HPLC Columns. Capture the Essence SHOWA DENKO K.K.

Transcription

1 HPLC Columns 0 0 Capture the Essence SHOWA DENKO K.K.

2 TM We provide a wide range of products to meet your analytical needs, from pretreatment and separation columns to calibration standards for size exclusion chromatography. Please visit the Shodex website to see application data and uses. Shodex website The following names are trademarks or registered trademarks of SHOWA DENKO K.K. Shodex, AFpak, Asahipak, AXpak, CLNpak, CXpak, HILICpak, MSpak, ODP, OHpak, ORpak, RSpak, SUGAR, USPpak [Caution]. Please read the operating manual included on the product carefully before use.. For improvement purposes, some specifications are subject to change without notice.. Provided to help you select the appropriate column, the figures and descriptions in this catalogue are not guaranteed and do not warrant suitability for your applications.. It is essential to take normal precautions when handling reagents and other chemical products even if the safety information is not included on the operating manual.. Products described in this brochure are not intended for medical use or medical applications including medical diagnosis.

3 Contents Column selection Analysis and preparative columns Sample pretreatment columns Notice of change of product name Information Index Reversed Phase, Hydrophilic Interaction and Normal Phase Chromatography Ligand Exchange Chromatography Ion Exclusion Chromatography Ion Chromatography Size Exclusion Chromatography Calibration Standards for SEC Ion Exchange Chromatography Columns for Special Separation Modes Types of Columns, Base Materials, Functional Groups and Ligands HPLC Separation Modes Column Selection by Sample Character and Separation Mode Column Selection (Application) Comparison of the Features Among Shodex Reverse Phase Chromatography (RPC) Columns Columns for Polymer-based Reversed Phase Chromatography (ODP HP) Columns for Polymer-based Reversed Phase Chromatography (Asahipak) Columns for Polymer-based Reversed Phase Chromatography (Rspak) Columns for Silica-based Reversed Phase Chromatography (ODS Columns) Columns for Silica-based Reversed Phase Chromatography (Other Columns) Columns for Silica-based HILIC and Normal Phase Chromatography Columns for Polymer-based Hydrophilic Interaction Chromatography (HILIC) (HILICpak) Columns for Polymer-based Hydrophilic Interaction Chromatography (HILIC) (Asahipak) Columns for Ligand Exchange Chromatography Columns for Ion Exclusion Chromatography Columns for Ion Chromatography (Anion Analysis) Columns for Ion Chromatography (Cation Analysis) Column Selection for Size Exclusion Chromatography (SEC) Precautions for Polar Polymer Analysis Aqueous SEC (GFC) Columns : Silica-based Aqueous SEC (GFC) Columns : Polymer-based Multimode Columns Aqueous/Organic SEC Columns Organic SEC (GPC) Columns (General Analysis) : THF Organic SEC (GPC) Columns (General Analysis) : Chloroform Organic SEC (GPC) Columns (General Analysis) : DMF Solvent-peak Separation Columns for Organic SEC (GPC) Organic SEC (GPC) Columns : Rapid Analysis Organic SEC (GPC) Columns : High Performance Analysis Organic SEC (GPC) Columns : For Preparative Organic SEC (GPC) Columns : Linear Calibration Type Organic SEC (GPC) Columns : High Temperature/Ultra High Temperature Analysis Organic SEC (GPC) Columns : HFIP Applicability of SEC (GPC) Columns to Solvent Replacement Calibration Standards for SEC Columns for Anion Exchange Chromatography Columns for Cation Exchange Chromatography Column for Hydrophobic Interaction Chromatography Columns for Affinity Chromatography Columns for Chiral Separation Column for High Temperature Reversed Phase Chromatography Columns for GPC Clean-up Pretreatment Columns for Column Switching Method Notice of Changes USP (Ver.) Column List Column Cleaning Procedures General Precautions for Column Handling Column Trouble Shooting HPLC System Trouble Shooting Index by Index by

4 Types of Columns, Base Materials, Functional Groups and Ligands Separation Type Base Material Reversed Phase & HILIC (Polymer-based) Reversed Phase & HILIC & Normal Phase (Silica-based) Ligand Exchange Ion Exclusion Ion Chromatography Aqueous SEC (GFC) Multimode Aqueous/Organic SEC Organic SEC (GPC) Ion Exchange Hydrophobic Interaction Affinity Chiral Separation GPC Clean-up Column Switching Pretreatment Asahipak Asahipak Asahipak RSpak RSpak RSpak RSpak RSpak HILICpak HILICpak Asahipak Silica Silica Silica Silica Silica Silica Silica Silica SUGAR SUGAR SUGAR RSpak SUGAR EP USPpak SUGAR RSpak IC IC IC IC IC PROTEIN OHpak OHpak Asahipak Asahipak MSpak IEC IEC IEC PIKESS Asahipak AXpak IEC IEC PIKESS IEC Asahipak CXpak HIC AFpak ORpak ORpak CLNpak CLNpak MSpak MSpak ODP HP ODP-0, ODP-0 CP-0 CP-0 RP-, DS DE DM- NN JJ-0 VG-0 VT-0 NHP ET-RP CM, CP C C CN NPE PYE SIL NH SC SP00 KS-00 DC- SZ SC0-F MN- SH KC- NI-, I-A SI YS-0 YK- Y-, T- KW-00 KW00 SB-00 HQ LB-00 GS-HQ GF-HQ GF-0 KF-00, K-00, KD-00, KF-00, KF-00HQ, LF, HT-00, UT-00, AT-0MS HFIP-00, HFIP-00 QA- DEAE- DEAEN DEAE-B ES-0N WA- SP- SP-0N SP-B CM- ES-0C P-S PH- Various CDBS- CRX- EV PAE PK GF-A Polyhydroxymethacrylate Polyvinyl alcohol Polyvinyl alcohol Polyvinyl alcohol Styrene divinylbenzene copolymer Polymethacrylate Polyhydroxymethacrylate Polyhydroxymethacrylate Polyvinyl alcohol Polyvinyl alcohol Polyvinyl alcohol Polyvinyl alcohol Polyvinyl alcohol Silica Silica Silica Silica Silica Silica Silica Silica Styrene divinylbenzene copolymer Styrene divinylbenzene copolymer Styrene divinylbenzene copolymer Styrene divinylbenzene copolymer Styrene divinylbenzene copolymer Styrene divinylbenzene copolymer Styrene divinylbenzene copolymer Styrene divinylbenzene copolymer Styrene divinylbenzene copolymer Polyhydroxymethacrylate Polyvinyl alcohol Polyvinyl alcohol Silica Styrene divinylbenzene copolymer Silica Silica Polyhydroxymethacrylate Polyhydroxymethacrylate Polyvinyl alcohol Polyvinyl alcohol Polyvinyl alcohol Styrene divinylbenzene copolymer Polyhydroxymethacrylate Polyhydroxymethacrylate Polyhydroxymethacrylate Polyhydroxymethacrylate Polyvinyl alcohol Polyhydroxymethacrylate Polyhydroxymethacrylate Polyhydroxymethacrylate Polyhydroxymethacrylate Polyhydroxymethacrylate Polyvinyl alcohol Styrene divinylbenzene copolymer Polyhydroxymethacrylate Polyhydroxymethacrylate Silica Polyhydroxymethacrylate Styrene divinylbenzene copolymer Polyvinyl alcohol Hydrophilic copolymers containing N-vinyl acetamide Polyvinyl alcohol Functional Group, Ligand Page Octadecyl Octyl Butyl Sulfo Quaternary ammonium Ao Quaternary ammonium Ao Octadecyl Octadecyl Octyl Butyl Cyanopropyl Nitrophenylethyl Pyrenylethyl Aopropyl Sulfo(Ca + ) Sulfo(Pb + ) Sulfo(Na + ) Sulfo(Na + ) Sulfo(Zn + ) Sulfo(Ca + ) Sulfo(Ca + ) Sulfo Sulfo Quaternary ammonium 0 Quaternary ammonium 0 Carboxyl Carboxyl Sulfo Hydrophilic polymer Hydrophilic polymer,, 0,,,, 0 Quaternary ammonium Diethylaoethyl Diethylaoethyl Diethylaoethyl Diethylaoethyl Diethylaoethyl Sulfopropyl Sulfopropyl Sulfopropyl Carboxymethyl Carboxymethyl Sulfo(Na + ) Phenyl Various ligand β-cyclodextrin derivative L-Ao acid derivative 0 0

5 HPLC Separation Modes Liquid chromatography (LC) uses liquid as mobile phase (eluent). It is an analytical method that separates a mixture of compounds based on their physical and chemical differences. High performance liquid chromatography (HPLC) is a method that introduces the mobile phase under high-pressure conditions resulting in rapid and high-performance separations. The various interactions between the analyte, stationary phase (packing material), and mobile phase are the key factors for the separation. A wide variety of separation modes can be achieved by using particular combinations of stationary and mobile phases. Separation mode Reversed Phase Chromatography (RPC) Hydrophilic Interaction Chromatography (HILIC) Normal Phase Chromatography (NPC) Ligand Exchange Chromatography (LEX) Ion Exclusion Chromatography (IEX) Ion Chromatography (IC) Size Exclusion Chromatography (SEC) Ion Exchange Chromatography (IEC) Hydrophobic Interaction Chromatography (HIC) Affinity Chromatography (AFC) Chiral Separation Chromatography (CS) Multimode Chromatography Separation is based on hydrophilic interaction. A high polarity stationary phase is used. Typically the mobile phase contains a mixture of organic solvents such as acetonitrile and aqueous solvents (water or buffer). Using the higher polarity mobile phase causes a faster elution. Applicable for the analysis of high polar substances. Separation is based on the partition equilibrium between the stationary phase and the mobile phase. The polarity of the stationary phase is higher than that of the mobile phase. Typically the mobile phase contains a mixture of organic solvents with different polarities such as hexane and isopropanol. Using the higher polarity mobile phase causes a faster elution. Separation is based on differences in analytes coordination complex. Stationary phase modified with metal sulfonate complex ion. Works in combination with size exclusion or HILIC modes. Separation is based on electrostatic interaction (repulsion) between the ion exchanger and ionic solutes. Dissociated ionic molecules elute faster than non-dissociated forms. Used mainly for the analysis of organic acids. Separation is based on electrostatic interaction (bonding) between the ion exchanger and ionic solutes. Has a relatively small ion exchange capacity. Electrical conductivity detector can be used with low-salt concentration mobile phase. Used mainly for the analysis of inorganic compounds. Network or pores on the surface of the packing material works as molecular sieve to separate molecules based on their sizes. To separate molecules solely based on their sizes, it requires an analytical condition without any analyte and packing gel interaction. The bigger the molecule size, the faster the elution sequence. Used for molecular weight or molecular distribution deteration of macromolecules and qualification of oligomers. Separation is based on electrostatic interactions between the ion exchanger and ionic solutes. The mobile phase of choice should have a sufficient buffering capacity at the ph that produces the largest charge differences between the analyte of interest. The elution position is optimized by varying the ph, salt concentration, and/or ionic strength of the mobile phase. Separation is based on hydrophobic interaction. Hydrophobic functional group is modified on the stationary phase. Adsorption of analytes generally occurs at a high salt concentration and they are released by lowering the salt concentration. Used mainly for the analysis of proteins. Separation is based on adsorption of the analyte to the specific biologically derived ligand pair. Highly selective. A buffer solution with the appropriate ph and ionic strength is selected based on the type of ligand, analytes, and their interaction. Used mainly for the purification and concentration of biological active substances. Separation of optical isomers using chiral selectors. Highly selective. Separation is based on the combination of different modes. Column Selection by Sample Character and Separation Mode Sample Solubility Aqueous soluble Sample MW,000,000 Separation Mode RPC LEX IEX SEC IEC HIC AFC RPC HILIC LEX IEX IC SEC IEC AFC CS Sample Solubility Organic soluble Sample MW,000,000 Characteristics Separation is based on the partition equilibrium between stationary phase and mobile phase. The polarity of the stationary phase is lower than that of the mobile phase. Typically the mobile phase contains a mixture of organic solvents (methanol, acetonitrile, or THF) and aqueous solvents (water or buffer). Using the lower polarity mobile phase causes a faster elution. Separation Mode SEC RPC NPC SEC RPC : Reversed Phase Chromatography HILIC : Hydrophilic Interaction Chromatography NPC : Normal Phase Chromatography LEX : Ligand Exchange Chromatography IEX : Ion Exclusion Chromatography IC : Ion Chromatography SEC : Size Exclusion Chromatography IEC : Ion Exchange Chromatography HIC : Hydrophobic Interaction Chromatography AFC : Affinity Chromatography CS : Chiral Separation Chromatography Types of Columns, Base Materials, Functional Groups and Ligands HPLC Separation Modes Column Selection by Sample Character and Separation Mode

6 Column Selection (Application) Pharmaceuticals, Cosmetics Foods Separation Mode Page Separation Mode Page Hydrophobic substances RPC HILIC, 0,,, 0, Monosaccharides Disaccharides Sugar alcohols HILIC LEX+SEC LEX+HILIC 0, Pharmaceuticals Metabolites Additives Hydrophilic substances Substances in bio-fluid (serum plasma urine) IEC+RPC LEX+SEC RPC SEC+RPC,,, Oligosaccharides Low molecular water-soluble dietary fiber HILIC LEX+HILIC SEC SEC 0,,, Polymer SEC,, 0, Polysaccharides SEC, RPC RPC, Moisturizers Emulsifiers Preservatives Optical active materials Polyalcohols Protein hydrolysates Mucopolysaccharides Surfactants Paraben Dehydroacetic acid LEX+SEC LEX+HILIC SEC RPC SEC SEC SEC+RPC SEC RPC CS, 0,, 0,,, Nutritional ingredients Organic acids Water-soluble vitas Fat-soluble vitas Fatty acids Nucleic acids (umami) IEX+RPC IC RPC IEC+RPC HILIC RPC NPC SEC RPC SEC IEC+SEC IEC+IEX+RPC 0, 0, 0, 0,,,,,, Ao acids IC IEC Food additives RPC HILIC 0,, 0, RPC Food safety Pesticides IEC+RPC HILIC 0 IC 0 Mycotoxin RPC Separation Mode (Page and Page ) Pretreatment of residual pesticides SEC GPC ( Clean-up ) 0 RPC : Reversed Phase Chromatography HILIC : Hydrophilic Interaction Chromatography NPC : Normal Phase Chromatography LEX : Ligand Exchange Chromatography IEX : Ion Exclusion Chromatography IC : Ion Chromatography SEC : Size Exclusion Chromatography IEC : Ion Exchange Chromatography HIC : Hydrophobic Interaction Chromatography AFC : Affinity Chromatography CS : Chiral Separation Chromatography New Materials Synthetic polymers Additives Oligomers Organic solvent soluble Polar organic solvent soluble High temperature/ Ultra high temperature Water-soluble Organic solvent soluble Polar organic solvent soluble Water-soluble Separation Mode SEC RPC SEC Page,,,,,, 0,,,,, 0,,,,,,,, 0,,,,

7 Biotechnology Genomics Nucleobases Nucleotides Nucleosides Oligo nucleic acids DNA/RNA Separation Mode RPC IEC+SEC IEC RPC IEC+SEC IEC SEC Page,, Environment Water quality Anions Oxyhalides Cyanide Cyanogen chloride Cations Surfactants Separation Mode IC IC IEC+HILIC IEX IC RPC SEC+RPC Page , Column Selection (Application) Ao acids RPC IEC+IEX+RPC IEC 0 Perchloric acids IC IEC+HILIC RPC 0 0,, Proteomics IEC+SEC RPC 0, Pesticides IEC+RPC HILIC 0 Peptides Proteins SEC IEC,,,, Anions IC IC 0 0 HIC Heavy metals IC RPC 0, Humic substances SEC Glycoproteins SEC IEC,,,, Soil Organic arsenic IEX+RPC RPC,, HIC AFC Pesticides IEC+RPC HILIC 0 Glycomics Sugar chains Monosaccharides HILIC AFC HILIC LEX+SEC 0, 0, Environmental hormones Pretreatment of Phthalates PCBs Benzo [a] pyrene IC SEC GPC ( Clean-up ) 0 0 Hormones Sialic acids Uronic acids Aldonic acids Aes LEX+HILIC IEX+RPC RPC IEC RPC, 0, 0 Bioethanols Monosaccharides Oligosaccharides Oligosaccharides Alcohols Furfural Saccharides Organic acids Alcohols Furfural HILIC LEX+SEC LEX+SEC IEX+RPC+SEC 0, Steroids HILIC SEC 0,, Hemicelluloses Celluloses SEC 0, Lipids Phospholipids Lipoproteins NPC SEC SEC AFC,, Biodiesels Cations Fatty acid glycerides Fatty acid methyl esters Organic acids IC SEC RPC IC 0

8 Comparison of the Features Among Shodex Reverse Phase Chromatography (RPC) Columns ODS columns are the most popular reversed phase columns that are packed with silica-based octadecyl group. Shodex provides not only ODS columns but also polymer-based reversed phase columns with different functional groups. You can choose a suitable column for your application based on the features listed below. Features ODP HP ODP-0 CP-0 CP-0 ODP-0 RP- DS- DS- DE DM- NN JJ-0 CM CP C C CN NPE PYE Provides a large theoretical plate number nearly twice as much as generally available polymer-based reversed phase columns Offers enhanced retention of high polar substances compared to ODS columns Suitable for the analysis of small molecules such as pharmaceuticals in the presence of protein matrix Ideal for LC/MS analysis of high polar compounds Corresponds to USP L9 Relatively large pore size is suitable for the analysis of ao acids, peptides, and proteins Usable in a wide ph range from ph to Usable in 00% water or buffer solution Best used for the analysis of basic substances ODP-0 corresponds to USP L Higher performance type of ODP-0 series Corresponds to USP L Large pore size is suitable for the analysis of proteins and peptides Corresponds to USP L Suitable for reversed phase analysis of highly hydrophilic substances, that are not well retained by ODS columns Corresponds to USP L Polymer-based columns, with similar polarity to that of ODS columns, can be used in general purposes Wide working ph range (form ph to ), usable in 00% water and buffer solutions Corresponds to USP L Suitable for the analysis of ao acids and water-soluble vitas Corresponds to USP L9 The packing material contains sulfo groups, and supports multimode (reversed phase and cation exchange) analysis Ideal for the analysis of complex samples containing neutral and ionic substances The packing material contains trace amounts of quaternary ammonium groups, and supports multimode (reversed phase and anion exchange) analysis Ideal for analysis of complex samples containing neutral and ionic substances Monomeric type ODS column, fully end capped high purity silica (99.99% or higher) Corresponds to USP L Polymeric type ODS column, fully end capped high purity silica (99.99% or higher) Excellent acid tolerance Advantageous for separating planar and nonplanar compounds from each other Corresponds to USP L Use when the retention capacity of C is too strong or that of C is too week Applicable to ion pair chromatography, because of its rapid mass transfer and equilibrium Corresponds to USP L Use when the retention capacity of C or C is too strong Corresponds to USP L Utilizes reversed phase interaction and π-electron interaction to separate regioisomers, which typically cannot be separated with ODS, C, or C columns Corresponds to USP L0 Utilizes several types of interactions based on π-electrons to separate structural isomers NPE corresponds to USP L

9 The interrelation between hydrophobicity and retention, and the interrelation between steric selectivity and retention were compared among Shodex columns for reversed phase chromatography. The retention factor (k ) of amylbenzene was used as the retention, the separation factor (α) between n-butylbenzene and n-ao benzene was used as the hydrophobicity. The separation factor between o-terphenil and triphenylene was used as the steric recognition. Lager separation factor means higher hydrophobicity and higher steric selectivity. Interrelation between hydrophobicity and retention Hydrophobicity NPE CN ODP HP C C PYE CP-0 CP-0 DE- Retention Column size :.mm I.D. x 0mm each Eluent : H O/CH OH=0/0 Flow rate :.0mL/ Detector : UV (nm) Column temp. : 0 C CM CP ODP-0 Comparison of separation of alkylalcohol due to different functional groups Steric selectivity ODP HP CN NPE n-buthylbenzene CP-0 C o-terphenyl PYE CP-0 DE- C Retention n-amylbenzene Triphenylene Interrelation between steric selectivity and retention Column size :.mm I.D. x 0mm each Eluent : H O/CH OH=0/0 Flow rate :.0mL/ Detector : UV (nm) Column temp. : 0 C ODP-0 CP CM Comparison of separation due to the difference in steric selectivity Comparison of the Features Among Shodex Reverse Phase Chromatography (RPC) Columns EG C C C C0 C C Sample : Ethyleneglycol, n-alkylalcohol Sample : µl. n-buthylbenzene. n-amylbenzene CP-0 (Butyl). o-terphenyl CM D. Triphenylene ODP-0 EG C C C CP-0 (Octyl) C0 C C Elution volume (ml) 0 0 Carbon number (n) CP-0 CP-0 Relationship between the carbon number of alkylalcohols and elution volume PYE D EG C C 0 C ODP-0 (Octadecyl) C0 C C 0 0 Column : Shodex Asahipak ODP-0 D Shodex Asahipak CP-0 D Shodex Asahipak CP-0 D Eluent : H O/CH OH=0/0 Flow rate : 0.mL/ Column temp. : 0 C Column : Shodex Silica CM D Shodex Silica PYE D Eluent : H O/CH OH=0/0 Flow rate :.0mL/ Detector : UV (nm) Column temp. : 0 C

10 Columns for Polymer-based Reversed Phase Chromatography (ODP HP) Please refer to Comparison of the Features Among Shodex Reverse Phase Chromatography (RPC) Columns on page and for features. Standard columns Functional Group Shipping Solvent F00 ODP HP-B, H O/CH CN=/ F00 ODP HP-D, H O/CH CN=/ F00 ODP HP-E, H O/CH CN=/ F00 ODP HPG-A. 0 H O/CH CN=/ F00 ODP HP-B, H O/CH CN=/ F00 ODP HP-D, H O/CH CN=/ F0 ODP HPG-A.0 0 H O/CH CN=/ Base Material : Polyhydroxymethacrylate mm I.D columns [Customized columns] Functional Group F00 ODP HP-B F00 ODP HP-D F0 ODP HPG-A.0 0 Base Material : Polyhydroxymethacrylate Tolerance of ODP HP for alkaline condition Comparison between the chromatograms obtained before and after passing alkaline eluent Before After 00 hr ODP HP-D (.mm I.D.x0mm) Sample : µl. Pyridine 00µg/mL. Phenol 0µg/mL 0 0 Before After hr ODS (.mm I.D.x0mm) 0 0 Correlation between alkaline eluent passing time and relative theoretical plate number Relative theoretical plate number for pyridine (%) ODP HP-D ODS Alkaline eluent passing time (hr) Analysis condition Column : Shodex ODP HP-D ODS from other manufacturer Eluent : H O/CH OH=0/0 Flow rate :.0mL/ Detector : UV (nm) Column temp. : 0 C Eluent passing conditions for alkali tolerance test Column : Shodex ODP HP-D ODS from other manufacturer Eluent : 0mM Sodium phosphate buffer (ph) /CH CN=/ Flow rate : 0.mL/ Column temp. : 0 C

11 Comparison between ODP HP and ODP-0 (existing) ODP HP-D N:,00 ODP-0 D (existing) Ratio of Pressure (%) * Sample : µl. Phenol 00mg/L. Methyl benzoate 0mg/L. Toluene 000mg/L. Naphthalene 0mg/L Column : Shodex ODP HP-D Eluent : H O/CH CN=/ Flow rate : 0.mL/ Detector : UV (nm) Column temp. : 0 C ODS (.0mm I.D. 0mm) ODP HP-B (.0mm I.D. 0mm) Number of Injection * Considering original pressure as 00% N:,00 Column : Shodex Asahipak ODP-0 D Eluent : H O/CH CN=/ Flow rate : 0.mL/ Detector : UV (nm) Column temp. : 0 C Influence of repeated protein injection on column pressure ODP HP columns are packed with gels with increased surface polarity and smaller pore size which prevent the adsorption of proteins. BSA was injected multiple times to both ODS and ODP HP columns. A significant column pressure increase was observed for the ODS column, while no considerable change was observed for the ODP HP column even after 0 injections. Sample : µl BSA.0mg/mL Column : Shodex ODP HP-B ODS from other manufacturer Eluent : mm Ammonium acetate aq./ch CN=90/0 Flow rate : 0.mL/ Detector : UV (0nm) Column temp. : 0 C Imidazoles. Sample : 0.% each, 0µL. Imidazole. -Methylimidazole. -Methylimidazole Column : Shodex ODP HP-E Eluent : 0mM Na HPO aq./ch CN=90/0 Flow rate : 0.mL/ Detector : UV (0nm) Column temp. : 0 C Anticonvulsant in serum Sample : 0µL. -Acetaophenol (I.S.) 0µg/mL. Zonisamide.0µg/mL. Phenobarbital 9.0µg/mL. Carbamazepine.µg/mL. Phenytoin 9.0µg/mL Column : Shodex ODP HP-E Eluent : mm Sodium phosphate buffer (ph.)/ch CN=0/0 Flow rate : 0.mL/ Detector : UV (0nm) Column temp. : 0 C Sample pretreatment : Mix the same volumes of serum and acetonitrile. Centrifuge the mixture at 000g for utes. Use the supernatant as sample. Data courtesy of Katsuko Hara.MT Yutaka Komiyama Ph.D., Department of Clinical Sciences and Laboratory Medicine, Kansai Medical University. Columns for Polymer-based Reversed Phase Chromatography (ODP HP) Barbital recovery rate comparison of ODP HP-B and ODS in the presence of BSA ODP HP Barbital 00ng/mL ODS Barbital 00ng/mL For the analysis of drugs in samples containing proteins in matrix using LC/MS, ODP HP columns show less matrix effects (ion suppression in this case) compared to ODS columns due to fact that proteins can not be retained on ODP HP columns and are eluted as a void. Barbital Barbital Peak Area : 00% Peak Area : 00% Barbital 00ng/mL in BSA.0mg/mL Barbital 00ng/mL in BSA.0mg/mL Barbital Barbital Peak Area : 99% Peak Area : % Column : Shodex ODP HP-B ODS from other manufacturer Eluent : 0mM Ammonium acetate aq. /CH CN=0/0 Flow rate : 0.mL/ Detector : ESI-MS (SIM Negative : m/z ) Column temp. : 0 C Injection vol. : 0µL 9

