Handbook of Size Exclusion Chromatography and Related ...

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Table 3 Comparison of Mn Results by Various Methods Sample By titration a By NMR (500MHz) By GPC b1 (PS) that a, the exponential constant in the Mark–Houwink–Sakurada equation of two polymers, is the same. Fortunately,the Mn is normally more important than Mw forlowMWmaterial.Aslightdeviationofthecalibrationcurveonthehigher MWsidecannormallybetolerated.TheMWs ofpolyolsamples calculated using this calibration curve agree well with other primary methods, such as NMR and titration (Table 3). It is important to note, during the above procedure, that the peak maximum MW (Mp) of MALDI/MS is different from that of SEC for two reasons: (1) the peak height in MALDI/MS is approximately proportional to the number of polymermolecules,insteadoftheconcentration,weightbyvolume,asinSEC;(2) thex-axisinMALDI/MSislinearinMW,insteadofroughlylog(MW)asinSEC. The Mp needs to be converted before creating the calibration curve. The dead volume between the RI detection cell and the outlet for collection, which is normally significantly large, should be adjusted for acorrect calibration. It is not uncommon that the MW distribution of alow MW material covers the solvent mismatch peaks. As mentioned earlier, the concentration effect is normallynotsignificantforlowMWsamples.Thesolventmismatchproblemcan be reduced by increasing sample load, which means higher sample concentration and larger sample volume. 5 SPECIAL SUBJECT:ANALYSIS OF POLYMER ADDITIVES USING SEC By GPC b2 (Polyol) Polyol-1A 974 966 1808 1079 Polyol-1B 1032 1088 1933 1121 Polyol-2 2370 2590 3283 2311 Polyol-4 3993 4520 4599 3573 a Determined using ASTM Standard Test Method D1957-86. b GPCconditions:seeFig.11.Mn results in column b1 were calculated using a PS calibration. Those in column b2 were calculated using the polyol absolute MW calibration. Owing to improved resolution in low MW SEC, many polymer additives, if they are soluble in acommon solvent with the polymer, can be directly analysed using SEC without tedious extraction steps. Acrawax C(N,N 0 -ethylene-bis-stearamide) as an additive in polyurethane is agood example, because of the difficulty in finding agood solvent for HPLC analysis. Both Acrawax Cand polyurethane are © 2004 by Marcel Dekker, Inc.
Figure 12 Chromatogram of Acrawax C in polyurethane. Column: Jordi DVB 100A ˚ , 250 10mm; mobile phase: benzyl alcohol, 250ppm BHT; flow rate: 1.0mL/min; temperature: 1508C; injection volume: 200mL. soluble in benzyl alcohol at 1508C. The concentration of Acrawax C can be analyzed using a high-temperature SEC with a small-pore (100A ˚ ) column. Acrawax C, which appears as a negative peak at 9.1min (Fig. 12), is well separated from the polyurethane and other additives. In this type of method, the viscosity of the polymer should not be too high. ACKNOWLEDGEMENTS The author expresses his appreciation to Noveon, Inc., for its permission to publish this article and for its support on all research work, to Dr CS Wu for his encouragment and discussion, and to D Hanshumaker for his help in preparation of this article. REFERENCES 1. JM Mays, N Hadjichristidis. In: HG Barth and JM Mays, eds. Modern Methods of Polymer Characterization. New York: Wiley-Interscience, 1991, Chs. 6, 7. 2. G Montaudo, MS Montaudo, F Samperi. In: G Montaudo and RP Lattimer, eds. Mass Spectroscopy. Boca Raton: CRC Press, 2002, ch. 10. © 2004 by Marcel Dekker, Inc.
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MARCEL DEKKER Handbook of Size Excl
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CHROMATOGRAPHIC SCIENCE SERIES A Se
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60. Modern Chromatographic Analysis
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Preface to the Second Edition Gel p
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Preface to the First Edition Molecu
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Contents Preface to the Second Edit
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17. Size Exclusion Chromatography o
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James F. Curry Analytical Departmen
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Iwao Teraoka, Ph.D. Othmer Departme
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The graphical data display typicall
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polymer molecule can elute. The tot
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individual states) allowed to them.
