LIT203B 6/17/98 12:00 PM Page A AG® 50W and AG MP-50 Cation Exchange Resins Instruction Manual
LIT203B 6/17/98 12:00 PM Page B Table of Contents Section 1 Introduction ..........................................1 Section 2 Technical Description...........................1 Section 3 Mechanism ............................................4 Section 4 Resin Conversion ..................................7 Section 5 Instructions for Use ............................10 5.1 5.2 Batch Method ...............................................10 Column Method............................................
LIT203B 6/17/98 12:00 PM Page 1 Section 1 Introduction AG 50W and AG MP-50 strong acid cation exchange resins are useful for single step purification methods, for concentrating cationic solutes, and for analytical determinations of various mixed cationic solutes. Section 2 Technical Description Strong acid cation exchange resin is available as Analytical Grade AG 50W resin, AG MP-50 macroporous resin, and Biotechnology Grade AG 50W resin.
LIT203B 6/17/98 12:00 PM Page 2 a styrene divinylbenzene copolymer lattice. The amount of resin crosslinking determines the bead pore size. A resin with a lower crosslinkage has a more open structure permeable to higher molecular weight substances than a highly crosslinked resin. It also has a lower physical resistance to shrinking and swelling, so that it absorbs more water and swells to a larger wet diameter than a highly crosslinked resin of equivalent dry diameter.
LIT203B 6/17/98 12:00 PM Page 4 Table 2. Summary of the Properties of AG 50 and AG MP 50 Resins Active Group (X8 Resin) Thermal Stability Solvent Stability R-SO3- Good to 150 °C Very good Resistance to Oxidizing Agents Resistance to Reducing Slowly oxidizes in Very good hot 15% HN03 Section 3 Mechanism ion with equivalent or lower affinity. Table 3 shows the relative selectivity of various counterions.
LIT203B 6/17/98 12:00 PM Page 6 Table 3. Relative Selectivity of Various Counterions Relative Selectivity Counterion for AG 50W-X8 Resin Relative Selectivity Counterion for AG 50W-X8 Resin Large mesh material (20-50 and 50-100 mesh) is used primarily for large preparative applications and batch operations where the resin and sample are slurried together. Medium mesh resin (100-200 mesh) is used primarily in column chromatography for analytical and laboratory scale preparative applications.
LIT203B 6/17/98 12:00 PM Page 8 remember that the resin may shrink, or it may swell as much as 100%, depending on the conversion. Conversions to ionic forms not listed in Table 5 can be achieved using the information supplied in Table 3, which lists relative selectivities of various counterions for AG 50 resin. To convert a resin to an ionic form with a higher selectivity, wash the resin with 2-3 bed volumes of a 1 M solution of the desired counterion.
LIT203B 6/17/98 12:00 PM Page 10 Section 5 Instructions for Use AG 50 and AG MP-50 resin may be used in either a batch method or a column method. The batch method consists of adding the resin directly to the sample and stirring. The column method requires preparing a column filled with resin, and passing the sample through. 5.1 Batch Method The batch method is performed by adding the resin directly into the sample and stirring. The resin should be in the correct ionic form prior to beginning. 1.
LIT203B 6/17/98 12:00 PM Page 12 4. Slurry and pour the resin into the column. Equilibrate the resin in the initial buffer using 3 bed volumes of buffer. Poorly equilibrated resin will not give reproducible results. Alternatively, equilibration can be done by the batch technique, prior to pouring the column. First, convert the resin to the appropriate form, then suspend it in the starting buffer. Check the pH with a pH meter while stirring continuously.
LIT203B 6/17/98 12:00 PM Page 14 Section 6 Sample Protocol for Cation Exchange Resins 6.1 Determination of Total Salts in Tap Water Approximately 85% of the Continental United States is afflicted with hard water (3 grains or greater/gal). The following is a rapid method of determining the total ionic content of tap water as well as a good illustration of the potential of ion exchange techniques.
LIT203B 6/17/98 12:00 PM Page 16 6.5 Notes The experimental error is only that inherent in the titration procedure. The error due to the ion exchange itself is less than that of the titration. To avoid the error due to interfering carbonate ions, neutralize alkaline tap water with 0.1 M HCl, one drop at at time, to the methyl orange end point. The column may be used several times before regeneration is necessary.