12 Columns for Polymer-based Reversed Phase Chromatography (Asahipak) Please refer to Comparison of the Features Among Shodex Reverse Phase Chromatography (RPC) Columns on page and for features. Standard columns Functional Group Column Size (mm) Shipping Solvent F00 Asahipak ODP-0 D,000 Octadecyl 0. 0 H O/CH CN=/ F00 Asahipak ODP-0 E,000 Octadecyl 0. 0 H O/CH CN=/ F000 Asahipak ODP-0 D 9,000 Octadecyl H O/CH CN=/ F000 Asahipak ODP-0 E,000 Octadecyl H O/CH CN=/ F000 Asahipak ODP-0G A Octadecyl.0 0 H O/CH CN=/ F00 Asahipak ODP-0 B,00 Octadecyl 0. 0 H O/CH CN=/ F000 Asahipak ODP-0 D 9,000 Octadecyl 0. 0 H O/CH CN=/ F000 Asahipak ODP-0 E,000 Octadecyl 0. 0 H O/CH CN=/ F00 Asahipak ODP-0G A Octadecyl. 0 H O/CH CN=/ F0009 Asahipak ODP-0 D,000 Octadecyl H O/CH CN=/ F00 Asahipak ODP-0G A Octadecyl.0 0 H O/CH CN=/ F000 Asahipak CP-0 D,000 Octyl 0. 0 H O/CH CN=/ F000 Asahipak CP-0 E,000 Octyl 0. 0 H O/CH CN=/ F000 Asahipak CP-0G A Octyl. 0 H O/CH CN=/ F000 Asahipak CP-0 D,000 Butyl 0. 0 H O/CH CN=/ F000 Asahipak CP-0 E 9,000 Butyl 0. 0 H O/CH CN=/ F000 Asahipak CP-0G A Butyl. 0 H O/CH CN=/ mm I.D columns [Customized columns] Base Material : Polyvinyl alcohol Functional Group F0 Asahipak ODP-0 B Octadecyl F0 Asahipak ODP-0 D Octadecyl F0 Asahipak ODP-0G A Octadecyl Semi-micro columns * The following semi-micro columns are made to order. Functional Group F0 ODP0-B Octadecyl F0 ODP0-D Octadecyl Preparative columns * Preparative columns are made to order. Standard Column F000 Asahipak ODP-0 0E 0, ODP-0, ODP-0 F00 Asahipak ODP-90 0F 9, ODP-0, ODP-0 F000 Asahipak ODP-0G B. 0 F000 Asahipak CP-0 0E, CP-0 F00 Asahipak CP-0G B. 0 0 F000 F00 Asahipak CP-0 0E Asahipak CP-0G B, CP-0

13 Tolerance of ODP-0 for alkaline condition hr 90hr 0hr 0hr ph 0 0 Scopolae m/z 0(+) HC N O OH O O Sample :. Acetophenone. Butyrophenone. Hexanophenone. Heptanophenone. Octanophenone Column : Shodex Asahipak ODP-0 D Eluent : 0mM NaOH aq. (ph.0)/ch CN=/ Flow rate : 0.mL/ Detector : UV (nm) Column temp. : 0 C LC/MS analysis of basic drugs Sample : 0ng/mL each, µl. Scopolae. Atropine Local anesthetics Dissociation of tertiary ao groups in basic drugs can be suppressed by making ph of the eluent higher than pka of the ao groups. This increases the relative hydrophobicity of the basic drugs, thereby allowing the column to retain the drugs stronger and provide baseline separation of them. Fat-soluble vitas ph 0 0 Column : Shodex AsahipakODP-0 D Eluent : mm Phosphate buffer/ch CN=0/0 Flow rate : 0.mL/ Detector : UV (nm) Column temp. : 0 C ph 0 0 Sample :. Benzocaine HN. Lidocaine CH CH. Tetracaine NHCOCHN COOCHCHN NH(CH)CH COOCH Sample : 0µL. Vita K.µg/mL. Vita A.0 IU/mL. Vita A acetate 0. IU/mL. Vita D.µg/mL. Vita D. IU/mL. Vita E acetate.µg/ml. Vita E.µg/mL. Vita K.µg/mL CH CH CH CH Columns for Polymer-based Reversed Phase Chromatography (Asahipak) Atropine m/z 90(+) N O O OH Column : Shodex ODP0-D Eluent : 0.0% Ammonia aq./ch CN=0/0 Flow rate : 0.mL/ Detector : ESI-MS (SIM Positive) Column temp. : 0 C Column : Shodex Asahipak ODP-0 E Eluent : CH CN/CH OH=0/0 Flow rate : 0.mL/ Detector : UV (0nm) Column temp. : 0 C Analysis of azithromycin according to JP method Macrolide antibiotics Gradient analysis of proteins and peptides Sample : µl. Azithromycin mg/ml.,'-bis(diethylao)benzophenone 0µg/mL Resolution (Azithromycin/Internal standard) Sample : 0.% each, 0µL. Erythromycin. Azithromycin 0%CHCN 9 0 Sample : 0µL No %CHCN Sample Lys-Bradykinin Bradykinin Met-Enkephalin Neurotensin Leu-Enkephalin Substance P Bacitracin Insulin Insulin B chain Lysozyme Mastoparan Myoglobin Recovery MW (%) Column : Shodex Asahipak ODP-0 E Column : Shodex Asahipak ODP-0 E Column : Shodex Asahipak ODP-0 D Eluent : 0mM K HPO aq. (ph.0 adjusted with potassium hydroxide aq.)/ch CN=0/0 Eluent Flow rate : 0mM Potassium phosphate buffer (ph.0)/ch CN=0/0 : 0.mL/ Eluent : (A); 0.0% TFA aq./ch CN=0/0 (B); 0.0% TFA aq./ch CN=0/0 Linear gradient; (A) to (B), 0 Flow rate :.0mL/ Detector : UV (nm) Flow rate :.0mL/ Detector : UV (nm) Column temp. : 0 C Detector : UV (0nm) Column temp. : 0 C Column temp. : 0 C

14 Columns for Polymer-based Reversed Phase Chromatography (RSpak) Please refer to Comparison of the Features Among Shodex Reverse Phase Chromatography (RPC) Columns on page and for features. Standard columns Functional Group Base Material Particle Size Pore Size Column Size (mm) Shipping Solvent F RSpak RP-,000 Styrene divinylbenzene copolymer 0. 0 H O/CH CN=/9 F09 RSpak RP-G Styrene divinylbenzene copolymer. 0 H O/CH CN/THF=0/0/0 F0000 RSpak DS-,00 Styrene divinylbenzene copolymer H O/CH CN/THF=0/0/0 F000 RSpak DS-,000 Styrene divinylbenzene copolymer H O/CH CN/THF=0/0/0 F000 RSpak DS-G Styrene divinylbenzene copolymer 0. 0 H O/CH CN/THF=0/0/0 F0000 RSpak DE-,000 Polymethacrylate.0 0 H O F0000 RSpak DE-,000 Polymethacrylate. 0 H O/CH CN=0/0 F00900 RSpak DE-L,000 Polymethacrylate. 0 H O/CH CN=0/0 F000 RSpak DE-G A (RSpak DE-G) Polymethacrylate 0. 0 H O F0000 RSpak DE-,000 Polymethacrylate.0 0 H O/CH CN=0/0 F00 RSpak DE-G A (RSpak DE-SG) Polymethacrylate.0 0 H O/CH CN=0/0 F0000 RSpak DM-,00 Polyhydroxymethacrylate mm H PO aq. F000 RSpak DM-G A (RSpak DM-G) Polyhydroxymethacrylate. 0 mm H PO aq. F000 RSpak NN- 9,000 Sulfo Polyhydroxymethacrylate M Sodium phosphate buffer (ph.0) F000 RSpak NN-,000 Sulfo Polyhydroxymethacrylate M Sodium phosphate buffer (ph.0) F000 RSpak NN-G Sulfo Polyhydroxymethacrylate M Sodium phosphate buffer (ph.0) F000 RSpak NN-,000 Sulfo Polyhydroxymethacrylate M Sodium phosphate buffer (ph.0) F000 RSpak JJ-0 D,00 Quaternary ammonium Polyvinyl alcohol H O/CH CN=0/0 F000 RSpak JJ-0 D,00 Quaternary ammonium Polyvinyl alcohol H O/CH CN=0/0

15 Semi-micro columns * The following semi-micro columns are made to order. F0 F0 F0 DE-B DE-E NN-D Functional Group Sulfo 0,000 Base Material Polymethacrylate Polymethacrylate Polyhydroxymethacrylate Preparative columns * Preparative columns are made to order. F0 F0090 F0 F009 RSpak DE-0 RSpak DE-G B (RSpak DE-LG) RSpak DE-0 RSpak DE-G 0C (RSpak DE-LLG) Standard Column DE-, DE- DE-, DE- DE-, DE- Columns for Polymer-based Reversed Phase Chromatography (RSpak) F0 RSpak DM-0, DM- F000 RSpak DM-G B (RSpak DM-LG).0 0 F0 RSpak DM DM- F00 RSpak DM-G 0C (RSpak DM-LLG)

16 Separation and recovery rate of standard proteins Alkylbenzenes Recovery (%). Ribonuclease A 9. Insulin 9. Cytochrome c 00. Lysozyme 00. BSA 9. Myoglobin 0. Ovalbu Sample : µl. m-cresol 0.%.,-Xylenol 0.%. Benzene 0.%. Toluene 0.%. Ethylbenzene 0.%. n-propylbenzene 0.% Column : Shodex RSpak RP- Eluent : (A); 0.% TFA aq./ch CN=99/ (B); 0.% TFA aq./ch CN=/9 Linear gradient; (B%) 0% to 0%, 0 Flow rate :.0mL/ Detector : UV (0nm) Column temp. : Room temp. 0 0 Column : Shodex RSpak DS- Eluent : H O/CH CN/THF=0/0/0 Flow rate :.0mL/ Detector : UV (nm) Column temp. : 0 C Fatty acid methyl esters Organic acids Sample : 0.% each, 0µL. Methyl linoleate. Methyl palmitate. Methyl oleate. Methyl stearate Sample : µl. Glyoxylic acid.mg/ml. Tartaric acid.9mg/ml. Malic acid.0mg/ml. Lactic acid µl/ml. Malonic acid.9mg/ml. Acetic acid µl/ml. Succinic acid.0mg/ml. Levulinic acid.9mg/ml 9. Propionic acid µl/ml Column : Shodex RSpak DS- Eluent : H O/CH CN/THF=//0 Flow rate :.0mL/ Detetor : RI Column temp. : 0 C Column : Shodex RSpak DE- Eluent : 0mM H PO aq. Flow rate :.0mL/ Column temp. : 0 C Food additives (Preservatives) Diols Sample : 0µL. Saccharin sodium 0.00%. p-hydroxybenzoic acid 0.00%. Sorbic acid 0.0%. Benzoic acid 0.0%. Methyl paraben 0.0%. Dehydroacetic acid 0.0%. Ethyl paraben 0.0%. Propyl paraben 0.0% n= Sample : % each,.µl HO(CH)nOH Column : Shodex RSpak DE- Eluent : 0mM KH PO + 0.% H PO aq. /CH CN=/ Flow rate :.0mL/ Detector : UV (0nm) Column temp. : 0 C 0 0 Column : Shodex RSpak DE- Eluent : H O Flow rate :.0mL/ Column temp. : 0 C

17 LC/MS analysis of organic acids 0 9 Ao acids Glyoxylic acid Pyruvic acid Glycolic acid Lactic acid Maleic acid Succinic acid Fumaric acid Adipic acid Sample : 0.% each, 0µL. Aspartic acid. Glycine. Alanine. Valine. Methionine. Isoleucine Sample : 0ng/mL each, 0µL m/z m/z m/z m/z 9 m/z m/z m/z m/z Column : Shodex RSpak DE- Eluent : (A); 0.% (v/v) Formic acid aq. (B); CH CN Linear gradient; B%; % (0) % () % (.) % (0) Flow rate : 0.mL/ Detector : ESI-MS (SIM Negative) Column temp. : 0 C Speciation of arsenic Standard arsenic compounds Carnitine Sample : 0µL. L-Carnitine 0.% N + OH O OH Column : Shodex RSpak NN- Eluent : 0.M H PO aq. Flow rate :.0mL/ Detector : UV (0nm) Column temp. : C Sample : Arsenic compounds, 0µL. Monomethylarsonic acid. Arsinic acid. Dimethylarsinic acid. Arsenobetaine. Tetramethylarsonium. Trimethylarsine oxide Columns for Polymer-based Reversed Phase Chromatography (RSpak) sec Urine from an arsenic poisoning patient 000 Urine of a healthy person Column : Shodex RSpak NN- Eluent : 0mM H PO aq. Flow rate :.0mL/ Column temp. : 0 C sec Column : Shodex RSpak NN- Eluent : mm HNO /mm NH NO aq. Flow rate : 0.mL/ Detector : ICP-MS (SIM m/z ) sec Source: Noriko Tsunoda, Pharmacia. 99, vol., No., p.- Phenylenediae isomers LC/MS analysis of haloacetic acids Sample : 00mg/L each, 0µL. p-phenylenediae. m-phenylenediae. o-phenylenediae Standard Sample : 0µL each (Standard) ng/ml each (Tap water). Chloroacetic acid. Dichloroacetic acid. Trichloroacetic acid (.µg/l) Tap water (0.µg/L) 0 0 Column : Shodex RSpak JJ-0 D Eluent : mm Ammonium acetate buffer (ph9.)/ch CN=0/0 Flow rate : 0.mL/ Detector : UV (nm) Column temp. : 0 C Column : Shodex RSpak JJ-0 D Eluent : mm Ammonium acetate aq. (ph9.)/ch CN=0/0 Flow rate : 0.mL/ Detector : Chloroacetic acid : ESI-MS (SIM Negative : m/z 9) Dichloroacetic acid : ESI-MS/MS (MRM Negative : m/z ) Trichloroacetic acid : ESI-MS (SIM Negative : m/z ) Column temp. : 0 C

18 Columns for Silica-based Reversed Phase Chromatography (ODS Columns) Please refer to Comparison of the Features Among Shodex Reverse Phase Chromatography (RPC) Columns on page and for features. Standard columns Functional Group Carbon Load (%) Shipping Solvent F000 Silica CM D 0,000 Octadecyl H O/CH OH=0/0 F00 Silica CM E,000 Octadecyl H O/CH OH=0/0 F00 Silica CM D 9,000 Octadecyl H O/CH OH=0/0 F00 Silica CP D 0,000 Octadecyl H O/CH OH=0/0 F00 Silica CP E,000 Octadecyl H O/CH OH=0/0 F00 Silica CP D 9,000 Octadecyl H O/CH OH=0/0 Base Material : Silica Preparative columns * Preparative columns are made to order. Standard Column F000 Silica CM 0E, CM F00 Silica CM 0E, CM

19 Columns for Silica-based Reversed Phase Chromatography (Other Columns) Please refer to Comparison of the Features Among Shodex Reverse Phase Chromatography (RPC) Columns on page and for features. Standard columns F00 F00 F00 F00 F00 F009 F00 F00 Silica C D Silica C E Silica C D Silica C E Silica CN D Silica CN E Silica NPE D Silica PYE D 9,000,000 9,000,000,000,000,000,000 Functional Group Octyl Octyl Butyl Butyl Cyanopropyl Cyanopropyl Nitrophenylethyl Pyrenylethyl Preparative columns * Preparative columns are made to order. F00 F00 F00 F00 Standard columns F000 F00 F000 F00 Silica SIL D Silica SIL E Silica NH D Silica NH E Silica C 0E Silica C 0E Silica C 0E Silica C 0E 9,000,000,000,000,000 Functional Group Aopropyl Aopropyl,000,000,000 Carbon Load (%) 0 0 Carbon Load (%) Columns for Silica-based HILIC and Normal Phase Chromatography Features SIL Uses high purity silica (99.99% or higher) Suitable for normal phase analysis using a nonpolar organic solvent Corresponds to USP L NH Preparative columns * Preparative columns are made to order. F Suitable for saccharides analysis by hydrophilic interaction chromatography (HILIC) Corresponds to USP L Silica SIL 0E, Shipping Solvent H O/CH OH=/ H O/CH OH=/ H O/CH OH=/ H O/CH OH=/ H O/CH OH=0/0 H O/CH OH=0/0 H O/CH OH=/ H O/CH OH=0/0 Base Material : Silica Standard Column C C C C Shipping Solvent C H /C H OH=9/ C H /C H OH=9/ H O/CH CN=/9 H O/CH CN=/9 Base Material : Silica Standard Column SIL Columns for Silica-based Reversed Phase Chromatography (ODS Columns) (Other Columns) Columns for Silica-based HILIC and Normal Phase Chromatography F00 Silica SIL 0E, SIL F000 Silica NH 0E, NH F00 Silica NH 0E, NH

20 Aflatoxins TFA method Sample : µg/l each, 0µL. Derived Aflatoxin G. Derived Aflatoxin B. Aflatoxin G. Aflatoxin B Sample : 0µL. Aflatoxin M µg/l Reference : Shoku-An No. 0- (August, 0, Japan) "Test Methods Related to Total Aflatoxin" in Notice Reference : Shoku-An No. 0- (July, 0, Japan) Column : Shodex Silica CM E Eluent : H O/CH CN/CH OH=0/0/0 Flow rate :.0mL/ Detector : Fluorescence (Ex. : nm, Em. : 0nm) Column temp. : 0 C Column : Shodex Silica CM E Eluent : H O/CH CN=/ Flow rate :.0mL/ Detector : Fluorescence (Ex. : nm, Em. : nm) Column temp. : 0 C Trichothecene mycotoxins Ochratoxin Sample : 0µL. Nivalenol (NIV). Deoxynivalenol (DON) * mg/l each of DON and NIV were added to wheat sample Sample : 0µL. Ochratoxin A 0µg/L Reference : Shoku-An No. 000 (July, 00, Japan) Test method of deoxynivalenol Column : Shodex Silica CM E Eluent : H O/CH CN/CH OH=90// Flow rate :.0mL/ Detector : UV (0nm) Column temp. : 0 C Column : Shodex Silica CM D Eluent : H O/CH COOH/CH CN=// Flow rate :.0mL/ Detector : Fluorescence (Ex. : nm, Em. : 0nm) Column temp. : 0 C Gingerol and shogaol Chlorogenic acid Sample : 0.mg/mL each, 0µL. -Gingerol. -Shogaol Sample : 0.mg/mL each, 0µL. Chlorogenic acid. Caffeine. Caffeic acid Column : Shodex Silica CM D Eluent : (A) ; H O (B) ; CH CN Linear gradient : (B%) 0% to 0% () Flow rate :.0mL/ Detector : UV (0nm) Column temp. : 0 C Column : Shodex Silica CM D Eluent : 0mM H PO aq. /CH OH=0/0 Flow rate :.0mL/ Detector : UV (0nm) Column temp. : 0 C

21 Catechins Sample : 0µg/mL each, 0µL. Epigallocatechin. Catechin. Epigallocatechingallate. Epicatechin. Epicatechingallate. Catechingallate 0 0 Dinitronaphthalene isomers 0 0 Column : Shodex Silica CP D Eluent : (A) ; 0mM H PO aq. (B) ; CH CN Linear gradient: (B%) 0% (0 to ), 0 to 0% ( to ), 0% ( to 0) Flow rate :.0mL/ Detector : UV (0nm) Column temp. : 0 C Coenzyme Q0 Column : Shodex Silica CP D Eluent : CH OH/C H OH=/ Flow rate :.ml/ Detector : UV (nm) Column temp. : C 0 Sample : µl. Coenzyme Q0 mg/ml Sample :. Naphthalene.,-Dimethylnaphthalene.,-Dinitronaphthalene.,-Dinitronaphthalene Estradiols Column : Shodex Silica CP D Eluent : H O/CH CN=/ Flow rate :.0mL/ Detector : UV (0nm) Column temp. : 0 C Benzylpyridine isomers Column : Shodex Silica PYE D Eluent : 0mM KH PO aq./ch OH=0/0 Flow rate :.0mL/ Detector : UV (nm) Column temp. : 0 C Sample : 0.mg/mL each, 0µL. Estriol. β-estradiol. Estrone Simultaneous analysis of vita E homologs Sample :. -Benzylpyridine. -Benzylpyridine. -Benzylpyridine Sample : 0µL. α-tocopherol µg/ml. α-tocotrienol 0µg/mL. β-tocopherol µg/ml. γ-tocopherol µg/ml. γ-tocotrienol 0µg/mL. δ-tocopherol µg/ml. δ-tocotrienol 0µg/mL Columns for Silica-based Reversed Phase Chromatography (ODS Columns) (Other Columns) Columns for Silica-based HILIC and Normal Phase Chromatography Column : Shodex Silica NPE D Eluent : H O/CH OH=0/0 Flow rate :.0mL/ Detector : UV (nm) Column temp. : 0 C Column : Shodex Silica SIL D Eluent : n-hexane/isopropanol/acetic acid=000// Flow rate :.0mL/ Detector : Fluorescence (Ex. : 9nm, Em. : nm) Column temp. : 0 C 9

22 Columns for Polymer-based Hydrophilic Interaction Chromatography (HILIC) (HILICpak) Features VG-0 Suitable for saccharides analysis by hydrophilic interaction chromatography (HILIC) High recovery ratio of reducing saccharides Polymer-based packing material provides excellent chemical stability and imum deterioration over extended time period Easily regenerated by washing in a alkaline solution Also suitable for evaporative light scattering detector, corona charged aerosol detector, and LC/MS VT-0 Suitable for anionic substances analysis by hydrophilic interaction chromatography (HILIC) Depends on the eluent selected, the column adds ion exchange mode Polymer-based packing material provides excellent chemical stability and imum deterioration over extended time period Suitable for LC/MS Standard columns VG-0 (Housing Material : SUS) Functional Group Shipping Solvent F000 HILICpak VG-0 D,00 Ao H O/CH CN=0/0 F000 HILICpak VG-0 E,00 Ao H O/CH CN=0/0 F00 HILICpak VG-0G A Ao H O/CH CN=0/0 Base Material : Polyvinyl alcohol (Housing Material : PEEK) Functional Group Shipping Solvent F000 HILICpak VG-0 D,00 Ao H O/CH CN=/ F00 HILICpak VG-0G A Ao H O/CH CN=/ Base Material : Polyvinyl alcohol VT-0 (Housing Material : PEEK) Functional Group Shipping Solvent F000 HILICpak VT-0 D,00 Quaternary ammonium mm HCOONH aq. /CH CN=/ F00 HILICpak VT-0G A Quaternary ammonium mm HCOONH aq. /CH CN=/ Base Material : Polyvinyl alcohol 0

23 Recovery of reducing sugar Sample : mg/ml each, µl. Fructose. Mannose. Glucose. Sucrose VG-0 D (.mm I.D. x 0mm) Silica-based ao column (company-a) (.mm I.D. x 0mm) Silica-based ao column (company-b) (.mm I.D. x 0mm) VG-0 D Silica-based Silica-based Column : Shodex HILICpak VG-0 D Ao Column Ao Column (company-a) (company-b) Silica based ao columns from other manufacturers Eluent : H O/CH CN=0/0 Flow rate : 0.mL/ (VG-0 D).0mL/ (Silica based ao column) Column temp. : 0 C Lactose, epilactose, and aactulose When an ao column is used for analyzing saccharides, the recovery ratio of reducing saccharides such as mannose, arabinose or xylose is low because the ao group forms Schiff base with reducing saccharides. HILICpak VG-0 is the ao column that the recovery ratio of reducing saccharides is improved. By the improvement of the recovery ratio, the sensitivity of the analysis gets higher. Recovery of Mannose (%) Sample : mg/ml each, µl. Lactulose. Epilactose. Lactose m/z 0 (+) m/z () Rare sugar 0 0 Column : Shodex HILICpak VG-0 E Eluent : H O/CH CN/CH OH=//0 Flow rate : 0.mL/ Column temp. : 0 C Sample : 0.% each, 0µL. L-Ribose. D-Psicose. D-Xylitol. D-Tagatose. D-Allose. L-Glucose 0 LC/MS analysis of pantothenic acid and vita C Sample : 00ng/mL each, 0µL. Vita B (Pantothenic acid) as Calcium pantothenate. Vita C (Ascorbic acid) Columns for Polymer-based Hydrophilic Interaction Chromatography (HILIC) (HILICpak) 0 0 Column : Shodex HILICpak VG-0 E Eluent : H O/CH CN/CH OH=//0 Flow rate :.0mL/ Column temp. : 0 C Column : Shodex HILICpak VT-0 D Eluent : 0mM HCOONH aq./ch CN=0/0 Flow rate : 0.mL/ Detector : ESI-MS (SIM) Column temp. : 0 C LC/MS analysis of glyphosate and glufosinate LC/MS analysis of phosphorylated saccharides AMPA (Aomethylphosphonic acid) Sample : µg/ml each, µl m/z 0 () Sample : µm each, µl. Glucose--phosphate (GP). Fructose--phosphate (FP). Glucose--phosphate (GP). Fructose--phosphate (FP) Glufosinate m/z (+) Glyphosate m/z () MPPA (-Methylphosphinicopropionic acid) m/z (+) Column : Shodex HILICpak VT-0 D Eluent : H O/% HCOOH aq./ch CN=0/0/0 Flow rate : 0.mL/ Detector : ESI-MS (SIM) Column temp. : 0 C Column : Shodex HILICpak VT-0 D Eluent : mm HCOONH aq./ch CN=0/0 Flow rate : 0.mL/ Detector : ESI-MS (SIM Negative : m/z 9) Column temp. : 0 C

