COSMOSIL HILIC. HPLC Column for Hydrophilic Interaction

Transcription

1 PLC Column for ydrophilic Interaction CSMSIL ILIC Triazole bonded stationary phase Enhanced hydrophilic interaction Excellent separation for organic acids Silica Gel Average Particle Size Average Pore Size Stationary Phase Main Interaction igh Purity Porous Spherical Silica approx. 120 Triazole ydrophilic Interaction Anion-exchange N N Triazole N Comparison with C18 Columns 18 deterioration. CSMSIL ILIC without Ion-pair Reagents CSMSIL 5C18-PAQ without 30 UV 210 nm 1. Glycine 2. Glycylglycine 2.0 μl (5.0 mg/ml) (0.125 mg/ml) with Ion-pair Reagents Acetonitrile : 10 mmol/l C3CN4 = 60 : mmol/l Phosphatase Buffer (p2.5) 20 mmol/l Phosphatase Buffer (p2.5) - 5 mmol/l Sodium 1-ctanesulfonate (Ion-pair Reagent) Different Interactions Separation of Anionic Compounds CSMSIL ILIC M company (ILIC column) T company (Amide column) W company (ILIC column) Acetonitrile : 10 mmol/l Phosphate Buffer (p7.0) = 50 : UV 210 nm 1. xamic Acid 2. xalic Acid 1.0 μl (0.4 mg/ml) (2.0 mg/ml) N 2 1. xamic Acid 2. xalic Acid

2 Different Interactions Separation by ydrophilic Interaction ydrophilic Interaction (Low) Ionic Interaction (Low) ydrophilic Interaction (Low) Ionic Interaction (igh) ydrophilic Interaction (igh) Ionic Interaction (Low) Suppress ionic interaction by increasing salt concentration Enhance hydrophilic interaction by increasing organic solvent concentration. Acetonitrile : 10 mmol/l C3CN4 = 50 : 50 Acetonitrile : 100 mmol/l C3CN4 = 50 : 50 Acetonitrile : 100 mmol/l C3CN4 = 80 : UV 245 nm 1. Isoascorbic AcidErythrbic Acid 2. Ascorbic Acid 2.0 μl (0.5 mg/ml) (0.5 mg/ml) 1. Isoascorbic Acid Erythorbic Acid 2. Ascorbic Acid Separation of Acidic, Basic and Neutral Compounds meso (Basic/ CSMSIL ILIC M companyilic column) (Basic/Neutral) = 1.48 (Acidic/Neutral) = 4.36 (Basic/Neutral) = 4.45 (Acidic/Neutral) = 1.71 Acetonitrile : 50mmol/l C3CN4 = 80 : ELSD 1. meso-erythritol 2. Tris 3. Glyceric Acid 3.0 μl (1.0 mg/ml) (1.0 mg/ml) (2.0 mg/ml) T company (Amide column) (Basic/Neutral) = 3.07 (Acidic/Neutral) = N C W company (ILIC column) (Basic/Neutral) = 5.03 (Acidic/Neutral) = meso-erythritol Neutral 2. Tris Basic 3. Glyceric Acid Acidic