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unsaturation while the IR detector
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Sampleconcentrationshouldbeminimize
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averagesdefinedintermsofthemolecula
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information regarding appropriate M
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Analytical. Two models, the KMX-6 a
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Figure 6 Overlay of time-sliced pea
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accurately for very large and very
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4 GENERAL REFERENCES The interested
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46. TA Chamberlin, HE Tuinstra. US
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materials are commercially availabl
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Table 1 Commercial Column Packing M
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necessarily comparable. For this re
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adjusted such that the pore size di
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narrowmolecular weight range, indiv
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Figure 8 Effect of particle size on
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Figure 9 SEC calibration using poly
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Table 4 Typical Eluent Systems for
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Figure 12 Analysis of poly-2-vinyl
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24. E Meehan, S O’Donohue. The ro
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into accepted laboratory techniques
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Table 1 Trace Metal Impurities in C
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mass and biological activity, for m
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Experimental efficiency vs. velocit
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Table 3 Properties of SEC Silica Ge
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operate at aflow rate that can easi
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Figure 3 Efficiency of amide-bonded
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mechanically stable packing materia
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Figure 5 Pore size distributions of
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Figure 6 Separation of polystyrenes
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60-120 A ˚ are large enough to be
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Table 6 Selected Silica-Based Colum
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Table 7 Separation Ranges for Polym
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the decline in permeability is perm
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sample components can be varied by
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Figure 10 Diol bonding reactions. I
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ecause of the higher relative molec
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valuesfork1,k2,anda,theunknownmolec
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water-soluble and detergent-soluble
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the importance of secondary retenti
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discussed in Table 8(33). Based on
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individual dispersion processes ins
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The detector time constant can dist
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5.3 Mobile Phase A mobile phase is
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less steep and still linear at high
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35. P Bristow, J Knox. Chromatograp
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112. K Gooding, K Lu, G Vanecek, F
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for branched polymers or copolymers
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Figure 1 “Bridge design” flow-r
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Figure 3 A light-scattering photome
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Figure 5 SEC universal calibration
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where hn is the hydrodynamic volume
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It should be noted that dn=dc also
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sensitivity for many samples compar
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that is, log[h] vs. logM,generated
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Figure 8 Effect of band broadening
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Table 1 Measurement of Molecular We
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Figure 10 Differential refractomete
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Figure 11 (A) Mark-Houwink plot of
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Table 3 Measurement of Molecular We
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size information is derived from un
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Table 5 Molecular Weight Distributi
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By measuring the scattered intensit
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8 APPENDIX: INSTRUMENT COMPANIES 8.
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53. G Marot, J Lesec. J Liq Chromat
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125. D Slootmaekers, JAPP Van Dijk,
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200. JG Bindels, GJJ Bessems, BM De
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possibly with hydrodynamic volume (
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An insurmountable limitation of SEC
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to sDR(V) when either nPS ¼nPB ¼c
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1,4-trans;and8%1,2-vynil.TheglobalC
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The goal is to find the MWD and CCD
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As expected, pS is closer to the no
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Figure 3 Example 3: MWD and BD of a
