TSKgel Size Exclusion Chromatography Columns

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1 Silica-based for protein analysis: TSKgel SW mab TSKgel SW TSKgel SWXL TSKgel SuperSW Polymer-based for desalting: TSKgel BioAssist DS Columns Polymethacrylate-based for water-soluble polymers analysis: TSKgel PW TSKgel PWXL TSKgel PWXL-CP TSKgel SuperMultiporePW Polymethacrylate-based for polar organic-soluble polymers analysis: TSKgel Alpha TSKgel SuperAW Polystyrene-divinylbenzene-based for organic-soluble polymers analysis: TSKgel HXL TSKgel HHR TSKgel SuperH TSKgel SuperHZ TSKgel SuperMultiporeHZ For more info visit:

2 About: TSKgel PW Size Exclusion Columns TSKgel PW columns are composed of spherical, hydrophilic polymethacrylate beads. Particle sizes range from µm for the smaller pore size columns to 7 µm for the larger pore size columns. Stable from ph to, TSKgel PW columns can be used in mobile phases of water or buffer (up to 0% methanol/80% aqueous) and can tolerate temperatures up to 80 C. The TSKgel PW column line consists of the following columns: TSKgel G000PW TSKgel G500PW TSKgel G000PW TSKgel G000PW TSKgel G5000PW TSKgel G6000PW TSKgel GMPW The mixed bed column, TSKgel GMPW, has an extended linear calibration range, suitable for samples with a broad molar mass distribution, as well as for unknown samples. The pore volume can be accessed by polymers ranging in molar mass from 500 to Da. By quickly categorizing the molar mass profile of an unknown sample, the column enables a fast selection of the best TSKgel PW column for routine analysis. Table : Product attributes TSKgel column Particle size (mean) Pore size (mean) G000PW µm.5 nm G500PW G000PW µm and 7 µm µm and 7 µm <0 nm 0 nm G000PW 7 µm 50 nm G5000PW 7 µm 00 nm G6000PW 7 µm >00 nm GMPW 7 µm mixed pore sizes Calibration range Up to,000 Da glycols and Up to,000 Da glycols and Up to Da glycols and Up to Da glycols and Up to Da glycols and Up to Da glycols and Da glycols and Attributes and Applications Product attributes of all eight TSKgel PW columns are shown in Table. All TSKgel PW columns have a base material of hydroxylated polymethacrylate, can be used in a maximum of 0% organic, and are shipped in water. The main application area for TSKgel PW columns is the analysis of water-soluble polymers, such as celluloses, acrylamides, glycols, dextrans, polyvinylalcohol, and oligosaccharides. TSKgel G000PW, the larger particle size equivalent of TSKgel G-ligo-PW, is most suitable for semipreparative and preparative isolation of oligosaccharides. Representative application examples for the PW columns are illustrated in Table. The calibration curve for polyethylene glycol and oxides for the TSKgel PW columns is shown in Figure. Table : Representative application examples for TSKgel PW columns Classification Examples. Synthetic polymers Nonionic Cationic Anionic. Polysaccharides and derivatives. Very large biopolymers Polynucleotides Viruses Proteins. Small molecules ligomers thers PEG, polyglycerin, polyacrylamide Polyethyleneimine, polyvinylpyrolidine Poly (sodium acrylate), Poly (sodium styrene sulfonate) Standard dextran, clinical dextran, pullulan, inulin, heparin, chitosan Carboxymethylcellulose DNA fragments TMV, SBMV, TBSV Lipoprotein (VLDL, LDL), apoferritin, gelatin, sea worm chlorocruorin oligosaccharides (dextran hydrolysate, cyclodoxtrin hydrolysate), cyclodextrins oligopeptides oligonucleotides 0 Call customer service: , technical service:

