TCI Dual ~Mixed-Mode Mode column~

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1 TCI Dual ~Mixed-Mode Mode column~ TCI Chromatography Department

2 Mixed-Mode Column Target compound is basic Choose DS + Cation-Exchange Type Target compound is acidic Choose DS + Anion-Exchange Type o ion-pair reagent is required Hydrophobic compounds also can be retained on mixmode columns The packing material has both ion-exchange and DS phases ot a mixture of packing materials of an ion-exchange and DS

3 Product Line DS+Cation-Exchange Type (For basic compound) DS+Anion-Exchange Type (For acidic compound) Column TCI Dual DS-CX0 TCI Dual DS-CX5 TCI Dual DS-CX0 TCI Dual DS-AX0 TCI Dual DS-AX0 Kaseisorb LC DS- SAX Ion-Exchange Strong Medium Weak Strong Medium Weak Usable mobile phase ph range.5~7.5.5~7.5.5~7.5.5~7.5.5~7.5.5~7.5 Ion-Exchange ph range.5~7.5.5~ ~7.5.5~7.5.5~7.5.5~7.5 Silicagel particle size and pore diameter 5μm, nm 5μm, nm 5μm, nm 5μm, nm 5μm, nm 5μm, nm

4 Choose Cation-Exchange Type Basic Compound TCI Dual DS-CX5 Elution Power Up Elution Power Down TCI Dual DS-CX0 TCI Dual DS-CX0 Retention Power of Basic Compound TCI Dual DS-CX0>TCI Dual DS-CX5>TCI Dual DS-CX0

5 Retention Power : Cation-Exchange Retention Power: TCI Dual DS-CX0>TCI Dual DS-CX5>TCI Dual DS-CX0 Column Size : 4.6 mmi.d. 50 mm Mobile Phase : CH 3 C / 0mmol/L KH P 4 (ph3.0) = 40/60 Flow Rate :.0 ml/min Detection : UV 30 nm Temperature : 5 uv Sample : Melamine TCI Dual DS-CX TCI Dual DS-CX TCI Dual DS-CX min

6 Choose Anion-Exchange Type Acidic Compound Kaseisorb LC DS-SAX Super Elution Power Up TCI Dual DS-AX0 Elution Power Up TCI Dual DS-AX0 Elution Power of Acidic Compound TCI Dual DS-AX0>TCI Dual DS-AX0>Kaseisoeb LC DS-SAX

7 Retention Power : Anion-Exchange Column Size :4.6mmI.D. 50mm Temperature:30 Flow Rate:.0 ml/min Mobile Phase:CH 3 C/50mmol/L HCH 4 (ph3.5 ) =50/50 Sample:.Benzene.Toluene 3.Ethylbenzene 4.Benzenesulfonic Acid 5.p-Toluenesulfonic Acid 6.4-Ethylbenzenesulfonic Acid Kaseisorb LC DS-SAX TCI Dual DS-AX0 TCI Dual DS-AX min

8 Mobile Phase Adjustment. Changing the rganic Solvent Ratio rganic solvent ratio UP Elution Power UP. Changing the Buffer Concentration Buffer concentration UP Elution Power of ionic compounds UP 3. Changing the ph Dissociate ph of ionic compounds Ionic interaction UP

9 .Changing the rganic Solvent Ratio Column:TCI Dual DS-AX0 4.6mmI.D. 50mm Temperature:5 Flow Rate:.0 ml/min. 5 -Adenylic Acid (Acidic compound). Methyl Benzoate (Hydrophobic compound) Methanol 30% Mobile Phase 0mmol/L Phosphate buffer 70% Time(min) Methanol 60% 0mmol/L Phosphate buffer 40%

10 .Changing the Buffer Concentration Column:TCI Dual DS-AX0 4.6mmI.D. 50mm Temperature:5 Flow Rate:.0 ml/min. 5 -Adenylic Acid (Acidic compound). Methyl Benzoate (Hydrophobic compound) Methanol 30% Mobile Phase 0mmol/L Phosphate buffer 70% Time(min) Methanol 30% 50mmol/L Phosphate buffer 70%

11 3.Changing the ph Column:TCI Dual DS-AX0 4.6mmI.D. 50mm Temperature:30 Flow Rate:.0 ml/min Mobile Phase:CH 3 C/50mmol/L Buffer=50/50 ph7.0:k HP 4 (ph7.0, H 3 P 4 ) ph5.0:ch 3 CH 4 (ph5.0,ch 3 CH) ph3.5:hch 4 (ph3.5, HCH) Sample:.Benzene.Toluene 3.Ethylbenzene 4.Benzenesulfonic Acid 5.p-Toluenesulfonic Acid 6.4-Ethylbenzenesulfonic Acid uv ph7.0 ph5.0 ph min 6

