Poly column

Capillary Tubes Figure 12.42 shows a cross section of a typical capillary tube. Most capillary tubes are made from fused silica coated with a 20-35-)J,m layer of poly-imide to give it mechanical strength. The inner diameter is typically 25-75 )J,m, which is smaller than that for a capillary GC column, with an outer diameter of 200-375 )J,m. 

Another type of synthetic polymer-based chiral stationary phase is formed when chiral catalyst are used to initiate the polymerisation. In the case of poly(methyl methacrylate) polymers, introduced by Okamoto, the chiraUty of the polymer arises from the heUcity of the polymer and not from any inherent chirahty of the individual monomeric subunits (109). Columns of this type (eg, Chiralpak OT) are available from Chiral Technologies, Inc., or J. T. Baker Inc. 

Alternatively a bonded poly(ethylene glycol) capillary column held at 35°C for 5 min and programmed to 190°C at 8°C/min may be employed to determine all components but water. The Kad-Eischer method for water gives inaccurate results. 

The tnmethylsilyl enol ether of 1 mdanone (3 2 mmol) in 2 mL of methylene chlonde is added to a mixture of xenon difluonde (4 mmol) and a catalytic amount of pyndimum poly (hydrogen fluonde) m 5 mL of methylene chlonde The mixture IS stured at 0 C for 2 h and poured into dilute sodium bicarbonate solution, tlie organic layer is separated and dned After concentration and column chromatogra phy (silica gel, hexanes), 2-fluoro-1 -mdanone (mp, 59 C) is obtamed in 87% yield... 

TSK-GEL H type columns are for gel-permeation chromatography (GPC) in organic solvents. They are packed with porous poly(styrene-divinylbenzene) resins that have a high degree of cross-linking. 

FIGURE 4.50 FVjrifIcation of mRNA on Toyopearl HW-65F. Column Toyopearl HW-65F. 25 mm X 90 cm. Sample 40 mg of poly(A) RNA/5 ml of formamide, sample from silkworm. Elution 20 mA1 sodium citrate, 5 mM EDTA, 0.5% SDS, 6 M urea, pH 3.5. Flow rate 24 ml/hr. Detection UV at 254, total mRNA activity (—), mRNA activity for major plasma proteins (------------). 

FIGURE 6.19 Polyamide. Column Shodex GPC HFIP-80S + HFIP-803, 8 mm i.d. x 300 mm x 2. Eluent S mM CFaCOONa/HFIP. Flow rate 1.0 mL/min. Detector Shodex Rl. Column temp. 40°C. Sample 0.05%. SOO /iL. Poly(hexamethylene adipamide) (Nylon 66). 

FIGURE 6.29 Poly(allylamine hydrochloride). Column Shodex Asahipak GF-7I0 HQ + GF-5I0 HQ + GF-310 HQ. 7.6 mm i.d. x 300 mm x 3. Eluenc LiCI aqueous solution. Flow rate 0.6 mL/min. Detector Shodex Rl. Column temp. 50°C. Sample I %, 50 /cL Poly(allylamine hydrochloride). 

FIGURE 9.22 Analysis of poly(vinyl pyrrolidone). Eluent 0.1 M Tris buffer, pH 7. Flow rate I ml/ min. Columns PSS Suprema 10 /itm, 100 + 1000, 8 x 300 mm. Oven temp 30°C. Detection Rl. Standards PSS polyvinylpyridin standards. 

FIGURE 9.24 Analysis of ultrahigh poly(ethylene oxide), MW about 37 million. Eluent O.OS M NaNOj. Flow rate 0.3 ml/min. Columns PSS Suprema 20 /tim, 30000, 8 x 300 mm. Oven temp 30°C. Detector Rl. Standards PSS PEO standards. 

FIGURE 9.28 Room temperature analysis of poly(amide-6). Columns PSS PFG 100 + 1000. Eluent TFE + 0.1 M NatFat. Temp 2S°C. Detection Rl. Calibration PSS PA-6 standards (broad). 

Smaller diameter columns are especially useful when expensive solvents are used. Figure 11.3 shows the analysis of poly (1,4-butylene terephthalate) using a Waters Alliance narrow-bore GPC system, quantitated against narrow polymethylmethacrylate standards. In this case, the solvent used is hexaflu-oro-2-isopropanol with 0.05 M sodium trifluoroacetic acid at a flow rate of... 

FIGURE I 1.3 Analysis of poly(l, 4-butylene terephthalate) usinga Waters Alliance narrow-bore GPC system. Columns 4.6 X 300 mm Styragel HR 2, HR 3, and HR 4. Mobile phase hexafluoroisopropanol. 0.35 ml/min at 30 C. (Chromatogram courtesy of Peter Alden, Waters Corp.)... 

The GBR resin works well for nonionic and certain ionic polymers such as various native and derivatized starches, including sodium carboxymethylcel-lulose, methylcellulose, dextrans, carrageenans, hydroxypropyl methylcellu-lose, cellulose sulfate, and pullulans. GBR columns can be used in virtually any solvent or mixture of solvents from hexane to 1 M NaOH as long as they are miscible. Using sulfonated PDVB gels, mixtures of methanol and 0.1 M Na acetate will run many polar ionic-type polymers such as poly-2-acrylamido-2-methyl-l-propanesulfonic acid, polystyrene sulfonic acids, and poly aniline/ polystyrene sulfonic acid. Sulfonated columns can also be used with water glacial acetic acid mixtures, typically 90/10 (v/v). Polyacrylic acids run well on sulfonated gels in 0.2 M NaAc, pH 7.75. 

As stated in Section I, columns should be selected so the low molar mass portions of the samples in question can be sufficiently separated from the elution interval of the system peaks. This task cannot always be accomplished, e.g., dimethylacetamide often replaces dimethylformamide as a GPC eluent the analyzed, mostly polar, samples require a neutral salt (e.g., FiBr) (7). The calibration is usually carried out with poly(methylmethacrylate) standards... 

Water-soluble polymers obtained through a radical polymerization [e.g., poly(acrylic acid) PAA] often contain sodium sulfate Na2S04 as a decomposition product of the initiator. The peak of Na2S04 is eluted before the dimer. In the interpretation of the chromatogram, a typical GPC program has to be truncated before the Na2S04 peak, or at a Mpaa value of about 200. The calibration curve in this region can be flattened by an additive small pore column as well, but the principle problem remains unsolved. 

The quaternized copolymer of vinylpyrrolidone and dimethylaminoethylmetha-crylate (poly-VP/DMAEMA) has been analyzed successfully with Ultrahydrogel columns and a mobile phase of a 0.1 M Tris pH 7 buffer with 0.3 or 0.5 M lithium nitrate (14). In this study, poor recovery of a poly-VP/DMAEMA sample was noticed when 0.2 M lithium nitrate was used for KB-80M, SB806-MHQ, and TSK GM-PWxl columns. Good recovery was achieved with 0.4 M lithium nitrate, and M of the poly-VP/DMAEMA were found to be 290,000, 300,000, and 320,000 for the respective columns. This demonstrates the equivalence of these columns for SEC of cationic polymers. 

TSK PWxL, Shodex OH-pak KB-80M, and SB806MHQ columns are equivalent for SEC of a quaternized poly-(VP/DMAEMA) in a pH 7 mobile phase. 

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