ACCLAIM oa - Cromlab

Bioscience ApplicationsProteins and Peptides350mAU0Cytochrome c from Three Species on Acclaim 300 C1821–1000 2 4 6 8 10 12Minutes3Column: Acclaim 300 C18, 3 µmDimensions: 4.6 × 150 mmMobile Phase: (A) 95/5/0.1 v/v water/acetonitrile/TFA(B) 95/5/0.1 v/v acetonitrile/water/TFAGradient: Time (min) % A % B–5 80 200 80 2015 45 5520 45 55Flow Rate: 1.0 mL/minInj. Volume: 10 µL (1 µg/µL)Detection: UV, 214 nmPeaks: 1. Yeast cytochrome c2. Equine cytochrome c3. Bovine cytochrome c19104Protein Mixture by Reversed-Phase HPLC on Acclaim 300 C181,000mAU14325–506 8 10 12 14 16Minutes67Column: Acclaim 300 C18, 3 µmDimensions: 4.6 × 150 mmMobile Phase: (A) 5 mM MSA in 95/5 H 2 O/acetonitrile(B) 5 mM MSA in 5/95 H 2 O/acetonitrileInj. Volume: 25 µLDetection: UV, 214 nmGradient: Time % A % B Flow0 100 0 1.03 100 018 0 10023 0 100Peaks: 1. RNAse A2. Impurity in carbonic anyhdrase3. Impurity in lysozyme4. Lysozyme5. Impurity in carbonic anhydrase6. Myoglobin7. Carbonic anhydrase20407Comparison of Acid Additives for Peptide Analysis on Acclaim 3002AU0ABC2 4 6 8 10 12MinutesColumn: Acclaim 300 C18, 3 µm,4.6 × 150 mmPump: Summit P680AMobile Phases: A: 70% acetonitrile v/vB: WaterC: Acid stock solutionGradient Times: –8 0 16%A 7 7 75%B 68 68 0%C 25 25 25Flow: 1.0 mL/minTemperature: TCC100 thermostat at 30 °CInjection: ASI-100 autosampler; 5 µLDetector: UVD 340U; UV at 214 nmAcid Additive: A. Phosphoric Acid, 0.15%B. Trifluoroacetic Acid, 0.1%C. Formic Acid, 0.1%Sample: Sigma H2016 Peptide MixPeaks: 1. Gly-Tyr2. Val-Tyr-Val3. Met-Enkephalin4. Angiotensin-II5. Leu-Enkephalin230641.2AUPeptide Test Mix on Acclaim 300 and PA2 with Formic Acid0AB2 4 6 8 10 12MinutesColumns: (A) Acclaim 300 C18, 3 µm,4.6 × 150 mm(B) Acclaim PA2, 3 µm,4.6 × 150 mmPump: Summit P680AMobile Phases: (A) 70% acetonitrile v/v(B) Water(C) 0.4% formic acid v/vGradient Times: –8 0 16%A 7 7 75%B 68 68 0%C 25 25 25Flow: 1.0 mL/minTemperature: TCC100 thermostat at 30 °CInjection: ASI-100 autosampler; 5 µLDetector: UVD 340U; UV at 214 nmSample: Sigma H2016 Peptide MixPeaks: 1. Gly-Tyr2. Val-Tyr-Val3. Met-Enkephalin4. Angiotensin-II5. Leu-Enkephalin23066APPLICATIONSwww.dionex.com - 47 -