LC ACQUITY UPLC H-Class Bio TUV Sigma, mg/ml 0.01 N 276 nm μl 0.4 ml/min L g/l / 99% / 65/15/20 v/v/v 10% 10% 0.01 N ACQUITY
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1 Ultra Performance Liquid Chromatography Stephan Koza, Paula Hong, and Kenneth J. Fountain Waters Corporation, 34 Maple Street, Milford, MA, USA & SEC USP EPHMW USP EP Volume HPLC SEC HMW USP L μm 7.8 mm x 30 cm EP R nm 5-10 μm 7.5 mm 30 cm μm BEH Waters Ultra Performance Liquid Chromatography UPLC HMW ACQUITY UPLC H-Class Bio ACQUITY UPLC BEH125 SEC, 1.7 μm Empower TM 3 UPLC USP EP 1
2 LC ACQUITY UPLC H-Class Bio TUV Sigma, mg/ml 0.01 N 276 nm μl 0.4 ml/min L g/l / 99% / 65/15/20 v/v/v 10% 10% 0.01 N ACQUITY UPLC BEH125 SEC, 1.7 μm, 4.6 x 150 mm P/N ACQUITY UPLC BEH125 SEC, 1.7 μm, 4.6 x 300 mm P/N ACQUITY UPLC BEH200 SEC, 1.7 μm, 4.6 x 300 mm P/N BioSuite TM 125 UHR SEC, 4 μm, 4.6 x 300 mm P/N Insulin HMWP HPLC, 10 μm, 7.8 x 300 mm P/N WAT Empower 3 Auto Blend Plus TM USP EP HMW ACQUITY UPLC BEH125 SEC, 1.7 μm, 4.6 x 300 mm P/N g/l L- 15% 20 % v/v HMW HMW ACQUITY UPLC H-Class Bio Empower 3 USP EP USP P/V HP HV EP USP 0.4% HMW 2.0 EP HMW nmusp ACQUITY UPLC BEH125 SEC, 1.7 μm 125 EP SEC 1 BEH125 SEC, 1.7 μm 200 ACQUITY UPLC BEH200 SEC, 1.7 μm HMW % 2 HMW % 0.4% P/V= P/V=37 P/V USP EP 0.4% HMW P/V= HMW HMW Ultra Performance Liquid Chromatography 2
3 ACQUITY UPLC BEH125, 1.7 µm Rs=3.37 HMW ACQUITY UPLC BEH200, 1.7 µm Rs=3.13 Rs=2.38 Rs= min min ACQUITY UPLC BEH ACQUITY UPLC BEH200 HMW SEC 1 2 EP SEC HMW SEC 5-10 μm USP 3-10 μm μm 3 SEC mm μm 4 μm 10 μm 35% 2 HMW P/V 3 EP USP 1.7 μm P/V=37 4 μm P/V=9 10 μm P/V=81.7 μm ~5 ml 4 μm ~6 ml 10 μm ~13 ml μm 10 μm 2 μm UPLC Ultra Performance Liquid Chromatography 3
4 ACQUITY UPLC BEH125, 1.7 µm Rs=3.37 HMW BioSuite125 UHR, 4 µm Rs=2.21 HMWP, 10 µm (7.8 x 300 mm) Rs=2.08 Rs=1.93 Rs=1.95 Rs=1.92 Rs= min min min μm 125HMW SEC 10 μmep 100 μl 0.5 ml/min 1 2 AU min 0.83% HMW ACQUITY UPLC BEH125 Insulin min min Waters HMWP (7.8 x 300 mm) AU % HMW min 3 2 ACQUITY UPLC BEH125 Waters HMWP Waters HMWP EP 100 μl 0.5 ml/min ACQUITY UPLC BEH Ultra Performance Liquid Chromatography 4
5 4.6 x 150 mm 4.6 x 300 mm ACQUITY UPLC BEH mm 300 mm 41-43% 41% mm 300mm EP USP HMW mm mm P/V=5 300 mm 150 mm 300 mm 2 ACQUITY UPLC BEH125, 1.7 µm Rs=3.37 ACQUITY UPLC BEH125, 1.7 µm (4.6 x 150 mm) Rs=2.35 HMW Rs=1.87 Rs= min min 4 ACQUITY UPLC BEH mm 150 mm 0.5 ml/min 300 mm 10 μl 150 mm 5 μl Ultra Performance Liquid Chromatography 5
6 Average Peak/Valley Average Rs P/V Rs USP/EP Limit (NLT 2) 5 0 ACQUITY UPLC BEH125, 1.7 µm, 300 mm ACQUITY UPLC BEH125, 1.7 µm, 150 mm HMWP, 10 µm, 300 mm Columns BioSuite BEH125, 4 µm, 300 mm ACQUITY UPLC BEH200, 1.7 µm, 300 mm HMWP/V USPRs EPUSP P/V HMW USP EP SEC 5 SEC μm BEH Waters Ultra Performance Liquid Chromatography UPLC SE- HPLC 1. Walsh G. Biopharmaceutical benchmarks Nat Biotechnol. 2010; 28: Insulin, Human, European Directorate for the Quality of Medicines (EDQM), Insulin, The United States Pharmacopoeia Convention, Inc., Rockville, MD, TEL FAX F TEL FAX Waters UPLC ACQUITY UPLC Waters Corporation Empower BioSuite Auto Blend Plus The Science of What s Possible Waters Corporation 2012 Waters Corporation. Printed in Japan JA PDF
Waters Sigma-Aldrich 1.0 mg/ml LC Waters Alliance HPLC ACQUITY UPLC H-Class Bio 30 cm Alliance HPLC UV ACQUITY UPLC TUV mm nm XBridge Protein B
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