Development of Master Method—Individual basic and disperse dye instrumental methods were combined and evaluated with the dye standards. A mixture of all dye standards, basic and purified disperse dyes, was injected into the column. The optimized parameters for the master method are listed in Table 2, and the chromatogram of the dye mixture can be seen in Fig. 7. After the individual methods were optimized, it was found that there was no overlap in their solvent gradients. Thus, their gradients could simply be added together to create a "master method." Mass spectrometer parameters for the first section of the method in Table 2, average values calculated during optimization, included a capillary voltage (CV) set to −3375 V, capillary exit voltage (CEV) set to 188 V, and the trap drive (TD) set to 60 V. After 30 min, where the method changes from the parameters for the basic dyes to those for the disperse dyes, the MS parameters change to a CV of −4000 V, CEV of 154 V, and TD of 60 V. The MS scanned a mass range of 50–1000 m/z in 200 ms or a target of 30,000 ions for the entire run. Evaluation of Table 2 shows that each method, Basic, Disperse, and the final "master" method, has several segments. For each method, it was necessary to adjust the slope of the gradient to resolve as many of the dyes as possible. In some instances, the slope had to be decreased to obtain adequate resolution. Lastly, the flow rate for each method was steeply increased toward the end of the run to facilitate cleaning the column of any late-eluting components. FIG. 7–Chromatogram of 15 basic dyes and 13 disperse dyes.
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