FLASH CHROMATOGRAPHY: AREA & APPLICATIONS

 

MODERN FLASH CHROMATOGRAPHIC TECHNIQUES

Pre-packed plastic cartridges[15]
In the modern Flash Chromatography system, the glass columns are replaced with pre-packed plastic cartridges which are much safer and also more reproducible.Solvent is pumped through the cartridge, which is much quicker and more reproducible. Systems may also be linked with detectors and fraction collectors providing automation.The introduction of gradient pumps means quicker separations, less solvent usage and greater flexibility.

Column characteristics:
1. Disposable plastic cartridges –save time and increase reproducibility
2. Cartridges of different size -easy scale-up
3. Solid sample module and injection valve - easy sample loading
4. Pressure up to 100 psi - fast separation
5. Narrow particle distribution - Low backpressure and higher efficiency

Advanced Detection Techniques for Flash  Chromatography[11]
UV detection is the traditional method used in Flash chromatography to monitor and fractionate peaks during the purification process. There are few detection options available in Flash  chromatography for compounds that lack chromophores, and thus cannot be detected by UV.  These invisible compounds may not be detected with UV due to the absence of UV chromophores, or their absorbance may be “lost” in that of the solvents used in Flash chromatography. In other cases, the compounds’ absorption spectrum may be unknown and or detection was at a  sub-optimal wavelength. Additionally, the UV absorption of the necessary mobile phase may interfere with the  λ-max of the compound in question. In other cases, the absorption spectrum of the compound of interest or co-eluting impurities may not be known and therefore not detected. These  advanced detection techniques allow users to easily fractionate compounds without the need  for follow-up TLC and subsequent staining to determine where the purified compound eluted.

Evaporative Light Scattering Detection (ELSD) has long been used for High Performance Liquid Chromatography, but has only  recently been employed for Flash chromatography. All-Wavelength Collection allows the collection of compound with unknown absorbance or collection in the presence of interfering solvent absorbance.[11]

Green Flash Chromatography[16]
Green Flash Chromatography is the ultimate flash chromatographic technologythat achieves most efficient sample purification.The sample run is always carried out with minimum eluting volume.It minimizes run time and solvent use while achieving a good separation.It is Eco-friendly. Optimum method will be developed automatically based on the true theory of the flash chromatography, with the simple input of the TLC results, which allows easy sample purification.

Features of Green Flash Chromatography

  • Optimal parameters for flow rate, run time, fraction volume, etc. will be calculated and set automatically upon selecting a column on “Green Flash” software. The default parameters will be shown in System Setting window.
  • Software provides the maximum sample load information for the selected column.
  • State-Of-The-Art Software Based On True Theory of Chromatography.
  • Sample Eluting Position and Resolution Can Be Fully Controlled for Systems.
  • Automatic Method Setup for Reverse Phase Chromatography.
  • Parallel Detection of UV Detector and RI Detector or ELSD.

FLASH CROMATOGRAPHYWITH TLC IMAGE READER.[14]
The system is equipped with a built-in UV light source and a camera. By shooting the TLC plate and clicking the target compound on the TLC plate, the Rf value of the target compound will be calculated and the optimized chromatography method will be developed automatically. The TLC plate is displayed on the screen during run. Compound spots on the TLC plate and the compound peaks both are displayed on the screen. Both the photographic image of the TLC plate and the purification are saved as a data file. Click the target compound and the nearest impurity on the TLC plate, and the maximum sample load for each column will be automatically calculated. Thus, enabling the chemists to choose the best suited column for their sample.

Advantages[14]
* Fast and economic methods for the synthesis laboratory.
* Ideal for the separation of compounds up to gram quantities.
* No expensive equipment required.
* In an ideal way transfers results from TLC to CLC.
* Automated changes between normal phase and reversed phase chromatography.

APPLICATIONS OF FLASH CHROMATOGRAPHY:-[12] [13]

Natural Products/Nutraceuticals Application:
1. Separation and Isolation of α-Santalol and β-Santalol from Sandalwood Extraction
2 .Isolation and Purification of Chromophoric and Nonchromophoric Compounds in Giant Knotweed Rhizome.
3.Isolation and Purification of Flavonoids from Ginkgo Biloba Leaves Extract.
4.Isolation and Purification of Ginsenosides from Red Panax Ginseng Extract.
5.Isolation and Purification of Catechins from Green Tea Extract .
6.In Purification of GallaChinensis.
7.Purification of Ferulic Acid in RhizomaChuanxiong Extract.

Carbohydrate Application:
1. Purification of Conjugated Quercetin and Rutinose.
2. Impurity Isolation of Valproic Acid from CyclodextrinvDuring Encapsulation.
3. Isolation ofAminosugar and Acarbose .
4. Flavanone Glycoside Purification.
5. Isolation of Aminoglycoside Antibiotics.

Lipids Application:
1. Purification of Fatty Acid Methyl Esters (FAMEs).
2. Purification of a Mixture of Glycerides, Mono-, Di-, and Tristearin.
3. Purification of Sterols.

Pharmaceutical/Small Molecules Application:
1.  Bile Acid Purification During Lead Generation in Drug Discovery.
2.  In Impurity Isolation During Drug Purification.
3.  Mestranol Purification During Chemical Synthesis.
4 .In Anti-malarial Drug Purification in Drug Discovery.

Conclusion:
Purification of drug is an important step in any branch of research.Preparative chromatography is used to separate the components of a mixture for more advanced use and is thus a form of purification. Flash Chromatography can be alternative to preparative HPLC as it saves time and solvent. Extrapolation of TLC results on preparative scale can be achieved by Flash chromatography. Modern Flash chromatography with disposable cartridges and advanced detection techniques is applicable to a wide range of compounds.

REFERENCES:
1.Still WC. Kahn M., Mitra A, Flash chromatography, J.Org.Chem. 43(14)1978; 2923-2925.
2.A. B. Roge*, S. N. Firke, R. M. Kawade, S. K. Sarje, and S. M. Vadvalkar,BRIEF REVIEW ON: FLASH CHROMATOGRAPHY,IJPSR (2011), Vol. 2, Issue 8,1930-1937.
3.William CSand Hill DC. General methods for flash chromatography using disposable column. Mol. Divers, 13(2), 2009, 247-252.
4.HetalChaudhari*, FalguniChaudhari, Madhavi Patel, P.K. Pradhan, U.M. Upadhyay,A REVIEW ON A FLASH CHROMATOGRAPHY,International Journal of Pharmaceutical Development & Technology,2 (2), 2012, 80-84.
5. chem.rochester.edu/how to flash.html
6. saiadsorbants.com
7. sorbeadindia.com
8. orgchem.colorado.edu.
9. Buchi Preparative Chromatography.
10.Dane Ganesh D*, Raka KC, Honde BS, Bhawal GS, Tajane PJ, REVIEW ON FLASH CHROMATOGRAPHY,Vol 3,Issue 1, 2013 ,45-49.
11.jsilver@teledyne.com
12.Dewick, P.; John Wiley & Sons, Inc., Hoboken, Medicinal natural products; a biosynthetic approach, 3rd edition ,New Jersey, 2009.
13. discoverysciences.com
14. pretech.nu/products
15. sigmaaldrich.com
16. yamazenusa.com

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