3 Figure : Polyethylene glycol and oxide calibration curves for TSKgel PW columns Log molar mass 0 6 G A D C B E F Polymers Sodium polyacrylate, an anionic polymer, is effectively separated on two TSKgel GMPW columns in Figure. The addition of 0.0 mol/l NaN results in normal elution and peak shape overcoming the ionic repulsion between the anionic sample and the resin. Figure : Effect of ionic strength on the elution of anionic polymers 0 H Column: Mobile phase: Retention volume (ml) A. G000PW B. G500PW C. G000PW D. G000PW E. G5000PW F. G6000PW G. GMPW all 7.5 mm ID 60 cm distilled H mol/l NaN 0.05 mol/l NaN 0.05 mol/l NaN 0. mol/l NaN ligosaccharides TSKgel PW columns are recommended for polysaccharide analysis due to their ability to separate a wide molar mass distribution. An effective separation of the anionic hydrophilic glucosaminoglycan, hyaluronic acid, is shown in Figure on a TSKgel G6000PW and TSKgel G000PW column in series with a 0. mol/l sodium chloride mobile phase. To obtain shorter analysis time and similar resolution, we recommend using TSKgel G000PWXL and G000PWXL columns in series. Column: TSKgel GMPW, 7 µm, 7.5 mm ID 60 cm Mobile phase: H, 0.0 mol/l, 0.05 mol/l, 0.05 mol/l or 0. mol/l NaN in H 0.5 ml/min 0.5 ml of % of the sodium salt of polyacrylic acid, an anionic polymer Figure : Analysis of polysaccharides Detector response (V). hyaluronic acid. hyaluronic acid. polyethylene oxide SE0 LS Columns: TSKgel G6000PW + G000PW, two 7.5 mm ID 60 cm columns in series Mobile phase: 0. mol/l NaCl 0.9 ml/min Temperature: 0 C hyaluronic acid, polyethylene oxide For more info visit:

4 About: TSKgel PWXL Size Exclusion Columns TSKgel PWXL columns are composed of spherical, hydrophilic polymethacrylate beads. The smaller particle size of TSKgel PWXL columns provide.7x higher resolution than their TSKgel PW columns counterpart, making TSKgel PWXL columns more suitable for analytical purposes. Four specialty columns are included in the TSKgel PWXL column line. The TSKgel G-DNA-PW column is designed for the separation of large polynucleotides such as DNA and RNA fragments of 500-5,000 base pairs. This column is a smaller particle size version of the TSKgel G6000PWXL column. The TSKgel G-ligo-PW column is designed for high resolution separations of aqueous nonionic and cationic oligomers, and oligosaccharides such as hydrolyzed cyclodextrins. Because of the presence of cationic groups on the gel matrix, this column is not suitable for separating anionic polymers. The TSKgel G-ligo-PW column has a PEG and PE calibration curve identical to that of the TSKgel G500PWXL column. The mixed-mode column, TSKgel GMPWXL, has an extended linear calibration range, suitable for samples with a broad MM distribution and unknowns. The TSKgel SuperligoPW column is designed for the determination of molar mass of aqueous oligomers, particularly oligosaccharides, and low molar mass aqueous polymers. The combination of the decreased particle size and semi-micro dimensions of the TSKgel SuperligoPW column enables high speed separation with high resolution and lowered solvent consumption. Since the packing material in the TSKgel SuperligoPW columns is more hydrophilic compared with TSKgel G-ligo-PW columns, an even wider range of water-soluble polymers can be analyzed without the need to add organic solvent to the eluent. The following TSKgel PWXL columns are offered: TSKgel G500PWXL TSKgel G000PWXL TSKgel G000PWXL TSKgel G5000PWXL TSKgel G6000PWXL TSKgel G-DNA-PW TSKgel GMPWXL TSKgel G-ligo-PW TSKgel SuperligoPW Attributes and Applications The main application area for TSKgel PWXL columns is the analysis of water-soluble polymers, such as celluloses, acrylamides, glycols, dextrans, polyvinylalcohol, and oligosaccharides. Because of the presence of cationic groups on the base bead of TSKgel G500PWXL, this column is not suited for separating anionic polymers. Product attributes of all of the TSKgel PWXL columns are shown in Table. All TSKgel PWXL columns have a base material of hydroxylated polymethacrylate, can be used in a maximum of 0% organic and are shipped in water. Figures 5-9 show the calibration curves for all of the TSKgel PWXL columns. Table : Product attributes TSKgel column Particle size (mean) Pore size (mean) G500PWXL 7 µm <0 nm G000PWXL 7 µm 0 nm G000PWXL 0 µm <50 nm G5000PWXL 0 µm 00 nm G6000PWXL µm >00 nm G-DNA-PW 0 µm >00 nm GMPWXL µm mixed pore sizes G-ligo-PW 7 µm.5 nm SuperligoPW µm.5 nm Calibration range <,000 Da <.0 0 Da, Da, Da Da Da, Da Up to,000 Da 00 -,000 Da (PE,PEG/H ) Call customer service: , technical service:

