ACCLAIM oa - Cromlab

Lot Qualification TestsPa r t Fo u r: AppendixAppendix 1: Lot Qualification Testsfor Acclaim-Bonded SilicaColumn Performance Test, Acclaim C18, C8, PA, PA2Polar SelectivityThis test protocol is part of the column performance test:column pressure, retention time, efficiency, and asymmetry forphenanthrene are specified for each column. This parameter alsois used to monitor batch-to-batch reproducibility as measured bythe selectivity of a polar analyte (dimethylphthalate) relative to anonpolar analyte (phenanthrene).Mobile Phase: 70/30 v/v acetonitrile/waterFlow Rate: 1.0 mL/minTemperature: 30 °CInj. Volume: 5 µLDetection: UV, 254 nm, 6-mm path lengthPeaks: 1. Uracil 0.015 mg/mL2. Dimethylphthalate 0.075 µL/mL3. Phenanthrene 0.015 mg/mLMetal Activity2,2’-Bipyridyl chelates with metals whereas 4,4’-bipyridyldoes not form chelation complexes. If metals are present, the 2,2’isomer elutes with a high asymmetry factor (tailing). The peakasymmetry ratio for the two analytes should be approximately 1.0if no metal contamination is present.Mobile Phase: 50/50 v/v methanol/waterFlow Rate: 1.0 mL/minTemperature: 30 °CInj. Volume: 5 µLDetection: UV, 254 nmPeaks: 1. 4,4’-Bipyridyl 50 ng/µL2. 2,2’-Bipyridyl 200 ng/µLBase AsymmetryBasic compounds exhibit poor peak shape when the silica surfaceis incompletely covered during the bonding process, or whenthe silica substrate is inadequately prepared. This test monitorssurface coverage with an application-based analysis.Mobile Phase: 80/20 v/v methanol/30 mM phosphate, pH 6.0Flow Rate: 1.0 mL/minTemperature: 30 °CIn. Volume: 5 µLDetection: UV, 220 nmPeaks: 1. Uracil 0.04 mg/mL2. Propranolol 0.06 mg/mL3. Toluene 0.08 µL/mL4. Amitriptyline 0.24 mg/mLHydrophobic Steric SelectivityThis parameter is used to monitor batch-to-batch bondingreproducibility as measured by the relative selectivity of two differentlyshaped aromatic analytes: o-terphenyl and triphenylene. Thistest is used to characterize all the C18 and polar-embedded phases.Mobile Phase: 90/10 v/v methanol/waterFlow Rate: 1.0 mL/minTemperature: 30 °CInj. Volume: 5 µLDetection: UV, 254 nmPeaks: 1. Toluene 2 µl/mL2. Phenanthrene 0.03 mg/mL3. o-terphenyl 0.14 mg/mL4. Triphenylene 0.03 mg/mLAPPENDIXAcid AsymmetryThis test is used to characterize polar-embedded phases.Acidic compounds exhibit poor peak shape when the bondedphase has basic sites due to degradation or contamination.Mobile Phase: 35/65 v/v acetonitrile/0.1% TFA in waterFlow Rate: 1.0 mL/minTemperature: 30 °CInj. Volume 5 µLDetection: UV, 240 nmPeaks: 1. 4-Hydroxybenzoic acid 50 ng/µL2. Benzoic acid 70 ng/µL3. Ethyl 4-hydroxybenzoate 50 ng/µLSome application-specific tests are used for lot validation. Readabout these tests in the chapters for Acclaim 300 and Acclaim OA.- 50 -www.dionex.com