LIT203B 6/17/98 12:00 PM Page 18 Table 7 (Continued) Table 7 (Continued) Application Resin Reference Metal removal AG 50W-X8 Siemer, D. D., Anal. Chem., 52, resin 1874 (1980). Cyclic nucleotide extraction AG 50W-X8 Schwartz, J. P., Morris, N. R. and resin Breckenridge, B. M., J. Biol. Chem., 248, 2699 (1973); Kuo, W., Hodgins, D. S. and Kuo, J. F., J. Biol. Chem., 248, 2705 (1973). Concentration of vitamin B-6 AG 50W-X8 Tryfiates, G. P. and Sattsangi, S., J. resin Chromatog., 227, 181 (1982).
LIT203B 6/17/98 12:01 PM Page 20 Table 7 (Continued) Application Resin Ammonia determination in plasma Table 7 (Continued) Reference Application Resin AG 50W-X8 Forman, D. T., Clinical Chem., 10, resin 497 (1964). L-Tryptophan purification AG 50W-X2 Yoshida, R., et al., J. Immunol., 141, resin 2819 (1988). Metal removal AG 50W-X8 Graf, E., J. Agric. Food Chem., 31, resin 851 (1983). Iron detection in wine AG 50W-X8 Ajlec, R. and Stupar, J., Analyst, resin 114, 137 (1989).
LIT203B 6/17/98 12:01 PM Page 22 Table 8 (Continued) Metals Table 8 (Continued) Recommended Eluant and Resin Eluted ions V, U, Sc, Y AG 50W-X8 resin V - 0.25 M H2SO4; U - 0.5 M H2SO4; Sc - 1 M H2SO4; Y 4 N HCI Reference Strelow, F. W. E., Rethemeyer, R. and Bothma,C. J. C., Anal. Chem., 37, 106 (1965). Be, Ba, Sr AG 50W X8 resin Be, Ba-9 M HCl04; Nelson, F., Murase, T. Sr-5 M HN03 and Kraus, K. A., J. Chromatog., 13, 503 (1984). K, Ti, Sc AG 50W X8 resin K-9 M HClO4; Ti-9 M HCI; Sc-4 M HCI, 0.
LIT203B 6/17/98 12:01 PM Page 24 Table 9. Cation Exchange Resins in Nucleic Acid Analysis Table 10. Separation of Organic Acids and Amines Application Resin Reference Application Resin Reference Separation of adenosine and riboflavin nucleotides AG 50W-X4 resin Brunius, G ., J. Chromatog., 170, 486 (1979). Separation of maleic and fumaric acids AG 50W-X4 resin Richards, M., J. Chromatog., 115, 259 (1975).
LIT203B 6/17/98 12:01 PM Page 26 Table 11. Cation Exchange Resins in Enzymatic Assays Section 10 Product Information Application Resin Reference Separation of acetylglutamate from glutamate AG 50W-X8 resin Alonso, E. and Rubio, V., Anal. Biochem., 146, 252 (1985). Adenylate cyclase assay AG 50W-X4 resin Marcus, R. and Orner, F., Endocrinol., 101, 1570 (1977). AG 50W-X2 Resin 142-1231 50-100 Hydrogen 500 g 0.6 300-1,180 0.70 Adenylate cyclase assay AG 50W-X4 resin Salomon, Y., et al.
LIT203B 6/17/98 12:01 PM Page 28 Catalog Number Mesh Size Ionic Form Capacity Density Pkg. (meq/ml) Diameter (g/ml) Size Nominal (µm) Nominal AG 50W-X12 Resin 142-1641 100-200 Hydrogen 500 g 2.1 106-250 0.85 142-1651 200-400 Hydrogen 500 g 2.1 053-106 0.85 Hydrogen 500 g 2.4 053-106 0.85 106-300 0.70 075-150 0.80 106-250 0.80 AG 50W-X16 Resin 142-1751 200-400 AG 50W-X2 Resin, Biotechnology Grade 143-5241 100-200 Hydrogen 100 g 0.
LIT203B 6/17/98 12:01 PM Page BC2 Bio-Rad Laboratories, 2000 Alfred Nobel Dr.
