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Exact(5)
False identifications were prevented by the user assessment for each peak or by the retention times and mass errors of the compounds.
Based on MS2 fragmentation patterns and exact mass determination (4-digit mass errors of < 1.6 ppm), these peaks were annotated as desulfoBGLS (dBGLS) and malonylated dBGLS (mdBGLS).
The MS spectra were internally calibrated using porcine trypsin autolytic products (m/z 842.509, m/z 1045.564, m/z 1940.935 and m/z 2211.104) resulted in mass errors of less than 30 ppm.
The calculated mass errors of the unmodified and HNE-adducted peptide were each within 1 part per million (ppm) of theoretical values, adding high confidence in the identification of the HNE-adducted peptide.
First, the single-scan FC-43 profile spectra in Figure 2A,B highlight the instrument's typical mass accuracy and precision, demonstrating externally calibrated mean mass errors of 0.24 ± 1.04 ppm for 38 routinely observed ions (errorrms = 1.06 ppm) (Supplemental Table S2 in the Supporting Information).
Similar(55)
mass error of each single spectrum 2000; desired mass error for the MSP 200; desired peak frequency 25%%; max.
To validate the MS data, the mass error of all the identified peptides were checked and the results showed that the distribution of mass error was near zero and most of them were less than 5 ppm (Fig. 2b), which means the mass accuracy of the MS data fits the requirement.
As for the N-oxide DVEN when its precursor ion at m/z 280.1910 (accurate mass error: –0.71 ppm) was isolated and fragmented, an ion at m/z 113.0966 was observed that could be interpreted as [C7H12O + H]+ with an accurate mass error of ± 0.00 ppm (Figure 7b).
The relevant search parameters were as follows: enzyme specificity was fully tryptic, a precursor mass error of 3.0 Da, a fragment mass error of 0.5 m/z, and a peptide mass range spanning 400 to 5100 Da.
Searches were performed allowing for complete carbamidomethylation (alkylation) of cysteine residues, partial oxidation of methionine residues, one missed cleavage and a mass error of 250 ppm.
Other parameters used in data analysis were: four allowed missing cleavages; mass error of 10 ppm for precursor ions and 0.5 Da for fragment ions.
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