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On the other hand, no need for internal standard since a simple procedure for sample treatment was applied.
The CCS method was studied in two procedures for internal standard calibration (CCSI) and external standard calibration (CCSE).
In this study, a new method for the absolute quantification of three proteases involved in insulin maturation, namely PC1/3, PC2 and CPE, was developed by coupling a stable isotope dimethyl labeling strategy for internal standard peptide preparation with SID-MRM-MS quantitative technology.
for internal standard and fragment size determination and for allelic designations.
The working solution was then diluted 1∶10 in a chloroform/methanol mixture (2∶1, v/v) for same-day use to the final concentration used for internal standard addition.
The transition m/z 257.3 to m/z 213.3 was monitored for internal standard.
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The retention times for internal standards were 3.27 min (LAM), 2.2 min (ZDV), and 2.85 min (NVP), respectively.
Deuterium labelled 25-hydroxyvitamin D2 and D3 were used for internal standards.
This eliminates the need for internal standards and makes measurements of both known and novel metabolites possible [52], [53].
Instrument variability was determined by calculating the median relative standard deviation (RSD) for internal standards that were added to each sample prior to injection into the mass spectrometers.
The sample was analyzed in targeted SIM mode with 2 Th windows for native congeners and full scan mode for internal standards (at 1 pg/μL).
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