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9. BH Zimm, WH Stockmayer. The dime
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45. RO Bielsa, GR Meira. Linear cop
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solubility properties. They are use
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determination of the absolute molec
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Figure 4 Result of the PBT analysis
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Figure 8 Result of natural spider s
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7 Size Exclusion Chromatography of
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As pointed out earlier, rubber must
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Table 2 Various Solvents and Operat
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Figure 2 Typical GPC calibrations w
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© 2004 by Marcel Dekker, Inc. Figu
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Natural and Synthetic Rubbe177 vari
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Another example, shown in Fig. 5 (3
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APPENDIX: SEC CONDITIONS FOR RUBBER
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Appendix (Continued) Polymer Column
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5. J West, E Rodriguez. Rubber Chem
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associate. Girdler (67) and Speight
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Table 1 Fractions Obtained Using Co
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Figure 1 SEC analyses of an aged as
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Apparently more polar compounds are
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40% was fractionated into four frac
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Figure 7 SEC analyses of an unaged
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Figure 9 Comparison of percentage L
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Figure 11 Comparison of SEC chromat
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Figure 12 Comparisons of SEC chroma
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percentage LMS at the other locatio
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the total area into up to 12 sectio
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attempted to develop an SEC method
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parameterisbasedontheinternalpressu
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5 MODIFIED ASPHALTS The addition of
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Oxidation of the rubber and asphalt
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Figure 20 SEC chromatogram for an a
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Figure 21 Comparisons of apparent m
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© 2004 by Marcel Dekker, Inc. S/
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PS/10 4 þ 10 3 þ 500 þ 100 THF/1
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Nevertheless, SEC of asphalts is es
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56. NW Garrick, RR Biskur. Trans Re
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Asphalts 235 122. M-S Lin, JM Chaff
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Table 1 Applications of Acrylamide
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(AM/AA), and acrylamide/dimethyldia
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polymer, a large size filter of 5,
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Figure 1 Size exclusion chromatogra
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Table3 MWandMWDofaBroad-MWDPAMStand
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Another commercially available MW d
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Table 5 Weight-Average Molecular We
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Figure 6 (a) Size exclusion chromat
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compared to anormal product. By com
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StudyingtheKineticsofaChemicalReact
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this information, the higher MW pea
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46. CJ Kim, A Hamielec, A Bendek. J
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A relatively new technique utilizin
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where ais the exponent of the Mark-
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Figure 2 TDS chromatograms for full
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Figure 5 Comparison of molecular we
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Table 3 Summary of TDS Molecular We
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thepartiallyhydrolyzedPVAwiththecol
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Table 5 Summary of Conformation and
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a¼0.627andlog(K) ¼23.249.Theseare
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The limits of detection of the PVA
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Figure 12 PVAc broad molecular weig
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4 SUMMARY Advances in SEC character
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2.1 Molecular Weight Grades of PVP
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exclusion chromatography with low-a
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weightandmolecularweightdistributio
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Figure 4 PEOcalibrationofUltrahydro
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Figure 6 Overlay of GPC chromatogra
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plus Ultrahydrogel 120 A ˚ ,Shodex
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Figure 10 SEC traces of PVP/AA copo
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Figure 11 Molecular weight distribu
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2 CHEMICAL, MACROMOLECULAR AND MORP
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Table 2 The Relative Composition of
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considered to be 3. After the DS ha
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een extensively studied by methods
