Poly packed columns

Figure 6.3 Conparlson of the separation of the octylphenol poly(ethylene glycol) ether, Triton X-16S on a packed column, left, and an open tubular column, right, using UV detection. For the packed column separation al0cmx2mmI.D. column packed with Nucleosil C g, d. 3 micrometers, temperature > 170 C, and mobile phase carbon dioxide (2 ml/min] and methanol (0.15 nl/rnin). pressure programmed from 130 to 375 bar in 12 min were used. For the open tubular column separation a 10 m x 50 micrometers I.O., SB-Biphenyl-30, temperature = 175°C, mobile phase carbon dioxide (0.175 ml/min) and 2-propanol (0.0265 ml/min) pressure programmed, 125 bar for 5 min, then ramped from 125 to 380 bar over 19.5 min, and held at 380 bar for 15 min. were used. (Reproduced with permission from ref. 57. Copyright Preston Publications, Inc.) ...

Although open tubular columns have been used for RPLC,42 most applications use packed columns. The most widely used column packings are formed by chemically bonding butyl (C4), octyl (C8), or octadecyl (C18) chains to a silica surface. Phenyl (Ph), cyano (CN), and amino (NH2) functionalities are also used. Polymeric packings which are finding broad acceptance include alkyl-grafted poly(methylmethacrylate) and alkyl-grafted or unmodified... 

Pack the matrix in a Poly Prep column (Bio-Rad) (see Note 5). 

The amount of protein that is bound to the column can be estimated by subtracting the quantity of IgG that is eluted. This is only an estimate, but generally sufficient for antibody purification purposes. Continue with the subsequent steps if no more than 20% of the applied protein concentration is found in the eluate. Poly Prep columns are convenient since they are unbreakable, disposable, can be capped easily and securely at both ends, and have graduation markings for measuring column volumes. After the column has been packed, it should be stored at 4°C. Do not let the column warm up again or dry out, since this will introduce air bubbles which can cause protein denaturation. All subsequent purification steps should be performed at 4°C. 

Preparation of 10% Poly A-[(lS,2S)-2-Benzyloxycyclopentyl]Maleimide on a Silica Gel Support in a Packed Column... 

Berger, T.A. Deye, J.F. Separation of benzene poly-carboxylic acids by packed column supercritical fluid chromatography using methanol-carhon dioxide mixtures with very polar additives. J. Chromatogr. Sci. 1991, 29, 141. 

Liu, Z. Zhao, S. Wang, R. Yang, G. Separation of poly-hydroxylflavonoids by packed-column supercritical fluid chromatography. J. Chromatogr. Sci. 1999, 37, 155-158. 

Characteristic properties of some liquid phases used in packed column gas chromatography PPE-5 = Poly(phenyl ether), EGS = poly(ethylene glycol succinate) and DECS = poly(diethylene glycol succinate), Carbowax 20M = poly(ethylene glycol) and FFAP = Carbowax 20M treated with 2-nitroterephthalic acid... 

Characteristic properties of some poly(siloxane) liquid phases used for packed column gas chromatography... 

The master retention equation of the solvation parameter model relating the above processes to experimentally quantifiable contributions from all possible intermolecular interactions was presented in section 1.4.3. The system constants in the model (see Eq. 1.7 or 1.7a) convey all information of the ability of the stationary phase to participate in solute-solvent intermolecular interactions. The r constant refers to the ability of the stationary phase to interact with solute n- or jr-electron pairs. The s constant establishes the ability of the stationary phase to take part in dipole-type interactions. The a constant is a measure of stationary phase hydrogen-bond basicity and the b constant stationary phase hydrogen-bond acidity. The / constant incorporates contributions from stationary phase cavity formation and solute-solvent dispersion interactions. The system constants for some common packed column stationary phases are summarized in Table 2.6 [68,81,103,104,113]. Further values for non-ionic stationary phases [114,115], liquid organic salts [68,116], cyclodextrins [117], and lanthanide chelates dissolved in a poly(dimethylsiloxane) [118] are summarized elsewhere. 

Finally, it is interesting to mention that capillary monolithic columns have also started to be used in gas adsorption chromatography [425, 426]. Poly-DVB monohth obtained in the presence of l.Svol of dodecanol-toluene mixtures possesses good separating power however, its efficiency (the theoretical plate height) still yields by a factor of 3-10 to that of traditional open capillary columns. On the other hand, the theoretical plate height for a similar monolith prepared for use in liquid chromatography proved to be comparable with that of conventional capillary silica-packed column [427]. 

One g of the polymer-coated glass beads were closely packed in a tubing of poly(vinyl chloride) (i.d. - 3 mm) equipped with a stopcock, and the packed column was primed with physiological saline for 24 h to exclude a liquid-air interface and to equilibrate the polymer surface with the physiological environment. 

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