24 Columns for Polymer-based Hydrophilic Interaction Chromatography (HILIC) (Asahipak) Features NHP NHP-0 Suitable for saccharides analysis by hydrophilic interaction chromatography (HILIC) Polymer-based packing material provides excellent chemical stability and imum deterioration over extended time period Easily regenerated by washing in a alkaline solution Also suitable for evaporative light scattering detector, corona charged aerosol detector, and LC/MS Corresponds to USP L Provides higher theoretical plate number compared to NHP-0 series Corresponds to USP L Standard columns Functional Group Shipping Solvent F000 Asahipak NHP-0 B,00 Ao CH CN F000 Asahipak NHP-0 D,00 Ao CH CN F000 Asahipak NHP-0 E,00 Ao CH CN F00 Asahipak NHP-0G A Ao. 0 CH CN F000 Asahipak NHP-0 D,00 Ao CH CN F000 Asahipak NHP-0G A Ao.0 0 CH CN F000 Asahipak NHP-0 E,00 Ao CH CN F000 Asahipak NHP-0G A Ao.0 0 CH CN F000 Asahipak NHP-0 B,000 Ao CH CN F0009 Asahipak NHP-0 D,00 Ao CH CN F000 Asahipak NHP-0 E,000 Ao CH CN F000 Asahipak NHP-LF (line filter) Ao.0 CH CN Base Material : Polyvinyl alcohol mm I.D columns [Customized columns] Functional Group F00 Asahipak NHP-0 B Ao F00 Asahipak NHP-0 D Ao Preparative columns * Preparative columns are made to order. Standard Column F000 Asahipak NHP-0 0E 0, NHP-0 F00 Asahipak NHP-90 0F 0, NHP-0 F00 Asahipak NHP-0G B. 0

25 Fructooligosaccharide syrup Cyclodextrins Stevioside and rebaudioside A 0 Sample : Fructooligosaccharide syrup,.%, 0µL. Fructose. Glucose. Sucrose. -Kestose. Nystose. -Fructofuranosyl-D-nystose 0 0 Column : Shodex Asahipak NHP-0 E Eluent : H O/CH CN=0/0 Flow rate :.0mL/ Column temp. : C Imidazole dipeptides Sample : 0µL. β-alanine 00µg/mL. -Methyl-L-histidine µg/ml. L-Anserine µg/ml. Histidine µg/ml. L-Carnosine µg/ml. Nitrate (derived from L-Anserine Nitrate) Sample : 0µg/mL each, 0µL. α-cyclodextrin. γ-cyclodextrin. β-cyclodextrin Column : Shodex Asahipak NHP-0 E Eluent : H O/CH CN=0/0 Flow rate :.0mL/ Column temp. : 0 C Simultaneous analysis of water-soluble vitas Sample : 0µL. Vita B 0µg/mL. Nicotinamide 0µg/mL. Vita B 0µg/mL. Nicotinic acid 0µg/mL. Folic acid 0µg/mL. Vita C 0µg/mL 0 0 Ascorbic acid and erythorbic acid O=C HOC O HOC HC HCOH CHOH Sample : 0.0% each, 0µL. Stevioside. Rebaudioside A Column : Shodex Asahipak NHP-0 E Eluent : H O/CH CN=/ Flow rate :.0mL/ Detector : UV (0nm) Column temp. : 0 C Sample : µg/ml each, 0µL. Erythorbic acid. L-Ascorbic acid O=C HOC O HOC HC HOCH CHOH Columns for Polymer-based Hydrophilic Interaction Chromatography (HILIC) (Asahipak) Column : Shodex Asahipak NHP-0 E Eluent : 0mM NaH PO aq./ch CN=0/0 Flow rate :.0mL/ Detector : UV (0nm) Column temp. : 0 C 0 Column : Shodex Asahipak NHP-0 E Eluent : 0mM H PO aq./ch CN=/ Flow rate :.0mL/ Detector : UV (nm) Column temp. : 0 C 0 0 Column : Shodex Asahipak NHP-0 E Eluent : 0mM NaH PO + 0mM H PO aq. /CH CN=0/0 Flow rate :.0mL/ Detector : UV (nm) Column temp. : 0 C Comparison of saccharides analysis using corona charged aerosol detector LC/TOF-MS analysis of -ao benzoic acid derivatized sugar chains Sample : 0µg/mL each, µl. Fructose. Glucose Silica-based amido column (.mm I.D. x 0mm) Silica-based ao column (.mm I.D. x 0mm) A corona charged aerosol detector measures effluents as particles. Accordingly, the baseline will be significantly influenced by the components eluted from the column. NHP series, the polymer based ao columns eliate the components originated from the column, resulting a stable baseline with lower noise level. Sample : µl Human IgG (µg protein) AA-labeled N-Glycan * AA : -Aobenzoic acid 0 0 NHP-0 E (.mm I.D. x 0mm) Column : Shodex Asahipak NHP-0 E Silica based ao column from other manufacturer Silica based amide column from other manufacturer Eluent : H O/CH CN=0/0 Flow rate :.0mL/ Detector : Corona charged aerosol Column temp : 0 C (NHP-0 E, Silica based ao column) 0 C (Silica based amide column) 0 0 Migration time () Column : Shodex Asahipak NHP-0 D Eluent : (A); 9% CH CN/0.% Formic acid aq. (B); % CH CN/0.% Formic acid aq. Linear gradient; (B%) 0% (0-.), 0-9% (.-0) Flow rate : 0.mL/ Detector : ESI-TOF MS (Polarity : Negative, Full MS range : 000 ) Column temp. : C Data provided by Michihiro Kinoshita Ph.D., Faculty of Pharmacy, Kinki University

26 Columns for Ligand Exchange Chromatography * A list of elution volume of saccharides for Shodex columns is available. Please refer to our website ( or technical notebook (No. and ). Features SC0 SC SP00 KS-0 to 0 KS-0 to 0 DC- SZ SC SC0-F Separates saccharides by the combination of ligand exchange and size exclusion modes Three types of counter ions are available: Ca +, Pb +, and Na+ Only water is required for the analysis of neutral sugars SC0 and SC correspond to USP L9 and L SP00 corresponds to USP L and L KS-0 and KS-0 correspond to USP L and L Suitable for separation of polysaccharides by size exclusion mode Can be used in combination with other columns e.g., KS-0 and KS-0 Only water is required for the analysis of neutral sugars Corresponds to USP L and L Separates by the combination of ligand exchange and HILIC modes DC- can analyze sugars without removing sodium salts in the sample SZ is recommended for the separation of disaccharides or trisaccharides SC is suitable for separation of sugar alcohols DC- corresponds to USP L and L SZ corresponds to USP L SC corresponds to USP L9 and L For the analysis of mannitol under the method of JP, USP and EP Ca-type ligand exchange chromatography column Only water is required for the analysis of neutral sugars Corresponds to USP L9 and L Standard columns Ligand exchange and size exclusion Functional Group (Counter Ion) Exclusion Limit (Pullulan) Shipping Solvent F0 SUGAR SC0,000 Sulfo (Ca + ), H O F0 SUGAR SC,000 Sulfo (Ca + ) 0, H O F00090 SUGAR SC-G B (SUGAR SC-LG) Sulfo (Ca + ) H O F0 SUGAR SP00,000 Sulfo (Pb + ), H O F000 SUGAR SP-G B (SUGAR SP-G) Sulfo (Pb + ) H O F00 SUGAR KS-0,000 Sulfo (Na + ), H O F00 SUGAR KS-0,000 Sulfo (Na + ) 0, H O F0 SUGAR KS-0,000 Sulfo (Na + ) 0, H O F0 SUGAR KS-0,000 Sulfo (Na + ) 00, H O F00 SUGAR KS-0 9,000 Sulfo (Na + ),000, H O F00 SUGAR KS-0 9,000 Sulfo (Na + ) *(0,000,000).0 00 H O F0000 SUGAR KS-G B (SUGAR KS-G) Sulfo (Na + ) H O F00 SUGAR KS-0,000 Sulfo (Na + ) *(00,000,000) H O F000 SUGAR KS-0G Sulfo (Na + ) H O Ligand exchange and HILIC *( ) Estimated value Base Material : Styrene divinylbenzene copolymer Functional Group (Counter Ion) Shipping Solvent F0000 RSpak DC-,00 Sulfo (Na + ) H O/CH CN=0/0 F000 RSpak DC-G A (RSpak DC-G) Sulfo (Na + ) 0. 0 H O/CH CN=0/0 F0000 SUGAR SZ,00 Sulfo (Zn + ).0 0 H O/CH CN=0/0 F000 SUGAR SZ-G Sulfo (Zn + ). 0 H O/CH CN=0/0 F0000 SUGAR SC,00 Sulfo (Ca + ) H O/CH CN=/ F000 SUGAR SCG A (SUGAR SC-G) Sulfo (Ca + ) 0. 0 H O/CH CN=/ Base Material : Styrene divinylbenzene copolymer

27 For the analysis of mannitol in conformity with JP and USP F900 F90 Preparative columns * Preparative columns are made to order. F000 F000 F0009 F000 F00 F00 F0000 EP SC0-F USPpak MN- Ligand exchange and size exclusion SUGAR KS-00 SUGAR KS-00 SUGAR KS-00 SUGAR KS-00 SUGAR KS-00 SUGAR KS-00 SUGAR KS-G B (SUGAR KS-LG) Ligand exchange and HILIC Functional Group (Counter Ion) Sulfo (Ca + ) Sulfo (Ca + ),000,000,000,000,000, See p. for USP (Ver.) Column List. Base Material : Styrene divinylbenzene copolymer Shipping Solvent H O H O Standard Column KS-0 KS-0 KS-0 KS-0 KS-0 KS-0 Columns for Ligand Exchange Chromatography Standard Column F0 RSpak DC-0, DC- F000 RSpak DC-G B (RSpak DC-LG) F0 RSpak DC DC- F00 RSpak DC-G 0C (RSpak DC-LLG) Elution volume of saccharides analysis with various columns [Partial list only; refer to our website for complete list] Elution Volume (ml) Elution Volume (ml) Substances SP00 SC0 KS-0 SZ NHP-0 SC E Substances SP00 SC0 KS-0 SZ NHP-0 SC E Arabinose D-Mannose D-Arabitol Melibiose Dulcitol Nystose * meso-erythritol Palatinit peaks peaks.90 peaks. peaks D(-)-Fructose Palatinose D(+)-Fucose Panose * D(+)-Galactose D(+)-Raffinose * Gentiobiose * Rhamnose Glucose D(-)-Ribose myo-inositol D(-)-Sorbitol Isomaltose * Sorbose Isomaltotriose * Stachyose...9. * -Kestose * Sucrose * Kojibiose * α-d-talose Lactitol Trehalose * Lactose Trehalulose Lactulose Xylitol Maltitol Xylobiose * Maltose * D(+)-Xylose Maltotriose * D-Xylulose Mannitol ( ) Not detected ( * ) Overlap with solvent peak ( ) Not detected ( * ) Overlap with solvent peak Column : SUGAR SP00, SC0, KS-0 Eluent : H O Flow rate :.0mL/ Column temp. : 0 C Column : SUGAR SC Eluent : H O/CH CN=/ Flow rate :.0mL/ Column temp. : 0 C Column : SUGAR SZ Eluent : H O/CH CN=/ Flow rate :.0mL/ Column temp. : 0 C Column : Asahipak NHP-0 E Eluent : H O/CH CN=/ Flow rate :.0mL/ Column temp. : 0 C

28 Anomer separation of saccharides Calibration curves for KS-00 series using pullulan Temperature can affect chromatograms when separating saccharide anomers. When using a SUGAR column to analyze saccharides, the analysis at high temperatures would suppress the influence of anomer separation, resulting in better chromatograms. Sample : 0.% each, 0µL Glucose Mannose Xylose Galactose Arabinose 0 0 KS-0 9 C Glucose Mannose Xylose Galactose Arabinose Molecular weight (Pullulan) 0 0 KS-0 KS-0 KS-0 KS-0 0 KS-0 0 C Elution volume (ml) Column : Shodex SUGAR SC0 Eluent : H O Flow rate : 0.mL/ Column temp. : 9 C, 0 C Column : Shodex SUGAR KS-00 series Eluent : H O Column temp. : 0 C Hydrolyzed starch Saccharides in wood (model) Ketohexoses Dp-0 Dp- Dp- Sample : 0µL Hydrolyzed starch % Sample : µl. Cellobiose.0%. Glucose.%. Xylose 0.%. Galactose 0.%. Arabinose 0.%. Mannose.0% Sample : 0.0% each, 0µL. Sorbose. Fructose. Tagatose. Psicose Column : Shodex SUGAR KS-0 x Eluent : H O Flow rate :.0mL/ Column temp. : 0 C Column : Shodex SUGAR SP00 Eluent : H O Flow rate : 0.mL/ Column temp. : C Column : Shodex SUGAR SP00 Eluent : H O Flow rate :.0mL/ Column temp. : 0 C Pinitol Oligosaccharides in soybean Saccharides related to raffinose biosynthesis Sample : 0.% each, 0µL. Sucrose. Glucose. Pinitol. Fructose. chiro-inositol. myo-inositol Sample : 0.% each, 0µL. Verbascose. Stachyose. Raffinose. Sucrose. Pinitol Sample : 0.% each, 0µL. Raffinose. Sucrose. Galactinol. Galactose. myo-inositol Column : Shodex SUGAR SP00 Eluent : H O Flow rate : 0.mL/ Column temp. : C Column : Shodex SUGAR KS-0 + KS-0 Eluent : H O Flow rate : 0.mL/ Column temp. : C Column : Shodex SUGAR SC0 Eluent : H O Flow rate :.0mL/ Column temp. : 0 C

29 Acesulfame K and sucralose Sample : 0µL. Acesulfame K 0.%. Sucrose 0.%. Glucose 0.%. Unknown from Acesulfame K. Fructose 0.%. Sucralose 0.% 0 0 Column : Shodex SUGAR SC0 Eluent : 0mM CaSO aq. Flow rate : 0.mL/ Column temp. : 0 C 0 Sucrose and turanose Sample : 0.% each, 0µL. Fructose. Glucose. Sucrose. Turanose. Lactose Column : Shodex SUGAR SZ Eluent : H O/CH CN=0/0 Flow rate : 0.mL/ Column temp. : 0 C Maltose and isomaltose Sample : 0.% each, 0µL. Glucose. Maltose. Isomaltose. Maltotriose. Isomaltotriose Column : Shodex SUGAR SZ Eluent : H O/CH CN=/ Flow rate :.0mL/ Column temp. : 0 C Columns for Ligand Exchange Chromatography Saccharides and sugar alcohols Saccharides and Ao Acids Sample : 0.% each, 0µL. Rhamnose. Xylose. Arabinose. Fructose. Glucose. Galactose. Arabitol. Xylitol 9. Mannitol 0. Sorbitol Sample : 0.% each, 0µL. Sucrose. Glucose. Fructose. Taurine. myo-inositol Column : Shodex SUGAR SZ Eluent : H O/CH CN=0/0 Flow rate :.0mL/ Column temp. : C Column : Shodex SUGAR SC Eluent : H O/CH CN=0/0 Flow rate : 0.mL/ Column temp. : 0 C Moisturizing components Mannitol and sorbitol Sample : 0.% each, 0µL.,-Propanediol. Propylene glycol. Ethylene glycol. Glycerin. meso-erythritol Analytical conditions of mannitol for United States Pharmacopeia (USP) were partially changed in USP -NF operated on December st, 0. Now USP, European Pharmacopeia (EP) and Japanese Pharmacopeia (JP) request the column whose resolution between mannitol and sorbitol is more than.0. SC0-F is the columns under the method of these pharmacopeias. Sample : mg/ml each, 0µL. Mannitol. Sorbitol Resolution (Mannitol/Sorbitol) Column : Shodex SUGAR SC Eluent : H O/CH CN=0/0 Flow rate : 0.mL/ Column temp. : 0 C Column : Shodex EP SC0-F Eluent : H O Flow rate : 0.mL/ Column temp. : C

30 Columns for Ion Exclusion Chromatography Features SH0 SH Columns for simultaneous analysis of saccharides and organic acids Separates neutral sugars in size exclusion mode and organic acids in ion exclusion mode Suitable for the analysis of uronic and aldonic acids Corresponds to USP L and L KC- Columns for the analysis of organic acids Ion exclusion mode (+ reversed phase mode) Highly selective detection with post column method KC- E is suitable for the analysis of cyanide ions and cyanogen chloride in accordance with the Japanese Water Supply Act Corresponds to USP L and L Standard columns For simultaneous analysis of saccharides and organic acids Functional Group Exclusion Limit (Pullulan) Shipping Solvent F00 SUGAR SH0,000 Sulfo, H O F0 SUGAR SH,000 Sulfo 0, H O F0000 SUGAR SH-G Sulfo H O Base Material : Styrene divinylbenzene copolyme For organic acids, cyanide ions and cyanogen chloride Functional Group Shipping Solvent F00 RSpak KC-,000 Sulfo % H PO aq. F0 RSpak KC- E,000 Sulfo % H PO aq. F0000 RSpak KC-G B (RSpak KC-G) Sulfo % H PO aq. F0000 RSpak KC-G B (RSpak KC-LG) Sulfo % H PO aq. * As a guard column, use KC-G B for samples with relatively high impurity and KC-G B for samples with relatively low impurity. Base Material : Styrene divinylbenzene copolyme Preparative columns * Preparative columns are made to order. Standard Column F00 RSpak KC-0, KC- F0000 RSpak KC-G B (RSpak KC-LG).0 0

31 Maltooligosaccharides, organic acids and ethanol Sample : 0.0% each, 0µL. Maltotetraose. Maltotriose. Maltose. Glucose. Lactic acid. Glycerol. Acetic acid. Methanol 9. Ethanol Column : Shodex SUGAR SH Eluent : 0.mM H SO aq. Flow rate : 0.mL/ Column temp. : C 9 Cello-oligosaccharides and furfurals Sample : 0.% each, 0µL. Cellopentaose. Cellotetraose. Cellotriose. Cellobiose. Glucose. Glyceric acid. Acetic acid. -HMF 9. Furfural Column : Shodex SUGAR SH Eluent : mm H SO aq. Flow rate : 0.mL/ Column temp. : 0 C 9 Glycerin and glyceric acid Sample : 0.% each, 0µL. Glyceric acid. Glycerin HOCH CH(OH)CH OH HOCHCH(OH)COOH Column : Shodex SUGAR SH0 Eluent : mm H SO aq. Flow rate : 0.mL/ Column temp. : 0 C Columns for Ion Exclusion Chromatography General organic acids Hydrophobic organic acids Glucronolactone and organic acids Sample :. Citric acid. Tartaric acid. Pyruvic acid. Malic acid. Succinic acid. Glycolic acid. Lactic acid. Fumaric acid 9. Acetic acid 0. Levulinic acid. Pyroglutamic acid. Propionic acid. Isobutyric acid. n-butyric acid 9 0 Eluent = mm HClO aq. 0 Sample :. Succinic acid. Lactic acid. Formic acid. Acetic acid. Propionic acid. Isobutyric acid. n-butyric acid. Isovaleric acid 9. n-valeric acid 0 0 To shorten analysis time 9 Sample : 0µL. Citric acid 0.0%. Malic acid 0.0%. Glucronolactone 0.0%. Glycerin 0.0%. Ethanol 0.0% Eluent = mm HClO aq./ch CN=90/ Column : Shodex RSpak KC- x Eluent : mm HClO aq. Flow rate :.0mL/ Detector : VIS (0nm) post column method Column temp. : 0 C 0 0 Column : Shodex RSpak KC-LG + KC- x Flow rate :.0mL/ Detector : VIS (0nm) post column method Column temp. : C 0 0 Column : Shodex RSpak KC- Eluent : mm HClO aq. Flow rate :.0mL/ Column temp. : 0 C Organic acids in sake Analysis of Cyanide ion and cyanogen chloride with post column method Sample : 00µL. Phosphoric acid etc.. Citric acid. Pyruvic acid. Malic acid. Succinic acid. Lactic acid. Fumaric acid. Acetic acid 9. Pyroglutamic acid Fine rice wine Fermented liquor Pure rice wine CN - Sample : 0µg/L each, 00µL CNCl Column : Shodex RSpak KC-LG + KC- x Eluent :.mm HClO aq. Flow rate :.0mL/ Detector : VIS (0nm) post column method Column temp. : C Column Eluent Reagent A Reagent B Flow rate : Shodex RSpak KC- E :.0mM H SO aq. : Chlorae T solution : -Pyridinecarboxylic acid-pyrazolone solution : (Eluent).0mL/ (Reagent) 0.mL/ each : VIS (nm) Detector Column temp. : 0 C Reaction temp. : (Reagent A) 0 C (Reagent B) 0 C 9

32 Columns for Ion Chromatography (Anion Analysis) Features NI- I-A SI-90 E SI-0 E SI- D SI- E Columns for anion analysis with non-suppressor method NI- supports simultaneous analysis of fluoride and phosphate ions Columns for anion analysis with suppressor method Suitable for the quantitative analysis of fluoride ion SI-0 separates target inorganic anions from organic acids Not interfered by the system peak derived from carbonate Columns for the analysis of oxyhalides with suppressor method SI- supports rapid analysis of oxyhalides and general inorganic ions SI- supports simultaneous analysis of oxyhalides and general inorganic ions Standard columns For anions (non-suppressor method) Functional Group Particle Size Column Size (mm) Shipping Solvent F99 F09 IC NI- IC NI-G,000 Quaternary ammonium Quaternary ammonium mm -Hydroxybenzoic acid +.mm Bis-Tris + mm Phenylboronic acid mM CyDTA aq. mm -Hydroxybenzoic acid +.mm Bis-Tris + mm Phenylboronic acid mM CyDTA aq. F990 IC I-A,000 Quaternary ammonium. 00.mM Phthalic acid aq. F0000 IC IA-G Quaternary ammonium. 0.mM Phthalic acid aq. For anions (suppressor method) Base Material : Polyhydroxymethacrylate Housing Material : SUS Functional Group Particle Size Column Size (mm) Shipping Solvent F99 IC SI-90 E,000 Quaternary ammonium mM Na CO +.mm NaHCO aq. F090 IC SI-90G Quaternary ammonium 9. 0.mM Na CO +.mm NaHCO aq. F99 IC SI-0 E 0,000 Quaternary ammonium.0 0.mM Na CO +.0mM NaHCO aq. F09 IC SI-0G Quaternary ammonium. 0.mM Na CO +.0mM NaHCO aq. For oxyhalides (suppressor method) Base Material : Polyvinyl alcohol Housing Material : PEEK Functional Group Particle Size Column Size (mm) Shipping Solvent F9990 IC SI- D,000 Quaternary ammonium..0 0.mM Na CO aq. F09 IC SI-9G Quaternary ammonium 9. 0.mM Na CO aq. F990 IC SI- E,000 Quaternary ammonium.0 0.mM Na CO aq. F09 IC SI-9G Quaternary ammonium 9. 0.mM Na CO aq. Base Material : Polyvinyl alcohol Housing Material : PEEK Line filters for IC Contents F000 IC FL- One holder and one filter 0 F000 IC FL- filter filters

33 Anions analysis with non-suppressor method (NI- and I-A) 0 NI- Sample : 0µL. HPO - 0mg/L. F - mg/l. CI - mg/l. NO - mg/l. Br - mg/l. NO - mg/l. SO - mg/l Column : Shodex IC NI- Eluent : mm -Hydroxybenzoic acid +.mm Bis-Tris + mm Phenylboronic acid mM *CyDTA aq. Flow rate :.0mL/ Detector : Non-suppressed conductivity Column temp. : 0 C *CyDTA : trans-,-diaocyclohexane-n,n,n',n'-tetra acetic acid, I-A Column : Shodex IC I-A Eluent :.mm Phthalic acid +.mm Tris(hydroxymethyl)aomethane aq. Flow rate :.ml/ Detector : Non-suppressed conductivity Column temp. : 0 C NI- is a high performance type of column offers an increased theoretical plate number twice as much as I-A. [Features of NI-] () Enables the separation of H PO - and F - which were difficult to separate with I-A. () The shape of each peak is sharper, and the separation balance is proper. Especially, the separation of Cl - and NO - is improved. Columns for Ion Chromatography (Anion Analysis) Anions analysis using SI-90 E (suppressor method) Perchloric acid analysis using SI-90 E (suppressor method) Anions analysis using SI-0 E (suppressor method) Sample : 0µL. F - mg/l. Cl - mg/l. NO - mg/l. Br - 0mg/L. NO - 0mg/L. HCO - 00mg/L. PO - mg/l. SO - mg/l Sample : 00µL. Cl - mg/l. NO - 0mg/L. SO - mg/l. ClO - 0mg/L SI-0 E is a high performance type of SI-90 E. Acetic acid, formic acid, and methacrylic acid eluted between F - and Cl -. The carbonate system peak appears between NO - and Br - peaks. Sample : 0µL. F - mg/l. Acetic acid 0mg/L. Formic acid mg/l. Methacrylic acid 0mg/L. Cl - mg/l. NO - mg/l. Br - 0mg/L. NO - 0mg/L 9. PO - mg/l 0. SO - mg/l. Oxalic acid mg/l Column : Shodex IC SI-90 E Eluent :.mm Na CO +.mm NaHCO aq. Flow rate :.ml/ Detector : Suppressed conductivity Column temp. : Room temp. ( C) Column : Shodex IC SI-90 E Eluent : mm Na CO aq. + 0% CH CN Flow rate :.0mL/ Detector : Suppressed conductivity Column temp. : C Column : Shodex IC SI-0 E Eluent :.mm Na CO +.0mM NaHCO aq. Flow rate : 0.mL/ Detector : Suppressed conductivity Column temp. : C Oxyhalides and anions analysis using SI- E (suppressor method) Oxyhalides and anions analysis using SI- D (suppressor method) Tricarboxylic acid (suppressor method) SI- E is a high resolution column offering,000 or higher theoretical plate number. It supports simultaneous analysis of oxyhalides and inorganic anions. The recommended analysis temperature is C. Sample : 0µL. F - mg/l. ClO - mg/l. BrO - mg/l. Cl - 0mg/L. NO - mg/l. Br - 0mg/L. ClO - mg/l. Dichloroacetic acid mg/l 9. NO - 0mg/L 0. PO - mg/l. SO - 0mg/L 9 0 Sample : 0µL. F - mg/l. ClO - mg/l. BrO - mg/l. Cl - 0mg/L. NO - mg/l. Br - 0mg/L. ClO - mg/l. NO - 0mg/L 9. PO - mg/l 0. SO - 0mg/L Sample : 0µL. Citric acid 0mg/L. Isocitric acid 0mg/L. trans-aconitic acid 0mg/L. cis-aconitic acid 0mg/L Column : Shodex IC SI- E Eluent :.mm Na CO aq. Flow rate : 0.mL/ Detector : Suppressed conductivity Column temp. : C Column : Shodex IC SI- D Eluent :.0mM Na CO +.mm NaHCO aq. Flow rate : 0.mL/ Detector : Suppressed conductivity Column temp. : C Column : Shodex IC SI- D Eluent : 9.0mM Na CO aq. Flow rate : 0.mL/ Detector : Suppressed conductivity Column temp. : C

