BETTER THAN SECs.ppt

BETTER THAN SEC’s Obligatory Equation GPC Simple SEC Osmometry: Real Science Light Scattering: Osmometer without the membrane LS adds optical effects ? Size SEC/MALLS SEC/MALLS Scattering Envelope for a Single Slice Universal Calibration Grubisic, Rempp Benoit, JPS Pt. B, 5, 753 (1967) Universal Calibration Equations Objectives Use a-helical rodlike homopolypeptides to test validity of universal calibration in GPC. Can GPC/Multi-angle Light Scattering arbitrate between disparate estimates of stiffness from dozens of previous attempts by other methods? Strategy Polymers Used Mark-Houwink Relations Polystyrene Standards: the Usual Universal Calibration Works for These Rods and Coils 2nd Virial Coefficient Equations 2nd Virial Coefficient (Excluded Volume Limit) is Another Universal Descriptor Persistence Length ap from Rg Persistence Length of Helical Polypeptides is “Very High” SEC/MALLS in the Hands of a Real Expert They were young when GPC was. Small Subset of GPC Spare Parts Other SEC Deficiencies 0.05 M salt at 10 am, 0.1 M salt at 2 pm? 45oC at 8 am and 50oC at noon? Non-size exclusion mechanisms: binding. Big, bulky and slow (typically 30 minutes/sample). Temperature/harsh solvents no fun. You learn nothing by calibrating. Must we separate ‘em to size ‘em?Your local constabulary probably doesn’t think so.  Sizing by Dynamic Light Scattering—a 1970’s advance in measuring motion, driven by need to measure sizes, esp. for small particles. Hot Ben Chu / Chi Wu Example Reptation: inspired enormous advances in measuring polymer speed…and predicts More favorable results for polymers in a matrix. Painting Molecules* Makes Life Easier Fluorescence Photobleaching Recovery FPR for Pullulan (a polysaccharide) FPR Chromatogram Separation ResultsPullulan M = 50/50 mix of 11,800 and 380,000 Better Resolution “Soon”? Simulation of FPR Results(Most Desirable Situation) Ultimate Goal: A Black Box for MWD Conclusions For a limited number of cases, thi