3 Application Data Melamine Acetonitrile : 10 mmol/l C3CN4 = 70 : UV 225 nm 1. Melamine (0.1 μg) 2. Ammeline (0.075 μg) 3. Cyanuric Acid (0.75 μg) 4. Ammelide (0.05 μg) Allantoins Acetonitrile : Phosphate Buffer (p7.0) = 70 : 30 UV 210 nm 1. Allantoin 2. Allantoic Acid BCAA (Amino Acid Branched-chain) Juice Components Acetonitrile : 10 mmol/l C3CN4 = 85 : ELSD (Atten = 8, Gain = 6) 1. Leucine (1.0 mg/ml) 2. Isoleucine (1.0 mg/ml) 3. Valine (1.0 mg/ml) 3.0 μl Acetonitrile : 10 mmol/l Phosphate Buffer (p7.0) = 50 : 50 UV 210 nm 1. Ascorbic Acid (0.5 mg/ml) 2. Malic Acid (1.0 mg/ml) 3.0 μl Phosphorylated Saccharides Water-soluble Vitamins Acetonitrile : 20 mmol/l Phosphate Buffer (p7.0) = 60 : 40 RI 1. Glucose (2 mg/ml) 2. α-d-glucose-1-phosphate (10 mg/ml) 3. D-Glucose-6-Phosphate (10 mg/ml) 5.0 μl 1. Nicotinamide 2. Pyridoxine (Vitamin B6) 3. Vitamin B2 (Riboflavin) 4. Nicotinic Acid 5. D-Pantothenic Acid 6. L(+)-Ascorbic Acid Acetonitrile : 100 mmol/l C3CN4 = 80 : 20 UV 220 nm 0.5 ml (0.25 mg/ml) (0.50 mg/ml) (0.50 mg/ml) (0.25 mg/ml) (6.25 mg/ml) (1.75 mg/ml) 0.5μl ydrophilic Drugs Acetonitrile : 10 mmol/l C3CN4 = 90 : 10 UV 254 nm Famotidine (0.25 mg/ml) 2.0 μl Natural Products Acetonitrile : 10 mmol/l C3CN4 = 80 : 20 UV 220 nm L-(+)-Ergothioneine (1.0 mg/ml) 1.0 μl

4 Combination with C18 Columns CSMSIL 5C18-AR-II A: Acetonitrile : 0.1%TFA = 10 : 90 B: Acetonitrile : 0.1%TFA = 30 : 70 B: 020 min Linear Gradient 30 UV 220 nm 3,4, CSMSIL ILIC Acetonitrile : 10 mmol/l Phosphate Buffer (p7) = 70 : UV 220 nm 1. Angiotensin II, [Sar1, Thr8] 2. Angiotensin II, [Sar1, Ala8] 3. Angiotensin II, Des-Asp1- [Ile8] 4. Angiotensin II, [Asn1, Val5] 5. Angiotensin II, [Sar1, Ile8] 6. Angiotensin II, [Val5] 7. Angiotensin II, (uman) Separate peak No.3-6 by CSMSIL ILIC (min) Selection of 3CN 3CN 3CN (not eluted) CSMSIL Chromatogram Index CSMSIL Chromatogram Index : Allantoin CAS No.: [ ] Molecular formula:c 4 6 N 4 3 Column: ILIC Ammonium acetate =90/10 Flow rate: : 30 C : UV220 nm Attenuation: aufs conc.: 1.0mg/ml Injection volume: 1.0ml Retention time: 8.75min Capacity factor: 2.02 N N N N 2 rdering Information Product Name Product Number CSMSIL ILIC Product Name Product Number CSMSIL ILIC CSMSIL ILIC TEL : +81-(0) FAX. 1205

5 Triazole CSMSIL Application Data Column: Column size: Ammonium Acetate = 80/20 Flow rate: : 40 C : UV249nm : 1; Creatinine (0.1mg/ml) 2; 2'-Deoxyguanosine (0.1mg/ml) 3; 8-ydroxy-2'-Deoxyguanosine (0.1mg/ml) 4; 8-ydroxy Guanosine (0.1mg/ml) Inj. Vol: 1.0µl ILIC (5m) (4.6mmI.D.-250mm) 2.5ILIC (2.5m) (3.0mmI.D.-100mm) Analysis time: 1/6 CSMSIL Application Data CSMSIL ILIC Competitor A Competitor B Competitor C Column: Ammonium Acetate = (CSMSIL) = 70/30 (thers) = 80/20 Flow rate: : 30 C : UV225nm (Triazole type) (Sulfobetaine type) (Silica type) (Amide type) : 1; Melamine (0.10mg/ml) 2; Ammeline (0.075mg/ml) 3; Cyanuric Acid (0.75mg/ml) 4; Ammelide (0.05mg/ml) Inj. Vol: 1.0µl Ionic Interaction (igh) + ydrophilic Interaction (Low) ydrophilic Interaction (igh) + Ionic Interaction (Low) Enhance hydrophilic interaction by increasing acetonitrile concentration : 1. Isoascorbic Acid Erythorbic Acid Suppress ionic interaction by decreasing salt concentration 2. Ascorbic Acid Acetonitrile:10 mmol/l C 3CN 4=50:50 Acetonitrile:100 mmol/l C 3CN 4=80:20