34 Columns for Ion Chromatography (Cation Analysis) Features YS-0 YK- High performance type of YK- Applicable to both suppressor and non-suppressor methods Peak shape is sharper, especially for divalent cation analysis Supports the analysis of alkylaes and transition metals Column for cation analysis with non-suppressor method Simultaneous analysis of monovalent and divalent cations Suitable for separation of alkylaes Y- Column for cation analysis with non-suppressor method For the separation of monovalent or divalent cations Corresponds to USP L and L T- Column for transition metal ion analysis Highly sensitive analysis is achieved by post column color reaction method Standard columns For cations Functional Group Base Material Particle Size Column Size (mm) Shipping Solvent F000 IC YS-0,00 Carboxyl Polyvinyl alcohol. H O F000 IC YS-G Carboxyl Polyvinyl alcohol. 0 H O F00 IC YK-,00 Carboxyl Silica. mm Tartaric acid + mm Dipicolinic acid +.g/l Boric acid aq. F090 IC YK-G Carboxyl Silica. 0 mm Tartaric acid + mm Dipicolinic acid +.g/l Boric acid aq. F990 IC Y-,000 Sulfo Styrene divinylbenzene copolymer. 0 mm HNO aq. F000 IC Y-G Sulfo Styrene divinylbenzene copolymer. 0 mm HNO aq. Housing Material : SUS For transition metal ions Functional Group Shipping Solvent F990 IC T-,000 Sulfo. 0 mm HNO aq. F00 IC T-G Sulfo. 0 mm HNO aq. Base Material : Styrene divinylbenzene copolymer Housing Material : PEEK Line filters for IC Contents F000 IC FL- One holder and one filter F000 IC FL- filter filters

35 Standard cations (YS-0 and YK-) 0 YS Column : Shodex IC YS-0 Eluent : mm Methanesulfonic acid aq. Flow rate :.0mL/ Detector : Non-suppressed conductivity Column temp. : 0 C 0µL inj. YK- µl inj. Sample :. Li + mg/l. Na + 0mg/L. NH + 0mg/L. K + 0mg/L. Mg + 0mg/L. Ca + 0mg/L YS-0 is a high performance type of column offers an increased theoretical plate number twice as much as YK-. In particular, the peak shape is improved. The quantitative performance for NH + in a sample containing high concentration Na + is also enhanced. Resolution (Na + and NH + ) TP Mg + Ca + Column : Shodex IC YK- Eluent : mm Tartaric acid + mm Dipicolinic acid +.g/l Boric acid aq. Flow rate :.0mL/ Detector : Non-suppressed conductivity Column temp. : 0 C YS-0. YS-0 YK-. YK-,900,000,00,000 Columns for Ion Chromatography (Cation Analysis) Effects of crown ether in eluent Simultaneous analysis for cations and ethylenediae Crown ether forms complex with cations. The elution of cations (particularly K + ) can be well controlled by the difference in complex formation rate. mm Sample : 0µL. Li + mg/l. Na + 0mg/L. NH + 0mg/L. K + 0mg/L. Mg + 0mg/L. Ca + 0mg/L Sample : 0µL. Li + mg/l. Na + 0mg/L. NH + 0mg/L. K + 0mg/L. Mg + 0mg/L. Ca + 0mg/L. Ethylenediae 0mg/L -Crown -ether conc. mm mm 0mM Column : Shodex IC YS-0 Eluent : mm Methanesulfonic acid + -Crown -ether aq. Flow rate :.0mL/ Detector : Non-suppressed conductivity Column temp. : 0 C Column : Shodex IC YS-0 Eluent : mm Nitric acid +.mm -Crown -ether aq. /CH CN=90/0 Flow rate :.0mL/ Detector : Non-suppressed conductivity Column temp. : 0 C Ao alcohols Alkylaes Alkaline earth metal ions Sample : 0µL. Monoethanolae 0µg/mL. Diethanolae 0µg/mL. N-Methylethanolae 0µg/mL. Triethanolae 0µg/mL. N-Methyldiethanolae 0µg/mL. N,N-Diethylethanolae 0µg/mL. N-(-Aoethyl)ethanolae 0µg/mL Sample : 0µL. NH + mg/l. Methylae mg/l. Dimethylae mg/l. Trimethylae 0mg/L. Ethylae 0mg/L. Propylae 0mg/L. Butylae 0mg/L Cu + Zn + Ni + Co + Mg + Fe +, Fe + Mn +, Pb + Cd + Ca + Sr + 0 Ba + Sample : 0µL. Ni + 0mg/L. Mg + 0mg/L. Mn + 0mg/L. Ca + 0mg/L Column : Shodex IC YK- Eluent : mm Nitric acid aq. Flow rate :.0mL/ Detector : Non-suppressed conductivity Column temp. : 0 C 0 0 Column : Shodex IC YK- Eluent : mm H PO aq./ch CN=90/0 Flow rate :.0mL/ Detector : Non-suppressed conductivity Column temp. : C Column : Shodex IC Y- Eluent : mm Tartaric acid + mm Ethylenediae aq. Flow rate :.0mL/ Detector : Non-suppressed conductivity Column temp. : 0 C

36 Column Selection for Size Exclusion Chromatography (SEC) Application Solvent Column Page Biological macromolecules (proteins, peptides, nucleic acids, etc.) Buffer etc. KW-00 series KW00 series High performance (solvent-saving) Aqueous SEC (GFC) Biological macromolecules (high MW range) Water-soluble polymers (polyacrylamide, polyethylenie, etc.) Polysaccharides Buffer etc. H O, Buffer Aqueous solution etc. SB-00 HQ series LB-00 series SB-00 HQ series LB-00 series Oligosaccharide, polysaccharides H O, Aqueous solution etc. KS-00 series GS-HQ series KF-00 series General polymers THF KF-00 series KF-00HQ series Rapid analysis (solvent-saving) High performance (solvent-saving) LF series High linearity of calibration curve Chloroform K-00 series Organic SEC (GPC) Polar polymers (polyimides, polyvinylpyrrolidones etc.) DMF KD-00 series 0 SB-00 HQ series LB-00 series Analysis at high temperature (polyethylene, polypropylene etc.) ODCB etc. HT-00 series UT-00 series AT-0MS Engineering resin analysis at room temperature [polyamide (Nylon), polyethylene terephthalate (PET) etc.] HFIP HFIP-00 series HFIP-00 series Rapid analysis (solvent-saving) High linearity of calibration curve 0 0 LF series Aqueous/Organic SEC GF-HQ series Guideline for column solvent usability High Polarity of solvent H O Methanol DMSO Acetonitrile DMF THF Chloroform Toluene Low KW-00 series, KW00 series SB-00 HQ series, LB-00 series GS-HQ series GF-HQ series KD series KF, K, LF series * See page for the solvent replaceability of organic solvent SEC (GPC) packed columns.

37 Precautions for Polar Polymer Analysis Size exclusion chromatography analysis of polar polymers can be influenced by unexpected interactions in the column. These interactions may change elution patterns and results in an invalid molecular weight calculation. It is important to reduce them in order to obtain the accurate molecular weight distribution. Interactions between the analyte and the packing materials Hydrophobic interaction The analyte is adsorbed into the packing material. This delays the analyte elution, and thus results in under estimation of its molecular weight (Figure B, D). Ionic interaction () Ion Exclusion The analyte is repelled from the packing material. This accelerates the analyte elution, and thus results in over estimation of its molecular weight (Figure A, C). () Ion Exchange The analyte is adsorbed onto the packing material. This delays the analyte elution, and thus results in under estimation of its molecular weight (Figure B, D). Interaction within and between the analyte Ionic repulsion effects observed within the multivalent macromolecules causes structure expansion This accelerates the analyte elution, and thus results in over estimation of its molecular weight (Figure A). Association between the molecules Associated molecule detected as a larger molecule (Figure A). Interactions between the analyte and the solvent The multivalent ion of the solvent works as a bridge to bind ionic molecules (analyte). Aqueous SEC (GFC) Interfering interactions likely to be observed Exclusion limit molecular weight log MW (C) Methods to reduce interactions Elution will be accelerated Exclusion limit volume (A) Elution will be retarded Organic solvent SEC (GPC) (B) Permeation limit volume (D) Column Selection for Size Exclusion Chromatography (SEC) Precautions for Polar Polymer Analysis Ionic Interaction Add salt Hydrophobic interaction Increase dissociation of the analyte Cationic polymer Lower the ph Anionic polymer Higher the ph Lower the eluent polarity (Example) Add acetonitrile or methanol Ionic Interaction Add salt (Example) Add LiBr to DMF Add CF COONa to HFIP Hydrophobic interaction Lower the polarity of the eluent (Example) Change the eluent from DMF to THF Hydrophillic interaction Increase the polarity of the eluent (Example) Change the eluent from THF to DMF

38 Aqueous SEC (GFC) Columns : Silica-based Features KW-00 KW00 Silica-based packed columns for aqueous SEC (GFC) analysis Suitable for the analysis of proteins and enzymes Corresponds to USP L0, L, and L9 Reducing particle size of the packing material enhances column performance Three- or four-fold higher sensitivity than KW-00 series KW0-F is applicable to samples with a molecular weight above,000,000 Corresponds to USP L0, L, and L9 Standard columns F99000 F990 F990 F00 PROTEIN KW-0. PROTEIN KW-0 PROTEIN KW-0 PROTEIN KW-G B (PROTEIN KW-G) High performance semi-micro columns F990 F990 F990 F990 F00 KW0.-F KW0-F KW0-F KW0-F KW00G-A,000,000,000,000,000,000,000 Semi-micro columns * The following semi-micro columns are made to order. F F F F KW0.-B KW0.-D KW0-B KW0-D Base Material : Silica Usable ph range : ph.0-. Use of the KW00 series with semi-micro type devices is recommended ,000, ,00, Base Material : Silica H O H O H O H O Usable ph range : ph Shipping Solvent Shipping Solvent H O H O H O H O H O Use of the KW00 series with semi-micro type devices is recommended. Preparative columns * Preparative columns are made to order. Standard Columns F000 PROTEIN KW-00., KW-0. F00 PROTEIN KW-00, KW-0 F00 PROTEIN KW-00, KW-0 F09 PROTEIN KW-G B (PROTEIN KW-LG).0 0 Target molecular weight range and Exclusion limit With Protein (eluent : phosphate buffer) With Pullulan (eluent : ultrapure water) Target Molecular Weight Range Exclusion Limit KW-0. KW-0 KW-0 KW0. KW0 KW0 KW0,000 00,000 0,000 00,000 0,000 *(,000,000),000 0,000 0,000 00,000 0,000 *(,000,000) 00,000 *(0,000,000) 0,000 *(,000,000) *(,000,000) 0,000 00,000 *(,000,000) *(0,000,000) * Please use the above table as a rough indication for the column selection. Target Molecular Weight Range Exclusion Limit KW-0. KW-0 KW-0 KW0. KW0 KW0 KW0,000 0,000,000 00,000 0,000 00,000,000 0,000,000 0,000 0,000 00,000 00,000 00,000 0,000 0,000 00,000 0,000 0,000 00,000,00,000 *( ) Estimated value * Please use the above table as a rough indication for the column selection.

39 Calibration curves for KW00 series and KW-00 series using protein Molecular weight (Protein) KW0-F KW0.-F KW0-F KW0-F Elution volume (ml) Molecular weight (Protein) KW-0 KW-0. Column : Shodex KW00-F series, Shodex PROTEIN KW-00 series Eluent : 0mM Sodium phosphate buffer + 0.M NaCl (ph.0) Flow rate : 0.mL/ (KW00).0mL/ (KW-00) Detector : UV (0nm) (small cell volume) (KW00) UV (0nm) (conventional type) (KW-00) Column temp. : 0 C KW Elution volume (ml) Recovery rate of proteins Protein γ - Globulin Bovine serum albu Ovalbu Myoglobin Cytochrome c Lysozyme α-chymotrypsinogen A Recovery (%) KW0.-F KW0-F Column : Shodex KW0.-F Shodex KW0-F Eluent : 0mM Sodium phosphate buffer + 0.M NaCl (ph.0) Flow rate : 0.mL/ Detector : UV (0nm) (small cell volume) Column temp. : C Aqueous SEC (GFC) Columns : Silica-based Comparison of KW0.-F and KW-0. KW00 series is a high performance type of semi-micro columns, offering approximately. times larger theoretical plate number and to times higher detection sensitivity (peak height) than KW-00 series columns does. Sample : 0µL. Blue dextran mg/mL. γ-globulin 0.mg/mL. Ovalbu 0.mg/mL. Myogrobin 0.mg/mL. Uridine 0.0mg/mL (A)KW0.-F N=,000 Whey in yogurt Lectins Sample : Whey, µl Sample : µl. Lectin from Soybean 0.mg/mL. Lectin from Arachis hypogaea.mg/ml. Lectin from Canavalia ensiformis (Con A) 0.9mg/mL. Lectin from Lens culinaris (LCA) 0.mg/mL. Lectin from Triticum vulgaris (WGA) 0.mg/mL (B)KW-0. N=, Column : Shodex KW0.-F Shodex PROTEIN KW-0. Eluent : 0mM Sodium phosphate buffer + 0.M NaCl (ph.0) Flow rate : (A) 0.mL/, (B).0mL/ Detector : UV (0nm) (small cell volume) Column temp. : C Column : Shodex KW0.-F + KW0-F Column : Shodex KW0.-F Eluent : 0mM Sodium phosphate buffer Eluent : 0mM Sodium phosphate buffer + 0.M NaCl (ph.0) + 0.M NaCl (ph.0) Flow rate : 0.0mL/ Flow rate : 0.mL/ Detector : UV (0nm) (small cell volume) Detector : UV (0nm) (small cell volume) Column temp. : 0 C Column temp. : 0 C Comparison of various GFC columns for separation of standard proteins Sample :. Thyroglobulin (bovine). γ-globulin (bovine). Ovalbu (chicken). Myoglobin (horse). Cyanocobala SB-0 HQ GF-0 HQ KW Three aqueous SEC columns (SB-0 HQ, GF-0 HQ, and KW-0) were compared for their separation performances. KW-0, silica-based column, showed the best separation performance for the analysis of protein standards. Molecular weight (Protein) 0 0 KW-0 GF-0 HQ SB-0 HQ 0 9 Elution time () Column : Shodex OHpak SB-0 HQ Shodex Asahipak GF-0 HQ Shodex PROTEIN KW-0 Eluent : 0.M Phosphate buffer (ph.9) Flow rate : 0.mL/ Detector : UV (0nm) Column temp. : 0 C Proteins in human blood serum Sample : 0.% each. Fibrinogen 0µL. α -Macroglobulin 0µL. IgG 0µL. Transferrin 0µL. Plasogen 0µL. Albu 00µL. Antitrypsin 00µL. Hemoglobin 00µL Column : Shodex PROTEIN KW-0 Eluent : 0mM Phosphate buffer + 0.M NaCl (ph.0) Flow rate :.0mL/ Detector : UV (0nm) Column temp. : Room temp.

40 Aqueous SEC (GFC) Columns : Polymer-based Features SB-00 HQ SB-0 HQ LB-00 Polymer-based packed columns for aqueous SEC (GFC) analysis Supports a wide range of molecular weight sample analysis The eluent can be replaced with DMF (except SB-0 HQ and SB-0 HQ), enabling the analysis of polar polymers Method using SB-0 HQ or SB-0 HQ for gelatin s mean molecular weight deteration is comparable with PAGI method (Ver. 0, Japan) Corresponds to USP L and L9 SB-0 HQ corresponds to USP L SB-0. HQ corresponds to USP L SB-0 HQ corresponds to USP L Column for the analysis of water-soluble ultra high molecular weight polymers Large particle size gel is packed to prevent shear degradation of polymers Corresponds to USP L and L9 Suitable for light scattering detector because of the controlled column bleed Polymer-based packed columns for aqueous SEC (GFC) analysis The eluent can be replaced with DMF enabling the analysis of polar polymers Corresponds to USP L and L9 Standard columns Shipping Solvent F900 OHpak SB-0 HQ, % NaN aq. F90 OHpak SB-0. HQ, % NaN aq. F90 OHpak SB-0 HQ, % NaN aq. F90 OHpak SB-0 HQ,000 0, % NaN aq. F90 OHpak SB-0 HQ,000, % NaN aq. F90 OHpak SB-0 HQ,000, % NaN aq. F90 OHpak SB-0M HQ,000, % NaN aq. F090 OHpak SB-G B (OHpak SB-G) % NaN aq. SB-0M HQ is a mixed-gel column capable of analyzing samples over a wide range of molecular weight distribution. Base Material : Polyhydroxymethacrylate Usable ph range : ph-0 For water-soluble ultra high molecular weight polymers Shipping Solvent F90 OHpak SB-0 HQ,00 0, H O F09 OHpak SB-0G.0 0 H O Base Material : Polyhydroxymethacrylate Usable ph range : ph-0 For light scattering Shipping Solvent F90 OHpak LB-0, H O F90 OHpak LB-0M,000, H O F09 OHpak LB-G B.0 0 H O Base Material : Polyhydroxymethacrylate Usable ph range : ph-0

41 Preparative columns * Preparative columns are made to order. F0 F0 F0 F0 F0 F0 F0 F09 OHpak SB-00 OHpak SB-00. OHpak SB-00 OHpak SB-00 OHpak SB-00 OHpak SB-00 OHpak SB-00M OHpak SB-G B (OHpak SB-LG) 9,000,000,000,000,000,000, Standard Column SB-0 HQ SB-0. HQ SB-0 HQ SB-0 HQ SB-0 HQ SB-0 HQ SB-0M HQ Aqueous SEC (GFC) Columns : Polymer-based Usable concentration of organic solvents The maximum usable concentration (%) Methanol Acetonitrile DMF SB-0 HQ SB-0. HQ, SB-0 HQ SB-0 HQ~SB-0M HQ SB-G B SB-0 HQ, SB-0G LB-0, LB-0M, LB-G B (Note) The maximum solvent tolerance of preparative type SB-00 HQ, SB-000 series, is 0% of methanol, acetonitrile, and DMF (SB-00 is not tolerant of organic solvents, similar to SB-0 HQ). Target molecular weight range and Exclusion limit With Pullulan (eluent : ultrapure water) With *PEG/PEO (eluent : DMF) Target Molecular Weight Range Exclusion Limit SB-0 HQ SB-0. HQ SB-0 HQ SB-0 HQ SB-0 HQ SB-0 HQ SB-0M HQ SB-0 HQ 00, ,000,000 00,000,000 00,000 00,000,000,000 00,000 *(0,000,000) 00 *(0,000,000) 00,000 *(00,000,000),000 0,000 00,000,000,000 *(,000,000) *(0,000,000) *(0,000,000) *(00,000,000) SB-0. HQ SB-0 HQ SB-0 HQ SB-0 HQ SB-0 HQ SB-0M HQ LB-0 LB-0M Target Molecular Weight Range 00, , ,000 0,000 00,000 0,000 **(0,000,000) 00 **(0,000,000) 00 0, **(0,000,000) LB-0 LB-0M * Please use the above table as a rough indication for the column selection.,000 00, *(0,000,000) 00,000 *(0,000,000) *( ) Estimated value * Please use the above table as a rough indication for the column selection. *PEG : polyethylene glycol *PEO : polyethylene oxide **( ) Estimated value 9

42 Calibration curves for SB-00 HQ series using pullulan (eluent : H O) Calibration curves for SB-00 HQ series using PEG/PEO (eluent : DMF) Carboxymethylcellulose 0 0 Sample : Carboxymethylcellulose 0.% each, 0µL,00-,000cP Molecular weight (Pullulan) SB-0 HQ SB-0 HQ SB-0. HQ SB-0 HQ SB-0 HQ SB-0 HQ SB-0M HQ Molecular weight (PEG/PEO) SB-0 HQ SB-0 HQ SB-0. HQ SB-0 HQ SB-0 HQ SB-0M HQ Mn : 0,00 Mw :,90,00 Mw/Mn : cP Mn : 0,00 Mw :,,000 Mw/Mn : cP Mn :,00 Mw : 0,000 Mw/Mn : Elution volume (ml) 9 0 Elution volume (ml) * Molecular weight was detered from the calibration curve of pullulan. Column : Shodex OHpak SB-00 HQ series Eluent : H O Flow rate :.0mL/ Column temp. : 0 C Column : Shodex OHpak SB-00 HQ series Eluent : DMF Flow rate :.0mL/ Column temp. : 0 C Column : Shodex OHpak SB-0M HQ x Eluent : 0.M NaCl aq. Flow rate :.0mL/ Column temp. : 0 C Effects of sodium nitrate in eluent on the analysis of polyallylae Gelatin For the analysis of cationic polymers, such as pollyallylae, undesired adsorption of the polymer is observed when low (0.0M) sodium nitrate eluent was used. By using higher concentration (> 0.M) salt, it suppresses the sample adsorption and enables to obtain accurate chromatograms. Sample : Polyallylae 0.%, 00µL 0. M 0.0 M Sample : 0.% each, 00µL Gelatin from bovine skin (Acid treatment, Gel strength : g) Gelatin from porcine skin (Alkali treatment, Gel strength : 90-00g) 0. M From bovine skin Mn :,00 Mw : 0,00 Mw/Mn : M From porcine skin Mn :,00 Mw :,00 Mw/Mn : Volume (ml) * Molecular weight was detered from the calibration curve of pullulan. Column : Shodex OHpak SB-0M HQ x Eluent : 0.M Acetic acid + NaNO aq. Flow rate :.0mL/ Column temp. : 0 C Column : Shodex OHpak SB-0M HQ x Eluent : 0.M KH PO aq./ 0.M Na HPO aq.=0/0 Flow rate :.0mL/ Column temp. : 0 C Polyacrylamide Cellulose acetate SB-0 HQ Sample : Polyacrylamide, 00µL Sample : Cellulose acetate 0.% each, 00µL.0x0 SB-0 HQ Mn~0,000 Molar Mass (g/mol).0x0.0x0 Mw:,00,000 Mn~0, x Volume (ml) 0 Column : Shodex OHpak SB-0 HQ, SB-0 HQ Eluent : 0.M NaCl aq. Flow rate : 0.mL/ MALS (Multi angle light scattering) Column temp. : 0 C Column : Shodex OHpak SB-0M HQ x Eluent : 0mM LiBr in DMF Flow rate :.0mL/ Column temp. : 0 C

43 Copovidones Sample : 00µL Poly(-vinylpyrrolidone-co-vinyl acetate) 0.% each Copolymer : Mn :,000 Mw :,00 Mw/Mn :.0 Copolymer : Mn :,00 Mw :,900 Mw/Mn : * Molecular weight was detered from Elution volume (ml) the calibration curve of PEG/PEO. Column : Shodex OHpak SB-0M HQ x Eluent : 0mM LiBr in DMF Flow rate :.0mL/ Column temp. : 0 C Calibration curves for LB-00 series using pullulan (eluent : H O) Molecular weight (Pullulan) LB-0 LB-0M Column : Shodex OHpak LB-00 series Eluent : H O Flow rate :.0mL/ Column temp. : 0 C Calibration curves for LB-00 series using PEG/PEO (eluent : DMF) Molecular weight (PEG/PEO) LB-0 LB-0M 9 0 Elution volume (ml) Column : Shodex OHpak LB-00 series Eluent : DMF Flow rate :.0mL/ Column temp. : 0 C Aqueous SEC (GFC) Columns : Polymer-based Comparison of the detection of pullulan using multiangle light scattering detector The multiangle light scattering (MALS) signal of low molecular weight substances is low with competitors columns due to the baseline noise. New product OHpak LB-00 series enables the detection by improving the baseline noise level. LB-0M (.0mm I.D. x 00mm) Sample : Pullulan (MW :,000) 0.%, 0µL 0 00 LB-0M (.0mm I.D. x 00mm) SB-0M HQ (.0mm I.D. x 00mm) SEC column from other manufacturer (.mm I.D. x 00mm) SB-0M HQ (.0mm I.D. x 00mm) Signal to noise ratio (S/N) SEC column from other manufacturer (.mm I.D. x 00mm) 0,0,00,000 0,00 Molecular weight (Pullulan) 0 0 Column : Shodex OHpak LB-0M Shodex OHpak SB-0M HQ SEC column from other manufacturer Eluent : 0.M NaNO aq. Flow rate :.0mL/ Detector : MALS (Multi angle light scattering) (90 ) Column temp. : 0 C Sodium alginate Sodium heparin Sample : Sodium alginate 0.%, 00µL Sample : Sodium heparin 0.%, 00µL.0x0.0x0.0x0.0x0 Molar Mass (g/mol).0x0.0x0.0x Molar Mass (g/mol).0x0.0x0.0x Mn :,90 Mw :,00 Mw/Mn : Mn :,00 Mw :,0 Mw/Mn :, Time () Time () Column : Shodex OHpak LB-0M x Eluent : 0.M NaNO aq. Flow rate :.0mL/ MALS (Multi angle light scattering) Column temp. : 0 C Column : Shodex OHpak LB-0M x Eluent : 0.M NaNO aq. Flow rate :.0mL/ MALS (Multi angle light scattering) Column temp. : 0 C

44 Multimode Columns Features GS-HQ SEC is the main separation mode Depends on the eluent selected, the column adds multimode features of reversed phase, HILIC, and ion exchange modes to SEC Suitable for the separation of peptides or nucleic acids with similar molecular weights Suitable for desalting samples or substituting buffer in protein analysis Standard columns Shipping Solvent F0000 Asahipak GS-0 HQ 9, H O/CH OH=0/0 F0000 Asahipak GS-0 HQ 9, H O/CH OH=0/0 F0000 Asahipak GS-0 HQ,000, H O/CH OH=0/0 F0000 Asahipak GS-0 HQ,000, H O/CH OH=0/0 F009 Asahipak GS-G B 9. 0 H O/CH OH=0/0 Semi-micro columns * The following semi-micro columns are made to order. Base Material : Polyvinyl alcohol Usable ph range : ph- (GS-0 HQ : ph-9) Usable concentration of methanol is up to 00% (GS-0 HQ : up to 0%) Usable concentration of acetonitrile is up to 0% F0 GS0A-D F0 GS0A-E F0 GS0A-B, Preparative columns * Preparative columns are made to order. Standard Column F00 Asahipak GS-0 0F, GS-0 HQ F00 Asahipak GS-0 0F, GS-0 HQ F009 Asahipak GS-0 0F, GS-0 HQ F000 Asahipak GS-0 0F, GS-0 HQ F00 Asahipak GS-0 0G, GS-0 HQ F00 Asahipak GS-0 0G, GS-0 HQ F00 Asahipak GS-0 0G, GS-0 HQ F00 Asahipak GS-0 0G, GS-0 HQ F00 Asahipak GS-0G B 0. 0 Target molecular weight range and Exclusion limit With Pullulan (eluent : ultrapure water) Target Molecular Weight Range Exclusion Limit GS-0 GS-0 00, ,000,000 0,000 GS-0 GS-0,000 00,000 0,000 00,000 00,000,000,000 * Please use the above table as a rough indication for the column selection.

