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Kromasil® Eternity HPLC columns come with particles down to 2.5 μm. EternityXT extends down to 1.8 μm to fit any UHPLC instrument for better efficiency and flexibility in the laboratory. Both can be used for reversed-phase separations and purifications that could demand harsh conditions, fast turnaround, easy method transfer and seamless scale-up from R&D to production.

Work fast across the board

With columns built on the Eternity platform, users can now easily develop and validate UHPLC methods for synthetic and natural products, even under tough pH conditions. method transfer to HPLC for characterization and quality control can be made seamlessly and, if required, scaled up directly for isolation and purification. our extensive assortment of slurry-packed columns, combined with the wide range of particle sizes from 1.8 μm to 10 μm for the Eternity platform, help businesses improve productivity by using one stationary phase type across the entire company.

High efficiency with small particles

When scientists need to get results fast and within an extended pH range, EternityXT columns can help achieve the desired laboratory efficiency.

Same selectivity on particles 5, 2.5 and 1.8 μm
With EternityXT columns you can maintain separation power across all dimensions and particle sizes. Here is an illustration of faster result turnaround with maintained resolution when using shorter columns with smaller particles.
  • Conditions
  • Columns: Kromasil EternityXT, C18
  •   5 μm 4.6 × 150 mm
  •   2.5 μm 4.6 × 75 mm
  •   1.8 μm 4.6 × 50 mm
  • Mobile phase: acetonitrile / water / formic acid [25/75/0.1]
  • Flow rate: 1 ml/min, 2 ml/min, 2.8 ml/min (respectively)
  • Temperature: 25 °C
  • Detection: UV @ 254 nm
  • Substances: 1 = uracil
  •   2 = sulfathiazole,
  •   3 = sulfamerazin,
  •   4 = sulfamethoxazole

Alternative selectivity

While C18 columns are the most commonly used for reversed-phase chromatography, PhenylHexyl is an alternative phase chemistry that provides additional interaction opportunities, especially when the analytes of interest contain an aromatic ring. available for both Eternity and EternityXT.

Separation of Xanthines on EternityXT PhenylHexyl
Separation of xanthines on Kromasil® EternityXT PhenylHexyl.
  • Conditions
  • Column: Kromasil EternityXT-5-PhenylHexyl, 4.6 × 250 mm
  • Mobile phase: acetonitrile / water/formic acid [40/60/0.1]
  • Flow rate: 1 ml/min
  • Temperature: 30 °C
  • Detection: UV @ 254 nm
  • Substances: 1 = theobromine
  •   2 = 1.7-dimethylxanthine,
  •   3 = theophylline,
  •   4 = caffeine

Consistent results between columns and batches

since Nouryon controls the entire manufacturing process of the Eternity platform, from the initial production steps of the stationary phase to the finished packed columns, batch-to-batch as well as column-to-column reproducibility is assured.

Column-to-column reproducibility
Batch-to-batch reproducibility
Comparisons of three columns showing column-to-column (A) and batch-to-batch (B) reproducibility.
  • Conditions
  • Column: Kromasil EternityXT, 2.5 μm, C18,
  •   A: 4.6 x 100 mm
  •   B: 2.1 x 100 mm
  • Mobile phase: acetonitrile / water: A: (70/30), B: (65/35)
  • Flow rate: A: 1.7 ml/min, B: 0.65 ml/min
  • Temperature: A: 25 °C, B: 35 °C
  • Detection: UV @ 254 nm
  • Substances: 1 = dimethyl phthalate
  •   2 = toluene,
  •   3 = biphenyl,
  •   4 = phenanthrene

Scale-up with ease

as it is fairly straightforward to scale HPlC up or down, having the reproducible Eternity platform phases available on a broad range of particle and column sizes gives the user the key tools to carry out method scaling efficiently.

Scalability: same selectivity over column diameters
  • Conditions
  • Columns: Kromasil EternityXT-10-C18 4.6 × 250 mm
  • Mobile phase: acetonitrile / 10 mm ammonium hydrogen carbonate, pH 10.5
  • Flow rate: 1 ml/min
  • Temperature: ambient
  • Detection: UV @ 230 nm
  • Substances: 1 = sotalol
  •   2 = nadolol,
  •   3 = pindolol,
  •   4 = metoprolol
The separation of β-blockers illustrates the possibility to scale up your separation developed in analytical scale to larger scale chromatography, essentially without any loss of performance. Use 4.6 mm ID or 10 mm ID columns for the method development, and use the data obtained for predicting the performance in larger scale. With dynamic axial columns (DAC) it is possible to reproduce the performance obtained in analytical columns even in very large scale.

Loadability increases at high pH

Loadability increased with higher pH
The loadability increase that can be obtained at high pH for basic compounds is illustrated in the adjoining chromatograms, where diphenhydramine is run at pH = 3.7 and 10.5, respectively. At low pH, the molecule is ionised, leading to a large band broadening even at very low loadings. The same loading at high pH (upper right chromatogram) produces a sharp peak without any tendency to broaden as a function of concentration overload. To obtain the same band broadening at high pH, the loading has to be increased more than 160 times. Hence, loading capacity is increased by a factor >160!
  • Conditions
  • Column: Kromasil EternityXT-10-C18, 4.6 x 250mm
  • Substance: diphenhydramine
  • Flow rate: 1 ml/min
  • Detection: UV @ 254 nm
  • Low pH, low loading
  • Loading: 30 μg
  • Mobile phase: acetonitrile / 25 mm ammonium format, pH 3.7 (35/65)
  •  
  • High pH, low loading
  • Loading: 30 μg
  • Mobile phase: acetonitrile / 25 mm ammonium hydrogen carbonate, pH 10.5 (70/30)
  • High pH, high loading
  • Loading: 5 mg
  • Mobile phase: acetonitrile / 25 mm ammonium hydrogen carbonate, pH 10.5 (70/30)

Swift application development in UHPLC mode

In research and development as well as in quality control, natural and synthetic peptides are regularly analysed using reversed-phase chromatography. Kromasil® EternityXT C8 can be the adequate moderate hydrophobic phase for the task. With small particles available, you can take advantage of any UHPLC equipment to develop your analysis with quick turn around.

Application: separation of a mix of peptides (1)
  • Conditions
  • Column: Kromasil EternityXT-1.8-C8 2.1 × 50 mm
  • Mobile phase: acetonitrile / water + 0.1% TFA
  • Gradient: 0 - 0.5 min: 5%, 5 min: 30% acetonitrile
  • Flow rate: 1.5 ml/min
  • Temperature: ambient
  • Detection: UV @ 220 nm
  • Substances: 1 = gly-tyr,
  •   2 = val-tyr-val,
  •   3 = met-enkephalin,
  •   4 = leu-enkephalin,
  •   5 = angiotensin II
Application: separation of a mix of peptides (2)
  • Conditions
  • Column: Kromasil EternityXT-1.8-C8 2.1 × 50 mm
  • Mobile phase: acetonitrile / water + 0.1% TFA
  • Gradient: 0 min: 8%, 3 min: 30% acetonitrile
  • Flow rate: 1.5 ml/min
  • Temperature: ambient
  • Detection: UV @ 220 nm
  • Substances: 1 = oxytocin,
  •   2 = bradykinin,
  •   3 = angiotensin II,
  •   4 = neurotensin,
  •   5 = angiotensin I

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