HPLC Columns

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RPP 1 - Macroporous Reversed Phase Polymer

RPP 1 is a hydrophobic copolymer based on polystyrene and divinylbenzene. The macroporous RPP 1 reversed phase resins are available as 6 µm particles within a very narrow size distribution (deviation <5%) and an exclusion limit of >106 Dalton.
The chemical inert polymer matrix guarantees chemical stability and allows applications in the pH-ranges from 0 to 14.
K' values measured on RPP 1 are rather high because of the very pure and uniform hydrophobic surface. The high efficiency (35000 plates/m) and high selectivity of the RPP 1 columns allows the separation of analysts within a few minutes. Even basic substances are separated very efficiently without any peak tailing.


RPP 1 Stationary Phase PS/DVB copolymer
Particle Size 6 µm
Pore Size 450 Å
Exclusion Limit 1.2 x 106 Dalton
Surface 100 m2/g
Pore Volume 1.3 ml/g
Max. Pressure 150 bar



Features of RPP 1
  • high selectivity
    		•
  • high resolution
    		•
  • no restriction in eluents
    		•
  • stable at low and high pH values
    		•
  • outstanding physical properties
    		•
  • chemical stability
    		•


    Stationary Phase Particle Size Pore Size Column Dimension Part No. Price in €
    RPP 1   PS/DVB 6 µm 450 Å 100 x 4.6 mm 1-100.46 290.--
    RPP 1   PS/DVB 6 µm 450 Å 100 x 2.0 mm 1-100.20 340.--



    Applications



    1 N,N-diethylaniline
    2 N,N-dimethylaniline
    3 N-methylaniline
    4 aniline    		 1 toluene
    2 ethylbenzene
    3 propylbenzene
    4 butylbenzene

    Column: RPP 1, 60 x 4.6 mm
    Mobile phase: CH3CN / H2O (85:15, v/v)
    Flow: 1.0 ml/min
    Detection: UV at 220nm





    Oligonucleotides

    With the forthcoming of the antisense technology an increasing demand of oligonucleotides is required. Separation by ion exchange chromatography requires a desalting step which follows up the purification.
    5´-DMT blocked oligonucleotides can be purified by reversed phase chromatography. No desalting step is necessary. The example demonstrates the rather effective separation of a partial protected 30 mer oligonucleotide from unblocked failure sequences.


    DMT-C TCC GTT TAC TAA GCA
    GAC CTA GGT TTT TT-3'

    Column: RPP 1, 100 x 4.6 mm
    A: 0.05 M TEAA / 10% CH3CN
    B: 0.05 M TEAA / 60% CH3CN
    Gradient: 20 min 0 - 25% B
                       25 min 50% B
                       30 min 100% B
    Flow: 0.8 ml/min
    Detection: UV at 220nm





    Basic Drugs

    Separation of basic substances on reversed phase material based on silica is often not very effective because of broad tailing and unspecific interactions with the stationary phase. RPP 1 columns show no adverse effects and the drugs shown in the figure are separated with high efficiency and high selectivity.


    1 Procainamide x HCl
    2 N-Propionyl procainamide
    3 Tropicamide
    4 Diazepam
    5 Clonazepam

    Column: RPP 1, 100 x 4.6 mm
    A: 0.1% TFA / water
    B: 0.085% TFA / CH3CN
    Gradient: 15 min 5 - 50% B
                       20 min 100% B
    Flow: 0.5 ml/min
    Detection: UV at 220nm





    Proteins and Peptides

    RPP 1 is special suited for high resolution separation of proteins and peptides.


    Protein MW Dalton pI
    1 Ribonuclease I 13,700 9.45
    2 Lysozyme 14,400 11.0
    3 Bovine Serum Albumin 67,000 4.9
    4 Myoglobin 17,800 7.3
    5 β-Lactoglobuline A + B 36,500    
    6 Ovalbumin 45,000 4.6

    Column: RPP 1, 100 x 4.6 mm
    A: 0.1% TFA / water
    B: 0.085% TFA / CH3CN
    Gradient: 10 min 5 - 35% B
                       25 min 50% B
                       35 min 100% B
    Flow: 0.5 ml/min
    Detection: UV at 220nm




    Protected peptides, used in segment condensation for synthesis of small proteins get more and more important. Nevertheless, these rather hydrophobic peptide derivatives often show poor recoveries. A completely protected crude synthetic peptide with MW 3734 Dalton is separated on RPP 1:


    Column: RPP 1, 250 x 4.6 mm
    A: 0.1% TFA / water
    B: 0.085% TFA / CH3CN
    Gradient: 40 min 5 - 65% B
                       50 min 95% B
    Flow: 1.0 ml/min
    Detection: UV at 220nm




    Methionine and cysteine containing peptides are sensitive to oxidation. The oxidized methionine derivative of synthetic VPG 2 from foot and mouth disease virus is separated on RPP 1:


    GPYAGPMERRQKPLKVKAKAPVVKE

    Column: RPP 1, 100 x 4.6 mm
    A: 0.1% TFA / water
    B: 0.085% TFA / CH3CN
    Gradient: 30 min 10 - 40% B
    Flow: 1.0 ml/min
    Detection: UV at 220nm





    Index

    Rapp Polymere GmbH
    Ernst-Simon-Str. 9
    D 72072 Tübingen
    Germany

    Phone: xx49 7071 763157
    Fax: xx49 7071 763158
    E-mail:     support@rapp-polymere.com