45 Calibration curves for GS-HQ series using pullulan (eluent : H O) Molecular weight (Pullulan) GS-0 HQ GS-0 HQ GS-0 HQ GS-0 HQ Peptides GS-HQ columns can be used not only for SEC (GFC) in an aqueous system, but also for multimodal analysis where hydrophobic interaction and ionic interaction are used together as separation criteria, under certain conditions of the eluent. This results in unprecedented separation analysis. GS-0 HQ columns are excellent in the performance for separating hydrophilic peptides, in particular, acidic or basic peptides, from each other. Glu-Ala-Glu Arg-Asp Gly-His-Lys Arg-Pro-Lys-Pro MW pl Σf Sample : 0µL. Glu-Ala-Glu 0.0%. Arg-Asp 0.0%. Gly-His-Lys 0.0%. Arg-Pro-Lys-Pro 0.0% Multimode Columns 0 Σf : Hydrophobic parameter pl : Isoelectric point 9 0 Elution volume (ml) Column : Shodex Asahipak GS-HQ series Eluent : H O Flow rate : 0.mL/ Column temp. : 0 C Column : Shodex Asahipak GS-0 HQ Eluent : 0mM Ammonium acetate buffer (ph.) Flow rate : 0.mL/ Detector : UV (0nm) Column temp. : 0 C Analysis of purine bases in beer Purine in foods is analyzed as purine base after a step of sample preparation; homogenization, freeze drying, hydolyzation with 0% perchloric acid, and neutralization. Example below shows the analysis of purin in regular beer and beer treated with guanase (an enzyme that degrades guanine to xanthine). The following data indicate that guanine was decreased and xanthine was increased by guanase. Purine bases in beer Normal beer Guanase treated beer adenine guanine guanine hypoxanthine xanthine xanthine uric acid xanthine Column : Shodex Asahipak GS-0 HQ Eluent : 0mM Sodium phosphate buffer (ph.) Flow rate : 0.mL/ Detector : UV (0nm) Column temp. : C Data provided by Kiyoko Kaneko Ph.D., Faculty of Pharmaceutical Sciences, Teikyo University Low molecular weight water-soluble dietary fiber Umami Lignosulfonic acid By using GS-0 HQ, monosaccharides, disaccharides, and sugar alcohols elute after indigestible component faction (indicated by an arrow on the chromatogram). This separation makes the method preferable for the quantification of low molecular weight water-soluble dietary fiber. Sample : 0µg/mL each, 0µL. IMP. GMP. AMP. Inosine. Hypoxanthine. Guanosine. Guanine. Adenosine 9. Adenine Sample : 00µL Lignosulfonic acid sodium salt 0.% Column : Shodex Asahipak GS-0 HQ x Eluent : H O Flow rate : 0.mL/ Column temp. : 0 C Column : Shodex Asahipak GS-0 HQ Eluent : 0mM NaH PO aq./0mm Na HPO aq. =000/ Flow rate :.0mL/ Detector : UV (0nm) Column temp. : 0 C Column : Shodex Asahipak GS-0 HQ x Eluent : 0mM Na HPO aq. Flow rate : 0.mL/ Detector : UV (nm) Column temp. : 0 C

46 Aqueous/Organic SEC Columns Features GF-HQ Polymer-based SEC columns exhibit high solvent durability Supports both aqueous and organic solvents Standard columns Shipping Solvent F00000 Asahipak GF-0 HQ 9, H O F0000 Asahipak GF-0 HQ 9, H O/CH OH=0/0 F0000 Asahipak GF-0 HQ 9,000, H O/CH OH=0/0 F0000 Asahipak GF-0 HQ, , H O/CH OH=0/0 F0000 Asahipak GF-M HQ, , H O/CH OH=0/0 F00 Asahipak GF-G B 9. 0 H O/CH OH=0/0 F0000 MSpak GF-0 B, H O F000 MSpak GF-0 D 0, H O F000 MSpak GF-0 E, H O F000 MSpak GF-0 D, H O GF-M HQ is a mixed-gel column capable of analyzing samples over a wide range of molecular weight. Base Material : Polyvinyl alcohol Usable ph range : ph-9 Semi-micro columns * The following semi-micro columns are made to order. F0000 Asahipak GF-0 D 0. 0 F000 Asahipak GF-0 E 0. 0 F0 GF0A-D, F0 GF0A-E, F0 GF0A-D 9 0, Preparative columns * Preparative columns are made to order. Standard Columns F000 Asahipak GS-0 0F, GF-0 HQ F00 Asahipak GS-0 0F, GF-0 HQ F00 Asahipak GS-0 0F, GF-0 HQ F00 Asahipak GSM-00 0F, GF-M HQ F00 Asahipak GS-0 0G, GF-0 HQ F009 Asahipak GS-0 0G, GF-0 HQ F000 Asahipak GS-0 0G, GF-0 HQ F00 Asahipak GSM-00 0G, GF-M HQ F000 Asahipak GS-0G B 0. 0 Target molecular weight range and Exclusion limit With Pullulan (eluent : ultrapure water) With *PEG/PEO (eluent : DMF) Target Molecular Weight Range GF-0 00,000 GF ,000 GF-0,000 00,000 GF-0 00,000 *(0,000,000) GF-M 00 *(0,000,000) * Please use the above table as a rough indication for the column selection. Exclusion Limit 9,000 0,000 00,000 *(0,000,000) *(0,000,000) GF-0 GF-0 GF-0 GF-0 GF-M Target Molecular Weight Range 00,000 00,000,000 00,000 0,000 **(0,000,000) 00 **(0,000,000) *( ) Estimated value * Please use the above *PEG : polyethylene glycol table as a rough indication *PEO : polyethylene oxide for the column selection. **( ) Estimated value

47 Usable solvents Solvent Water 0-0.M sodium concentration Methanol Ethanol Acetonitrile THF DMF GF-0 GF-0 GF-0 GF-0 GF-M * When replacing acetonitrile, ethyl acetate or chloroform with water, replace with methanol first and then replace with water. * When replacing water with ethyl acetate or chloroform, replace with methanol first and then replace with the required eluent condition. (Note) The usable solvents for preparative columns of GF-0 HQ, GS-0 0F and 0G, are water and methanol. GSM-00 0F or 0G is recommended when other solvents are required. Sodium polystyrene sulfonates MW:0, MW:, Polymers having both hydrophobic and hydrophilic functional groups may exhibit hydrophobic interactions with packing materials. When analyzing such polymers, the hydrophobic interaction can be suppressed by adding organic solvents to the eluent. Sample : mg/ml each, 0µL Sodiumn polystyrene sulfonate CH CH SONa n Aqueous/Organic SEC Columns Acetone Chloroform Ethylacetate DMSO 0~0% 0 MW:, Column : Shodex Asahipak GF-0 HQ Eluent : 0mM LiCl aq./ch CN =0/0 Flow rate : 0.mL/ Detector : UV (nm) Column temp. : 0 C Calibration curves for GF-HQ series using pullulan (eluent : H O) Calibration curves for GF-HQ series using PEG/PEO (eluent : DMF) Polyacrylonitrile 0 0 Sample : Polyacrylonitrile 0.%, 00µL 0 0 Molecular weight (Pullulan) GF-0 HQ GF-0 HQ GF-0 HQ GF-0 HQ GF-M HQ Molecular weight (PEG/PEO) GF-0 HQ GF-0 HQ GF-0 HQ GF-M HQ GF-0 HQ Elution volume (ml) 9 0 Elution volume (ml) Column : Shodex Asahipak GF-HQ series Eluent : H O Flow rate : 0.mL/ Column temp. : 0 C Column : Shodex Asahipak GF-HQ series Eluent : DMF Flow rate : 0.mL/ Column temp. : 0 C Column : Shodex Asahipak GF-0 HQ x Eluent : 0mM LiBr in DMF Flow rate : 0.mL/ Column temp. : 0 C Comparison of two GF columns for the separation performance of maltooligosaccharides Comparison of polystyrenes separation using various solvents GF-0 HQ demonstrates an improved separation of low molecular substances. The sample below shows that the peaks obtained by GF-0 HQ are separated with deeper notches compared to peaks obtained by GF-0 HQ. GF-0 HQ is capable of separating oligosaccharides (trisaccharides to hexasaccharides) while those oligosaccharides were eluted all together when analyzed by GF-0 HQ. GF-0 HQ x GF-0 HQ x ~ Sample : 0.% each, 0µL. Maltohexaose. Maltopentaose. Maltotetraose. Maltotriose. Maltose. Glucose (A) THF Sample : Polystyrene mg/ml each, 0µL. Mw :,090,000. Mw : 90,000. Mw :,900. Mw : 9,00. Mw :,90. Mw : 00 (B) Chloroform (C) DMF 0 0 Column : Shodex Asahipak GF-0 HQ x Shodex Asahipak GF-0 HQ x Eluent : H O Flow rate : 0.mL/ Column temp. : 0 C Column : Shodex Asahipak GF-0 HQ + GF-0 HQ Eluent : (A); THF, (B); Chloroform, (C); DMF Flow rate : 0.mL/ Detector : (A),(B) UV (nm), (C) UV (0nm) Column temp. : 0 C

48 Organic SEC (GPC) Columns (General Analysis) : THF Features KF-00 Standard organic solvent SEC (GPC) column Supports a wide range of applications from low to high molecular weight compounds Corresponds to USP L Standard columns KF-00 series : Shipping solvent Tetrahydrofuran (THF) F000 GPC KF-0, F000 GPC KF-0, F00 GPC KF-0., F000 GPC KF-0, F000 GPC KF-0L, F000 GPC KF-0,000, F000 GPC KF-0L,000, F000 GPC KF-0,000 0, F000 GPC KF-0L,000 0, F000 GPC KF-0, , F0090 GPC KF-0M, , F000 GPC KF-0L, , F000 GPC KF-0,000 0, F000 GPC KF-0L,000 0, F0000 GPC KF-G A (GPC KF-G). 0 F090 GPC KF-00D solvent-peak separation column * The columns with L or M at the end of column names are mixed-gel column capable of analyzing samples over a wide range of molecular weight distribution. * See page 0 for details of the solvent-peak separation columns. * See pages and for details preparative columns. * See page for applicability of SEC (GPC) columns to solvent replacement. Base Material : Styrene divinylbenzene copolymer Target molecular weight range and Exclusion limit With Polystyrene (eluent : THF) Target Molecular Weight Range Exclusion Limit Target Molecular Weight Range Exclusion Limit KF ,00 KF-0 0,000,000,000,000,000 KF-0 00,000,000 KF-0L 00,000,000,000,000 KF-0. 00,000 0,000 KF-0 0,000 *(0,000,000) *(0,000,000) KF-0,000 0,000 0,000 KF-0M,000 *(0,000,000) *(0,000,000) KF-0L 00 0,000 0,000 KF-0L 00 *(0,000,000) *(0,000,000) KF-0,000 00,000 00,000 KF-0 00,000 *(00,000,000) *(00,000,000) KF-0L 00 00,000 00,000 KF-0L 00 *(00,000,000) *(00,000,000) * Please use the above table as a rough indication for the column selection. *( ) Estimated value

49 Calibration curves for KF-00 series using polystyrene Molecular weight (Polystyrene) KF-0 KF-0 KF-0 KF-0 KF-0 KF-0. KF-0 KF-0 KF-0M 9 0 Elution volume (ml) Column : Shodex GPC KF-00 series Eluent : THF Flow rate :.0mL/ Column temp. : 0 C Calibration curves for KF-00L (linear type) series using polystyrene Molecular weight (Polystyrene) KF-0L KF-0L KF-0L KF-0L KF-0L 9 0 Elution volume (ml) Column : Shodex GPC KF-00L series Eluent : THF Flow rate :.0mL/ Column temp. : 0 C Fat-soluble vitas Sample : 00µL. Vita A palmitate 00µg/mL. Vita E 0µg/mL. Vita D 0µg/mL. Vita K 0µg/mL 0 0 Column : Shodex GPC KF-0 x Eluent : THF Flow rate :.0mL/ Detector : UV (0nm) Column temp. : 0 C Organic SEC (GPC) Columns (General Analysis) : THF Standard polystyrenes Sample : 0µL. Polystyrene (Mw :,000) 0.%. Polystyrene (Mw :,00) 0.%. Polystyrene (Mw : 9,00) 0.%. Polystyrene (Mw :,0) 0.%. Polystyrene (Mw :,00) 0.%. Polystyrene (Mw : 0) 0.%. Toluene (Mw: 9) 0.% Sample : 0µL. Polystyrene (Mw : 9,,000) 0.0%. Polystyrene (Mw :,0,000) 0.%. Polystyrene (Mw :,000) 0.%. Polystyrene (Mw :,0) 0.%. Polystyrene (Mw : 0) 0.%. Toluene (Mw : 9) 0.% Column : Shodex GPC KF-0L x Eluent : THF Flow rate :.0mL/ Detector : UV (nm) Column temp. : 0 C Column : Shodex GPC KF-0L x Eluent : THF Flow rate :.0mL/ Detector : UV (nm) Column temp. : 0 C Styrene isoprene ABA block copolymer Polycarbonate resin Raw rubber Sample : Styrene isoprene ABA block copolymer Sample : Polycarbonate resin 0.%, 00µL Sample : Rubber 0.%, 00µL CH O C O C CH O n Column : Shodex GPC KF-0M x Eluent : THF Flow rate :.0mL/ Column temp. : 0 C Column : Shodex GPC KF-0L x Eluent : THF Flow rate :.0mL/ Column temp. : 0 C Column : Shodex GPC KF-0M x + KF-0 Eluent : Toluene Flow rate :.0mL/ Column temp. : Room temp.

50 Organic SEC (GPC) Columns (General Analysis) : Chloroform Features K-00 Standard organic solvent SEC (GPC) column Supports a wide range of applications from low to high molecular weight compounds Corresponds to USP L Standard columns K-00 series : Shipping solvent Chloroform F00 GPC K-0, F00 GPC K-0, F0 GPC K-0., F00 GPC K-0, F09 GPC K-0L, F00 GPC K-0,000, F09 GPC K-0L,000, F00 GPC K-0,000 0, F09 GPC K-0L,000 0, F00 GPC K-0, , F090 GPC K-0M, , F09 GPC K-0L, , F00 GPC K-0,000 0, F09 GPC K-0L,000 0, F000 GPC K-G A (GPC K-G). 0 F090 GPC K-00D solvent-peak separation column * The columns with L or M at the end of column names are mixed-gel column capable of analyzing samples over a wide range of molecular weight distribution. * See page 0 for details of the solvent-peak separation columns. * See pages and for details preparative columns. * See page for applicability of SEC (GPC) columns to solvent replacement. Base Material : Styrene divinylbenzene copolymer Target molecular weight range and Exclusion limit With Polystyrene (eluent : Chloroform) Target Molecular Weight Range Exclusion Limit Target Molecular Weight Range Exclusion Limit K ,00 K-0 0,000,000,000,000,000 K-0 00,000,000 K-0L 00,000,000,000,000 K-0. 00,000 0,000 K-0 0,000 *(0,000,000) *(0,000,000) K-0,000 0,000 0,000 K-0M,000 *(0,000,000) *(0,000,000) K-0L 00 0,000 0,000 K-0L 00 *(0,000,000) *(0,000,000) K-0,000 00,000 00,000 K-0 00,000 *(00,000,000) *(00,000,000) K-0L 00 00,000 00,000 K-0L 00 *(00,000,000) *(00,000,000) * Please use the above table as a rough indication for the column selection. *( ) Estimated value

51 Epoxy resin Sample : Epikote TM 00 0.%, 00µL 0 0 Column : Shodex GPC K-0L x Eluent : Chloroform Flow rate :.0mL/ Detector : UV (nm) Column temp. : Room temp. Silicone oil Low molecular polydimethylsiloxanes Sample : 0.% each, 00µL. Polydimethylsiloxane (MW:0). Polydimethylsiloxane (MW:). Hexamethyldisiloxane (MW:) 0 0 Column : Shodex GPC K-0 x Eluent : Chloroform Flow rate :.0mL/ (polarity : -) Column temp. : 0 C 0 Bionolle TM (Polybutylene succinate/adipate) Sample : Silicone oil 0.%, 00µL Sample : Polybutylene succinate/adipate 0.%, 00µL Organic SEC (GPC) Columns (General Analysis) : Chloroform Silicone oil Column : Shodex GPC K-0M x Eluent : Toluene Flow rate :.0mL/ Column temp. : C Column : Shodex GPC K-0M x + K-0 Eluent : Chloroform Flow rate :.0mL/ Column temp. : 0 C Recycling fractionation of lauryl stearyl thiodipropionate Sample : %, 00µL. Distearyl stearyl thiodipropionate. Lauryl stearyl thiodipropionate. Dilauryl thiodipropionate CYCLE CYCLE CYCLE CYCLE0 Column : Shodex GPC K-LG + K-00 Eluent : Chloroform Flow rate :.0mL/ (preparative type) Column temp. : 0 C * See page for K-00 9

52 Organic SEC (GPC) Columns (General Analysis) : DMF Features KD-00 Standard organic solvent SEC (GPC) column Supports a wide range of applications from low to high molecular weight compounds Corresponds to USP L Standard columns KD-00 series : Shipping solvent Dimethylformamide (DMF) F00 GPC KD-0, F00 GPC KD-0, F0 GPC KD-0., F00 GPC KD-0, F00 GPC KD-0,000, F00 GPC KD-0,000 0, F00 GPC KD-0, , F090 GPC KD-0M, , F00 GPC KD-0,000 0, F00 GPC KD-G A (GPC KD-G). 0 * The columns with L or M at the end of column names are mixed-gel column capable of analyzing samples over a wide range of molecular weight distribution. * See page for applicability of SEC (GPC) columns to solvent replacement. Base Material : Styrene divinylbenzene copolymer Target molecular weight range and Exclusion limit With *PEG/PEO (eluent : DMF) Target Molecular Weight Range Exclusion Limit Target Molecular Weight Range Exclusion Limit KD-0 KD-0 KD-0. KD-0 KD-0 Features KF-00D K-00D 00,00 00, ,000,000 0,000,000 00,000,00,000 0,000 0,000 00,000 KD-0 KD-0 KD-0M KD-0 0,000 **(,000,000) 0,000 **(0,000,000),000 **(0,000,000) 0,000 **(00,000,000) * Please use the above table as a rough indication for the column selection. Solvent-peak Separation Columns for Organic SEC (GPC) **(,000,000) **(0,000,000) **(0,000,000) **(00,000,000) *PEG : polyethylene glycol *PEO : polyethylene oxide **( ) Estimated value Use this column in combination with a linear column By shifting the elution of monomers, polymer additives, and the solvent-peak in low molecular region, it reduces interferences in the calculation of the molecular weight distribution of polymers and oligomers Solvent-peak separation columns Column Combination Shipping Solvent F090 GPC KF-00D KF-0L, 0L, 0M, 0L THF F090 GPC K-00D K-0L, 0L, 0M, 0L Chloroform 0 Base Material : Styrene divinylbenzene copolymer

53 Calibration curves for KD-00 series using PEG/PEO Molecular weight (PEG/PEO) Molecular mass Celluloses 0 KD-0 KD Elution volume (ml) 9 0 Elution volume (ml) Column : Shodex GPC KD-0M Eluent : % LiCl in *DMI Flow rate : 0.mL/, MALS (Multi angle light scattering) Column temp. : 0 C * DMI,-dimethyl--imidazolidinone Potato starch KD-0 KD-0 Sample : 00µL Potato starch in DMSO 0.% * solved at 0 C KD-0 KD-0 KD-0M KD-0. KD-0 Column : Shodex GPC KD-00 series Eluent : DMF Flow rate :.0mL/ Column temp. : 0 C Polyvinylpyrrolidones Sample : Polyvinylpyrrolidone 0.% each (A) K-0 00µL (B) K-0 00µL (C) K-90 00µL (A) (B) (C) Column : Shodex GPC KD-0M x Eluent : 0mM LiBr in DMF Flow rate :.0mL/ Column temp. : 0 C Sample : 00µL Cellulose ca. 0.0% each Cellulose is known to be difficult to dissolve. A cellulose solution is prepared by repeating solvent replacement. It is reported that it requires a long time to dissolve ( to 0 days), depending on the solvent type, crystallinity and molecular weight. Data provided by Dr. Masahiko Yanagisawa, Isogai group, Graduate School of Agricultural and Life Sciences, The University of Tokyo Effects of solvent-peak separation column Sample : Poly(vinyl chloride) DOP (Dioctyl phthalate) Melae formaldehyde resin 0 Phenoxy resin Sample : 0µL Melae-formaldehyde resin % 0 Column : Shodex GPC KD-0 x Eluent : 0mM LiBr in DMF Flow rate :.0mL/ Column temp. : 0 C Sample : 0µL Phenoxy resin 0.% 0 Column : Shodex GPC KD-0M x Eluent : 0mM LiBr in DMF Flow rate :.0mL/ Column temp. : 0 C Organic SEC (GPC) Columns (General Analysis) : DMF Solvent-peak Separation Columns for Organic SEC (GPC) KF-0L 0 0 Molecular weight (Polystyrene) KF-0L KF-0L+KF-00D KF-0L + KF-00D Elution volume (ml) Column : Shodex GPC KD-0M Eluent : 0mM LiBr in DMSO/DMF=/ Flow rate :.0mL/ Column temp. : 0 C 0 0 Column : Shodex GPC KF-0L Shodex GPC KF-0L + KF-00D Eluent : THF Flow rate :.0mL/

54 Organic SEC (GPC) Columns : Rapid Analysis, High Performance Analysis Features KF-00 KF-00HQ Approximately half of the analysis time compared with standard columns The amount of solvent used is reduced to about a third Improved applicability of solvent replacement Corresponds to USP L About. times better separation performance than standard columns, obtains higher resolution About times better sensitivity than that of standard columns, supports high sensitivity analysis The amount of solvent used is reduced to about a third Improved applicability of solvent replacement Corresponds to USP L Rapid analysis downsized columns KF-00 series F009 F009 F009 F009 F009 F009 F009 F009 F0099 F0000 GPC KF-0 GPC KF-0 GPC KF-0. GPC KF-0 GPC KF-0 GPC KF-0 GPC KF-0 GPC KF-0M GPC KF-0 GPC KF-G A (GPC KF-G),000,000,000,000,000,000,000,000,000 Use of the KF-00 series with semi-micro type devices is recommended ,00,000 0,000 0,000 0, High performance semi-micro columns KF-00HQ series F0 F0 F0 F0 F0 F09 F0 F0000 GPC KF-0HQ GPC KF-0HQ GPC KF-0.HQ GPC KF-0HQ GPC KF-0HQ GPC KF-0LHQ GPC KF-0LHQ GPC KF-G A (GPC KF-G) Use of the KF-00HQ series with semi-micro type devices is recommended.,000,000,000,000,000 0,000 0, ,00,000 0, [KF-00 series and KF-00HQ series] * The columns with L or M at the end of column names are mixed-gel column capable of analyzing samples over a wide range of molecular weight distribution. * See page for applicability of SEC (GPC) columns to solvent replacement. [KF-00 series and KF-00HQ series] Base Material : Styrene divinylbenzene copolymer Shipping Solvent : Tetrahydrofuran (THF) Target molecular weight range and Exclusion limit With Polystyrene (eluent : THF) Target Molecular Weight Range Exclusion Limit Target Molecular Weight Range Exclusion Limit KF ,00 KF-0HQ 00 00,00 KF-0 00,00,000 KF-0HQ 00,00,000 KF ,000 0,000 KF-0.HQ 00 0,000 0,000 KF ,000 0,000 KF-0HQ 00 0,000 0,000 KF-0,000 00,000,000,000 KF-0HQ,000 00,000,000,000 KF-0 0,000,000,000,000,000 KF-0LHQ 00,000,000,000,000 KF-0 0,000 *(0,000,000) *(0,000,000) KF-0LHQ 00 *(0,000,000) *(0,000,000) KF-0M KF-0,000 *(0,000,000) 00,000 *(00,000,000) *(0,000,000) *(00,000,000) * Please use the above table as a rough indication for the column selection. *( ) Estimated value

