhs peg12 biotin, by Thermo Fisher Scientific (2 mentions)
Mcp 150 polarimeter, by Anton Paar (1 mentions)
J 815, by Jasco (1 mentions)
Cosmosil 5c18 ms 2 reversed phase column, by Nacalai Tesque (1 mentions)
Ultimate 3000 series uhplc, by Thermo Fisher Scientific (1 mentions)
Esi ion trap ms, by Bruker (1 mentions)
Avance 3 500 spectrometer, by Bruker (1 mentions)
Agilent 1200 infinity series hplc uv system, by Agilent Technologies (1 mentions)
Acquity uplc beh c18 column, by Waters Corporation (1 mentions)
241 spectrophotometer, by PerkinElmer (1 mentions)
12 protocols using esi tof ms
1
Comprehensive Spectroscopic Analysis of Compounds
2
Multi-Instrumental Analysis of Metabolites
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An Agilent 1200 series HPLC-UV system (Santa Clara, CA, USA) with an ESI-TOF-MS (MaXis, Bruker, Bremen, Germany) [column 2.1 × 50 mm, 1.7 µm, C18 Acquity UPLC BEH (Waters, Eschborn, Germany)], with deionized water + 0.1% formic acid (solvent A) as well as acetonitrile + 0.1% formic acid (solvent B) and a gradient of 5% B for 0.5 min increasing to 100% B in 19.5 min, maintaining 100% B for 5 min, flow rate: 0.6 mL/min, UV detection 200–600 nm, was used to obtain the HRESIMS data. NMR spectra were acquired on a Bruker Avance III 700 MHz spectrometer (Bremen, Germany) equipped with a 5 mm TCI cryoprobe (1H NMR (700 MHz), 13C NMR (175 MHz)). Optical rotations of the metabolites were measured with an MCP 150 polarimeter (Anton Paar, Graz, Austria). The CD spectra were obtained from a JASCO spectropolarimeter, model J-815 (Pfungstadt, Germany). The UV spectra were recorded on a Shimadzu UV–vis spectrophotometer UV-2450 (Duisburg, Germany).
3
Biotin-PEG Synthesis and HPLC Purification
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Reagents and solvents were purchased from standard providers and used without further refinement. The EZ-Link NHS-PEG12-Biotin was obtained from Thermo Scientific, USA (No. 21312). Analytical High Performance Liquid Chromatography (HPLC) was performed using a COSMOSIL 5C18-MS-II reversed phase column (4.6 × 150 mm, Nacalai Tesque) in 0.1% Trifluoroacetic acid (TFA) in water with CH3CN as eluent at 1.0 ml/min, and a linear gradient elution of 0−100% CH3CN over 20 or 40 min with detection at 254 nm. The HPLC purification was performed with a COSMOSIL 5C18-MS-II reversed phase column (10 × 150 mm, Nacalai Tesque) in 0.1% TFA in water with CH3CN as the eluent. The final products were analyzed by ESI-TOF-MS (Bruker). The complete PIP synthesis procedure is provided in Supplementary Data.
4
Characterization of Dendrons by ESI-TOF MS
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The AMT-competent G0- and G1- dendrons as well as the RMT-competent AEP-dendron were characterized by electrospray/ionization-time of flight (ESI-TOF MS) (Bruker Daltonics, UK) at high voltage (4 kV). The samples were dissolved in methanol, then filtered using a 0.22 μm filter and injected into the spectrometer. In electrospray/ionization mode, sample mass (m/z) gave rise to multiple charged ions labeled with a number of charges (n) as (MW+nH)n+, where H is the mass of a proton (1.008 Da).
5
NMR and Mass Spectrometry Analysis
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1D and 2D-NMR spectra were recorded on a Bruker Avance III 500 MHz spectrometer (Bremen, Germany) with a BBFO(plus) SmartProbe (1H 500 MHz, 13C 126 MHz), and a Bruker Avance III 700 MHz spectrometer (Bremen, Germany) with a 5 mm TCI cryoprobe (1 H 700 MHz, 13 C 175 MHz).
