Macherey-Nagel Chromabond® and Chromafix®



Alumina Amino (NH2) C2 C4 C8 C18 C18 Hydra C18ec (endcapped) C18ec f (endcapped, fast flow) C18 f (fast flow) Cyano Diol (OH) Easy Florisil HR-Xpert Phenyl (C6H5) Polyamide 6 Silica Gel
PCA PSA SA SB XTR Liquid-to-Liquid Extraction

Ultra-high performance Chromabond SPE Cartridges are available in polypropylene (standard & LV) and glass tubes from 1 to 70ml sizes precision packed with a wide array of sorbents: SiOH, Alumina (A, B,N), C18, C8, C4, C2, Cyclohexyl, Phenyl, NitroPhenyl, Amino, DMA, CN, Diol, PCA, SA, PSA, SB, PS-RP, PS-OH, PS-H+, PS-Ag+, PS-Ba2+ , Florisil®, Polyamide, Phases for Environmental, and specialty adsorbents. Phases are also packed in Chromafix (luer connect) and 96 Well Plates.

Benefits of Chromabond® and Chromafix®

  • High recovery rates
  • Excellent reproducibility
  • Improves selectivity of the analysis
  • Reduces consumption of organic solvents
  • Speeds analysis time
  • Provides variety and selection for even the most specific application
  • Over 40 Phases available
  • All SPE products sealed to maintain quality and shelf life
  • Cartridges available in Polypropylene, Glass, LV (large volume), Luer Connect, and 96 well plates

Selection Guide for SPE Phases and Solvents
Sample Solubilitysoluble in water
/--------------------\
soluble in organic solvents
/----------|----------\
not ionic
|
ionic
|
|
|
|
|
|
|
Solvent aqueous
/----------------|----------------\
aqueous
/------------\
aqueous
|
organic
|
organic
|
Sample Polarity nonpolar moderately
polar
polar cationic anionic nonpolar moderately
polar
polar
Phases Recommended for AdsorptionHR-X
Easy
HR-P
C18 ec
C18
C18 Hydra
C8
C4
C6H5
CN
SiOH
NH2
CN
OH
PA
DMA
NH2
PCA
SA
PSA
SB
NH2
DMA
HR-X
Easy
HR-P
C18 ec
C18
C18 Hydra
C8
C4
C6H5
CN
SiOH
NH2
CN
OH
PA
DMA
NH2
Solvents Recommended for Elution (Selection)hexane
CH2Cl2
acetonitrile
alcohols
CHCl3
CH2Cl2
ethyl
acetate
alcohols
water
CHCl3
CH2Cl2
ethyl
acetate
alcohols
water
acids
salts
solutions
buffers
acids
salts
solutions
buffers
hexane
CH2Cl2
acetonitrile
alcohols
CHCl3
CH2Cl2
ethyl
acetate
alcohols
CHCl3
CH2Cl2
ethyl
acetate
alcohols

Molecular interactions in SPE
SPE adsorbents are most commonly categorized by the nature of their primary interaction mechanism with the analyte of interest. The three most common extraction mechanisms used in SPE are reversed phase (RP), normal phase (NP) and ion exchanger.

Typical Extraction Mechanisms

  • Reversed Phase Extraction of hydrophobic or polar organic analytes from aqueous matrix
  • Normal Phase Extraction of polar analytes from non-polar organic solvents
  • Ion Exchanger Extraction of charged analytes from aqueous or non-polar organic samples


Types of Retention Mechanisms

Nonpolar interactions
  • Silica Based: C18 ec, C18, C18 Hydra, C8,…
  • Polymer-based: HR-X, HR-P, Easy, PS-RP
  • Interactions: Hydrophobic
  • Sample: Mostly aqueous
  • Elution: Solvents with lower polarity (compared to water) MeOH, CH2Cl2, CHCl3,… hexane

Polar interactions
  • Silica Based: SiOH, CN, NH2, OH (diol), C6H5,…
  • Other: Alox, Florisil ®
  • Interactions: Hyrdogen bonds, dipole-dipole, and π-π interactions
  • Sample: Mostly organic
  • Elution: Polar solvents (compared to sample solvent) (nonprotic) ethers, ketones (MTBE, THF, acetone,…) CH2Cl2, CHCl3

