Recent Developments in Polymer Characterization And how we may have to modify them for nanoparticles Obligatory Equation SEC = GPC = GFC Size Exclusion Chromatography Gel Permeation Chromatography Gel Filtration Chromatography A riddle After a hurricane, many trees fall over and bend into a river. Also, a cow and a dog fall into a flooded river. Which one reaches Moo! the ocean first, cow or dog? Woof! GPC •Solvent flow carries molecules from left to right; big ones come out first while small ones get caught in the pores. •It is thought that particle volume controls the order of elution. •But what about shape? Simple SEC c log10M c DRI Ve c degas pump injector log10M Osmometry: Real Science h pV = n R T n = g/M c = g/V 1 p = c R T ( A2c ...) M Semipermeable membrane: stops polymers, passes solvent. Light Scattering: Osmometer without the membrane 100,000 c x 2p q 1 p Is cRT ( 2 A2c ...) M c T , p 1 q 4 πn o sin( / 2) LS adds optical effects Size q = 0 in phase Is maximum 2 q > 0 out of phase, Is goes down Is 1 q R 3 2 g SEC/MALLS MALLS DRI DRI degas pump injector SEC/MALLS 3D Plot - PBLG Scattered intensity 6 7 16 15 14 13 12 11 10 9 8 5 4 Scattering Envelope for a Single Slice 140000 120000 R/Kc 100000 80000 60000 c = 0.044 mg/mL M = 130000 g/mol 40000 20000 0 0.0 0.2 0.4 0.6 sin ( /2) 2 0.8 1.0 SEC/MALLS in the Hands of a Real Expert Macromolecules, 29, 7323-7328 (1996) ap 15 nm Much less than PBLG Midpoint The new power of SEC/Something Else experiments is very real. SEC is now a method that even the most skeptical physical chemist should embrace. For example, our results (not shown) favor higher rather than lower values for persistence length of one polymer (PBLG). This helps to settle about 30 years of uncertainty. So, SEC is good enough for physical measurements, but is it still good enough for polymer analysis? What about nanoparticles, especially large ones, in GPC? They were young when GPC was. Small Subset of GPC Spare Parts To say nothing of unions, adapters, ferrules, tubing (low pressure and high pressure), filters and their internal parts, frits, degassers, injector spare parts, solvent inlet manifold parts, columns, pre-columns, pressure transducers, sapphire plunger, and on it goes… 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. Atlanta, Georgia I-85N at Shallowford Rd. Sat. 1/27/01 4 pm Sizing by Dynamic Light Scattering—a 1970’s advance in measuring motion, driven by need to measure sizes, esp. for small particles. Large, molecules Small, slow fast molecules Is t It’s fluctuations again, but now fluctuations over time! DLS diffusion coefficient, inversely proportional to size. kT Rh 6πηo D Molecular Weight Distribution by DLS/Inverse Laplace Transform--B.Chu, C. Wu, &c. Where: G() ~ cMP(qRg) = q2D q2kT/(6phRh) Rh = XRg g (t ) G ( ) exp( t )d g(t) ILT G() log10t log10D q2D 1/2 c MAP CALIBRATE log10M M Hot Ben Chu / Chi Wu Example Macromolecules, 21, 397-402 (1988) MWD of PTFE Special solvents at ~330oC Problems: •Only “works” because MWD is broad •Poor resolution. •Low M part goofy. •Some assumptions required. Reptation: inspired enormous advances in measuring polymer speed…and predicts More favorable results for polymers in a matrix. There once was a theorist from France Who wondered how molecules dance. "They're like snakes," he observed, "as they follow a curve, the large ones can hardly advance."