Mass spectra of D-AP1, D-AP2, and D-AP3 were recorded using an Agilent InfinityLab LC/MSD system to quantify the ratios of D-AP.
Infinitylab lc msd system
Manufactured by Agilent Technologies
Sourced in United States
The InfinityLab LC/MSD System is a liquid chromatography-mass spectrometry (LC/MS) instrument designed for analyzing and identifying compounds in complex samples. It provides sensitive and accurate detection of a wide range of molecules. The system combines high-performance liquid chromatography (HPLC) with a mass spectrometer to enable efficient separation, identification, and quantification of analytes.
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5 protocols using infinitylab lc msd system
1
Quantification of D-AP Ratios
2
NMR and Mass Spectrometry Analyses
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NMR experiments were performed on a Bruker Avance II 600 spectrometer (Karlsruhe, Germany) at 600.19 (1H) and 150.91 MHz (13C). The pure isolated compounds were dissolved in deuterated methanol from Euriso-Top (Saint Aubin, France). An Agilent InfinityLab LC/MSD System (Santa Clara, CA, USA) was used to measure the low-resolution mass spectra in positive mode at a capillary energy of 4000 V and nebulizer gas of 40.0 psi (dry gas), at 10.0 L/min and at a temperature of 300 °C; the recorded scan range was 100–1500 m/z. The high-resolution mass spectra were recorded on a micrOTOF-Q II mass spectrometer (Bruker-Daltonics, Bremen, Germany). The experiments were conducted in the positive as well as negative ESI mode by applying the following parameters: capillary energy, 4500 V for positive mode and 3500 V for negative mode; nebulizer gas, 6.4 psi; dry gas, 4.0 L/min at a temperature of 180 °C; and recorded scan range of 100–600 m/z.
3
Quantitative Amino Acid Analysis by LC-MS
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LC-MS data recorded following Marfey’s method were measured with an Agilent InfinityLab LC/MSD System (Santa Clara, CA, USA), comprising of an Agilent HPLC 1260 HPLC equipped with binary pump, autosampler, column oven, and photodiode array detector.
4
Synthesis and Characterization of Peptide Derivatives
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The peptide derivatives Rh‐GFFYE, Ac‐GFFYE, and Nap‐GFFYE were prepared similarly to our previous study.[19 ] Briefly, they were synthesized by standard Fmoc solid‐phase peptide synthesis (SPPS), which used 2‐chlorotrityl chloride resin and the corresponding N‐Fmoc protected amino acids with side chains suitably protected by a tert‐butyl group or tert‐butyloxycarbonyl group. The resulting crude products were purified by high‐performance liquid chromatography (HPLC, LUMTECH, German) and lyophilization. These compounds were characterized by 1H‐NMR (Bruker ARX 500) and MS. MS was conducted at the Agilent InfinityLab LC/MSD system.
5
High-Resolution Mass Spectrometry Analysis
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High-resolution mass spectra were measured with an Exploris 120 Orbitrap mass spectrometer (Thermo, MA, USA). The experiments were performed in positive ESI mode with the following parameters: capillary energy: 3500 V; sheath gas: 2 U; aux gas: 2 U and a vaporizer temperature of 250 °C; the recorded scan range was 100–600 m/z and the Orbitrap resolution was set to 15,000. Low-resolution mass spectra were recorded on an Agilent InfinityLab LC/MSD System. It comprised an Agilent 1260 HPLC, equipped with a binary pump, autosampler, column oven and photodiode array detector (Santa Clara, CA, USA).
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