PEG 400 by UHPLC with corona ultra

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Corona ® ultra Charged Aerosol DetectorApplication noteMethod Parameters - UHPLCSystem:Waters Acquity UPLCColumn:Acquity UPLC BEH C182.1 x 50 mm, 1.7 µmMobile Phase A: DI Water / Acetonitrile (93 : 7)Mobile Phase B: AcetonitrileGradient: HPLC - Table 2Injection Volume: 1 µL of a ~2 mg / mL standardin DI waterDetector: Corona ultraFilter:MediumRange:200pAResults and DiscussionFigure 1 shows a chromatogram from the UHPLCmethod overlaid onto the HPLC chromatogram witha full 20 minute time scale. The resolution for thethird major peak and seventh major peak observedin the cluster for each was calculated, and the comparisonis shown in Table 3. The traditional HPLCapproach did achieve superior resolution for bothpeaks, however the resolution for the UHPLCmethod was within 70% of the HPLC method with afour-fold reduction in analysis time and solventconsumption.TimeFlow Rate% B(min)(mL/min)0 3 115 30 117 3 1Table 1 - HPLC Gradient.TimeFlow Rate% B(min)(mL/min)0 3 0.051 10 12.4 20 13 30 13.4 3 0.5Table 2. UHPLC Gradient.Peak 3 Peak 7HPLC UPLC HPLC UPLCRetention Time (min) 8.67 1.35 12.42 2.24Resolution 7.8 5.6 4.1 3.4Table 3. Comparison of Resolution Between HPLCand UHPLC for Two Selected Analytes.Figure 2. Overlay of Four Individual Analytical Runs on the UHPLC Corona ultra System.
Corona ® ultra Charged Aerosol DetectorApplication noteThe UHPLC method reproducibility is shown Figure2, with overlays of four injections of a 2 µg PEG 400standard on column with the calculated %RSD foreach peak is represented in Table 4. A responsecurve was then prepared from ~2 to 2000 µg/mL. Alimit of quantification for the PEG 400 cluster wasobserved at ~18 ng total mass on column, which isshown in Figure 3. The prepared standards wereanalyzed, and the peak area versus the total masson column is plotted in Figure 4. Assuming that theaverage %-area for each peak from the 2 µg oncolumn injections can be used to adjust for themass on column for each peak over the entirecalibration range, a new adjusted response curvewas calculated and is plotted in Figure 5. This wasdone to illustrate the uniformity of response of theCorona CAD for the different MW fractions of PEG400.Retention TimeResponse(min)(peak area)% RSD1.11 1124 1.68%1.34 2596 1.73%1.56 2282 1.09%1.76 3050 1.32%1.94 1010 0.46%2.09 1730 0.95%2.24 1941 1.48%2.37 649 0.76%2.49 669 1.35%2.60 320 1.29%Table 4. Reproducibility of Ten PEG 400Analytes (n=4).0.200.00-0.20-0.40-0.60mV-0.80-1.00-1.20-1.40-1.600.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 3.60 3.80 4.00 4.20 4.40 4.60 4.80 5.00MinutesFigure 3. High Sensitivity Analysis of PEG 400 on the UHPLC Corona ultra System(~18 ng total, on column).
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PEG 400 by UHPLC with corona ultra

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