5 Figure 5: Polyethylene glycol and oxide calibration curves for TSKgel PWXL columns Figure 7: Double stranded DNA calibration curves for TSKgel G-DNA-PW column Log molar mass F E C A B D Base Pairs 8,000,000,000, Column: Mobile phase: Retention volume (ml) A. G500PWXL B. G000PWXL C. G000PWXL D. G5000PWXL E. G6000PWXL F. GMPWXL all 7.8 mm ID 0 cm distilled H Retention volume (ml) Column: TSKgel G-DNA-PW, 0 µm, 7.8 mm ID 0 cm Mobile phase: 0. mol/l NaCl in 0. mol/ltris-hcl, ph 7.5, + mmol/l EDTA 0.5 ml/min 60 nm Eco and Bst NI-cleaved pbr DNA, void volume determined with l-dna Figure 6: Protein calibration curves for TSKgel PWXL columns 0 7 Figure 8: ligosaccharide calibration curves for TSKgel G-ligo-PW column,000 Log molar mass a b c d e f g Log molar mass, Retention volume (ml) Column:. TSKgel G000PWXL. TSKgel G000PWXL. TSKgel G5000PWXL. TSKgel G6000PWXL 5. TSKgel GMPWXL all 7.8 mm ID 0 cm Mobile phase: 0. mol/l phosphate buffer, ph nm a. thyroglobulin ( Da) b. g-globulin ( Da) c. albumin (6.7 0 Da) d. ovalbumin (. 0 Da) e. b-lactoglobulin (.6 0 Da) f. myoglobin (.69 0 Da) g. cytochrome C (. 0 Da) 6 8 Retention volume (ml) Column: TSKgel G-ligo-PW, 7 µm, 7.8 mm ID 0 cm Mobile phase: distilled H 60 nm hydrolyzed b-cyclodextrin For more info visit:

6 Figure 9: Polyethylene glycol, oxide and ethylene glycol calibration curve for TSKgel SuperligoPW column Log molar mass ligosaccharides Column: TSKgel SuperligoPW, 6.0 mm ID 5 cm Mobile phase: H 0.60 ml/min Temperature: 5 C PE, PEG and ethylene glycol Figure 0 demonstrates the high speed analysis of maltose oligomers using a TSKgel SuperligoPW column compared to a TSKgel G-ligo-PW column. The faster analysis time is due to the semi-micro dimensions (6.0 mm ID 5 cm) and the small particle size ( μm) of the TSKgel SuperligoPW column compared to the 7.8 mm ID 0 cm size and 7 μm particle size of the TSKgel G-ligo-PW column. Figure 0: Analysis of maltose oligomers A B Large DNA fragments For the separation of large DNA fragments greater than,000 base pairs, a four column system is typically required. Baseline resolution of DNA fragments up to 7,000 base pairs can be achieved, provided there is a two-fold difference in the chain length of the fragments. Figure A shows the elution of double stranded DNA fragments, obtained from pbr DNA cleaved by both Eco and BstNI, on four TSKgel G-DNA-PW columns in series. The eluted peaks were collected and subjected to polyacrylamide gel electrophoresis, which showed almost complete separation of the 060, 857, and 6 base pair fragments. Although lower flow rates typically yield better separations of most fragments, the resolution of the 857 and 6 base pair fragments was slightly greater at the higher flow rate, as shown in Figure B. Figure A and B: Analysis of large DNA fragments Detector response (AU) A. 0.5 ml/min B. 0.5 ml/min Detector response (AU) Column: TSKgel G-DNA-PW, 0 µm, 7.8 mm ID 0 cm Mobile phase: 0. mol/l NaCl in 0. mol/l Tris-HCl, ph 7.5, + mmol/l EDTA Flow Rate: A. 0.5 ml/min B. 0.5 ml/min 60 nm 60 µl of Eco and Bst NI - cleaved pbr DNA, base pairs: & Columns: A. TSKgel SuperligoPW, µm, 6.0 mm ID 5 cm B. TSKgel G-ligo-PW, 7 µm, 7.8 mm ID 0 cm Mobile phase: H A: 0.6 ml/min B: Temperature: 0 C Injection vol.: A: 0 µl B: 50 µl. maltoheptose. maltohexose. maltopentose. maltotetraose 5. maltotriose 6. maltose 7. glucose Call customer service: , technical service:

7 ligomers The TSKgel G-ligo-PW column is designed for high resolution separations of nonionic and cationic oligomers. Figure demonstrates excellent resolution of chitooligosaccharides obtained by using the smaller, 6 μm particle size packing in the TSKgel G-ligo-PW column. Figure : Analysis of large chitooligosaccharides Complex Polymers An example on the influence of pore size on the separation of complex polymers is shown in Figure. While on the large pore TSKgel G6000PWXL column, gelatin elutes in one narrow peak, on the G000PWXL column the peak is much broader and the shoulder nearly separated from the main peak. This allows better determination of M w /M n and M z /M w. Figure : Separation of gelatin 5 A B C Column: TSKgel G-ligo-PW, 7 µm, 7.8 mm ID 0 cm Mobile phase: distilled H. chitohexaose. chitopentaose. chitotetraose. chitotriose 5. chitobiose Columns: A. TSKgel G6000PWXL B. TSKgel G5000PWXL C. TSKgel G000PWXL; all 7.8 mm ID 0 cm Mobile phase: 0. mol/l phoshpate buffer, ph 6.0 gelatin For more info visit: 5

8 Small Peptides Figure demonstrates that the separation of small peptides is possible on a TSKgel G000PWXL column under denaturing conditions. Using an aqueous eluent containing 5% acetonitrile and 0.% trifluoroacetic acid, the peptides were retained on the column using a size exclusion mechanism. An advantage of this method is that the eluent is volatile. Molar Mass Pullulan standard samples with a narrow molar mass distribution are commercially available. The molar mass of pullulan was analyzed by GFC/LALLS using a TSKgel GMPWXL column (Figure 5). Figure 5: Analysis of pullulan Figure : Analysis of small peptides Column: Mobile phase: TSKgel G000PWXL, 6 µm, 7.8 mm ID 0 cm 0.% TFA / 5% CH CN peptides. aprotinin. insulin β-chain. α-msh. bradykinin potentiator C 5. glutathione 5 Detector response (V) LS Column: TSKgel GMPWXL, µm, 7.8 mm ID 0 cm Mobile phase: 0. mol/l sodium chloride Temperature: 0 C LS Injection vol: 500 µl. pullulan P00. pullulan P00. pullulan P00. pullulan P50 6 Call customer service: , technical service:

9 Nucleic Acids Desalting of nucleosides can be accomplished using the TSKgel G500PWXL as depicted in Figure 6. Clearly, adenosine elutes after the void volume in the un-buffered water mobile phase. Figure 6: Desalting of nucelosides Sodium Polystrene Separation of sodium polystyrene sulfonate standards by GFC requires the addition of at least 0% acetonitrile or methanol to a 0. mol/l Na S mobile phase. Figure 7 shows chromatograms for sodium polystyrene sulfonate standards using a TSKgel GMPWXL column. Peak shapes for sodium polystyrene sulfonate samples obtained by adding 0% acetonitrile to a 0. mol/l Na S mobile phase remained constant upon addition of more acetonitrile. Detector response (AU) Figure 7: Separation of sodium polystyrene sulfonate standards LS Column: TSKgel G500PWXL, 7 µm, 7.8 mm ID 0 cm Mobile phase: distilled H 60 nm. 0.5 mol/l NaCl. uridine. adenosine Detector response (V) Lot. Lot. 6 Lot Column: TSKgel GMPWXL, µm, 7.8 mm ID 0 cm Mobile phase: ACN/0. mol/l sodium sulfate = 0/90 LS Temperature: 0 C Injection vol: 500 µl sodium poly(styrene sulfonates) For more info visit: 7

