Concentrations of major active compounds, i.e., cyanidin-3-O-glucoside, cyanidin-3-O-rutinoside, cyanidin-3-O-xylosyl-rutinoside and cyanidin-3-sambubioside, for both powder and extract samples were measured by HPLC–MS. The lyophilized samples were mixed with 80:20 (water: sample). The samples were filtered and then injected into an LC–MS system (Agilent 6538 UHD-QTOF equipped with Agilent 1290 infinity UPLC). Upon extracting the chromatograms based on the reported m/z, calculations were performed by integrating the peak to obtain the area. Anthocyanin standards were purchased from Extrasynthese S.A.S. (Lyon, France).
6538 uhd q tof
Manufactured by Agilent Technologies
The 6538 UHD Q-TOF is a high-resolution quadrupole time-of-flight mass spectrometer designed for accurate mass measurements and high-sensitivity detection. It features a ultra-high mass resolution and a wide mass range for comprehensive analysis of complex samples.
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3 protocols using 6538 uhd q tof
1
Quantification of Anthocyanins in Berry Extracts
2
Detailed Spectroscopic Characterization Protocol
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IR spectra recorded on a Bruker Tensor 37 (FT-IR) spectrophotometer. 1H-NMR spectra recorded on Bruker Avance 400 (400 MHz) spectrometer at 295 K in CDCl3; chemical shifts (δ in ppm) and coupling constants (J in Hz) reported in standard fashion with reference to either internal standard tetramethylsilane (TMS) (δH = 0.00 ppm) or CHCl3 (δH = 7.25 ppm). 13C-NMR spectra recorded on Bruker Avance 400 (100 MHz) spectrometer at RT in CDCl3; chemical shifts (δ in ppm) are reported relative to CDCl3 [δC = 77.00 ppm (central line of the triplet)]. In the 13C-NMR, the nature of carbons (C, CH, CH2 and CH3) was determined by recording the DEPT-135 spectra, and is given in parentheses and noted as s = singlet (for C), d = doublet (for CH), t = triplet (for CH2) and q = quartet (for CH3). In the 1H-NMR, the following abbreviations were used throughout: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet. The assignment of signals confirmed by 1H, 13C CPD and DEPT spectra. High-resolution mass spectra (HR-MS) were recorded on an Agilent 6538 UHD Q-TOF using multimode source. Reactions were monitored by TLC on silica gel coated on alumina plate or glass plate using a mixture of petroleum ether and ethyl acetate as eluents. Reactions carried out under oxygen atmosphere.
3
Rapid LC-MS Analysis of Analytes
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Samples were prepared by adding 25 μL of ultrapure water to 75 μL solutions of analytes in acetonitrile + 0.1% TFA. A PLRP-S column (Agilent) was equilibrated to an 85:15 ratio of solvent A (ultrapure water + 0.1% formic acid) to solvent B (acetonitrile + 0.1% formic acid). Liquid chromatography separations were achieved by linear gradient elution, transitioning from 15% to 95% solvent B over 6 min followed by a 2 min hold at 95% B. The column was re-equilibrated to 15% solvent B for 2 min in between injections of the same sample (two technical replicates run per sample, 2 μL injection volume, 600 μL/min flow rate, 50°C). Two blank runs were implemented between samples to ensure against column holdover of analytes. Electrospray ionization mass spectrometry analysis was carried out in positive mode with a capillary voltage of 2Hz (Agilent 6538 UHD Q-TOF).
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