1 hplc Bonded Phases Phase Description Structure Phase Description Structure Si Silica NH2 APS, Amino, Amino Propyl Silyl Classic normal phase material. Suitable Si OH Reversed phase, normal phase, or weak for separating polar non-ionic organic anion exchange material. compounds. Reversed phase: useful for separating C1 TMS, SAS, Trimethyl carbohydrates. Normal phase: alternative selectivity to Si CH2CH2CH2NH2. Reversed phase material. Unique selectivity Si CH3. silica, not deactivated by small amounts for polar and multifunctional compounds. of water. C2 RP-2, Dimethyl Ion exchange: weak anion-exchanger when Reversed phase material. Si C2H5 used with buffers; separates anions and organic acids.
2 C3 Propyl NO2 Nitro Reversed phase material. Used in Hydro- Si C3H7 Normal phase material. Separates aromatic phobic Interaction Chromatography (HIC) of Si NO2. compounds and compounds with double proteins and peptides. bonds. C4 Butyl OH Diol, Glycerol Reversed phase material. Useful for HIC Reversed phase or normal phase material. and ion-pairing chromatography. Offers less Used for Gel Filtration Chromatography (GFC) Si CH2CH2CH2 OCH2 CHCH2. retention than C8 and C18 Phases for non- Si C4H9 of proteins and peptides. Normal phase: polar solutes. When Bonded to 300 silica, it similar selectivity to silica, not deactivated by OHOH. is an ideal phase for analyzing and purifying small amounts of water.
3 Large proteins and macromolecules. Phenyl C6H5. C5 Pentyl Reversed phase material. Useful for analyz- Si CH2CH2CH2. Reversed phase material. Useful for reversed ing aromatic compounds. When Bonded to phase separation of hydrophobic proteins Si C5H11 300 silica, this phase is useful for HIC. when Bonded to 300 silica. More hydrolyti- Phenyl-Ether C6H5(C3H6O Linker). cally stable than C4. Reversed phase material. Used to separate Si CH2CH2CH2O. C6 Hexyl extremely polar aromatic compounds. Alter- Reversed phase material. Useful for ion- Si C6H13 native selectivity to Phenyl-hexyl and classic pairing chromatography. phenyl Phases . C8 MOS, RP-8, LC8, Octyl Phenyl-Hexyl C6H5 (C6H12 Linker).
4 Reversed phase material. Similar selectivity Si C8H17 Reversed phase material. Uses a patented to C18 but less retentive. Wide applicability hexyl linker as opposed to traditional propyl. Si CH2CH2CH2CH2CH2CH2. ( , pharmaceuticals, nucleosides, steroids). Offers selectivity of a phenyl phase with C12 Dodecyl greatly enhanced stability. Reversed phase material. Shorter chain PFP C6F5 F F. length provides higher Bonded phase cover- Si C12H25 Reversed phase material. Useful for age than most C18 columns for sharp peak analyzing substituted aromatic compounds. Si CH2CH2CH2 F. shapes of non-polar and moderately polar Alternative selectivity to phenyl-hexyl, classic compounds. F F. phenyl Phases , and alkyl Phases .
5 C18 ODS, RP-18, LC18, Octadecyl SAX Q, Quaternary amine, Strong Base Classic reversed phase material. Most Ion-exchange material. Strong anion- retentive for non-polar solutes. Excellent for Si CH2CH2CH2N+(CH3)3. exchangers (basic) are useful for separating ion-pairing chromatography. Wide applicabil- nucleotides, nucleosides, and organic acids. ity ( , nucleosides, nucleotides, steroids, Si C18H37. SCX S, Sulfonic Acid, Strong Acid pharmaceuticals, vitamins, fatty acids, envi- ronmental compounds). When Bonded to 300 Ion-exchange material. Strong cation- Si CH2CH2CH2SO2OH. silica, this phase is perfect for separating exchangers (acidic) are useful for separating small proteins and macromolecules.
6 Organic bases. CN CPS, PCN, Cyano, Cyanopropyl, Nitrile WAX DEAE, Diethylaminoethyl, Weak Base, PEI, Polyethyleneimine Si CH2CH2N(CH2CH3)2. Reversed phase or normal phase material. Slightly polar, unique selectivity for polar Ion-exchange material. Weak anion- compounds in both reversed phase and nor- exchangers (acidic) are useful for analyzing Si CH2CH2CH2CN Si CH2CH=NH. mal phase modes. Equilibrates very rapidly, and purifying acidic proteins and peptides. suitable for gradient separations. Useful for many pharmaceutical applications ( , WCX CM, Carboxymethyl, Weak Acid tricyclic antidepressants). Ion-exchange material. Weak cation- Si CH2 COOH. exchangers (basic) are useful for analyzing and purifying basic proteins and peptides.