55 Calibration curves for KF-00 series using polystyrene Molecular weight (Polystyrene) Elution volume (ml) Column : Shodex GPC KF-00 series Eluent : THF Flow rate : 0.mL/ (small cell volume) Column temp. : 0 C Comparison of standard, rapid analysis, and high performance type columns Standard type KF-0. x 0µL injection KF-0 KF-0 KF-0. KF-0 KF-0 KF-0 KF-0 KF-0 KF-0M ml Calibration curves for KF-00HQ series using polystyrene Molecular weight (Polystyrene) KF-0HQ KF-0HQ KF-0LHQ KF-0HQ KF-0.HQ KF-0HQ KF-0LHQ Elution volume (ml) Column : Shodex GPC KF-00HQ series Eluent : THF Flow rate : 0.mL/ (small cell volume) Column temp. : 0 C Sample : EPON00 0.% By using KF-0., the analysis time is reduced to less than half of that using KF-0.. Thus KF-00 series enables rapid analysis. On the other hand, KF-0.HQ has a theoretical plate number. times more than that of the standard column, thereby improving resolution especially in the analysis of molecules that have a small to medium molecular weight. Rapid analysis and high performance type columns use less than one third of solvent per analysis compared to standard type columns do. Organic SEC (GPC) Columns : Rapid Analysis, High Performance Analysis High performance type.ml KF-0.HQ x 0µL injection Rapid analysis type KF-0. x 0µL injection.0ml Column : Shodex GPC KF-0. x Shodex GPC KF-0.HQ x Shodex GPC KF-0. x Eluent : THF Flow rate :.0mL/ (KF-0.) 0.mL/ (KF-0.HQ) 0.mL/ (KF-0.) (conventional type) (KF-0.) RI (small cell volume) (KF-0.HQ, KF-0.) Column temp. : 0 C Styrene acrylonitrile copolymer Liquid paraffin Phthalates Sample : Styrene-Acrylonitrile (0:0) copolymer Sample : Liquid paraffin %, µl Sample : 0µL. Dioctyl phthalate 0.%. Dibutyl phthalate 0.%. Dipropyl phthalate 0.%. Diethyl phthalate 0.%. Dimethyl phthalate 0.%. Propylbenzene 0.%. Ethylbenzene 0.%. Toluene 0.% 9. Benzene 0.% Column : Shodex GPC KF-0M x Eluent : 0mM LiBr in DMF Flow rate : 0.mL/ (small cell volume) Column temp. : 0 C Column : Shodex GPC KF-0HQ Eluent : Chloroform Flow rate : 0.mL/ (small cell volume) Column temp. : 0 C Column : Shodex GPC KF-0HQ x Eluent : THF Flow rate : 0.mL/ Detector : UV (nm) (small cell volume) Column temp. : 0 C

56 Organic SEC (GPC) Columns : For Preparative Preparative columns * Preparative columns are made to order. KF-000 series : Shipping solvent Tetrahydrofuran (THF) Standard Columns F00 GPC KF-00, KF-0 F00 GPC KF-00, KF-0 F0 GPC KF-00., KF-0. F00 GPC KF-00, KF-0 F00 GPC KF-00, KF-0 F00 GPC KF-00 0, KF-0 F00 GPC KF-00 0, KF-0 F009 GPC KF-00M 0, KF-0M F000 GPC KF-G B (GPC KF-LG).0 0 * See page for GPC KF-00 series. Base Material : Styrene divinylbenzene copolymer K-000 series : Shipping solvent Chloroform Standard Columns F00 GPC K-00, K-0 F0 GPC K-00, K-0 F0 GPC K-00., K-0. F00 GPC K-00, K-0 F00 GPC K-00, K-0 F00 GPC K-00 0, K-0 F00 GPC K-00 0, K-0 F009 GPC K-00M 0, K-0M F000 GPC K-G B (GPC K-LG).0 0 * See page for GPC K-00 series. Base Material : Styrene divinylbenzene copolymer

57 Preparative columns * Preparative columns are made to order. H-000 series : Shipping solvent Chloroform F000 F000 F00 F000 F000 F000 GPC H-00 GPC H-00 GPC H-00. GPC H-00 GPC H-00 GPC H-00,000,000,000,000,000, Standard Columns K-0 K-0 K-0. K-0 K-0 K-0 Organic SEC (GPC) Columns : For Preparative F000 GPC H-00, K-0 F0009 GPC H-00M, K-0M F000 GPC H-G B (GPC H-G).0 0 * See page for GPC K-00 series. Base Material : Styrene divinylbenzene copolymer KF-000 series : Shipping solvent Tetrahydrofuran (THF) [Customized columns] Standard Columns F000 GPC KF KF-0 F000 GPC KF KF-0 F00 GPC KF KF-0. F000 GPC KF KF-0 F000 GPC KF KF-0 F000 GPC KF-G 0C (GPC KF-LLG) * See page for GPC KF-00 series. Base Material : Styrene divinylbenzene copolymer K-000 series : Shipping solvent Chloroform [Customized columns] Standard Columns F0900 GPC K K-0 F0900 GPC K K-0 F090 GPC K K-0. F0900 GPC K K-0 F0900 GPC K K-0 F0009 GPC K-G 0C (GPC K-LLG) * See page for GPC K-00 series. Base Material : Styrene divinylbenzene copolymer

58 Organic SEC (GPC) Columns : Linear Calibration Type Features LF Employs a special packing material with a wide pore size distribution (multi pore type) Highly linear calibration curve without inflection points Molecular weight distribution can be detered with high precision Enables analysis over a broad range of molecular weights Column for rapid analysis (LF-0) and column for high performance analysis (LF-0) enabling reduction in solvent use are also available Corresponds to USP L Standard columns F00 GPC LF-0,000, F09 GPC LF-G. 0 * See page for applicability of SEC (GPC) columns to solvent replacement. Rapid analysis downsized column Base Material : Styrene divinylbenzene copolymer Shipping Solvent : Tetrahydrofuran (THF) Use of the LF-0 with semi-micro type devices is recommended. F00 GPC LF-0 9,000, F09 GPC LF-G. 0 * See page for applicability of SEC (GPC) columns to solvent replacement. High performance semi-micro column Base Material : Styrene divinylbenzene copolymer Shipping Solvent : Tetrahydrofuran (THF) Use of the LF-0 with semi-micro type devices is recommended. F00 GPC LF-0,000, F09 GPC LF-G. 0 * See page for applicability of SEC (GPC) columns to solvent replacement. Target molecular weight range and Exclusion limit With Polystyrene (eluent : THF) Base Material : Styrene divinylbenzene copolymer Shipping Solvent : Tetrahydrofuran (THF) LF-0 LF-0 Target Molecular Weight Range 00,000,000 00,000,000 Exclusion Limit,000,000,000,000 LF-0 00,000,000,000,000 * Please use the above table as a rough indication for the column selection. Schematic diagram of linear calibration type packing Connecting linear calibration type columns (LF series) The linear calibration type column covers a broad range of molecular weights with only one kind of packing (column). Connecting mixed-gel columns (KF-0L, etc.) Connecting different grades columns (KF-0 + KF-0 + KF-0, etc.)

59 Calibration curve for LF-0 using polystyrene Molecular weight (Polystyrene) Xylan 9 0 Elution volume (ml) Column : Shodex GPC LF-0 Eluent : THF Flow rate :.0mL/ Column temp. : 0 C Calibration curves for LF-0 and LF-0 using polystyrene Molecular weight (Polystyrene) LF-0 LF Elution volume (ml) Column : Shodex GPC LF-0, LF-0 Eluent : THF Flow rate : 0.mL/ (LF-0) 0.mL/ (LF-0) (small cell volume) Column temp. : 0 C Polyamide (Nylon/) Polyurethane Sample : Polyurethane 0.%, 0µL Column : Shodex GPC LF-0 x Eluent : THF Flow rate : 0.mL/ (small cell volume) Column temp. : 0 C Polymethyl methacrylate Organic SEC (GPC) Columns : Linear Calibration Type Sample : Xylan 0.%, 00µL Sample : Nylon TM / 0.%, 0µL Sample : 00µL Polymethyl methacrylate Column : Shodex GPC LF-0 Eluent : 0mM H PO + 0mM LiBr in (DMSO/DMF=0/0) Flow rate : 0.mL/ Column temp. : 0 C Column : Shodex GPC LF-0 Eluent : mm CF COONa in HFIP Flow rate : 0.mL/ (small cell volume) Column temp. : 0 C Column : Shodex GPC LF-0 x Eluent : Methyl ethyl ketone Flow rate :.0mL/ Column temp. : 0 C Polyamic acid Sample : Poly(pyromellitic dianhydride-co-,'-oxydianiline), 00µL Comparison of polystyrenes separation with different numbers of LF-0 LF-0 x Sample : 0µL. Polystyrene (Mw :,00,000). Polystyrene (Mw :,000). Polystyrene (Mw :,000). Polystyrene (Mw :,000). Polystyrene (Mw :,00). Polystyrene (Mw : 0). Ethylbenzene LF-0 x Column : Shodex GPC LF-0 x n Eluent : THF Flow rate : 0.mL/ (small cell volume) Column temp. : 0 C Column : Shodex GPC LF-0 x Eluent : 0mM LiBr + 0mM H PO in NMP Flow rate : 0.mL/ Column temp. : 0 C LF-0 x

60 Organic SEC (GPC) Columns : High Temperature/Ultra High Temperature Analysis Features HT-00 Varied product lineup to support a wide range of molecular weights Corresponds to USP L UT-00 Dedicated to SEC analysis at high/ultra high temperatures with a maximum usable temperature of 0 C Suitable for the analysis of ultra high molecular weight polymer containing samples Corresponds to USP L Standard columns Usable Temperature ( C) F000 GPC HT-0, ~ F00 GPC HT-0, ~ 0, F00 GPC HT-0, ~ 0, F00 GPC HT-0, ~ 0 0, F00 GPC HT-0M, ~ 0 0, F00 GPC HT-0, ~ 0 0, F090 GPC HT-G 00 ~ F000 GPC UT-0.,00 00 ~ F00 GPC UT-0M,00 00 ~ 0 0 0, F00 GPC UT-0,00 00 ~ 0 0 0, F0900 GPC UT-G 00 ~ F090 GPC AT-0MS,000 *Ta ~ 0 0, F000 GPC AT-G *Ta ~ * The columns with M at the end of column names are mixed-gel column capable of analyzing samples over a wide range of molecular weight distribution. Base Material : Styrene divinylbenzene copolymer Shipping Solvent : Toluene *Ta : Ambient temperature Target molecular weight range and Exclusion limit With Polystyrene (eluent : THF) HT-0 HT-0 HT-0 HT-0 HT-0M HT-0 UT-0. UT-0M UT-0 Target Molecular Weight Range,000 0,000,000 00,000 0,000,000,000 0,000 *(0,000,000),000 *(0,000,000) 00,000 *(00,000,000) 00 0,000,000 *(0,000,000) 00,000 *(00,000,000) Exclusion Limit 0,000 00,000,000,000 *(0,000,000) *(0,000,000) *(00,000,000) 0,000 *(0,000,000) *(00,000,000) AT-0MS,000 *(0,000,000) *(0,000,000) * Please use the above table as a rough indication for the column selection. *( ) Estimated value

61 Calibration curves for HT-00 series using polystyrene Molecular weight (Polystyrene) HT-0 HT-0 HT-0 HT-0 Elution volume (ml) Molecular weight (Polystyrene) Elution volume (ml) Effects of gel particle size in high temperature GPC columns High temperature SEC columns are suitable for the analysis of high molecular weight polymers that are difficult to be dissolved in ambient temperature solvents; examples of such polymers are polyethylene and polypropylene. The GPC UT-00 series packed with large particle size (0 um) are recommended for the analysis of macromolecules. The large particle size prevents potential molecular shear degradation of the sample. Column : UT-0M x (Particle size : 0µm) HT-0 HT-0M Column : Shodex GPC HT-00 series Eluent : THF Flow rate :.0mL/ Column temp. : 0 C Calibration curves for UT-00 series using polystyrene UT-0. UT-0M Column : AT-0MS x (Particle size : µm) UT-0 Column : Shodex GPC UT-00 series Eluent : THF Flow rate :.0mL/ Column temp. : 0 C Calibration curve for AT-0MS using polystyrene Molecular weight (Polystyrene) Column : HT-0M x (Particle size : µm) 9 0 Elution volume (ml) Column : Shodex GPC AT-0MS Eluent : THF Flow rate :.0mL/ Column temp. : 0 C Sample :. Polystyrene (MW : 0,000,000). High density polyethylene (HDPE-A) Organic SEC (GPC) Columns : High Temperature/Ultra High Temperature Analysis Vo Shear degradation Vo Vo Shear degradation 0 Column : Shodex GPC UT-0M Shodex GPC HT-0M Shodex GPC AT-0MS Eluent : 0.% BHT in o-dichlorobenzene Flow rate :.0mL/ Column temp. : 0 C High density polyethylene Low density polyethylene Sample : High density polyethylene (HDPE-B) 0 Sample : Low density polyethylene (LDPE-C) 0 Molecular weight 0 RI Column : Shodex GPC UT-0M x Eluent : 0.% BHT in o-dichlorobenzene Flow rate :.0mL/, MALS (Multi angle light scattering) Column temp. : C Column : Shodex GPC HT-0M x Eluent : 0.% BHT in o-dichlorobenzene Flow rate :.0mL/ Column temp. : 0 C 9

62 Organic SEC (GPC) Columns : HFIP Features HFIP-00 HFIP-00 Columns exclusively for use with hexafluoroisopropanol (HFIP) Corresponds to USP L Rapid analysis, solvent saving type Corresponds to USP L Standard columns HFIP-00 series F00 GPC HFIP-0, F00 GPC HFIP-0,000, F00 GPC HFIP-0 0,000 0, F00 GPC HFIP-0 0, , F090 GPC HFIP-0M 0, , F00 GPC HFIP-0,000 0, F0000 GPC HFIP-G B (GPC HFIP-LG).0 0 * The columns with M at the end of column names are mixed-gel column capable of analyzing samples over a wide range of molecular weight distribution. Base Material : Styrene divinylbenzene copolymer Shipping Solvent : Hexafluoroisopropanol (HFIP) Rapid analysis downsized columns HFIP-00 series Use of the HFIP-00 series with semi-micro type devices is recommended. F000 GPC HFIP-0, F000 GPC HFIP-0,000, F000 GPC HFIP-0,000 0, F000 GPC HFIP-0, , F000 GPC HFIP-0M, , F000 GPC HFIP-0,000 0, F00 GPC HFIP-G A (GPC HFIP-G). 0 * The columns with M at the end of column names are mixed-gel column capable of analyzing samples over a wide range of molecular weight distribution. Base Material : Styrene divinylbenzene copolymer Shipping Solvent : Hexafluoroisopropanol (HFIP) Target molecular weight range and Exclusion limit With *PMMA (eluent : HFIP) Target Molecular Weight Range Exclusion Limit Target Molecular Weight Range Exclusion Limit HFIP-0,000 0,000 0,000 HFIP-0,000 0,000 0,000 HFIP-0 0,000 00,000 00,000 HFIP-0 0,000 00,000 00,000 HFIP-0 0,000 00,000,000,000 HFIP-0 0,000 00,000,000,000 HFIP-0 0,000 **(,000,000) **(,000,000) HFIP-0 0,000 **(,000,000) **(,000,000) HFIP-0M,000 **(,000,000) **(,000,000) HFIP-0M,000 **(,000,000) **(,000,000) 0 HFIP-0 0,000 **(0,000,000) **(0,000,000) HFIP-0 0,000 **(0,000,000) * Please use the above table as a rough indication for the column selection. **(0,000,000) *PMMA : Polymethylmethacrylate **( ) Estimated value

63 Calibration curves for HFIP-00 series using PMMA Molecular weight (PMMA) HFIP-0 HFIP-0 HFIP-0 HFIP-0 HFIP-0 HFIP-0M 9 0 Elution volume (ml) Calibration curves for HFIP-00 series using PMMA Molecular weight (PMMA) HFIP-0 HFIP-0 HFIP-0 HFIP-0 HFIP-0 HFIP-0M Elution volume (ml) Polyethylene terephthalate (PET) Sample : Polyethylene terephthalate 0.%, 0µL Mw=9, Organic SEC (GPC) Columns : HFIP Column : Shodex GPC HFIP-00 series Eluent : HFIP Flow rate :.0mL/ Column temp. : 0 C Column : Shodex GPC HFIP-00 series Eluent : HFIP Flow rate : 0.mL/ (HFIP-0, 0) 0.mL/ (HFIP-0, 0, 0M, 0) Detctor : RI (small cell volume) Column temp. : 0 C Column : Shodex GPC HFIP-0M x Eluent : mm CF COONa in HFIP Flow rate : 0.mL/ (small cell volume) Column temp. : 0 C Polyamide (effects of salt addition) Polybutylene terephthalate (PBT) Sample : Polycaprolactum (Nylon ) Sample : Polybutylene terephthalate 0.0%, 00µL HFIP mm CFCOONa in HFIP In SEC analysis using HFIP, some samples may yield abnormal peaks as a result of ionic interaction. In this case, ionic interaction can be suppressed by adding sodium trifluoroacetate to HFIP Column : Shodex GPC HFIP-0M x Eluent : HFIP (solid line), mm CF COONa in HFIP (broken line) Flow rate :.0mL/ Column temp. : 0 C Column : Shodex GPC HFIP-0 + HFIP-0 Eluent : mm CF COONa in HFIP Flow rate :.0mL/ Column temp. : 0 C Polyamides (Nylon 0 and Nylon ) Polyacetal Sample : Nylon 0 0.%, 0µL Sample : Nylon 0.%, 0µL Sample : Polyacetal 0.%, 0µL Mw=9,00 Mw=,00 Mw=, Column : Shodex GPC HFIP-0M x Eluent : mm CF COONa in HFIP Flow rate : 0.mL/ (small cell volume) Column temp. : 0 C Column : Shodex GPC HFIP-0M x Eluent : mm CF COONa in HFIP Flow rate : 0.mL/ (small cell volume) Column temp. : 0 C

64 Applicability of SEC (GPC) Columns to Solvent Replacement Shipping Solvent : THF Shipping Solvent : DMF Solvent KF-0 KF-0 KF-0. KF-0L KF-0L KF-0 KF-0 KF-0 KF-0 KF-0 KF-0M KF-0L KF-0L KF-0L KF-0 KF-0 KF-0. KF-0 KF-0 KF-0 KF-0 KF-0 KF-0M LF-0 LF-0 LF-0 KD-0 KD-0 KD-0. KD-0 KD-0 KD-0 KD-0 KD-0 KD-0M Shipping Solvent : Chloroform Shipping Solvent : THF K-0 K-0 K-0 K-0. K-0L K-0L K-0 K-0 K-0 K-0 K-0M K-0L K-0L K-0L KF-0HQ KF-0HQ KF-0.HQ KF-0HQ KF-0HQ KF-0LHQ KF-0LHQ Tetrahydrofuran (THF) Chloroform Carbon tetrachloride Benzene Toluene p-xylene o-dichlorobenzene (ODCB) Trichlorobenzene (TCB) Dioxane Diethyl ether Ethyl acetate Acetone Methyl ethyl ketone Dimethylformamide (DMF) Dimethylacetamide (DMAc) Hexafluoroisopropanol (HFIP) m-cresol o-chlorophenol Quinolin N-Methylpyrrolidone (NMP) Dimethylsulfoxide (DMSO) 0% m-cresol/chloroform 0% o-chlorophenol/chloroform 0% HFIP/Chloroform Hexane Acetonitrile Methanol Water : Solvent replacement possible : Solvent replacement possible, but this may cause column performance to deteriorate slightly : Usable at 0 C or higher : Solvent replacement not possible

65 Calibration Standards for SEC [ Polystyrene (PS)] Features For organic solvent SEC (GPC) Less branched polystyrene with anionic polymerization Easily soluble in tetrahydrofuran (THF), chloroform, toluene, and o-dichlorobenzene (ODCB) Kit type F00 F00 F00 SL-0 Std. No. Mp Mw/Mn S-9 S- S-9.9 S-.9 S-. S-. S-. S-. S-0. S-0. 9,00,00 9,90,90,0,0,0, STANDARD SL-0 STANDARD SM-0 STANDARD SH S-00 S-90 S- S- S- S- S- S-.9 S-.0 S-.,00,000,90,000,000,000,000,00,00,90,90,0 Contents 0.g x 0 kinds 0.g x 0 kinds 0.g x kinds SM-0 Std. No. Mp Mw/Mn [ Polymethylmethacrylate (PMMA)] Features Contents F00 STANDARD M- 0.g x kinds [Pullulan] Features S-0 S-90 S-990 S-0 S-0 S-99 S-9 MW Range 0 9,00,0,00,000 9,000,0,000 SH- Std. No. Mp Mw/Mn,0,000,90,000,990,000,0,000,0,000 99,000 9,000 For organic solvent SEC (GPC) Narrow molecular weight distribution range Easily soluble in hexafluoroisopropanol (HFIP) and dimethylformamide (DMF) Kit type MW Range,0,00,000 (Note) Molecular weights (Mp, Mw/Mn) of each kit may vary depending on production lots Structural formula of S series M-00 M-9 M- M-9 M- M-. M-.9 C H 9 CH CH (Note) Molecular weights (Mp, Mw/Mn) of each kit may vary depending on production lots. Std. No. Mp Mw/Mn,00,000 9,000,000,00,00,0,0 n H Applicability of SEC (GPC) Columns to Solvent Replacement Calibration Standards for SEC For aqueous SEC (GFC) Unbranched pullulan standard High solubility in water eliates the possibility of recrystallization Kit type Contents MW Range Structural formula of P series F00000 STANDARD P- Single type 0.g x kinds Contents Mp,00,000 Mw/Mn O CH H H H O OH OH H H OH O H CH OH H H CH OH H H O H O OH H O OH H H OH H OH n F0000 F0000 F0000 F0000 F0000 F0000 F0000 F0000 STD P-00 STD P-00 STD P-00 STD P-00 STD P-0 STD P-0 STD P-0 STD P- 0.g 0.g 0.g 0.g 0.g 0.g 0.g 0.g,000,000 9,000 0,000,00,00 9,00, Std. No. Mp Mw/Mn STD P-00 STD P-00 STD P-00 STD P-00 STD P-0 STD P-0 STD P-0 STD P-,000,000 9,000 0,000,00,00 9,00, (Note) Molecular weights (Mp, Mw/Mn) of each kit may vary depending on production lots.