Chemical shifts were referenced to the solvents: chloroform-d (1H, δ = 7.27 ppm; 13C, δ = 77.0 ppm) and acetone-d6 (1H, δ = 2.05 ppm; 13C, δ = 29.3 ppm). HRMS mass spectra were measured on the Agilent 1200 series HPLC-UV system (Santa Clara, CA, USA) combined with ESI-TOF-MS (Maxis, Bruker, Bremen, Germany), scan range 100−2500 m/z, capillary voltage 4500 V, temperature 200 °C, (column 2.1 × 50 mm, 1.7 µm, C18 Acquity UPLC BEH [Waters, MZ-Analysetechnik, Mainz, Germany], solvent A: 95% 5 mM ammonium acetate buffer [pH 5.5, adjusted with 1 M acetic acid] with 5% acetonitrile; solvent B: 95% acetonitrile with 5% 5 mM ammonium acetate buffer); gradient: 10% solvent B increasing to 100% solvent B within 30 min, continuing at 100% B for further 10 min, RF = 0.3 mL min−1, UV detection 200–600 nm. UV spectra were recorded using a Shimadzu UV-vis spectrophotometer UV-2450 (Shimadzu, Duisburg, Germany). Optical rotation was determined using a PerkinElmer 241 polarimeter (PerkinElmer LAS, Rodgau Jürgesheim, Germany).
Chemical shifts were referenced to the solvents: chloroform-d (1H, δ = 7.27 ppm; 13C, δ = 77.0 ppm) and acetone-d6 (1H, δ = 2.05 ppm; 13C, δ = 29.3 ppm). HRMS mass spectra were measured on the Agilent 1200 series HPLC-UV system (Santa Clara, CA, USA) combined with ESI-TOF-MS (Maxis, Bruker, Bremen, Germany), scan range 100−2500 m/z, capillary voltage 4500 V, temperature 200 °C, (column 2.1 × 50 mm, 1.7 µm, C18 Acquity UPLC BEH [Waters, MZ-Analysetechnik, Mainz, Germany], solvent A: 95% 5 mM ammonium acetate buffer [pH 5.5, adjusted with 1 M acetic acid] with 5% acetonitrile; solvent B: 95% acetonitrile with 5% 5 mM ammonium acetate buffer); gradient: 10% solvent B increasing to 100% solvent B within 30 min, continuing at 100% B for further 10 min, RF = 0.3 mL min−1, UV detection 200–600 nm. UV spectra were recorded using a Shimadzu UV-vis spectrophotometer UV-2450 (Shimadzu, Duisburg, Germany). Optical rotation was determined using a PerkinElmer 241 polarimeter (PerkinElmer LAS, Rodgau Jürgesheim, Germany).
6
Quantitative Analysis of Natural Extracts
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The extracts obtained were dissolved in Acetone:MeOH (1:1) and adjusted to a concentration of 4.5 mg mL−1. An injection volume of 2 µL was applied to an UltiMate® 3000 Series uHPLC (Thermo Fisher Scientific®, Waltman, MA, USA). Mass spectrometry was performed with a connected amaZon® speed ESI Iontrap MS (Amazon, Bruker). HRESIMS measurements were performed with sample concentrations of 1 mg mL−1 with an Agilent 1200 series HPLC-UV system in combination with an ESI-TOF-MS (Maxis, Bruker). The conditions were identical to the methods described before [26 (link)].
7
Characterization of Dendrimer-based Drug Delivery
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The dried powder of FP-loaded dendrimeric delivery systems were characterised by electrospray/ionisation-time of flight (ESI-TOF MS) (Bruker Daltonics, Coventry, UK) at high voltage (4 kV). In electrospray/ionisation mode, samples mass sample mass (m/z) gives rise to multiple charged related-ions typically labelled with a number of charges (n) as (MW + nH) n+, in which H = mass of proton (1.008 Da).