6 CSMSIL Application Data CSMSIL Application Data Column: 2.5ILIC Column size: 2.0mmI.D.-150mm Phosphate buffer(p7.0) = 50/50 Flow rate: 0.4 ml/min : 40 C : UV210nm : 1; Glutamic Acid (2.5mg/ml) 2; Succinic Acid (4.0mg/ml) 3; Inosine-5'-monophosphate (0.25mg/ml) 4; Guanosine-5'-monophosphate (0.25mg/ml) Inj.Vol.: 1.0µl Column: 2.5ILIC Column size: 2.0mmI.D.-50mm Mobile phase: Acetonitrile/ 20mmol/l Phosphate buffer(p7.0) = 50/50 Flow rate: 0.4 ml/min : 40 C : UV260nm : 1; Adenosine-5'-monophosphate (0.25mg/ml) 2; Adenosine-5 -diphosphate (0.50mg/ml) 3; Adenosine-5 -triphosphate (0.50mg/ml) Inj.Vol.: 0.5µl CSMSIL Application Data CSMSIL Application Data Column: 2.5ILIC Column size: 3.0mmI.D.-100mm Ammonium Acetate = 85/15 Flow rate: : 30 C : ELSD Column: 2.5ILIC Column size: 2.0mmI.D.-150mm Ammonium Acetate = 50/50 Flow rate: 0.4 ml/min : 40 C : UV210nm : 1; Taurine (1.0mg/ml) 2; ypotaurine (1.0mg/ml) Inj.Vol.: 2.0µl : Inj.Vol.: 1; Trichloroacetic Acid (1.0mg/ml) 2; Dichloroacetic Acid (2.0mg/ml) 3; Chloroacetic Acid (5.0mg/ml) 0.5µl

7 I. PLC Columns II. UPLC Columns III. Preparative Packing Materials IV. Related Products V. Applications VI. Technical Notes CSMSIL ILIC ILIC Comparison with C18 Column: Mobile phase: (ILIC) Acetonitrile/10mmol/l Ammonium Acetate = 60/40 (C 18 -PAQ) 20mmol/l Phosphate buffer(p2.5) -(Ion pair)5mmol/l Sodium 1-ctanesulfonate Flow rate: : 30C : UV210nm : 1; Glycine (5.0mg/ml) 2; Glycylglycine (0.125mg/ml) Inj.Vol: 2.0l ILIC 5C 18 -PAQ without Ion pair reagent 5C 18 -PAQ with Ion pair reagent VII. Index 36

8 CSMSIL Application Data Column: ILIC Ammonium Acetate = 70/30 Flow rate: : 30C : UV225nm : 1; Melamine (0.1g) 2A 2; Ammeline (0.075g) 075 3; Cyanuric Acid (0.75g) 4; Ammelide (0.05g) - CSMSIL Chromatogram Index : Allantoin CAS No.: [ ] Molecular formula: C 4 6 N 4 3 Column: ILIC Ammonium acetate =90/10 Flow rate: : 30C Dt : ti UV220 nm Attenuation: aufs conc.: 1.0mg/ml Injection volume: 10l 1.0l Retention time: 8.75min Capacity factor: 2.02 rdering Information AP-1076 CSMSIL Application Data Column: ILIC Ammonium Acetate = 85/15 Flow rate: : 30C : ELSD : 1; L-Leucine (3.0g) 2; L-Isoleucine I l i (3.0g) 3; L-Valine (3.0g) CSMSIL Chromatogram Index : L-(+)-Ergothioneine CAS No.: [ ] Molecular formula: C 9 15 N 3 2 S Column: ILIC Ammonium acetate =80/20 Flow rate: : 30C Dt : ti UV220 nm Attenuation: aufs conc.: 1.0mg/ml Injection volume: 10l 1.0l Retention time: 10.29min Capacity factor: * AP I. PLC Columns II. UPLC Columns III. Preparative IV. Related Products V. Applications VI. Technical Notes VII. Index Packing Materials