10 About: TSKgel PWXL-CP Size Exclusion Columns TSKgel PWXL-CP columns were specifically developed for the analysis of water-soluble cationic polymers. Composed of polymethacrylate beads, cationic groups are introduced on the surface of the TSKgel PWXL-CP packing material to prevent adsorption of cationic polymers and allow elution under low salt conditions. These columns show high theoretical plate numbers, linear calibration curves, and high durability because the base resin is the same as that used in the TSKgel PWXL columns. Three columns are available within the TSKgel PWXL-CP series, each with a different particle size, separation range, and exclusion limit, allowing polymers within a wide molar mass range to be separated and characterized. TSKgel G000PWXL-CP TSKgel G5000PWXL-CP TSKgel G6000PWXL-CP Attributes and Applications: Table 5 shows the product attributes for each of the three TSKgel PWXL-CP columns. Figure 8 shows calibration curves produced with standard polyethylene oxide and polyethylene glycol in a 0. mol/l aqueous solution of sodium nitrate. Table 5: Product attributes TSKgel column Base material G000PWXL-CP G5000PWXL-CP G6000PWXL-CP polymethacrylate polymethacrylate polymethacrylate Particle size 7 µm 0 µm µm Pore size 0 nm 00 nm >00 nm Exclusion limit Separation range (PE, PEG) Theoretical plates Da Da Da 00 ~ Da 00 ~ Da,000 ~ Da 6,000 0,000 7,000 Figure 8: Polyethylene glycol and oxide calibration curves for TSKgel PWXL-CP columns Log molar mass Cationic Polymers TSKgel G000PWXL-CP TSKgel G5000PWXL-CP TSKgel G6000PWXL-CP Columns: TSKgel G000PWXL-CP, 7 µm, 7.8 mm ID 0 cm TSKgel G5000PWXL-CP, 0 µm, 7.8 mm ID 0 cm TSKgel G6000PWXL-CP, µm, 7.8 mm ID 0 cm Mobile phase: 0. mol/l NaN Flow Rate: ml/min Temperature: 5 C polyethylene oxides (PE) standards polyethylene glycols (PEG) standards Various cationic polymers with different functional groups and molar masses were injected on the three TSKgel PWXL- CP columns (TSKgel G6000PWXL-CP, G5000PWXL-CP, and G000PWXL-CP) connected in series. Figure 9 demonstrates that these SEC columns can be utilized for the analysis of a wide variety of cationic polymers. Figure 9: Analysis of cationic polymers PAA (MM:8 kda) PAA (MM:5 kda) PEI (MM:66 kda) P(DADMACI) (MM:0 kda) PAS (MM:7800 Da) PAS (MM:87 kda) cationic dextran (MM: kda) chitosan (MM:. kda) -0 Columns: TSKgel G000PWXL-CP, 7 µm, 7.8 mm ID 0 cm TSKgel G5000PWXL-CP, 0 µm, 7.8 mm ID 0 cm TSKgel G6000PWXL-CP, µm, 7.8 mm ID 0 cm Mobile phase: 0. mol/l NaN Flow Rate: ml/min Temperature: 5 C Sample Load: g/l, 00 µl 8 Call customer service: , technical service:

11 PAA The TSKgel PWXL-CP columns eliminate ionic adsorption onto the particle by incorporating a cationic functionality on the particle surface. This is demonstrated in Figure 50 below. PAA [poly(acrylic acid)] was injected onto a TSKgel G5000PWXL-CP column. Each chromatogram, from the first injection (red) to the fifth injection (black), showed similar elution profiles without any adsorption of the polymer. Figure 50: Analysis of PAA Column: TSKgel G5000PWXL-CP, 0 µm, 7.8 mm ID 0 cm Mobile phase: 0. mol/l NaN Temperature: 5 C polyallylamine-hcl (PAA) Sample load: g/l, 00 µl Small Molar Mass Cationic Polymers Small molar mass cationic polymers were analyzed on two TSKgel G000PWXL-CP columns in series. As Figure 5 shows, these narrow molar mass cationic polymers eluted in order of their molar masses. Figure 5: Elution profiles of PAA and PEI polymers PEI ( kda) PEI (8,000 Da) PEI (,000 Da) PAA (6 kda) Column: TSKgel G000PWXL-CP, 7 µm, 7.8 mm ID 0 cm Mobile phase: 0. mol/l NaN Temperature: 5 C polyethyleneimine (PEI) polyallylamine-hcl (PAA) For more info visit: 9

