Kromasil technical evidence

analysis

You are working with HPLC for the separation and identification of analytical substances in laboratories. Challenge the selectivity of Kromasil for your most sensitive analysis.

development

You need to quickly purify substances for further studies in the development of pharmaceuticals. Kromasil will help you achieve your goals with ease and efficiency.

You are using preparative chromatography for high efficiency purification in a pilot or up to industrial-scale production. The robustness of Kromasil is available in ton quantities to meet your process demands.

Scalability

Kromasil packings are manufactured in large batches and then fractionated in very narrow particle sizes, all with the same characteristics. This means that you can easily make your scale-up work from analytical scale to process scale. You will obtain the same column efficiency, selectivity and retention time when using Kromasil packed in an analytical column, or a 600 mm I.D. column. Often the efficiency in a large diameter column exceeds that of an analytical column.

Kromasil can be efficiently packed in all commercial industrial HPLC, SFC and SMB systems.

In figure 1, the packing of Kromasil in three commercial preparative HPLC systems is shown.

Fig. 1 - Efficiency of three different preparative axial compressed columns packed with Kromasil 10 µm C8. Plate numbers >40 000 p/m can easily be obtained with 10 µm particles.
Conditions:
Column I.D.: 50 mm
Packing material: Kromasil 100 Å 10 µm C8
Packing pressure: 100 bar
Flow rate: 60 ml/min.
Sample: acetophenone
Eluent: acetonitrile/water (70/30)

To exemplify how easy the scalability with Kromasil phases is, three different applications are shown in figures 2, 3 and 4 respectively: insulin, three chiral separations and the purification of fish oils EPA and DHA.

Fig. 2 - Large scale purification of insulin.
Conditions, purity analysis of raw product and fractions:
Purity: 90%
Column: Kromasil 100 Å 3.5 µm C4, 4.6 x 120 mm
Detection: UV 214 nm
Buffer: 0.05 M NaH ₂PO ₄, 0.1M NaClO ₄/HClO ₄, pH = 2.5
Gradient slope: 30 – 36% acetonitrile / 0 – 55 min.
Flow rate: 1.0 ml/min
Conditions, preparative injection:
Loading: 6 g/l column volume
Column: 50 x 250 mm, DAC
Packing material: Kromasil 100 Å 10 µm C8
Flow rate: 60 ml/min

Fig. 3 - Large scale purification of three racemates.
Analytical conditions:
Column: Kromasil Chiral 5 µm, DMB, 4.6 x 250 mm
Flow rate: 1 ml/min
Preparative conditions:
Column: Kromasil Chiral 10 µm, DMB, 50 x 250 mm
Flow rate: 118 ml/min
Eluents (analytical and preparative)
Ibuprofen: Hexane/MTBE/HAc (90/10/0.1)
2-(octylsulfinyl)benzoic acid: Hexane/THF/HAc (80/20/0.1)
Binaftol: Hexane/MTBE (70/30)

Fig. 4 - Large scale purification of fish oils EPA and DHA as ethyl esters.
Analytical conditions:
Column size: 4.6 x 250 mm · Loading: analytical · Flow rate: 0.85 ml/min
Preparative conditions:
Column size: 50 x 250 mm · Loading: 2.2 g (7 mg/g) · Flow rate: 100 ml/min
Conditions, purity check, fraction I:
Column size: 4.6 x 250 mm · Loading: analytical · Flow rate: 0.85 ml/min
Conditions, purity check, fraction II:
Column size: 4.6 x 250 mm · Loading: analytical · Flow rate: 0.85 ml/min
Common conditions:
Stationary phase: Kromasil 100 Å 10 µm C18 · Elution gradient: MeOH/H ₂O (95/5): 0 – 16 min, MeOH (100): 16 – 30 min.

· Eka Chemicals, Separation Products · SE–445 80 Bohus, Sweden · Phone +46 31 58 70 00 · Fax +46 31 58 77 27 ·
· E-mail kromasil@akzonobel.com ·