8
Comprehensive Analytical Characterization
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Ultraviolet-Visible (UV/Vis) spectra were acquired using a UV-Vis spectrophotometer UV-2450 (Shimadzu, Kyoto, Japan). Optical rotations were recorded in methanol (MeOH) solution on a MCP 150 polarimeter at 20 °C (Anton-Paar Opto Tec GmbH, Seelze, Germany). ECD spectra were obtained on a J-815 spectropolarimeter (JASCO, Pfungstadt, Germany). High-resolution electrospray ionization mass spectra (HR-ESI-MS) were acquired with an Agilent 1200 Infinity Series HPLC-UV system (Agilent Technologies, Santa Clara, CA, USA) utilizing a C18 Acquity UPLC BEH column (2.1 × 50 mm, 1.7 µm: Waters, Milford, MA, USA), solvent A: H2O + 0.1% formic acid; solvent B: acetonitrile (ACN) + 0.1% formic acid, gradient: 5% B for 0.5 min increasing to 100% B in 19.5 min, maintaining 100% B for 5 min, flow rate 0.6 mL min−1, UV/Vis detection 190–600 nm) connected to an time-of-flight mass spectrometer (ESI-TOF-MS, Maxis, Bruker, Billerica, MA, USA) (scan range 100–2500 m/z, rate 2 Hz, capillary voltage 4500 V, dry temperature 200 °C). NMR spectra were recorded with an Avance III 500 spectrometer (Bruker, Billerica, MA, USA, 1H-NMR: 500 MHz, and 13C-NMR: 125 MHz).
9
Spectroscopic Analysis of Organic Compounds
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Optical rotation values were determined using a Perkin-Elmer (Überlingen, Germany) 241 spectrophotometer. NMR spectra were recorded on a Bruker (Bremen, Germany) 500 MHz Avance III spectrometer with a BBFO (plus) SmartProbe (1H 500 MHz, 13C 125 MHz) and Bruker Ascend 700 spectrometer equipped with 5 mm TXI cryoprobe (1H-700 MHz, 13C-175 MHz) spectrometers, locked to the deuterium signal of the solvent. NMR spectra were measured in acetone-d6; chemical shifts were referenced to the solvent signals. HR-ESI-MS mass spectra were recorded using a Bruker (Bremen, Germany) Agilent 1260 series HPLC-UV/Vis system (column 2.1 × 50 mm, 1.7 m, C18 Acquity UPLC BEH (waters); solvent A: H2O + 0.1% formic acid; solvent B: AcCN + 0.1% formic acid, gradient: 5% B for 0.5 min, increasing to 100% B in 19.5 min and then maintaining 100% B for 5 min, flow rate 0.6 mL/min, UV/Vis detection at 200–600 nm combined with ESI-TOF-MS (Maxis, Bruker, Bremen, Germany) with scan range 100–2500 m/z, capillary voltage 4500 V, dry temperature 200 °C.
10
Characterization of CrtTIR-APAZ, CrtAgo and CrtSPARTA complexes
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Reaction mixtures with a volume of 50 μL were prepared with the following final concentrations: 0.5 μM CrtTIR-APAZ, CrtAgo or CrtSPARTA complex, 500 μM NAD+, 10 mM HEPES pH 7.5 and 125 mM KCl. The mixture was incubated for 1h at 37 °C, after which they were transferred to ice. 150 μL MilliQ H2O was added to the mixture and the mixture was subsequently filtered using centrifugal filters (Pall, 3 kDa MWCO) according to the instructions provided by the manufacturer. The flow through was collected and analyzed by LC-MS using an G1311A HPLC pump (Agilent) with G1367B WP autosampler (Agilent) and an Alltima HP C18 3μm reverse phase column (Avantor) connected to a microTOF benchtop ESI-TOF MS (Bruker Daltonics). Results were analyzed using Compass (Bruker Daltonics, V1.2).
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