12 About: TSKgel SuperMultiporePW Size Exclusion Columns Figure 5: Polyethylene glycol, oxide, and ethylene glycol calibration curves for TSKgel SuperMultiporePW columns The innovative multi-pore particle synthesis technology*, pioneered by Tosoh scientists, is incorporated into TSKgel SuperMultiporePW columns for water-soluble polymer analysis. Three semi-micro columns varying in linear range are available within this series, enabling high speed and high resolution analysis with lowered solvent consumption. The base material of each TSKgel SuperMultiporePW column is polymethacrylate. A wide molar mass range can be analyzed with the three different TSKgel SuperMultiporePW columns, from high molar mass water-soluble polymers to oligomers. The packing material in the TSKgel SuperMultiporePW columns is more hydrophilic than that of TSKgel PWXL series columns, which further reduces the chance of adsorption of hydrophilic polymers. Log Molar Mass 0 7 TSKgel SuperMultiporePW-N TSKgel SuperMultiporePW-M 0 6 TSKgel SuperMultiporePW-H TSKgel SuperMultiporePW-N TSKgel SuperMultiporePW-M TSKgel SuperMultiporePW-H *Using this proprietary technology, Tosoh can manufacture particles, each containing a broad range of pore sizes. This innovative approach essentially creates a linear calibration curve within each particle. As a result, columns with an extended linear calibration curve can now be prepared without mixing particles of different pore sizes. Attributes and Applications: Columns: Mobile phase: Temperature: 5 C TSKgel SuperMultiporePW-N, 6.0 mm ID 5 cm TSKgel SuperMultiporePW-M, 6.0 mm ID 5 cm TSKgel SuperMultiporePW-H, 6.0 mm ID 5 cm H 0.60 ml/min polyethylene oxides (PE) standards polyethylene glycols (PEG) standards ethylene glycol (EG) standards Table 6 shows the product attributes for each of the three TSKgel SuperMultiporePW columns. Figure 5 shows polyethylene glycol, oxide and ethylene glycol calibration curves for each of the TSKgel SuperMultiporePW columns. Table 6: Product attributes TSKgel column SuperMultipore PW-N SuperMultipore PW-M SuperMultipore PW-H Base material polymethacrylate Particle size µm* 5 µm* 8 µm* Pore size 0 nm 00 nm >00 nm Exclusion limit (PE, PEG/H ) Separation range Theoretical plates/5cm column Da Da 00 ~ Da 500 ~ Da,000 ~ Da >6,000 >,000 >7,000 * Particle size distribution is monodisperse. - 0 Call customer service: , technical service:

13 Comparison with Conventional GPC Columns A mixture of polyethylene oxide (PE) and polyethylene glycol (PEG) was analyzed on a semi-micro TSKgel SuperMultiporePW-M column and on conventional-sized TSKgel G000PWXL and TSKgel G5000PWXL columns in series. As shown in Figure 5, the analysis using the TSKgel SuperMultiporePW-M column was completed in ½ the time and with higher resolution than the analysis performed using the TSKgel G000PWXL and TSKgel G5000PWXL columns. This is due to the semi-micro dimensions (6.0 mm ID 5 cm) and the smaller particle size ( μm) of the TSKgel SuperMultiporePW-M column compared to the 7.8 mm ID 0 cm size and 7 and 0 μm particle size of the TSKgel G000PWXL and TSKgel G5000PWXL columns respectively. PVP Figure 5 demonstrates the lower hydrophobicity of the TSKgel SuperMultiporePW columns compared to the conventional TSKgel PWXL columns. Hydrophobic interaction causes partial adsorption of PVP-5 polymer on the TSKgel G000PWXL and TSKgel G500PWXL columns, while the absence of adsorption on the TSKgel SuperMultiporePW-N column suggests that the internal particle surface is more hydrophilic than the conventional columns. Figure 5: Analysis of a PVP-5 polymer 55 Figure 5: Comparison of analysis A B (MM = 9 kda) (MM = 879 kda) (MM = 9 Da) (MM =,000 Da) Resolution TSKgel PWXL TSKgel SuperMultiporePW-M Peak /Peak.5.5 Peak /Peak.9.7 Peak /Peak.0.9 Columns: A: TSKgel SuperMultiporePW-M, 6.0 mm ID 5 cm B: TSKgel G5000PWXL + G000PWXL, each 6.0 mm ID 5 cm Mobile phase: H 0.6 ml/min Temperature: 5 C Injection vol.: A: 0 µl B: 00 µl mixture of PE and PEG 5 5 B A Columns: A. TSKgel SuperMultiporePW-N, 6.0 mm ID 5 cm x B. TSKgel G000PWXL+G500PWXL, 6.0 mm ID 5 cm x Mobile phase: 00 mmol/l NaN Flow Rate: 0.60 ml/min Temperature: 0 C Injection vol.: 0 µl PVP(K-5) Columns: A: TSKgel SuperMultiporePW-N, 6.0mm ID x 5cm x B: TSKgel G000PW XL +G500PW XL, 6.0mm ID x 5cm x Mobile phase: 00mmol/L NaN 0.60mL/min Temperature: 0 C Injection vol.: 0µL PVP(K-5) Columns: A: TSKgel SuperMultiporePW-M, 6.0mm ID x 5cm B: TSKgel G5000PW XL + G000PW XL, each 6.0mm ID x 5cm Mobile phase: H 0.6mL/min Temperature: 5 C For Injection more info vol.: visit: A: 0µL B: 00µL mixture of PE and PEG