12 Sample pka Acetic Acid CH 3 CH CH 3 C - + H + ph.75 Increase hydrophobicity pka:4.75 ph6.75 Increase ionicity Choose Reversed Phase Mode Choose Mixed-Mode Glucuronic Acid pka:3. Polar compound Increase ionicity Inapplicable Reversed Phase Mode Choose Mixed-Mode

13 Gradient Elution Change organic solvent ratio Serotonin Metabolites Column : TCI Dual DS-CX5 4.6 mmi.d. 50 mm Mobile phase : A) CH 3 C / 50mmol/L HCH 4 (ph3.5,hch) = 0 / 90 B) CH 3 C / 50mmol/L HCH 4 (ph3.5,hch) = 70 / 30 A to 50% B in 30min. CH Flow Rate :.0 ml/min.. CH 3. H H H Temperature : 5 H Detection : UV 54 nm H H Sample : H 4. CH 5. H. 5-Hydroxy-L-tryptophan 6. H H. 5-Hydroxy-3-acetic Acid CH 3 3. Acetylserotonin H 90 H H L-Tryptophan Serotonin Melatonin H H min

14 Gradient Elution Change buffer concentration Adenosine Phosphates Column : TCI Dual DS-AX0 (4.6 mmi.d. X 50 mm) Mobile phase : A) CH 3 H / 0mmol/L K HP 4 (ph7.0) = 0 / 80 B) CH 3 H / 50mmol/L K HP 4 (ph7.0) = 0 / % B (0-30min) mau Flow rate :.0 ml/min Detection : UV 54 nm Temperature : 40 C Sample :. AMP. camp ADP 4. ATP min

15 Sialic Acids Column size : 4.6 mmi.d. 50 mm Mobile phase : CH 3 C / 0mmol/L AcH 4 (AcH, ph5.0) = 5/95 Flow rate :.0 ml/min uv Kaseisorb LC DS-SAX Super Detection : UV 0 nm Temperature : Sample : Glycolylneuramic acid(eu5gc) H HH C H H H H H CH. -Acetylneuramic acid(eu5ac) H H H H H H CH TCI Dual DS-AX0 TCI Dual DS-AX0 Kaseisorb LC DS min

16 Galacturonic Acid, L-Ascorbic Acid Column : 4.6 mmi.d. 50 mm Mobile Phase : CH 3 H / 0mmol/L K HP 4 (ph7.0, H 3 P 4 ) = 0/90 Flow Rate :.0 ml/min Detection : UV 0 nm Temperature : 40 Sample :. Galacturonic Acid Kaseisorb LC DS-SAX Super TCI Dual DS-AX0. L-Ascorbic Acid H, TCI Dual DS-AX0 H Kaseisorb LC DS 000 H H min

17 Column Size : 4.6 mmi.d. X 50 mm Mobile phase : CH 3 C / H / H 3 P 4 = 5 / 95 / 0. Detection : UV 0 nm Flow rate :.0 ml/min Temperature : 5 Sample :. L-Glutamic Acid GABA TCI Dual DS-CX5 H H H. γ-aminobutyric acid (GABA) Kaseisorb LC DS 000 H H min

18 Basic Drugs Column : TCI Dual DS-CX0 4.6 mmi.d. 50 mm Mobile phase : CH 3 C / 50mmol/L CH 3 CH 4 (ph5.0,ch 3 CH) = 80 / 0 Flow Rate :.0 ml/min Detection : UV 0 nm Temperature : 40 Sample :. icardipine. Verapamil 3. Alprenolol 4. Clomipramine 5. Chlorpromazine mau H 3 C H 3 C H CH 3 H CH 3 H 3 C H 3 C C CH 3 H 3 C CH 3 CH 3 CH H H Cl S Cl min

19 Amino Acids Column : TCI Dual DS-CX5(4.6 mmi.d. 50 mm) Mobile Phase : A) CH 3 C / 5 mmol/l KH P 4 (ph3.0, H 3 P 4 ) = 5/95 B) CH 3 C / 50 mmol/l KH P 4 (ph3.0, H 3 P 4 ) = 40/ %B(5-0min.) uv Flow Rate :.0 ml/min Detection : UV 0 nm 0000 Temperature : Aspartic Acid Aspartic Acid Alanine Alanine S H Methionine H Methionine Phenylalanine Phenylalanine Arginine Arginine min