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The silylation was performed in LiC
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observed using column materials hav
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Table 6 (Continued) Packing materia
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4.5 Other Cellulose Derivatives Bes
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Table 8 SEC Conditions for Characte
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dextrans swell too much in cadoxen
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stirring at room temperature for an
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Figure 2 MMDofunbleached(HP)andblea
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Figure 4 MMDofbirchkraftpulpdegrade
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adequate when evaluating the influe
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differential refractive index (DRI)
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30. HA Krässig. Effects of structu
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67. E Sjöholm, K Gustafsson, A Col
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104. B Fleury, J Dubois, C Léonard
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136. D Miller, D Senior, R Sutcliff
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166. SS Cutié, CG Smith. Determina
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200. R Berggren, F Berthold, E Sjö
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13 Molar Mass and Size Distribution
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measurements, and vapor pressure os
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The calibration of SEC columns by c
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The division according to eluent ty
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Apolymer produced by UV irradiation
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Because DMAC/LiCl is also agood sol
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compoundsandwasexplainedbyadsorptio
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isolated lignins because they are e
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The Separon HEMA and Separon HEMA B
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Figure 4 GPC elution curves of the
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liquor is highest throughout the co
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topological anisotropy in the polym
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29. M Ristolainen, R Alen, J Knuuti
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eds. Proc Int Symp 1998. Canton, Pe
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92. L Jurasek. Towards a three dime
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Table 1 Selected Industries Connect
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into C3-metabolites, which then may
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Table 4 Key Enzymes in the Biosynth
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Table 5 Cloned Mutants of Starch En
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Figure 3 Modeled x-ray diffraction
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In the native environment (plant ce
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Figure 6 (a) Large starch granules
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number of branching points); crossl
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Both absolute approaches are extrao
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Table 7 (Continued) Approach Experi
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structures, excluded volumes, and t
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Figure 10 (a) SEC elution profile o
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Figure 12 Wheat glucans: DRI elutio
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Figure 13 Wheat glucans. Normalized
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Figure 16 Wheat glucan. Normalized
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Figure 18 Wheat glucan.Elution prof
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Figure 20 Wheat glucans. Absolute m
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Figure 22 Wheat glucan. (a) Intrins
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Figure 24 Wheat PA-glucan. Elution
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an Ubbelohde-viscometer for concent
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Rheological investigations of stabi
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shifts to applied laser light, whic
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Table 8 Characteristic Parameters f
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Table 8 (Continued) and H2O. A part
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10. SG Ball, MHBJ van de Wal, RGF V
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52. W Praznik, R Beck. J Chromatogr
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Page 455 and 456:
3 PROTEIN PARTITIONING IN SEC 3.1 G
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Figure 1 Plot of Kav vs. log M for
Page 459 and 460: yconsideringtheproteinsolutestobesp
Page 461 and 462: globular proteins and selected flex
Page 463 and 464: silanol groups (52-54); even capped
Page 465 and 466: most proteins to be maximally stabl
Page 467 and 468: Figure 4 Chromatograms of BSA and P
Page 469 and 470: concentrated) steps of protein puri
Page 471 and 472: Table 3 Characteristics of Dextran-
Page 473 and 474: preparative SEC. The capabilities o
Page 475 and 476: 37. GR Noll, N Nagle, JO Baker, DJ
Page 477 and 478: 16 Size Exclusion Chromatography of
Page 479 and 480: Figure 2 Separation of total E. col
Page 481 and 482: Figure 3 Separation of HaeIII-cleav
Page 483 and 484: The recovery of DNA fragments has b
Page 485 and 486: Endotoxin should also be removed fr
Page 487 and 488: Table 3 Best Columns for the Separa
Page 489 and 490: Figure 10 Dependence of HETP on flo
Page 491 and 492: Appendix (Continued ) Polymer Colum
Page 493 and 494: REFERENCES 1. CT Wehr, SR Abbott. J
Page 495 and 496: MALDI-MS has been applied to determ
Page 497 and 498: Figure 1 Calibration curvesof SEC c
Page 499 and 500: Figure 3 Chromatograms of epoxy res
Page 501 and 502: wherenandn0 aretheRIsofthesample an