* D ~ M-2 deGennes More generally, we could write D ~ M- where increases as entanglements strengthen *With apologies to Walter Stockmayer Matrix Diffusion/Inverse Laplace Transformation Goal: Increase magnitude of Difficult in DLS because matrix log10D Solution: 1/2 D D Matrix: log10M Stretching scatters, except special cases. Difficult anyway: dust-free matrix not fun! Still nothing you can do about visibility of small scatterers DOSY not much better Replace DLS with FPR. Selectivity supplied by dye. Matrix = same polymer as analyzed, except no label. No compatibility problems. G() ~ c (sidechain labeling) G() ~ n (end-labeling) Painting Molecules* Makes Life Easier *R. S. Stein Small Angle Neutron Scattering Forced Rayleigh Scattering Fluorescence Photobleaching Recovery Index-matched DLS match solvent & polymer refractive index can't do in aqueous systems Depolarized DLS works for optically anisotropic probes works for most matrix polymers Fluorescence Photobleaching Recovery C t C (0)e t B 10 9 8 6 5 4 3 2 1 0 0 50 100 150 200 t/s DK 2 0.40 0.35 0.30 Dapp < Dapp -1 0.25 /s C(t) 7 3. An exponential decay is produced by monitoring the amplitude of the decaying sine wave. Fitting this curve produces , from which D can be calculated. 0.20 Dapp 0.15 0.10 0.05 0.00 0.0 0.5 1.0 1.5 2 2.0 5 2.5 3.0 -2 K / 10 cm 1. An intense laser pulse photobleaches a striped pattern in the fluorescently tagged sample. 2. A decaying sine wave is produced by moving the illumination pattern over the pattern written into the solution. FPR for Pullulan (a polysaccharide) 1 10 5 10 4 M -7 Dapp / 10 cm s 2 -1 10 0.1 NaN3(aq) solution ( = 0.537 ± 0.035) 5% Matrix solution ( = 0.822 ± 0.018) 10% Matrix solution ( = 0.907 ± 0.038) 15% Matrix solution ( = 0.922 ± 0.037) 0.01 4 10 10 5 0.1 1 10 -7 M Probe Diffusion: Polymer physics 2 Dapp / 10 cm s -1 Calibration: polymer analysis FPR Chromatogram Pullulan, 5%w/w Dextran Matrix, 50/50 mix of 380K and 11.8K 45 40 CONTIN Analysis Exponential Analysis Exponential Analysis 35 FArbitrary Units 30 25 20 15 10 5 0 1000 10000 100000 M 1000000 Indicates targeted M. GPC vs. FPR for a Nontrivial Case 0.9 20 0.8 18 0.7 16 Mw = 1,400 PDI = 1.08 Total Amplitude % = 20.7 14 0.6 % Amplitude Relative Concentration 1.0 0.5 0.4 0.3 Mw = 52,000 PDI = 1.04 Total Amplitude % = 46.5 Mw = 15,000 PDI = 1.03 Total Amplitude % = 27.2 12 Mw = 250,000 PDI = 1.03 Total Amplitude % = 5.5 10 8 6 0.2 4 0.1 2 0 0.0 4 10 M / g mol -1 5 10 PL Aquagel 40A & 50A 3 10 4 10 5 M 10 6 10 User-chosen CONTIN 25% Matrix 20,000 & 70,000 Dextran Pullulan, 8% HPC Solution, M=12,200 and 48,000 1.0 FArbitrary Units 0.8 CONTIN Exponential Exponential 0.6 0.4 0.2 0.0 1000 10000 100000 M 1000000 Indicates targeted M. M = 10,000 and 20,000 Examples of Separation Results from Simulation Data 2.0 FArbitrary Units 1.5 CONTIN 2 Exponential 1.0 0.5 0.0 1000 10000 M = 10,000 and 160,000 100000 M 2.0 M = 10,000 and 57,000 CONTIN 2 Exponential 1.5 FArbitrary Units 1.5 FArbitrary Units 2.0 CONTIN 2 Exponential 1.0 0.5 1.0 0.0 1000 0.5 10000 100000 1000000 M 0.0 1000 10000 M 100000 Indicates targeted M. What about separating cows and elephants? Either will float around the trees. How do you separate Moo! them then? Eeee! Field Flow Fractionation, that’s how! In FFF, large nanoparticles are made to flow between plates. One plate is porous, and a crossflow is arranged. What happens? Little nanoparticles come out first! Potential Advantages of FFF Handles a wider range of particles. May be easier for some aggressive solvents. Conclusion GPC is essential in any Nano Lab GPC may eventually get replaced. Matrix FPR FFF Thank you! L S U