Separation of Fullerenes with Liquid Crystal Bonded Silica Phases in Microcolumn High Performance Liquid Chromatography

Yoshihiro Saito, Hatsuichi Ohta, Hideo Nagashima, Kenji Itoh, and Kyokatsu Jinno

A report by L. Lao, R. Lopez, K.M.P. Palmario, and S. Sibug

What are Fullerenes?

third allotropic form of carbon material (after graphite and diamond) molecules composed entirely of carbon, in the form of a hollow sphere, ellipsoid, or tube similar in structure to graphite, which is composed of a sheet of linked hexagonal rings, but they contain pentagonal (or sometimes heptagonal) rings that prevent the sheet from being planar Also very large PAHs

Polycyclic Aromatic Hydrocarbons

Potent atmospheric pollutants that consist of fused aromatic rings and do not contain heteroatoms or carry substituents (example: naphthalene) Produced as by-products of fuel burning; found in cooked food; found in meteorites Lipophilic Formed from incomplete combustion ofcarbon-containing fuels

Microcolumn HPLC
Advantages
Lesser reagent and starting material Greatly enhanced sensitivity. Dramatic solvent savings, without altering the resolution and retention values. Ideal for applications in which very small quantities of samples are available for analysis. Favorite choice for applications in LC/MS. Better choice for applications in drug discoveries and screening. Ideal for applications in genomics and proteomics.

Octadecylsilane-Bonded Silica Stationary Phases

Commonly used as stationary phase for separation of fullerenes May be monomeric or polymeric have different mol shape and size recognition capability
Retention Resolution Elution order

Liquid Crystal Bonded Silica Phases

Liquid crystals generally of HMW and low volatility; ordered fluid mesophase of a long chain molecule with both solid-like molec. order and liquid-like character Intermediate between solid and true isotropic liquid Advantage: Reduces or eliminates columnbleed

Objectives of the Study

To be able to separate C60 and C70 fullerenes using liquid crystal silica bonded phases in HPLC To be able to perform micro-column HPLC

Methodology
Syntheses and basic characteristics of the bonded phases.
Determination of surface coverage values of these bonded phase used specific area of the original silica gel and carbon content of these bonded phases.

Methodology

Methodology
Chromatographic measurements.
the synthesized bonded phases were packed using a slurry method. The mobile phases were prepared from guaranteed reagent grade n hexane, toluene,and dichloromethane. Chromatographic measurements were done at least 5 times.

Methodology
PAH sample
used for evaluation in this investigation were commercially available.
Except TBN and PhPh.

Results and Discussion

Separation of C60and C70 with two liquid crystal bonded silica phases
n-hexane as the mobile phase for the evaluation of the basic separation performance of the liquid crystal phases.

Results and Discussion

Effect of mobile phase composition on fullerene Separation

Using nhexane/dichloromethane and n-hexane/toluene as mobile phase
Smaller capacity factors with increasing dichloromethane and toluene

Better peak shapes of chromatogram observed using n-hexane/toluene

Due to increased solubility of C60 and C70 in the mobile phase

Effect of mobile phase composition on fullerene Separation

Liquid crystal phase and a mobile phase with toluene up about to 42%
Can be used to obtain the same separation factor as ODS-5 (1.84)

Advantageous feature
Better solubility greater sample loadability

Other good solvents having greater dissolution power can be used as mobile phase

Effect of column temperature

PAHs: - k decreases with increasing temperature - Normal temperature behavior observed fullerenes: - k increases with increasing temperature - Selectivity slightly decreases with increasing temperature

Effect if column temperature

Effects of column temperature

Values for enthalpy of transfer suggest that the PAHs and fullerenes have different retention mechanisms

Molecular shape recognition

Previous study (Saito, 1994) evaluated molecular shape recognition capabilities of liquid crystal bonded phase with PAHs as sample probes Results indicated:
Liquid crystal phase had better planarity and shape recognition power than ODS Selectivity significantly decreased with increasing temperature

Molecular shape recognition

Results indicate decrease in planar recognition capability of liquid crystal phase 1 with increasing

Molecular shape recognition

Results indicate that more elongated PAHs show larger decrease in retention with increasing temperature

Molecular shape recognition

At elevated column temperature, phase ordering of liquid crystals reduced by their thermal mobility slot-like structure (ligand interval that is smaller than molecular sizes of fullerenes) At elevated temperatures, slot-like structure is lost

Proposed interaction model

Planar, rod-like Nonplanar Fullerenes

Conclusion
At low temp, planar rod-like PAHs interact effectively with the bonded phases = ordered structure
Nonplanar molecules cannot interact as well

Fullerenes C60 have longer retention than PAHs at higher temp

Ligand interval increases as temp increases
Interaction between C60 and bonded phase is easier

Results indicate possible effective