20 Anti-diabetic agent Column : TCI Dual DS-CX5 4.6 mmi.d. 50 mm Mobile phase : CH 3 C / 50mmol/L KH P 4 (ph3.0,h 3 P 4 ) = 50 / 50 Flow Rate :.0 ml/min Detection : UV 30 nm Temperature : 40 mau Sample : 450.Isobutyl 4-Hydroxybenzoate H C CH 3 CH CHCH Metformin Hydrochloride H H H CH 3 H HCl CH min

21 Column : TCI Dual DS-CX0 4.6 mmi.d. 50 mm Mobile Phase : CH 3 H / 0mmol/L K HP 4 (ph6.0,h 3 P 4 ) = 0/90 Flow Rate :.0 ml/min Detection : UV 0 nm Temperature : 30 Sample :. Allantoin Allantoin, D-Panthenol H H H H. D-Panthenol H 3 C CH 3 H H H H time(min)

22 Levulinic Acid, Methyl Levulinate Column : TCI Dual DS-AX0 4.6mmI.D. 50mm Mobile Phase : CH 3 C / 0 mmol/l K HP 4 (ph7.0,h 3 P 4 ) = 0 / 80 Flow Rate :.0 ml/min Detection : UV 0 nm Temperature : 5 Sample :.Levulinic Acid H 3 C.Methyl Levulinate H H 3 C CH min

23 ucleotides Column Size : 4.6 mmi.d. 50 mm Mobile Phase : CH 3 H / 0mmol/L K HP 4 (ph 7.0,H 3 P 4 ) = 0 / 80 Flow Rate :.0 ml/min Detection : UV 54 nm Temperature : 40 Sample :. CMP. UMP 3. IMP 4. GMP 5. AMP 3, time(min) TCI Dual DS-AX time(min) TCI Dual DS-AX0

24 Halide Ions Coulmn : Kaseisorb LC DS-SAX Super 4.6mmI.D. 50 mm Mobile Phase:CH 3 C/ 50mmol/L CH 3 CH 4 (ph5.00, CH 3 CH) = 50/50 Detection Flow Rate : UV 0 nm, Corona CAD :.0 ml/min Temperature :40 Sample Size :5 ul Sample: Potassium cloride Potassium bromide Potassium Iodide K + Cl - Br - I - Corona CAD min

25 Alkali Halides CI - Li + Coulmn : TCI Dual DS-CX5 4.6 mmi.d. 50 mm Mobile Phase : CH 3 C/ 50mmol/L CH 3 CH 4 (ph5.00, CH 3 CH) = 50/50 Detection : UV 0 nm, Corona CAD Temperature : 40 Flow Rate :.0 ml/min Sample Size : 5uL Sample : Lithium chloride Sodium chloride Potassium chloride a + K min

26 Ionic Liquid Coulmn : TCI Dual DS-CX0 4.6mmI.D. 50 mm Mobile Phase : CH 3 C/ 50mmol/L CH 3 CH 4 (ph5.00, CH 3 CH) = 0/90 Detection : UV 0 nm, Corona CAD Flow Rate :.0 ml/min Temperature : 40 Sample Size : 5uL Sample : -Butyl-3-methylimidazolium chloride Cl - + (CH ) 3 CH min

27 Product Type Column DS+Cation-Exchange For Basic compounds analysis DS+Anion-Exchange For Acidic compounds analysis Retention Power Strong Medium Weak Strong Medium Weak.0 50mm S3705 S3765 S375 S3735 S375 S8.0 00mm S3707 S3767 S377 S3737 S377 S mm S370 S376 S37 S373 S37 S mm S3703 S3763 S373 S3733 S373 S mm S3704 S3764 S374 S3734 S374 S mm S3709 S3769 S379 S3739 S379 S mm S3700 S3760 S370 S3730 S370 S mm S370 S376 S37 S373 S37 S mm S3706 S3766 S376 S3736 S376 S mm S3708 S3768 S378 S3738 S378 S836 Cartridge type guard column [Holder:S450 ] 3.0 5mm TCI Dual DS-CX0 TCI Dual DS-CX5 TCI Dual DS-CX0 TCI Dual DS-AX0 TCI Dual DS-AX0 Kaeisorb LC DS-SAX Super S3770 S377 S377 S3775 S3774 S3773

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