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P. 6

Analysis





                     Ensures high-throughput method development and scaling up




                         Automatically performs a variety of method scouting processes                                                      Method scouting for optimization of separation conditions and scaling up
                                                                                                                                            to preparative size
                     The high-speed performance of SFC can shorten the time required for method scouting. The system automatically
                     generates a large number of methods by utilizing combinations of up to 12 columns, four modifiers, and different
                                                                                                                                        For high-purity isolation,  target peaks need to be adequately separated, for which the user must determine
                     ratios of modifiers to mobile phase. With the UHPLC / SFC Switching system, screening by UHPLC and SFC can be
                                                                                                                                        optimum column and separation parameter settings (method scouting). The Nexera UC chiral screening system and
                     carried out in the same batch. Moreover, Method Scouting Solution, optional software for method scouting, makes it
                                                                                                                                        Method Scouting Solution can be used to screen columns more quickly and accurately (Step 1). Once the optimum
                     easy to set up multiple conditions.
                                                                                                                                        column has been identified, smoothly scale up to preparative scale flow rates, preserving the peak separation
                                                                                                                                        while increasing the mass load (Step 2).
                                                                                                                                        *Preparative SFC: page 10 and following.
                                                                                                                                          Step 1   Method scouting at analytical scale

                                                                                                                                        Method  scouting is  easy even for first-time  users—simply  execute  the  batch  table generated  automatically  by Method
                                                                                                                                        Scouting Solution. The system can automatically switch between different settings to run the scouting process continuously
                                                                                                                                        day or  night, even for multiple modifiers and columns.  Various types of data can be displayed  in the data browser and
                                                                                                                                        multi-data reports generated with resolution values for all data to assist in evaluating separation conditions.
                                                                                                                                          Step 2   Scaling up                4.6 mm I.D. column (Nexera UC)
                                                                                                                                                                             mAU (×1,000)
                                                                                                                                                                                                           Column: Shim-pack UC-PBr 4.6 × 250 mm
                                                                                                                                        Using a Shim-pack UC series column enables you                     Flow rate: 3.0 mL/min
                                                                                                                                        to increase mass load while maintaining                            Injection volume: 20 µL
                                                                                                                                        separation performance. The optimum                                Fraction volume: 10 mg
                                                                                                                                        preparative column determined from Step 1 can
                                                                                                                                        be used to scale up the column size, flow rate
                                                                                                                                                                             0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 min
                                                                                                                                        and injection volume based on the desired
                                                                                                                                        fraction volumes.
                                                                                                                                                                             20 mm I.D. column (Nexera UC Prep)
                                                                                                                                                                             mAU (×1,000)
                                          A screen shot of Method Scouting Solution for Nexera UC user interface.                                                                                          Column: Shim-pack UC-PBr 20 × 250 mm
                                                                                                                                                                                                           Flow rate: 56.7 mL/min
                                                                                                                                                                                                           Injection volume: 500 µL
                                                                                                                                                                                                           Fraction volume: 250 mg
                         Chiral analysis with “Nexera UC Chiral Screening System”

                     CHIRALPAK  Series and CHIRALCEL  Series columns (Daicel Corporation) for chiral analysis are capable of resolving                                       0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 min
                                                ®
                              ®
                     a wide variety of compounds by showing complementary separation targets. The combination of the Nexera UC
                     Chiral Screening System and these columns simplifies method scouting for chiral analysis.                               Selecting an SFC Column
                                                                                                                                        The HPLC mobile phase for reverse phase analysis is very different from the one used for normal phase analysis. In contrast,
                                                                                                                                        SFC uses a mixture of supercritical CO2 and a modifier (an organic solvent such as methanol) regardless of the stationary phase
                                                                             mAU
                                                                                                                                        used. Therefore, the same mobile phase composition can be used for serial analysis through all columns. Column scouting is
                                                                             300  Indapamide
                                                                                            RT = 2.644                                  effective by using the following set of six columns, each providing a different separation selectivity.
                                                                                            Rs = 2.66
                                                                             200
                                                                                                                                                                                    6 columns set
                                                                                                                                                    UC-ODS        UC-Sil II   UC-Diol II    UC-PolyVP    UC-PolyBT      UC-PBr
                                                                             100
                                                                                                                                                                                                                            Br  Br
                                                                                                                                                                                      OH
                                                                              0                                                                                                      OH                                    Br
                                                                                                                                                                                                                               Br
                                                                                                                                         Chemistry                                   O                                     O  Br
                                                                               0.0  1.0  2.0  3.0  4.0  5.0  min
                                                                                                                                                                                  Si                                    Si
                                                                                  Columns: CHIRALPAK  AD-3
                                                                                                ®
                                                                                Modifier: Acetonitrile/Ethanol=7/3                                                               OH
                                                                                                                                                 The separation mode is  This is excellent for retention  The separation mode is   A favorable peak shape   This is excellent for   With ODS, separation of
                                                                                                                                                 reverse phase. Retention   of basic compounds and   normal phase. This   is obtained even   resolving aromatic   poorly retained
                                                                                                                                         Features
                                                                                                                                                  is provided through   recognition of their tertiary   inhibits non-specific   without acid-base   compounds through π-π  compounds is improved.
                                                                                                                                                 hydrophobic interaction.  structures.  interactions.  additives.  interactions.
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