7 hplc Bonded Phases . |. Appendices Phenomenex 379. a selection of hplc Material Sorbent Characteristics This selection is, neither in terms of manufacturers nor in terms of their products, a complete list and the accuracy of data is not guaranteed. Phenomenex Sorbents (cont'd). Calculated*. Particle Pore Pore Surface Bonded Phase Shape/Size Size Volume Area Carbon Load Coverage End pH USP. Packing Material ( m) ( ) (mL/g) (m2/g) % ( mole/m2) Capping Range Packing PolymerX RP-1 Spher. 3, 5, 7, 10, 15 100 410 0 N/A No 0-14 L21. PrimeSphere Si Spher. 5, 10, 15 110 0 0 No L3. PrimeSphere C8 Spher. 5, 10, 15 110 10, Monomeric Yes L7. PrimeSphere C18-MC Spher. 5, 10 110 13, Monomeric Yes L1.
8 PrimeSphere C18-HC Spher. 5, 10, 15 110 20, Monomeric Yes L1. PrimeSphere C4 Spher. 5, 10 300 110 2, Monomeric Yes L26. PrimeSphere C18-MC Spher. 5, 10 300 110 7, Monomeric Yes L1. Prodigy ODS(2) Spher. 5 150 310 , Monomeric Yes L1. Prodigy C8 Spher. 5 150 310 , Monomeric Yes L7. Prodigy ODS (3) Spher. 3, 5, 10 100 450 , Monomeric Yes L1. Prodigy Phenyl (PH-3) Spher. 5 100 450 , Polymeric No L11. SphereClone Silica Spher. 3, 5, 10 80 200 - No L3. SphereClone C6 Spher. 3, 5, 10 80 200 6 Yes L15. SphereClone C8 Spher. 3, 5, 10 80 200 6 Yes L7. SphereClone ODS (1) Spher. 3, 5, 10 80 200 7 Partial L1. SphereClone ODS (2) Spher. 3, 5, 10 80 200 12 Yes L1. SphereClone NH2 Spher. 3, 5, 10 80 200 2 No L8.
9 SphereClone Phenyl Spher. 3, 5, 10 80 200 3 Partial L11. SphereClone SAX Spher. 3, 5, 10 80 200 No . Synergi Fusion-RP Spher. 100 400 12 Yes L1. Synergi Max-RP Spher. 100 400 17 Yes . Synergi Hydro-RP Spher. 100 400 19 Proprietary L1. Synergi Polar-RP Spher. 100 400 11 Proprietary . Synergi Fusion-RP Spher. 4, 10 80 475 12 Yes L1. Synergi Max-RP Spher. 4, 10 80 475 17 Yes . Synergi Hydro-RP Spher. 4, 10 80 475 19 Proprietary L1. Synergi Polar-RP Spher. 4, 10 80 475 11 Proprietary L11. Ultracarb C8 Spher. 5 60 550 14, Monomeric Yes L7. Ultracarb ODS (20) Spher. 3, 5, 7 90 370 22, Monomeric Yes L1. Ultracarb ODS (30) Spher. 5, 7 60 550 31, Monomeric Yes L1. This selection is, neither in terms of manufacturers nor in terms of their products, a complete list and the accuracy of data is not guaranteed.
10 Other Sorbents Particle Shape/Size Pore Size Pore Volume Surface Area Carbon Load Calculated* Bonded Phase Packing Material ( m) ( ) (mL/g) (m2/g) % Coverage ( mole/m2) End Capping hplc Material Sorbent Characteristics Agilent Technologies (HP). Zorbax ODS Spher. 3,5,7 70 300 20 Yes Zorbax C8 Spher. 3,5,7 70 300 12 Yes Zorbax CN Spher. 3,5,7 70 300 7 Yes Zorbax Phenyl Spher. 3,5,7 70 300 12 Yes Zorbax TMS Spher. 5 70 300 4 n/a Zorbax NH2 Spher. 5,7 70 300 4 No Zorbax Eclipse Plus C18 Spher. , , 5 95 160 8 Yes, double Zorbax Eclipse Plus C8 Spher. , , 5 95 160 8 Yes, double Zorbax Eclipse XDB-C18 Spher. , , 5, 7 80 180 10 Yes, double Zorbax Eclipse XDB-C8 Spher. , , 5, 7 80 180 Yes, double Zorbax Eclipse XDB-Phenyl Spher.