Calibration of size exclusion chromatography

Nave, R., Weber, K., and Potschka, M., Universal calibration of size-exclusion chromatography for proteins in guanidinium hydrochloride including the high-molecular-mass proteins titin and nebulin,. Chromatogr. A, 654, 229, 1993. 

Wu, C., Simultaneous calibration of size exclusion chromatography and dynamic light-scattering for the characterization of gelatin, Macromolecules, 26, 5423, 1993. 

Calibration of size-exclusion chromatography columns based on the finding that the retention volume of a molecular or particulate species is usually a single-valued function of an appropriate size parameter of this molecule or particle, irrespective of its chemical nature and structure. 

Herman, D.P. Field, L.R. Calibration of size exclusion chromatography columns for molecular weight determination of polyacrylonitrile and poly(vinylpyrrolidone) in N,N-dimethylformamide. J. Chromatogr. Sci. 1981, 79, 470. 

Berrueco, C, Venditti, S., Morgan,T.I, Alvarez, R, Millan, M., Herod, A.A., Kandiyoti, R. (2008) Calibration of size exclusion chromatography columns with NMP/chloroform mixtures as eluent applications to petroleum derived samples. Energy Fuels, 22,3265-3274. 

The calibration of size exclusion chromatography measurements for the determination of the molecular weights of oligolactone macromonomers was carried out using PPG and PS as standards. Results are compared with those obtained by end-group analysis with proton NMR14 refs. 

Dubin, P. L. and Principi, J. M., Failure of universal calibration for size exclusion chromatography of rodlike macromolecules vs. random coils and globular proteins, Macromolecules, 22, 1891, 1989. 

Comparison of Size Exclusion Chromatography Calibration Techniques... 

The calibration standards included sodium form polystyrene sulfonates obtained from Pressure Chemical Co., Pittsburgh, Pa., and sodium toluene sulfonate. Measurements were taken at 0.5 to I.Oml/mln flow rates. The logarithm of the molecular weight of the standards was linear it suggests a framework for approaching an interpretion of the structure of the scission products. This application of size exclusion chromatography measurements must be viewed as a first approximation because of the unmeasured differences between the chromatographic behavior of the linear standards and the expected branched structure of the scission products. 

Stogiou, M. Kapetanaki, C. latrou, H. Examination of the universality of the calibration curve of size exclusion chromatography by using polymers having complex macromolecular architectures. Int. J. Polym. Anal. Charact. 2002, 7, 273-283. 

MALDI (matrix assisted laser desorption and ionization) combined with size-exclusion chromatography is increasingly being used to study the microstructure of polymers [542-544], MALDI allows the desorption and ionization of macromolecules with molecular mass up to hundreds of kilodaltons with little or no fragmentation. The MALDI mass spectra provides structural information for identification of the polymer repeat units or copolymer sequence, end group mass and mechanism of polymerization for polymers with a narrow polydispersity (< 1.25). For samples of high polydispersity MALDI fails to provide reliable information. The separation of polydisperse samples by size-exclusion chromatography provides fractions suitable for characterization by MALDI. MALDI can also be used for mass calibration of size-exclusion columns. 

Dubin, P.L. (ed.). Aqueous Size Exclusion Chromatography, Elsevier, New York, 1988. Grubisic, Z., Rempp, P., and Benoit, H., The Universal Calibration in Size Exclusion Chromatography of Polymers,. Polym. Sci, 5, 753 (1%7). 

Figure 2.2 Typical setup of size-exclusion chromatography equipped with triple detection for determination of the moiecuiar weight of polymers without the need for prerequisite calibration. DRi, differential refractive index detector DV, differentiai viscometer I, injector MALS, multi-angle light-scattering detector (depending on the model, the scattered light is measured at 3 or 18 different angles) P, pump. The central unit includes a computer and software.

Fig.l. Calibration curve for the mechanism of size-exclusion chromatography under ideal conditions ... 

Mlynar, J., and Sarkanen, S., 1996, Renaissance in Ultracentrifugal Sedimentation Equilibrium Calibrations of Size Exclusion Chromatographic Elution Profiles. In Strategies in Size Exclusion Chromatography (M. Potschka and P. L. Dubin, eds.), ACS Symposium Series No. 635, American Chemical Society, Washington, D.C., pp. 379-400. 

Calibration curve for the determination of formula weight by size-exclusion chromatography. 

Most size exclusion chromatography (SEC) practitioners select their columns primarily to cover the molar mass area of interest and to ensure compatibility with the mobile phase(s) applied. A further parameter to judge is the column efficiency expressed, e.g., by the theoretical plate count or related values, which are measured by appropriate low molar mass probes. It follows the apparent linearity of the calibration dependence and the attainable selectivity of separation the latter parameter is in turn connected with the width of the molar mass range covered by the column and depends on both the pore size distribution and the pore volume of the packing material. Other important column parameters are the column production repeatability, availability, and price. Unfortunately, the interactive properties of SEC columns are often overlooked. 

A new direct method for using size exclusion chromatography (SEC) to evaluate polymer intrinsic viscosity [n] is discussed. Sample viscosity information is obtained by combining SEC elution curve data and calibration data using direct SEC-[n] calibration procedures without involving polymer molecular weight calculations. The practical utility, convenience and the expected precision of the proposed method are illustrated. 

Size exclusion chromatography (SEC) separates molecules of a polymer sample on the basis of hydrodynamic volume. When the chromatograph is equipped only with a concentration-sensitive detector, i.e. conventional SEC, a molecular weight distribution (MWD) can be obtained from the chromatogram only through use of a calibration function relating molecular weight and elution volume V (2). 

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