66 Columns for Anion Exchange Chromatography Features QA- DEAE- DEAEN-T DEAE-B ES-0N C WA- Suitable for analysis of relatively high molecular weight compounds: proteins, peptides, DNA, and RNA Usable in a wide ph range from ph to QA- corresponds to USP L Non-porous base material For rapid analysis Non-porous base material Supports UHPLC (available under hyperbaric conditions up to 0 MPa) Compared to IEC series columns, polyvinyl alcohol is used as base material and this offers different separation pattern Low hydrophobic interaction of proteins allows analysis under mild conditions Suitable for anion exchange analysis of low molecular weight compounds such as nucleotides Standard columns Strong anion exchange resin [ Functional Group : Quaternary ammonium] Ion Exchange Capacity (meq/g) Base Material Particle Size Column Size (mm) Shipping Solvent F00 IEC QA- 0. Polyhydroxymethacrylate, mM Na SO aq. Weak anion exchange resin [ Functional Group : Diethylaoethyl] Ion Exchange Capacity (meq/g) Base Material Particle Size Column Size (mm) Shipping Solvent F IEC DEAE- 0. Polyhydroxymethacrylate, mM Na SO aq. F00 IEC DEAEN-T 0. Polyhydroxymethacrylate.. H O F000 Asahipak ES-0N C 0. Polyvinyl alcohol 9, mM,-Diaopropane + 0mM NaCl (ph0.0) F0 AXpak WA-. Polyhydroxymethacrylate 0, M Sodium phosphate buffer (ph.0)/ch CN =0/0 F00 AXpak WA-G Polyhydroxymethacrylate M Sodium phosphate buffer (ph.0)/ch CN =0/0 Weak anion exchange resin [Functional Group : Diethylaoethyl] : For UHPLC column Ion Exchange Capacity (meq/g) Base Material Particle Size Column Size (mm) Shipping Solvent F0 PIKESS DEAE-B 0. Polyhydroxymethacrylate..0 0 H O Semi-micro columns * The following semi-micro columns are made to order. Ion Exchange Capacity (meq/g) Base Material F90 DEAE9A-D 0. Polyvinyl alcohol 9, Preparative columns * Preparative columns are made to order. Standard columns F000 IEC QA QA- F00 IEC QA QA- F090 IEC QA-LG F00 IEC DEAE DEAE- F0 IEC DEAE DEAE- F090 IEC DEAE-LG F000 F00 Asahipak ES-0N 0C Asahipak GS-0G B ES-0N C

67 Proteins in human serum Sample : Human serum 0.%, 00µL. IgG. Transferrin. (Unknown). Serum albu 0 0 Column : Shodex IEC QA- Eluent : (A); 0mM Tris-HCl buffer (ph.) (B); (A) + 0.M NaCl Linear gradient; 00% (A) to 0% (B), 0 Flow rate :.0mL/ Detector : UV (0nm) Column temp. : Room temp. Effects of eluent ph for DEAE- ph ph ph ph Sample : 00µL. Conalbu 0.%. Transferrin 0.%. Ovalbu 0.%. Trypsin inhibitor 0.%, ph Column : Shodex IEC DEAE- Eluent : (A); 0mM Piperazine-HCl buffer (ph.0), 0mM Bis-Tris-HCl buffer (ph.0) 0mM Tris-HCl buffer (ph.0), 0mM Ethanolae-HCl buffer (ph9.0) 0mM,-Diaopropane-HCl buffer (ph0.0) (B); (A) + 0.M NaCl Linear gradient; (A) to (B), 0 Flow rate :.0mL/ Detector : UV (0nm) Column temp. : C Columns for Anion Exchange Chromatography Comparison of porous DEAE- and non-porous DEAEN-T for protein separation IEC DEAEN-T is a weak anion exchange column, having diethylaoethyl functional group modified on non-porous gel. The non-porous gel enables rapid analysis of proteins and peptides. DEAEN-T is also suitable for the analysis of small-volume samples, as it provides sharp peaks even with small injection volume. porous gel DEAE- non-porous gel DEAEN-T Gibberellin Isomers. GA CH HO HOOC COOH Sample : Gibberellin : Diethylaoethyl : Diethylaoethyl. endo GA CH Sample :. Conalbu 0.mg/mL. Ovalbu.mg/mL. Trypsin inhibitor.0mg/ml HO HOOC COOH Column Eluent Flow rate Detector Column temp. : Room temp. Injection vol. : 00µL : Shodex IEC DEAE- : (A); 0mM Piperazine-HCl buffer (ph.0) (B); (A) + 0.M NaCl Linear gradient; (A) to (B), 0 :.0mL/ : UV (0nm) Column Eluent Flow rate Detector Column temp. : Room temp. Injection vol. : 0µL : Shodex IEC DEAEN-T : (A); mm Piperazine-HCl buffer (ph.0) (B); (A) + 0.M NaCl Linear gradient; (A) to (B), 0 :.ml/ : UV (0nm) Column : Shodex Asahipak ES-0N C Eluent : Acetic acid/h O/CH OH =0./0./99. Flow rate :.ml/ Detector : UV (0nm) Column temp. : 0 C Data was provided by Prof. Yamaguch, Faculty of Agliculture, University of Tokyo. Mercaptoalbu and non-mercaptoalbu Nucleotides Sulfide ion and cyanide ion Sample : Normal control serum. Globulin. Uric acid. HMA (Human mercaptoalbu). HNA (Human nonmercaptoalbu) Sample : 0µL. 'CMP 0µg/mL. 'UMP 0µg/mL. 'TMP 0µg/mL. 'CMP 0µg/mL. 'ADP 00µg/mL. 'UMP 0µg/mL. 'GMP 0µg/mL. 'AMP 0µg/mL 9. 'AMP 0µg/mL 0. 'GMP 0µg/mL 9 Sample : 0µL. HS - 0.mg/L. CN - 0.0mg/L Column : Shodex Asahipak ES-0N C Eluent : 0mM N-methylpiperazine-HCl buffer (ph.) + 00mM Na SO + 0.% C H OH Flow rate :.0mL/ Detector : UV (0nm) Column temp. : C Column : Shodex AXpak WA- Eluent : 0.M CH COOH aq. /0.M CH COONH aq.=0/00 Flow rate :.0mL/ Detector : UV (0nm) Column temp. : 0 C Column : Shodex IEC DEAE- Eluent : 0mM Na CO + mm Ethylenediae aq. + 0% CH OH Flow rate :.0mL/ Detector : Electrochemical (Electrode; Silver, 0mV SCE) Column temp. : C

68 Columns for Cation Exchange Chromatography Features SP- CM- SP-0N SP-B ES-0C C P-S Suitable for analysis of relatively high molecular weight compounds: proteins, peptides, DNA, and RNA Usable in a wide ph range from ph to Non-porous base material For rapid analysis Non-porous base material Supports UHPLC (available under hyperbaric conditions for up to 0 MPa) Compared to IEC series columns, polyvinyl alcohol is used as base material offering different separation pattern Low hydrophobic interaction with proteins allows analysis under mild conditions Column for ao acids analysis by cation exchange mode Supports simultaneous analysis of different ao acids Corresponds to USP L and L Standard columns Strong cation exchange resin [ Functional Group : Sulfopropyl] Ion Exchange Capacity (meq/g) Base Material Particle Size Column Size (mm) Shipping Solvent F0 F000 IEC SP- IEC SP-0N Polyhydroxymethacrylate Polyhydroxymethacrylate., mM Na SO aq. 0mM Sodium acetate buffer + 0.M Na SO (ph.0) Strong cation exchange resin [Functional Group : Sulfopropyl] : For UHPLC column Ion Exchange Capacity (meq/g) Base Material Particle Size Column Size (mm) F0 PIKESS SP-B 0. Polyhydroxymethacrylate..0 0 Shipping Solvent 0mM Sodium acetate buffer + 0.M Na SO (ph.0) Weak cation exchange resin [ Functional Group : Carboxymethyl] Ion Exchange Capacity (meq/g) Base Material Particle Size Column Size (mm) Shipping Solvent F000 F000 IEC CM- Asahipak ES-0C C Polyhydroxymethacrylate Polyvinyl alcohol 9,000, mM Na SO aq. 0.M Sodium phosphate buffer (ph.) For ao acids [Functional Group : Sulfo (Na + )] Base Material Particle Size Column Size (mm) Shipping Solvent F CXpak P-S,00 Styrene divinylbenzene copolymer. 0 H O F000 CXpak P-G Styrene divinylbenzene copolymer. 0 H O Preparative columns * Preparative columns are made to order. Standard columns F00 IEC SP SP- F0 IEC SP SP- F090 IEC SP-G B (IEC SP-LG) F00 IEC CM CM- F0 IEC CM CM- F090 IEC CM-LG F000 F00 Asahipak ES-0C 0C Asahipak GS-0G B ES-0C C

69 Protein separation using cation exchange columns ( I ) CM- (Weak cation exchange) 90µL injection 0 0 ( II ) SP- (Strong cation exchange) 0µL injection Column : (I) Shodex IEC CM-, (II) Shodex IEC SP-, (III) Shodex IEC SP-0N Eluent : (A); 0mM Sodium phosphate buffer (ph.0) (B); (A) + 0.M NaCl (I,II) Linear gradient; (A) to (B), 0 (III) Linear gradient; (A) to (B), 0 Flow rate : (I,II).0mL/ (III).mL/ Detector : UV (0nm) Column temp. : Room temp. ( III ) SP-0N (Strong cation exchange) non-porous type gel Sample :. Myoglobin. Trypsinogen. Ribonuclease A. α-chymotrypsinogen A. Cytochrome c. Lysozyme Rapid analysis of proteins using UHPLC 0.0 Sample : µl (mg total protein).myoglobin. Trypsinogen. Ribonuclease A. α-chymotrypsinogen A. Cytochrome c. Lysozyme Column : Shodex PIKESS SP-B Eluent : (A); 0mM Sodium phosphate buffer (ph.0) (B); (A) + 0. M NaCl Linear gradient; 00% (A) to 0% (B),. Flow rate :.ml/ Detector : UV (0nm) Column temp. : C Columns for Cation Exchange Chromatography Catecholaes Angiotensins Paraquat and diquat Sample : 00µg/mL each, 0µL Sample : 0µL. Angiotensin II (human). DOPA Asp-Arg-Val-Tyr-lle-His-Pro-Phe HO CH CH NH HO COOH. Angiotensin I (human) Asp-Arg-Val-Tyr-lle-His-Pro-. Adrenaline PheHis-Leu HO HO CH CH NH OH CH. Noradrenaline HO CH CH NH HO OH. Angiotensin III (human) Arg-Val-Tyr-lle-His-Pro-Phe Paraquat HC N + + N CH Cl - Diquat Sample : 0µL + N + N Br -. Dopae HO HO CH CH NH Column : Shodex Asahipak ES-0C C Eluent : 0mM Sodium malonate buffer + 0.M NaCl (ph.0) Flow rate :.0mL/ Detector : UV (0nm) Column temp. : 0 C Column : Shodex Asahipak ES-0C C Eluent : 0mM Sodium malonate buffer (ph.0) /CH CN=0/0 Flow rate :.0mL/ Detector : UV (0nm) Column temp. : 0 C Column : Shodex Asahipak ES-0C C Eluent : 0mM Sodium phosphate buffer + 0mM NaCl (ph.0) Flow rate :.0mL/ Detector : UV (nm) Column temp. : 0 C Analysis of pochonicine and its analogues in filamentous fungi culture extract Standard ao acids Pochonicine Sample : µl Fraction purified from the culture extract of Pochonia suchlasporia var. suchlasporia TAMA (Fungi culture provided by Dr. Toru Okuda, Tamagawa University Research Institute) 0 9 Sample : 0.µM each, 00µL. Asp 0. Met. Thr. Ile. Ser. Leu. Glu. Tyr. Pro. Phe. Gly. Lys. Ala. NH. Cys. His 9. Val. Arg Analogue A Analogue B Column : Shodex Asahipak ES-0C C Eluent : 0mM Ammonium bicarbonate aq. Flow rate : 0.mL/ Detector : UV (0nm) Column temp. : 0 C Data provided by Dr. Teruhiko Nitoda, Faculty of Agriculture, Okayama University Column Eluent Reagent Flow rate Detector Column Temp. : C Reaction Temp. : 0 C : Shodex CXpak P-S : MCI Buffer L-00-PH Kit (Mitsubishi Chemical Corporation) Low pressure gradient: 0; PH-, 0.; PH-,.; PH-,.; PH ; PH-RG : Ninhydrin Coloring Solution Kit for HITACHI (Wako Pure Chemical Industries, Ltd.) 0-; R:R=0:0 : (Elunt) 0.mL/ (Reagent) 0.mL/ : VIS (0nm)

70 Columns for Special Separation Modes Column for Hydrophobic Interaction Chromatography Features PH- Separates proteins without denaturation Applicable to samples obtained after ammonium sulfate fraction treatment Standard columns Product Code Functional Group Shipping Solvent F000 HIC PH- Phenyl 0,000.0 H O Base Material : Polyhydroxymethacrylate Columns for Affinity Chromatography Features AFpak Standard columns Rigid polymer-based packing materials enable high speed analysis Functional group is modified with chemically stable ligand (spacer) Minimum detachment of functional groups ensures highly reproducible analysis Product Code Ligand Ligand Load/Gel (g) Shipping Solvent F9 AFpak ADS-9 Dextran sulfate 0mg.0 0 0mM Sodium phosphate buffer + 0.0% NaN (ph.) F9 AFpak AHR-9 Heparin mg.0 0 0mM Tris-HCl buffer + 0mM NaCl + 0.0% NaN (ph.) F9 AFpak APA-9 Protein A mg M Sodium phosphate buffer + 0.M NaCl + 0.0% NaN (ph.0) F00 AFpak APG-9 Protein G mg.0 0 0mM Sodium phosphate buffer + 0.M NaCl + 0.0% NaN (ph.) F99 AFpak AWG-9 Wheat germ agglutinin (WGA) mg M Tris-HCl buffer + 0.M NaCl + 0.M N-Acetylglucosae + 0.0% NaN (ph.) F9 AFpak ACH-9 Choline oxydase, Acetylcholine esterase. 0 0mM Phosphate buffer +.0M NaCl (ph.) Base Material : Polyhydroxymethacrylate Columns for Chiral Separation Features CDBS- Separates optical isomers by using their conformational compatibility differences Versatile column for chiral separation Corresponds to USP L CRX- Separates optical isomers by using the differences in metal complex formation capacities of functional group and metal ion in eluent and optical isomers Suitable for ao acids, hydroxyl acids, and their derivatives Standard columns Product Code Functional Group Base Material Shipping Solvent F00 ORpak CDBS- β -Cyclodextrin derivative Silica. 0.0% Acetic acid + 0.M NaCl aq. /CH CN=0/0 F000 ORpak CRX- L-ao acid derivative Polyhydroxymethacrylate mM CuSO aq. F0900 ORpak CRX-G L-ao acid derivative Polyhydroxymethacrylate. 0 0.mM CuSO aq. Column for High Temperature Reversed Phase Chromatography Features ET-RP Standard columns Capable of high temperature analysis up to 0 C High temperature analysis improves column efficiency and enables rapid analysis Corresponds to USP L Product Code Functional Group Shipping Solvent F00 ET-RP D,000 Octadecyl 0. 0 H O/CH CN=/ Base Material : Polyvinyl alcohol

71 Protein separation by hydrophobic interaction chromatography 0 Sample : 0µL. Cytochrome c 0.0%. Myoglobin 0.0%. Ribonuclease A 0.%. Ovalbu 0.%. Lysozyme 0.0%. α-chymotrypsinogen 0.0% Column : Shodex HIC PH- Eluent : (A);.M Ammonium sulfate + (B) (B); 0.M Phosphate buffer (ph.0) Linear gradient; (A) to (B), 0 Flow rate :.0mL/ Detector : UV (0nm) Column temp. : Room temp. Choline and acetylcholine 0 0 Sample : 0µL. Choline mg/l. Ethylhomocholine 0mg/L. Acetylcholine 0mg/L After choline and acetylcholine are separated using DE- polymer-based reversed phase column, solutes are passed through ACH-9 to generate hydrogen peroxide. The resulting hydrogen peroxide is detected using an electrochemical detector to enable highly sensitive analysis. * See page for DE- Column : Shodex RSpak DE- Post column : Shodex AFpak ACH-9 Eluent : 0.M H PO + 00mg/L Sodium -decansulfonate + mg/l Tetramethylammonium chloride (ph.0 adjusted by.0m NaOH) Flow rate :.0mL/ Detector : Electrochemical (Electrode : Pt, 0mV SCE) Column temp. : C Lipoproteins in plasma LDL Eluent A Eluent B HDL Eluent A Eluent B VLDL Eluent A Eluent B Sample : Lipoproteins(Sigma) Column : Shodex AFpak ADS-9 Eluent : (A); 0mM Sodium phosphate buffer (ph.) (B); (A) +.0M NaCl Step gradient; (A) to (B) Flow rate :.0mL/ Detector : UV (0nm) Column temp. : Room temp. Columns for Special Separation Modes Chiral separation of epinephrines Chiral separation of warfarin Chiral separation of lactic acids Sample : Epinephrine 0µg/mL, 0µL Sample : Warfarin 0µg/mL, 0µL Sample : Lactic acid 0µg/mL, 0µL. L-Lactic acid. D-Lactic acid HO N H CH O O CHCH COCH OH OH OH Column : Shodex ORpak CDBS- Eluent : 0.0% Acetic acid + 0.M NaCl aq. /CH CN=9/ Flow rate : 0.mL/ Detector : UV (nm) Column temp. : 0 C Column : Shodex ORpak CDBS- Eluent :.0% Acetic acid + 0.M NaCl aq. /CH CN=0/0 Flow rate : 0.mL/ Detector : UV (0nm) Column temp. : C Column : Shodex ORpak CRX- Eluent : 0.mM CuSO aq. Flow rate :.0mL/ Detector : UV (0nm) Column temp. : 0 C Chiral separation of mandelic acids Sample : Mandelic acid 00µg/mL, 0µL. D-Mandelic acid. L-Mandelic acid Comparison of ET-RP s column efficiencies (theoretical plate height) observed at high and normal temperature conditions high temp. (0 C).mL/ normal temp. (0 C) 0.mL/ Sample :. Uracil. Pyridine. Acetophenone. Benzene Sample Acetophenone Benzene Reduced plate height * 0 C 0 C.... * Plate height / particle diameter of the packed resin Column : Shodex ORpak CRX- Eluent : 0.mM CuSO aq. Flow rate :.0mL/ Detector : UV (0nm) Column temp. : 0 C Column : Shodex ET-RP D Eluent : (Left) H O/CH CN=0/0 (Right) H O/CH CN=/ Detector : Photodiode array (0nm) Column Oven : Polaratherm 9000 Series (SandraSelerity Technologies, Inc) Note : The eluent was introduced into the column after being preheated and was cooled after column elution, then introduced into the detector. Data provided by Research Institute for Chromatography bvba 9

72 Columns for GPC Clean-up Features EV Suitable for fractionation of residual pesticides in foods EV-000 AC is used in Shoku-An No (October rd, 00, Japan) of the Pharmaceutical and Food Safety Bureau, MHLW, Section Simultaneous GC/MS (LC/MS) Analyses of Agricultural Chemicals in Livestock and Marine Products. EV000AC-F is used in Shoku-An No. 0 (February th, 0, Japan) of the Pharmaceutical and Food Safety Bureau, MHLW, Section LC/MS Analyses of Agricultural Chemicals in Livestock and Marine Products. PAE Suitable for cleaning up high-moisture samples such as blood and bottom sediment Highly effective for fractionation of endocrine disruptors in environmental samples Eluent Column No Ethylacetate/Cyclohexane EV-000, EV-00 Appropriate columns for GPC Clean-up Does the sample contain water? Acetone/Cyclohexane Acetonitrile EV-000 AC, EV000AC-F PAE-000, PAE-00 Yes Acetone PAE-000 AC, PAE-00 AC GPC Clean-up for residual pesticides in foods, etc. Shipping Solvent F09000 CLNpak EV000AC-F Acetone/Cyclohexane=/ F09000 CLNpak EV-G ACC.0 00 Acetone/Cyclohexane=/ F09000 CLNpak EV-000 AC Acetone/Cyclohexane=/ F09000 CLNpak EV-G AC Acetone/Cyclohexane=/ F09000 CLNpak EV Ethylacetate/Cyclohexane=/ F09000 CLNpak EV-G Ethylacetate/Cyclohexane=/ F09000 CLNpak EV Ethylacetate/Cyclohexane=/ Base Material : Styrene divinylbenzene copolymer GPC Clean-up for phthalic acid esters in sediments, biological samples, blood, etc. Shipping Solvent F00 CLNpak PAE Acetonitrile F00 CLNpak PAE-G Acetonitrile F000 CLNpak PAE Acetonitrile F00 CLNpak PAE-000 AC Acetone F00 CLNpak PAE-G AC Acetone F000 CLNpak PAE-00 AC Acetone Base Material : Polyvinyl alcohol 0

73 Sample preparation outline for simultaneous GC/MS and LC/MS analysis of agricultural chemicals in livestock and marine products (part ) [Outline] Extraction Purification Liver, Kidney Fraction- [*] [*] Acetone/n-Hexane (/) GPC Fraction- Muscle, Fat, Fish Milk, Egg Honey Agricultural chemicals fraction GPC Agricultural chemicals fraction Acetonitrile Salting-out [*] [*] Preparaion range of agricultural chemicals using EV-000 AC Tricyclazole (mg/l) (marker agricultural chemicals) Acrinathrin (mg/l) (marker agricultural chemicals) muscle, fat, fish, milk, egg liver, kidney agricultural chemicals fraction fraction- fraction- Columns for GPC Clean-up Test sample GPC column : Shodex CLNpak EV-000 AC + EV-G AC * Purification with ethylenediae-n-propylsilyled silica gel i-column * Purification with silica gel i-column Column : Shodex CLNpak EV-G AC + EV-000 AC Eluent : Acetone/Cyclohexane=/ Flow rate :.0mL/ Detector : UV (nm) Column temp. : 0 C Injection vol. : ml Sample preparation outline for simultaneous LC/MS analysis of agricultural chemicals in livestock and marine products (part ) [Outline] Extraction Other than honey Honey Acetone/n-Hexane (/) under acidic conditions with acetic acid Preparaion range of agricultural chemicals using EV000AC-F Acrinathrin (mg/l) (marker agricultural chemicals) Tricyclazole (mg/l) (marker agricultural chemicals) GPC Clean-up of fatty tuna and beef tallow Sample : ml Beef tallow Fatty tuna. Acrinathrin mg/l. Tricyclazole mg/l Purification Macroporous diatomaceous earth column chromatography GPC Agricultural chemicals fraction [*] [*] Fluvalinate (mg/l) fraction range GPC Clean-up of beef tallow GPC Clean-up of fatty tuna [*] [*] fraction range (when purification is insufficient) Test sample GPC column : Shodex CLNpak EV000AC-F + EV-G ACC * Purification with trimethyl aopropylsilyled silica gel i-column * Purification with ethylenediae- N-propylsilyled silica gel i-column Column : Shodex CLNpak EV-G ACC + EV000AC-F Eluent : Acetone/Cyclohexane=/ Flow rate :.0mL/ Detector : UV (nm) Column temp. : C Injection vol. : ml Column : Shodex CLNpak EV-G AC + EV-000 AC Eluent : Acetone/Cyclohexane=/ Flow rate :.0mL/ Detector : UV (nm) Column temp. : 0 C Injection vol. : ml Separation of PSB and insulating oil using PAE-00 Sample : 0µL PCB std. KC-mixture Insulating oil dissolved by Hexane 00mg/L,000mg/L GPC Clean-up of carcinogens in diesel dust measured using PAE-00 AC Benzo[a]fluoranthene,-Dinitropyrene Bisphenol A Benzo[a]pyrene Hydroxybenzo[a]pyrene mixture Elution positions of phthalic acid esters using PAE-00 AC RI Corn oil Sulfer H O CO Insulating oil PCB RI Bis(-ethylhexyl) adipate methanol RI Phthalic acid esters acetone hexane Retention time () Column : Shodex CLNpak PAE-00 Eluent : Methanol, Acetone, Hexane Flow rate : 0.mL/ Detector : Photodiode array (09nm) Column temp. : 0 C Source: Mr. Tetsuya Sawatsubashi (Mitsubishi Heavy Industries, Ltd.) et al., Search of Liquid Chromatographic Clean-up Materials for Rapid PCB Analysis and Evaluation of Their Separation Characteristics. Journal of Environmental Chemistry, 00, Vol., No., p Compound Rt() Compound Rt() Anthracene Chrysene Fluoranthene Benzo[a]fluoranthene Benzo[b]fluoranthene Benzo[k]fluoranthene Pyrene -Hydroxypyrene Benzo[a]pyrene Hydroxybenzo[a]pyrene compound Bisphenol A,-Dinitronaphthalene -Nitropyrene,-Dinitropyrene,-Dinitropyrene,-Dinitropyrene Column : Shodex CLNpak PAE-00 AC Eluent : Acetone Flow rate : 0.mL/ Detector : UV (0nm) Column temp. : Room temp. Data provided by Kazuichi Hayakawa Ph.D., Faculty of Pharmaceutical Sciences, Kanazawa University. RI RI UV (0nm) UV (0nm) -Nonylphenol DDT Bisphenol A Anthracene Column : Shodex CLNpak PAE-00 AC Eluent : Acetone Flow rate : 0.mL/ Detector : UV (0nm), RI Column temp. : Room temp.

