水溶性聚合物中定性定量分析检测方案(凝胶色谱仪)

智能文字提取功能测试中

ANALYSISTOSOH BIOSCIENCETOSOHF：+49(0)711 113257-89 INFO.TBG@TOSOH.COMTOSOH BIOSCIENCEGMBH ZETTACHRING6WWW.TOSOHBIOSCIENCE.DE Regina Roemling1， Shinji Sato 2， Kuniyuki Tokunaga 2， Teruhiko Tsuda & Hiroyuki Moriyama 2 1： Tosoh Bioscience GmbH， Stuttgart， Germany 2： Separation Center， TOSOH Corporation， Yamaguchi， Japan INTRODUCTION Size exclusion chromatography (SEC) is a well established method for the determination of molecular mass distribution and average molecular mass of polymers. Recentadvances in SEC comprise of semi-micro SEC and the design of linear columns providing wide molecular weight separation ranges and near-linear calibrations. Multi-poreparticle technology is the most elegant way to achieve near-linear SEC calibration curves. It solves the known problem of peak disturbances/inflection points， whichtypically occur due to a mismatch of pore sizes when columns with different molecular weight ranges are coupled. Particles prepared by multi-pore technology contain abroad range of pore sizes in a single polymeric bead. This innovative approach essentially creates a linear calibration curve within each particle. The multi-pore particle synthesis technology (Figure 1) has been successfully applied to polystyrene divinylbenzene based TSKgel GPC columns for organic SEC since2001. Now the same strategy was applied to design multi-pore particles of the polymethacrylate based TSKgel PW packing for aqueous SEC. In order to not only improvepeak shape and calibration but also shorten analysis time the new polymeric multi-pore phases are packed in semi-micro SEC columns of 6 mm i.d. x 15 cm length. Theresulting TSKgel SuperMultiporePW series comprises of three column types covering different molecular weight ranges. We report on the basic properties of these new SEC columns and show elution profile comparisons of synthetic polymers on the newly developed columns andconventional columns. The latest development in SEC stationary phase design， the multi-poretechnology can provide a real solution to avoid inflection points in SEC. Particlesprepared by multi-pore technology contain a broad range of pore sizes in a singlepolymeric bead. This essentially creates a linear calibration curve within eachparticle. This technology was applied to create multi-pore particles of the poly-methacrylate based TSKgel PW packing for aqueous SEC. The new TSKgelSuperMultiporePW columns are packed with spherical mono-disperse particles，each containing a wide range of pore sizes. The series comprises of three columntypes covering different molecular weight ranges (PW-N； PW-M， PW-H). Theyexhibit high resolution and good linearity of the calibration curves across a widerange of molecular mass of PEO and PEG standards in aqueous eluent (Figure 2). 三FIGURE 1 ....... ...... Conventional and multi-pore SEC column technology FIGURE 2....... Calibration curves and SEM photographs of TSKgel SuperMultiporePW columns Due to their small particle sizes it is possible to reach high theoretical plates athalf of the length of conventional SEC columns of 30 cm length. Withdimensions of 6 x 150 mm SuperMultiporePW columns belong to the semi-microtype of SEC columns. When analyzed on a series of conventional SEC columns a PVP K-30 sampleshowssaninflection point，： whichi does nottappearon1theTSKgelSuperMultiporePW-M column. In addition analysis is much faster and moresensitive due to the smaller particle size and the semi-micro column dimensions(Figure 3). Figure 4 shows the analysis of several water soluble polymers. ........ ....... 多 FIGURE 3Analysis of Polyvinylpyrrolidone (PVP) K-30. TSKgel SuperMultiporePW-M column(red)compared with a series of conventional columns (blue). FIGURE 4....... ..... SEC elugrams of various water-soluble polymers on TSKgel SuperMultiporePW-MGERMANY T：+49(0)711 13257-0 EcoSEC Semi-Micro SEC System MATERIAL AND METHODS TSKgel Column Particle Size MW Range (PEO & PEG) Min. Theor. Plates SuperMultiporePW-N 4 um3x102-5x104 > 16000 SuperMultiporePW-M 5 um 5x102-1x106 > 12000 SuperMultiporePW-H 8 um1x 103-1x107 > 7000 Instrument： EcoSEC compact， semi-micro SEC system Chemicals/Reagents： Water for eluent and sample preparation was purified with a Milli-Q" water purification system. PEO， PEG and synthetic polymers were obtained from Tosoh， Wako Pure Chemical (Osaka) and other suppliers. PEO and PEG standards were dissolved in water at concentrations of 0.5- 1.0 g/L. Synthetic polymers were dissolved in eluent at concentrations of 3.0-5.0 g/L prior to use. Calibration Curves： Column： TSKgel SuperMultiporePW-N/M/L (6 mm ID x 15cm L) Flow rate 0.6 mL/min Mobile phase： H9 Detection： RI @25℃ Sample： PEO & PEG standards Analysis of Water Soluble Polymers： Flow rate： 0.6 mL/min Mobile phase： 0.1 M NaNO. Detection： RI @ 25℃ Injection volume： 35 pL Polyvinylpyrollidone (Figure 3) Columns： [A] TSKgel SuperMultiporePW-M， 6 mm ID x15 cm L， 5 pm [B] TSKgel G3000PWXL & G5000PWXL， 7.8 mm ID x30 cm L each Various polymers (Figure 4) Column： TSKgel SuperMultiporePW-M， 6 mm ID x 15cmL Samples： 1：polyvinylpyrrolidone (K-30)， 2： polyvinylpyrrolidone(K-15)， 3： hydroxypropylcellulose (200 K) 4： hydroxypropylcellulose (100 K)， 5： chondroitin sulfate sodium salt， 6： carboxymethyl cellulose， 7： arabic gum， 8：poly(sulfopropyl methacrylate-co-acrylic acid)， 9： dextran (200K) CONCLUSION TSKgel SuperMultiporePW columns are packed with spherical monodisperse polymethacrylateparticles， each containing a wide range of pore sizes. They exhibit a good linearity of thecalibration curves across a wide range of molecular mass of PEO and PEG standards inaqueous eluent. Chromatograms obtained on TSKgel SuperMultiporePW columns show highresolution and smooth peak shapes without shoulders or inflection points. This improvesaccuracy and reproducibility when determining the molecular mass distribution of watersoluble polymers. Various polymers were analyzed on the TSKgel SuperMultiporePW semii-micro columns. Compared with conventional SEC columns， the rnew columns deliveredsuperior results at shorter run times and lower solvent consumption.