Figure 1-10. Gradient method transfer from regular high-pressure liquid chromatography (HPLC) to ultrahigh-pressure liquid chromatography (UHPLC). Separation of a pharmaceutical mixture containing the main product (6) and 11 impurities in gradient mode with HPLC and UHPLC systems: (A) original HPLC method: column: XBridge C18 150 x 4.6 mm, 5 um; flow rate: 1000 uL/min; injected volume: 20 uL; total gradient time: 45 min. (B) Transferred UHPLC method: column: Acquity bridged ethylsiloxane/ silica hybrid (BEH) C18 50 x 2.1 mm, 1.7 um; flow rate: 610 uL/min; injected volume: 1.4 uL; total gradient time: 5.1 min. (C) Transferred and optimized UHPLC method: column: Acquity BEH C18 50 x 2.1 mm, 1.7 um; flow rate: 1000 uL/min; injected volume: 1.4 uL; total gradient time: 3.1 min.
Adapted from Guillarme, D., Nguyen, D.T.T., Rudaz, S., Veuthey, J.L., 2008. Method transfer for fast liquid chromatography in pharmaceutical analysis: Application to short columns packed with small particles - Part II, Gradient separation. Eur. J. Pharm. Biopharm. 68, 430-440..
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