74 Pretreatment Columns for Column Switching Method Features PK GF-A Effective for both hydrophilic and hydrophobic substances The high protein removal rate enables efficient pretreatment Higher protein removal rate than PK columns * GF-A column removes proteins well but is not suitable for trapping hydrophilic substances. Use PK columns for this purpose. Cartridge columns and holder for column switching method Shipping Solvent Pcs/box F00000 MSpak PK-A p H O F000 MSpak PK-A p H O F0000 MSpak HLD (Holder for PK) Base Material : Hydrophilic copolymers containing N-vinyl acetoamide * PK series are cartridge columns and thus should be installed in a column holder "MSpak HLD" before use. Column for column switching method Shipping Solvent F000 MSpak GF-A H O Base Material : Polyvinyl alcohol

75 System diagram for column switching Eluent Pump Injector Pretreatment column Proteins etc. in sample Analysis column Pump Eluent Detector Protein removal rate using pretreatment columns Column PK-A *GF-A Company-A Protein removal rate 99.% 99.0% 9.% Residual BSA (µg) PK-A GF-A Company-A (Pretreatment) Eluent : 0mM Ammonium acetate buffer (ph.0) Flow rate : 0.mL/ Column temp. : Room temp. (Analysis) Column : Shodex PROTEIN **KW-0S Eluent : 0.% TFA in (H O/CH CN=0/0) Flow rate : 0.mL/ Detector : UV (0nm) *GF-A : Custom-made column of Column temp. : Room temp. GF-A (.0mmI.D. x 0mm) Switching time : **KW-0S is phase-out product. Pretreatment Columns for Column Switching Method Recovery rate of medical compounds using PK-A Sample Recovery (%) Sample Recovery (%) Sample Recovery (%) Acetaophen Cloxazolam 9 Mianserin 9 Acetylpheneturide 9 Desiprae 09 Nimetazepam 90 Aconitine 0 Diazepam 9 Nitrazepam 9 Alprazolam 99 Diphenhydrae 9 Nortriptyline Amitriptyline 9 Estazolam 0 Oxazepam 9 Amobarbital 9 Ethenzamide 9 Oxazolam 99 Barbital 9 Etizolam 0 Pentobarbital 9 Benzoylaconitine 9 Fludiazepam 9 Perfenazine Benzoylhypaconine Flumazenil 9 Phenacetin 0 Biperiden 99 Flunitrazepam 9 Phenobarbital 9 Bromazepam 0 Flurazepam 0 Phenytoin 99 Bromocriptine 0 Glutethimide 9 Primidone 9 Bromperidol 9 Haloperidol 99 Promethazine 9 Bromvalerylurea 9 Haloxazolam 9 Propericiazine 90 Brotizolam 9 -Hydroxymethyltriazolam 90 Propranolol 9 Caffeine 0 -Hydroxytriazolam 9 Secobarbital 9 Carbamazepine 9 Hydroxyzine 99 Sildenafil citrate 9 Carpiprae 99 Hypaconitine 9 Thioridazine 9 Chlordiazepoxide Imiprae 9 Timiperone Chlormezanone 9 Indomethacin 9 Triazolam 9 Chlorphenirae Levomepromazine 9 Trihexyphenidyl 9 Chlorpromazine Lofeprae Trimethadione Clocaprae 9 Maprotyline 90 Trimiprae 0 Clofedanol 9 Medazepam 9 Warfarin Clomiprae 9 Mephobarbital 99 Zotepine 9 Clonazepam 9 Mesaconine Clotiazepam 9 Metharbital 9 (Adsorption) Eluent : 0mM Ammonium acetate buffer (ph.0) Flow rate : 0.mL/ (Elution) Eluent : 0mM Ammonium acetate buffer (ph.0)/ch CN Flow rate : 0.mL/ Detector : UV (0nm) Switching time :

76 Notice of Changes Some of Shodex guard columns have been renamed. There is no change of product code. Renamed products list Page New Former RSpak DE-G A RSpak DE-G F000 RSpak DE-G A RSpak DE-SG F00 RSpak DM-G A RSpak DM-G F000 RSpak DE-G B RSpak DE-LG F0090 RSpak DE-G 0C RSpak DE-LLG F009 RSpak DM-G B RSpak DM-LG F000 RSpak DM-G 0C RSpak DM-LLG F00 SUGAR SC-G B SUGAR SC-LG F00090 SUGAR SP-G B SUGAR SP-G F000 SUGAR KS-G B SUGAR KS-G F0000 RSpak DC-G A RSpak DC-G F000 SUGAR SCG A SUGAR SC-G F000 SUGAR KS-G B SUGAR KS-LG F0000 RSpak DC-G B RSpak DC-LG F000 RSpak DC-G 0C RSpak DC-LLG F00 RSpak KC-G B RSpak KC-G F0000 RSpak KC-G B RSpak KC-LG F0000 PROTEIN KW-G B PROTEIN KW-G F00 PROTEIN KW-G B PROTEIN KW-LG F09 OHpak SB-G B OHpak SB-G F090 9 OHpak SB-G B OHpak SB-LG F09, GPC KF-G A GPC KF-G F0000 GPC K-G A GPC K-G F000 0 GPC KD-G A GPC KD-G F00 GPC KF-G B GPC KF-LG F000 GPC K-G B GPC K-LG F000 GPC H-G B GPC H-G F000 GPC KF-G 0C GPC KF-LLG F000 GPC K-G 0C GPC K-LLG F GPC HFIP-G B GPC HFIP-LG F GPC HFIP-G A GPC HFIP-G F00 IEC SP-G B IEC SP-LG F090

77 USP (Ver.) Column List No. Packing Material Recommended Column Page L L L L L0 L L L9 L0 L L L L L L L L L L9 L L L9 L L L Octadecyl silane chemically bonded to porous or non-porous silica or ceramic micro-particles,. to 0µm in diameter, or a monolithic rod Porous silica particles,. to 0µm in diameter, or a monolithic silica rod. Octylsilane chemically bonded to totally or superficially porous silica particles,. to 0µm in diameter, or a monolithic silica rod. An essentially monomolecular layer of aopropylsilane chemically bonded to totally porous silica gel support,. to 0µm in diameter Nitrile groups chemically bonded to porous silica particles,. to 0µm in diameter, or a monolithic silica rod. Phenyl groups chemically bonded to porous silica particles,. to 0µm in diameter, or a monolithic silica rod. Strong cation-exchange resin consisting of sulfonated cross-linked styrene-divinylbenzene copolymer in the hydrogen form, to µm in diameter Strong cation-exchange resin consisting of sulfonated cross-linked styrene-divinylbenzene copolymer in the calcium form, about 9µm in diameter Dihydroxypropane groups chemically bonded to porous silica or hybrid particles,. to 0µm in diameter, or a monolithic silica rod. A rigid, spherical styrene-divinylbenzene copolymer, to 0µm in diameter A cation-exchange resin made of porous polystyrene gel with sulfonic acid groups, about 0µm in size An anion-exchange resin made of porous polymethacrylate or polyacrylate gel with quartenary ammonium groups, -µm in size Packing having the capacity to separate compounds with a molecular weight range from (as detered by polyethylene oxide), applied to neutral, anionic, and cationic water-soluble polymers. A polymethacrylate resin base, cross-linked with polyhydroxylated ether (surface contained some residual carboxyl functional groups) was found suitable Butyl silane chemically bonded to totally porous silica particles,. to 0µm in diameter Packing having the capacity to separate dextrans by molecular size over a range of,000 to 00,000 Da. It is spherical, silica-based, and processed to provide ph stability Strong cation-exchange resin consisting of sulfonated cross-linked styrene-divinylbenzene copolymer in the lead form, about to 9µm in diameter Packing having the capacity to separate proteins by molecular size over a range of,000 to 0,000 Da. It is a polymethacrylate gel A methacrylate-based size-exclusion packing for water-soluble samples A hydrophilic polyhydroxymethacrylate gel of totally porous spherical resin Beta cyclodextrin, R,S-hydroxypropyl ether derivative, bonded to porous silica particles, to 0µm in diameter Strong cation-exchange resin consisting of sulfonated cross-linked styrene-divinylbenzene copolymer in the sodium form, about to 0µm diameter Packing for the size-exclusion separations of proteins (separation by molecular weight) over the range of to 000 kda. It is spherical (.-0µm), silica or hybrid packing with a hydrophilic coating. Porous vinyl alcohol copolymer with a C alkyl group attached to the hydroxyl group of the polymer, to 0µm in diameter A rigid, spherical polymetacrylate, to µm in diameter Polyae chemically bonded to cross-linked polyvinyl alcohol polymer, -µm in diameter Silica CM Silica CP Silica SIL Silica C Silica NH Silica CN Silica NPE SUGAR SH0 SUGAR SH RSpak KC- IC Y- SUGAR SC0 SUGAR SC SUGAR SC EP SC0-F USPpak MN- PROTEIN KW-00 series KW00 series RSpak RP- RSpak DS- RSpak DS- GPC KF,K,KD,LF,HT,UT,AT,HFIP series SUGAR SC0 SUGAR SC SUGAR SP00 SUGAR KS-00 series RSpak DC- SUGAR SZ SUGAR SC EP SC0-F USPpak MN- SUGAR SH0 SUGAR SH RSpak KC- IC Y- CXpak P-S IEC QA- OHpak SB-0 HQ OHpak SB-0. HQ Silica C PROTEIN KW-00 series KW00 series SUGAR SP00 OHpak SB-0 HQ OHpak LB-0 OHpak SB-00 HQ series OHpak LB-00 series ODP HP RSpak DM- OHpak SB-00 HQ series OHpak LB-00 series ORpak CDBS- SUGAR KS-00 series RSpak DC- CXpak P-S PROTEIN KW-00 series KW00 series Asahipak ODP-0 Asahipak ODP-0 ET-RP RSpak DE- RSpak DE- RSpak DE- Asahipak NHP-0 Asahipak NHP-0,, 0,,,, Notice of Changes USP (Ver.) Column List

78 Column Cleaning Procedures Change in peak shapes, elution tig, and the elevated column pressure may be resolved by cleaning the column. This section describes general indications of column deterioration and column cleaning procedures. For details of column cleaning procedures, refer to each column s specific operation manual. Typical indicators of column deterioration possibility. Elevated column pressure. Abnormal peak shapes (broadening, leading, or tailing) and split peaks. Change in retention time. Unstable baseline Selection guide to the cleaning solvent Solvents capable of dissolving the adsorbed substances. Solvents with high eluting power (variable depending on separation mode) *Use the solvent specified in the operation manual. Standard cleaning procedures For an efficient cleaning, reverse the direction and reduce the flow rate to / of the regular flow. Columns for reversed phase chromatography Columns for sugar analysis chromatography Columns for aqueous SEC(GFC) chromatography Columns for ion exchange chromatography Clean the columns with solvent containing higher concentration of organic solvent such as methanol, acetonitrile, or THF. (In case of using buffer as a mobile phase, miscibility of the buffer solution and the organic solvents need to be checked) [Ligand exchange columns (SUGAR series)] In case of counter ion detachment Flush or inject solvent containing the salt corresponding to the modified counter-ligand. [Polymer-base ao columns (NHP series)] In cases where an acidic substance has been bound to the ao functional group Flush with solvents in the following sequence: water, 0.M perchloric acid (aq.), water, 0.M NaOH (aq.), water, and mobile phase. In cases where an ionic substance has been adsorbed Use a solvent with higher salt concentration or solvent with different ph from the mobile phase. In cases where a hydrophobic substance has been adsorbed Use a solvent containing organic solvent. (In case of using buffer as a mobile phase, miscibility of the buffer solution and the organic solvents need to be checked) In cases where an ionic substance has been adsorbed Use a solvent with higher salt concentration or solvent with different ph from the mobile phase. In cases where a hydrophobic substance has been adsorbed Use a solvent containing organic solvent. (In case of using buffer as a mobile phase, miscibility of the buffer solution and the organic solvents need to be checked) In cases where protein have been adsorbed Inject - ml of 0. M NaOH (aq.) or 0% (v/v) acetic acid (aq.) several times. Columns for hydrophobic interaction chromatography In cases where protein have been adsorbed Inject - ml of 0. M NaOH (aq.) or 0% (v/v) acetic acid (aq.) several times. *The volume of the cleaning solvent required is -0 times the column volume. *Avoid pressure elevation during the cleaning. *The cleaning is limited and does not guarantee the full regeneration of the column to its original condition. For your information One typical cause of the column pressure elevation is the clogging of solid substances at the inlet filter of the column. In this case, reverse the direction and reduce the flow to / of the regular flow rate. This may remove the solid substance causing the elevated pressure. *Use the solvent specified in the operation manual.

79 General Precautions for Column Handling For the best performance of the column, please follow the instructions given below. Column mounting Before mounting the column, replace the eluent within all the HPLC system with the mobile phase used for the analysis. *If the mobile phase of the choice is not miscible with the eluent already in the system, use solvent that is miscible with both solvents first to clean the system. *Buffer or salt solution may precipitate when mixed with organic solvent of different concentrations. Attach the column in the direction as indicated by arrow marked on the column. Gradually increase the flow rate of the solvent introduced to the column. When heating the column, be sure to pump the eluent at a low flow rate until the specified temperature is reached, and then gradually increase the flow rate up to the requirement after the column has been heated sufficiently. Column dismounting If the column is heated, turn off the heater while keeping the flow rate at / of the regular flow. Turn off the pump when the column is cooled to room temperature. Remove the column from the system securely tighten the end caps. Column storage For long-term storage, replace the solvent with shipping solvent and securely tighten the end caps. Column Cleaning Procedures General Precautions for Column Handling Store the column in a location with stable temperature. For long-term storage of SEC columns, immersion method is recommended. *Please refer to the immersion method on the operation manual. Other Avoid physical shock on the column. Be cautious not to drop the column from a high position. Do not bend the column. Avoid opening the column s end-fitting, it can cause alteration of column s performance. * Read the operation manual before using the column.

80 Column Trouble Shooting Common causes for abnormal chromatograms Abnormal peak shapes Main cause Solution Frequent problem Peak tailing Selection of column and/ or eluent appropriate N O YES Strong sample retention Void existing at top of column Change column and/or eluent Replace column Peak split, shoulders Adsorption of impurities in column Channeling in column Wash column Check column performance Replace column Replace column Peak leading Leading caused by sample properties (ionic and micellar properties) Change column and/or eluent Broad peak Problem solved by reducing injection volume N O YES Adsorption of impurities in column Deterioration of column Sample overload Wash column Check column performance Replace column Reduce injection volume Ghost peak Contaation Impurities existing in eluent Prepare sample again Wash flow line of HPLC Replace eluent No peak System trouble Eluent not flowing Injector at wrong position (load/inject) Detector deactivated Check system Sample adsorption in column Repeat sample injection Column equilibration insufficient Pressure elevation Pump A B C Column Detector Main cause START Injector Solution Frequent problem Salt precipitated is observed in eluent YES Pressure still high after washing N O Salting out Change eluent composition Wash system before analysis N O YES Pressure still high when C is disconnected N O Detector piping blocked Clean up detector Replace piping YES Pressure still high when B is disconnected YES N O Column blocked Pressure under maximum column pressure YES N O Screen eluent and sample through 0.µm filter YES N O Deterioration of column Column end-fitting blocked Check column performance Replace column Replace column Pressure still high when A is disconnected N O Injector blocked Clean up injector Replace injector YES Used line filter YES N O Pump piping blocked Replace filter in pump Clean up pump Replace pump piping Line filter blocked Wash line filter Replace line filter

81 Unstable baseline values START Column Pressure normal YES Baseline stable when pump stopped YES Leakage observed YES HPLC System Trouble Shooting N O N O N O Common causes for abnormal chromatograms YES Monotonic drift N O Main cause Solution Malfunction of check valve Temperature fluctuation Deterioration of detector lamp Insufficient grounding Malfunction of recorder Bubbles in flow cell Contaation of flow cell Insufficient eluent replacement (with different solvent, salt or others) Frequent problem Replace check valve Clean up check valve Warm up detector sufficiently Shield detector from wind Check column oven Check detector temperature control Replace lamp Establish sufficient grounding Repair recorder Wash inside of detector Wash inside of detector Wash flow line of HPLC Column Trouble Shooting HPLC System Trouble Shooting Flow rate fluctuation Contaation of column Pulsation of pump Wash column Check column performance Replace column Use damper Tighten connections Deterioration of eluent Use eluent containing stabilizer (for THF, use THF containing BHT) Noise Main cause Solution Frequent problem START Insufficient grounding Establish sufficient grounding Baseline stable when pump stopped N O Irregular noise Deterioration of detector lamp Replace detector lamp YES Malfunction of recorder Repair recorder Shodex DEGAS used N O Insufficient degassing eluent Use Shodex DEGAS YES Spike noise YES OK Spike noise found from beginning N O YES Air in column Problem solved by applying back pressure to column(*) (*) Do not apply back pressure to detector Column temperature exceeding eluent boiling point N O Check column performance Replace column Reduce temperature of column oven and/or post-column reaction coil Noise shape leading N O Contaation of flow cell Wash flow cell YES Bubbles in flow cell Repair detector 9

82 Index by Columns are listed in alphabetical order under the product name excluding series name. [Series name] AFpak Asahipak AXpak CLNpak CXpak EP GPC HIC HILICpak IC IEC MSpak OHpak ORpak PIKESS PROTEIN RSpak Silica STANDARD SUGAR USPpak A ACH-9 ADS-9 AHR-9 APA-9 APG-9 AT-0MS AWG-9 C CM CP CP-0 C C CP-0 CDBS- CM CN CRX- D DC DE DEAE DM DS E ET-RP D ES-0C ES-0N EV F 0 0,,, 0 FL-, FL- filter 0, G GF-0 GF-0 GF-A GF-0 GF-0 GF-M HQ GS-0 GS-0 GS-0 GS-0 GS-0 GS-0 GS-0 GSM-00 H H-000 HFIP-00 HFIP-00 HLD HT-00 I 0 0 I-A 0 J JJ-0 K K-000 K-000 K-00 K-00D KC- KD-00 KF-000 KF-000 KF-00HQ KF-00 KF-00D KF-00 KS-000 KS-00 KW-000 KW00 KW-00 L LB-00 LF M M- MN- N NH NHP NI- NN NPE D O ODP HP ODP, 0 0, 0 0, 0 P P-S P- PAE PH- PK PYE D Q 0 QA- R RP- S SB-000 SB-00 HQ SC0, SC SC0-F SC SH0, SH SH- SI- D SI-0 E SI- E SI-90 E SIL SL-0 SM-0 SP00 SP SZ T T- U UT-00 V VG-0 VT-0 W 0 0 WA- Y Y- YK- YS-0 0

83 F000 F000 F00 F00 F00 F000 F000 F000 F000 F000 F000 F000 F000 F000 F000 F000 F00 F000 F000 F000 F000 F000 F0090 F009 F009 F009 F009 F009 F009 F009 F009 F0099 F00 F0 F0 F0 F0 F0 F09 F00 F0 F0 F00 F00 F00 F00 F00 F090 F09 F09 F09 F09 F09 F00 F00 HFIP-0 HFIP-0 LF-0 LF-0 LF-0 HFIP-0 HFIP-0 HFIP-0 HFIP-0M KF-0L KF-0L KF-0L KF-0L KF-0L KF-0 KF-0 KF-0. KF-0 KF-0 KF-0 KF-0 KF-0 KF-0M KF-0 KF-0 KF-0. KF-0 KF-0 KF-0 KF-0 KF-0M KF-0 K-0 KF-0HQ KF-0HQ KF-0.HQ KF-0HQ KF-0HQ KF-0LHQ K-0 KF-0LHQ K-0. K-0 K-0 K-0 K-0 K-0 K-0M K-0L K-0L K-0L K-0L K-0L KD-0 KD Page F0 F00 F00 F00 F00 F00 F090 F00 F00 F00 F00 F00 F090 F09000 F09000 F09000 F09000 F09000 F09000 F09000 F000 F000 F000 F000 F000 F0009 F00 F00 F00 F00 F00 F00 F009 F0 F0 F00 F00 F00 F00 F00 F00 F009 F0 F000 F000 F00 F000 F000 F000 F000 F00 F00 F0 F000 F0 KD-0. KD-0 KD-0 KD-0 KD-0 KD-0 KD-0M HFIP-0 HFIP-0 HFIP-0 HFIP-0 HFIP-0 HFIP-0M EV-000 EV-G EV-000 AC EV-G AC EV-00 EV000AC-F EV-G ACC H-00 H-00 H-00 H-00 H-00 H-00M H-00. K-00 K-00 K-00 K-00 K-00 K-00M K-00 K-00. KF-00 KF-00 KF-00 KF-00 KF-00 KF-00 KF-00M KF-00. KF-00 KF-00 KF-00. KF-00 KF-00 CM- PH- QA- DEAEN-T DEAE-B SP-0N SP-B Page F0 F F090 F000 F00 F00 F000 F00 F00 F00 F00 F00 F F0 F00 F00 F0 F00 F0 F0 F00 F00 F00 F00 F0 F0 F0 F0 F90 F900 F900 F90 F90 F90 F90 F90 F90 F90 F90 F90 F000 F000 F0009 F000 F00 F00 F00 F000 F00 F00 F0 F0 F0 F0 F0 SP- DEAE- AT-0MS UT-0. UT-0M UT-0 HT-0 HT-0 HT-0 HT-0 HT-0M HT-0 P-S WA- KS-0 KS-0 KS-0 KC- KC- E KS-0 KS-0 KS-0 KS-0 SH0 SH SC0 SC SP00 MN- SC0-F SB-0 HQ SB-0. HQ SB-0 HQ SB-0 HQ SB-0 HQ SB-0 HQ SB-0M HQ SB-0 HQ LB-0 LB-0M KS-00 KS-00 KS-00 KS-00 KS-00 KS-00 KC-0 KW-00. KW-00 KW-00 DE-0 DE-0 DC-0 DM-0 DC-0 Page Index by Index by Index by

84 Page Page Page F0 F0 F0 F0 F0 F0 F0 F0 F000 F00 F00 F00 F00 F0 F0 F0 F000 F00 F00 F00 F00 F00 F000 F00 F00 F00 F00 F00 F00 F009 F000 F00 F00 F00 F0000 F0000 F0000 F000 F0000 F0000 F000 F00090 F000 F000 F00 F00 F000 DM-0 SB-00 SB-00. SB-00 SB-00 SB-00 SB-00 SB-00M QA-0 DEAE-0 SP-0 CM-0 QA-0 DEAE-0 SP-0 CM-0 CM D CM E CM D CP D CP E CP D SIL D SIL E C D C E C D C E CN D CN E NH D NH E NPE D PYE D KS-G B (KS-LG) KC-G B (KC-LG) KS-G B (KS-G) KS-0G KC-G B (KC-G) SH-G SP-G B (SP-G) SC-G B (SC-LG) SZ-G SCG A (SC-G) KW-G B (KW-G) KW00G-A DS-G F000 F00 F000 F00 F000 F00 F0090 F009 F000 F000 F00 F000 F0000 F000 F0000 F000 F000 F000 F000 F000 F000 F0009 F00 F00 F0000 F000 F00 F000 F0900 F090 F0900 F090 F090 F09 F09 F090 F09 DE-G A (DE-G) DE-G A (DE-SG) DM-G A (DM-G) DM-G 0C (DM-LLG) DC-G A (DC-G) DC-G 0C (DC-LLG) DE-G B (DE-LG) DE-G 0C (DE-LLG) P-G Y-G WA-G AT-G KF-G A (KF-G) H-G B (H-G) IA-G K-G A (K-G) DC-G B (DC-LG) DM-G B (DM-LG) KF-G B (KF-LG) K-G B (K-LG) KF-G 0C (KF-LLG) K-G 0C (K-LLG) KD-G A (KD-G) T-G HFIP-G B (HFIP-LG) NN-G HFIP-G A (HFIP-G) YS-G CRX-G KF-00D UT-G HT-G SB-G B (SB-G) SB-0G LB-G B K-00D SB-G B (SB-LG), ,0,0 9 F09 F09 F090 F090 F090 F090 F090 F09 F090 F09 F09 F09 F09 F000 F000 F000 F000 F00 F00 F00 F009 F000 F00 F00 F00 F000 F000 F00 F00 F00 F000 F00 F00 F00 F00 F00 F00 F0 F0 F0 F00 F00 F009 F000 F00 F00 F000 F00 F00 F00 F00 F00 KW-G B (KW-LG) RP-G QA-LG DEAE-LG SP-G B (SP-LG) CM-LG YK-G NI-G SI-90G LF-G SI-0G SI-9G SI-9G ODP-0G A CP-0G A CP-0G A ODP-0G B NHP-0G A NHP-0G B GF-G B GS-G B GS-0G B GS-0G B ODP-0G A ODP-0 B NHP-0G A NHP-LF VG-0G A VG-0G A VT-0G A NHP-0G A ODP-0G A CP-0G B CP-0G B PAE-G PAE-G AC ODP HPG-A ODP HPG-A ODP-0G A ODP HPG-A GS-0 0F GS-0 0F GS-0 0F GS-0 0F PAE-000 PAE-000 AC GS-0 0F GS-0 0F GS-0 0F GSM-00 0F GS-0 0G GS-0 0G ,,

85 Page F00 F00 F00 F009 F000 F00 F000 F000 F000 F00 F000 F00 F000 F000 F99000 F990 F990 F990 F990 F990 F990 F990 F990 F99 F99 F99 F990 F990 F9990 F0000 F0000 F0000 F0000 F0000 F0000 F000 F0000 F0000 F000 F000 F000 F000 F000 F F00900 F00 F9 F9 F99 F9 F00 F000 F000 F00 F000 GS-0 0G GS-0 0G GS-0 0G GS-0 0G GS-0 0G GSM-00 0G ODP-0 0E CP-0 0E CP-0 0E ODP-90 0F NHP-0 0E NHP-90 0F ES-0C 0C ES-0N 0C KW-0. KW-0 KW-0 KW0.-F KW0-F KW0-F KW0-F Y- I-A NI- SI-90 E SI-0 E T- SI- E SI- D DS- DM- DC- DE- DE- DE- DS- SZ SC NN- NN- NN- JJ-0 D JJ-0 D RP- DE-L APG-9 APA-9 ADS-9 AWG-9 ACH-9 YK- YS-0 CRX- CDBS- CM 0E Page F00 F000 F00 F00 F00 F000 F00 F00 F00 F00000 F0000 F0000 F0000 F0000 F0000 F0000 F0000 F0000 F000 F000 F000 F0000 F000 F000 F0000 F000 F000 F000 F000 F000 F000 F000 F000 F000 F0009 F00 F00 F0 F0 F00 F00 F00 F00 F00 F00 F00 F00 F000 F000 F000 F000 F000 F000 F0009 F000 CM 0E SIL 0E SIL 0E C 0E C 0E NH 0E NH 0E C 0E C 0E GF-0 HQ GF-0 HQ GF-0 HQ GF-0 HQ GF-M HQ GS-0 HQ GS-0 HQ GS-0 HQ GS-0 HQ GF-0 E PAE-00 PAE-00 AC GF-0 B GF-0 D GF-0 D GF-0 D GF-0 E ODP-0 E ODP-0 D ODP-0 E ODP-0 D CP-0 E CP-0 D CP-0 E CP-0 D ODP-0 D ODP-0 D ODP-0 E ODP-0 B ODP-0 D ODP HP-B ODP HP-D ODP HP-E ODP HP-B ODP HP-D ODP HP-B ODP HP-D ET-RP D NHP-0 E NHP-0 D NHP-0 B NHP-0 D NHP-0 E NHP-0 B NHP-0 D NHP-0 E Page F00 F00 F000 F000 F000 F000 F000 F000 F0 F0 F0 F0 F0 F0 F F F F F0 F0 F0 F0 F0 F90 F00000 F000 F000 F00 F00 F00 F00 F00000 F0000 F000 F000 NHP-0 B NHP-0 D VG-0 E VG-0 D VG-0 D VT-0 D ES-0C C ES-0N C GS0A-E GS0A-D GS0A-B GF0A-E GF0A-D GF0A-D KW0.-D KW0.-B KW0-D KW0-B ODP0-D ODP0-B DE-E DE-B NN-D DEAE9A-D P- FL- FL- filter SL-0 SM-0 SH- M- PK-A P HLD PK-A P GF-A , 0, Index by

86

87 You can receive support by Shodex worldwide. Languages other than English are available now. Please contact Shodex website of your area. Showa Denko Europe GmbH Service area : Europe Africa Middle East Russia Showa Denko America, Inc. Service area : North America Latin America Correspondence language : German English Correspondence language : English French Spanish Portuguese URL URL Shoko Co., Ltd. Service area : Japan Correspondence language : Japanese URL Shodex China Co., Ltd. Service area : China Hong Kong Macau Showa Denko K.K. Service area : Taiwan Correspondence language : Chinese (Simplified Chinese character) Correspondence language : Japanese English URL URL Shoko Korea Co., Ltd. Service area : Korea Correspondence language : Korean (Hangul alphabet) URL Showa Denko Singapore (Pte) Ltd. Service area : Southeast Asia India Oceania Correspondence language : English URL