Page 503 and 504: Figure 5 SEC chromatogram of n-alka
Page 505 and 506: Figure 7 Refractive indexes of comm
Page 507 and 508: Another detector recently applied i
Page 509: Figure 11 Absolute MW calibration p
Page 513 and 514: 18 Two-Dimensional Liquid Chromatog
Page 515 and 516: largerispressuredropandresultingexp
Page 517 and 518: elations between molar masses and s
Page 519 and 520: Figure 2 (Continued) chapter, one o
Page 521 and 522: where Nis the column efficiency exp
Page 523 and 524: composition/architecture/molar mass
Page 525 and 526: solvent. Here again, solvent-solven
Page 527 and 528: or repulsive interactions between m
Page 529 and 530: macromolecules may strongly differ
Page 531 and 532: temperature. The efficacy of an ads
Page 533 and 534: however, aliphatic bonded groups ma
Page 535 and 536: Figure 10 Schematic representation
Page 537 and 538: packing materials for SEC of synthe
Page 539 and 540: Quantitative relations between sila
Page 541 and 542: Both strength and thermodynamic qua
Page 543 and 544: where Cand Dare constants for apart
Page 545 and 546: ThisisimportantwhenLCCCisappliedtoc
Page 551 and 552: However, strong adsorption of macro
Page 553 and 554: elution step may be sufficient for
Page 555 and 556: Figure 16 Block scheme of a full ad
Page 557 and 558: processing of 2D-HPLC data. Knowled
Page 559 and 560: Figure 20 Set of full retention-elu
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distributions of complex polymers a
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constituents were not discriminated
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(Sec. 7) and therefore they can be
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2. Identify sample solvents. First
Page 569 and 570:
1 Segmental interaction energy para
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58. BG Belenkii. Pure Appl Chem 51:
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19 Methods and Columns for High-Spe
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Table 1 Summary of Methods for Incr
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not savealot of time and solvent, d
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Figure 3 Conventional SEC chromatog
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chromatogram at 4mL/min, which is f
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Sincethereductionoftheinternaldiame
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Table 3 Summary of Column Performan
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Figure 11 Dependence of column perf
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. Two-dimensional chromatography ru
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The standard deviations for the mol
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Figure 15 Comparison of time requir
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Figure 17 Accuracy and precision of
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polymer analysis. Samples are “ru
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just ignoring the fractionation of
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20 Automatic Continuous Mixing Tech
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out at low enough concentration tha
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educed viscosity, h r, to be comput
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Figure 2 Raw ACOMP signals from mul
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Figure 4 Mw vs. monomer conversion,
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decreases smoothly and monotonicall
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Figure 7 Effects of fluctuating rea
Page 617 and 618:
Finally, the use of a light-scatter
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Figure 8 Linear and cubic increases
Page 621 and 622:
where r(t) is now given by 2 6 r(t)
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Figure 11 Light scattering from sol
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3.1 ASimple System: Equilibrium Cha
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Figure 13 (a) ACM applied to the ch
Page 629 and 630:
Also of note in Fig. 13b is how hig
Page 631 and 632:
15. JM Starita, CL Rohn. Rheologica
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52. M Hulden. Hydrophobically modif
Page 635 and 636:
21 Light Scattering and the Solutio
Page 637 and 638:
therefore, is to remove any lingeri
Page 639 and 640:
is often found in the literature wh
Page 641 and 642:
Figure 2 Configuration of an SEC se
Page 643 and 644:
5 THE IMPORTANCE OF SOFTWARE Like a
Page 645 and 646:
standard deviations of the measured
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These include the differential weig
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Figure 8 Data of Fig. 7corrected fo
Page 651 and 652:
Figure 11 Data of Fig. 10 with spik
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Figure 13 Fitstodatacollectedatasli
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Table 1 Errors (in %) Characteristi
Page 657 and 658:
homogeneous copolymer, that is, tak
Page 659 and 660:
process may be used to identify the
Page 661 and 662:
fractionated sample. They represent
Page 663 and 664:
Figure 16 An example of poor SEC ch
Page 665 and 666:
on) methods, whereby the average di
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23. WH Stockmayer, LD Moore Jr, M F
Page 669 and 670:
condition, as is explained in this
Page 671 and 672:
Figure 2 Partition coefficients KL
Page 673 and 674:
4 COLUMNS FOR HOPC Detailsaregiveni
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and concomitant clogging of columns
Page 677 and 678:
Figure 5 Concentration dependence o
Page 679 and 680:
Table 1 Solution Volume, Polymer Ma
Page 681 and 682:
in Fig. 4a. The middle fractions (8
Page 683 and 684:
Figure 10 SEC chromatograms for som
Page 685 and 686:
appropriate for the early fractions
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23 Size Exclusion/ Hydrodynamic Chr
Page 689 and 690:
Figure 2 Calibration curve for the
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Figure 5 Elutionbehaviorofpolystyre
Page 693 and 694:
Table 1 Comparison Between SEC and
Page 695 and 696:
egular SEC column in which the pore
Page 697:
Such an ideal separation can be obt
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