Agilent HPLC Column Selection Guide

Columns for Bioanalytical ChromatographyCapillary Columns for HPLC Analyseswith UV and MS Detection100UVColumn:Mobile Phase:Flow Rate:Detector:MS Conditions:Sample:ZORBAX 300SB-C185064-82630.3 x 150 mm, 5 µm5-55% B in 50 min, to 85% B from 55-57 minA: 0.1% Formic Acid in WaterB: 0.1% Formic Acid in ACN5.5 µL/min206 nmLC/MS: Pos. Ion ESI withLC/MSD trap-Vcap 4000 VDrying Gas Flow: 7 L/minDrying Gas Temperature: 250°CNebulizer: 15 psiCapillary Exit Volt: 50 VMax Accum Time: 300 msTotal Averages: 3Isolation Width: 3 m/zFrag Ampitude: 1.0 V100 nLBeta Casein Digest (4 pmol)A ZORBAX 300SB-C18 capillary column (0.3 mm ID) is used forthe separation of the protein digest. Detection is by both UV andElectrospray MS. MS detection can be used for identification ofpeptide fragments.% Relative AbundanceUV at 206 nm806040200x100.80.60.40.20.010MS10151520 25 30 35 40 45 50Time (min)20 25 30 35 40 45 50Time (min)LCSB007ZORBAX Nano Columns For High SensitivityProtein Digest Analysis by LC/MSColumn: ZORBAX 300SB-C185065-99110.075 x 150 mm, 3.5 µmMobile Phase: A: Water + 0.1% Formic Acid,B: ACN + 0.1% Formic AcidFlow Rate: 600 nl/minGradient:2% B to 52% B in 25 minDetector:Positive Ion Nano Electrospray MSSample:100 fm (1 µl) Digest of 8 ProteinsIntens.1.51.00.5A ZORBAX nano HPLC column, 0.075 mm ID, is used for highsensitivity LC/MS analysis of a protein digest sample.2.5 5.0 7.5 10.0 12.5 15.0 17.5 20.0 22.5Time (min)LCSB008Order online at www.agilent.com/chem/store115