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CAMAG Automated Multiple Development (AMD 2)

The CAMAG Automated Multiple Development procedure allows High-Performance Thin-Layer Chromatography to be utilized for difficult analytical tasks that cannot be solved by isocratic HPTLC.

 

Only the AMD 2 procedure can be successfully employed for reproducible gradient elution with silica gel as the stationary phase. In column liquid chromatography, gradient elution is common on reversed-phases only because a silica gel column would call for a time consuming reconditioning or be irreversibly degraded, which is not acceptable in a technique depending on multiple use of the stationary phase. In High-Performance Thin-Layer Chromatography this is not relevant.

The principle of the CAMAG AMD procedure

  • The HPTLC plate is developed repeatedly in the same direction.
  • Each successive run extends over a longer solvent migration distance than the one before.
  • Between runs, the solvent is completely removed from the developing chamber and the layer is dried under vacuum.
  • Each successive run uses a solvent of lower elution strength than that of the one used before. In this way, a stepwise elution gradient is formed.
  • The combination of focusing effect and gradient elution results in extremely narrow bands. Their typical peak width is about 1 mm. This means that, with the available separation distance of 80 mm, up to 40 components can be completely resolved, i.e. with base line separation.
  • With visionCATS the AMD 2 can be IQ/OQ qualified and used in a cGMP environment. Operated with the mentioned software, the AMD 2 supports compliance with 21 CFR Part 11.

The AMD 2, like other software-controlled CAMAG instruments, communicates with visionCATS. The gradient, made from up to 5 solvent bottles, is defined by input into a table in visionCATS. Gradient and solvent migration distance for each run can be shown graphically for verification. Then all individual runs of the developing program are performed fully automatic and monitored by visionCATS.

Key features

  • Multiple development in the same direction with increasing solvent migration distancesEnhanced separation capacity with baseline separation of up to 40 components
  • Software-controlled with visionCATS
  • Utilizing time also after working hours

Example

Separation of various rhubarb samples by AMD 2

Detection: UV 366 nm
AMD gradient in 10 steps: methanol – dichloromethane (40:60) to (10:90) in 9 steps, 40 mm solvent migration distance, then one isocratic step methanol – dichloromethane (10:90) over 70 mm solvent migration distance

HPTLC chromatogram of Rhubarb under UV 366 nm, derivatized with Natural Product reagent

Note: for operating the AMD 2 system, an external supply of compressed nitrogen or air (4 bars) is required; consumption: about 10 L/h.

If the AMD 2 system is to be operated with a different type vacuum pump, note that a minimum average pumping speed of 3 m3/h according to PNEUROP is required and an ultimate partial pressure of < 10 mbar has to be reached.

Ordering information AMD 2



022.8860

CAMAG® AMD 2 System Automated Multiple Development
Chromatogram developing module,
accessories include all electrical and pneumatic connecting elements,
supply of sealing materials, HPTLC plate positioning device and small
ancillaries, RS 232, auto-range power supply (100-240V).

022.8880

Oil-sealed rotary vane vacuum pump RV3 (Edwards);
including gas ballast/oil return, oil vapor filter, flexible vacuum hose 1 m, ready to connect to the CAMAG AMD 2 System, 100 - 120 V or 220 - 240 V

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