Cation Exchangers
  • Silica Based: SSA (SCX), PCA (WCX), PSA
  • Polymer-based: HR-XC, HR-XCW, PS-H+
  • Interactions: Between charged analytes and functional groups of cation exchanger
  • Sample: Aqueous (pH 3-5)
  • Elution: Acidic- pH 2 (e.g. HCl, or 20% AcPH in MeOH/acetonitrile)
    Basic- pH 8-9 (e.g. 5% NH3 in MeOH/acetonitrile)
    Solvents or buffers with higher ionic strength and counter ions with high selectivity (e.g. Ca2+)

Anion Exchangers
  • Silica Based: SB (SAX), NH2, DMA
  • Polymer-based: HR-XA, HR-XAW, PS-OH-,…
  • Interactions: Between charged analytes and functional groups of anion exchanger
  • Sample: Aqueous (pH 8-9)
  • Elution: Basic- pH 10 (e.g. 20% NH3 in MeOH/acetonitrile)
    Acidic- pH 4-5 (e.g. HCl, or 5% AcOH in MeOH/acetonitrile)
    Solvents or buffers with higher ionic strength and counter ions with high selectivity (e.g. Citrate)

It should be noted, that in SPE the interactions described above are not found in pure form, but in combination. For example, modified silicas, unless they have been subjected to endcapping (silanisation of residual silanol groups with short-chain silanes), still possess free silanol groups, which can enter into secondary interactions.

Sample Pretreatment
For direct extraction with adsorbents the sample matrix (sample environment) has to fulfill three conditions:

  • The matrix has to be liquid, if possible with low viscosity
  • Solids should be removed from the liquid matrix
  • The matrix (sample environment) should be suitable for retention of the analyte

For solid samples there are different methods to convert the sample into a suitable matrix:

  • Dissolution of the solid sample in a suitable solvent
  • Lyophilisation of the sample and dissolution in a suitable solvent
  • Extraction of the solid sample with a suitable solvent
  • Homogenization of the sample in a suitable solvent

In order to find the suitable solvent, one has to consider all desired sample components. Also, the suitable solvent should enhance retention of the analyte. For example, samples with large contents of solids are often homogenised in nonpolar solvents like hexane, while for samples with high water content dissolution in acids, bases, buffers or very polar solvents such as methanol are recommended.

Additionally, SPE allows one to alter the properties of the sample matrix. If, for example, natural products are extracted with methanol or acetone, the polarity of the extracts can be increased by dilution with water, in order to enhance nonpolar solid phase extraction on the C18 material.

Chromabond® Polypropylene Columns

  • PP columns with PE filter elements
  • Different sizes from 1, 3, 6 up to 150 ml
  • Adsorbent weights from 20 mg to 50 g
  • Male luer tip as exit
  • Compatible with most robots (e. g. Gilson ASPEC™, Caliper AutoTrace®, …)

Chromabond® Glass Columns

  • Glass columns with chemically very inert glass fibre
  • Filter elements (nominal pore size 1 µm)
  • Two different sizes: 3 and 6 ml
  • Available with all CHROMABOND® phases
  • Excludes any influence from the column material (e. g. plasticizers, …)

Chromafix® Cartridges

  • PP cartridges with PE filter elements
  • Three different sizes with different adsorbent weights: Small (0.4 ml), Medium (0.8 ml), Large (1.8 ml)
  • Female Luer tip at the inlet, male Luer tip as exit
  • Offers alternative way of handling using positive pressure by syringes or peristaltic pumps
  • Especially suited for convenient solid phase extraction of small sample volumes

Chromabond® LV Columns

  • Large volume PP columns with PE filter elements
  • Three different adsorbent weights (100, 200 and 500 mg)
  • funnel-shaped reservoir with 15 ml volume
  • Especially for clinical samples-the whole sample (e. g. urine, serum, blood) can be applied to the column in one step
  • Can be directly used in the Zymate® lab robots of Zymark

Chromabond® Multi 96- SPE in 96-Well format

  • 96-well polypropylene plates with PE filter elements
  • Cavity volume 1.5 ml
  • Adsorbent weights from 25 to 100 mg
  • Supplied with any CHROMABOND® SPE adsorbents
  • For simultaneous preparation of 96 samples
  • Easy method transfer from CHROMABOND® columns or CHROMAFIX® cartridges to CHROMABOND® MULTI 96
  • Readily adaptable to all common automated/robotic handling systems