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An optimized HPLC method gave a baseline separation of the two glucuronides.
Applying these conditions, the baseline separation of the analytes was obtained in about 10 min.
The longer retention times and better peak shapes gave baseline separation of the product from other reaction components.
Experimental design strategies were applied in order to reach baseline separation of the compounds in a short analysis time.
Possibly, the UPLC parameters to achieve baseline separation of the two glucuronides can be applied to other cases of difficult to separate glucuronides.
A chromatogram is displayed in Additional file 1 with baseline separation of the two glucuronides and UV spectra of the three peaks.
Similar(41)
The baseline separation of these amino acid derivatives was obtained on a C8 column with a binary gradient elution using the mobile phase of 20 mmol/L mixed acid buffer solution (pH 6.40) and methanol with 2.5% tetrahydrofuran (THF).
The baseline separation of these derivatives was carried out on a C8 column with methanol tetrahydrofuran-50 methanol tetrahydrofuran-50(55/5/40, v/v/v) as a mMbile pHase.
Baseline separation of these compounds was not possible under the UPLC conditions tested.
When the longer, 48 cm, separation capillary (effective length: 33 cm) was connected to the CAs-CHIP instead of 12 cm capillary, however, baseline separation of these species after deproteinization was successfully achieved (resolution: R=5.7).
Baseline separation of these analogs was achieved by CE using β-cyclodextrin as additive and they were monitored at 214 nm.
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