Increasing loadability for SEC-applications:
Adjusting injection volume or concentration?
Separation of IgG (human) and Albumin (human)
YMC-Pack Diol-300 + Diol-200, 300 x 4.6 mm ID x 2
Specified resolution: Rs > 1.50
0.1 M KH2PO4-K2HPO4 (pH 7.0) containing 0.2 M NaCl
UV at 280 nm
Increasing injection volume for higher loading
Increasing concentration for higher loading
Summary: If the injection volume is increased resolution and peakshape are adversely affected as shown above. In order to increase the loading it is far better to increase the concentration per injection. This is shown by the resolution and peak shape being unaffected.
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A new benchmark for chemical stability of preparative media
Will YMC-Triart withstand more CIP cycles than silica-based media? Answer: Yes, it will!
CIP, or “cleaning in process”, is an important step to increase the column lifetime in preparative and industrial chromatography. For many applications this CIP step is performed using alkaline solution containing 0.1 M NaOH of pH = 13. In tests a silica based material withstood such treatment between 3 and 5 times before it had to be replaced.
Now, for the first time, a pH stable preparative grade material, in production scale lot sizes of up to several hundred kg gets available. The innovative multi-layer hybrid material, sets a new benchmark for RP phases with respect to chemical stability. The material named YMC-Triart is based on a newly developed particle, produced with flow reaction technology and will withstand more CIP cycles than conventional silica-based materials. This material is well suited for production scale applications, as it is available up to multi-ton scale.
The huge advantage for the user is its superior chemical stability which leads to more CIP cycles to be endured, when compared with conventional silica based material.
To demonstrate this, a durability test - study plan as outlined above - was performed. Bovine insulin was mixed with human serum, to simulate a real life sample containing a high amount of proteins and peptides other than insulin.
The result of one simulated production and CIP cycle is shown in the figure on the right. It shows the effect of 20 injections of the test mixture onto the column. As expected, the peak shape becomes distorted and the retention time decreases with time due to compounds from the sample binding to the column.
After 20 injections regeneration of the column with 0.1 M NaOH/acetonitrile (50/50) (3 column volumes) was performed. After thorough washing, the retention behaviour returned back to normal. The washing procedure has been effective in regaining good chromatographic behaviour. So, again, the question is:
Can YMC-Triart withstand more CIP cycles than silica-based media? Answer: Yes, it can!
To obtain the answer, the cycle of 20 injections followed by the CIP step was repeated 10 times. The results are shown in the figure below:
As can be seen from the figure on the left, the chromatographic behaviour could be restored for at least 10 cycles of 20 sample injections followed by the complete CIP run. Even after 10 alkali CIP steps little, or no change in peak shape and retention time was observed. This result shows that, using YMC-Triart material instead of conventional silica based material, the life time of a process column can be extended by several times.
This extension in life time will save not only the cost of bulk packing material, but the time involved for less frequent column packing. This gives a smoother process and contributes to a more cost efficient production process.
YMC-Triart materials with a pH stability from pH = 1 - 12 are available in C18 and C8 modification with pore sizes of 12 and 20 nm and particle sizes of 10 and 15 μm (50 μm expected early 2012). With these new and innovative media YMC leaps to the forefront of suppliers for preparative scale RP media. This gives a strong impulse to preparative and industrial scale chromatography applications.
At the same time YMC-Triart material has been established as a trusted phase for analytical applications with many customers world wide. It is available for analytical and UPLC applications in particle sizes of 5, 3 and 1.9μm.
If you would like to obtain more information on our YMC-Triart stationary phases or to apply for a sample to test its impressive abilities in your own lab, please contact:
Available materials (further modifications possible, please inquire):
* expected for beginning of 2012
YMC Europe GmbH
Phone: +49 (0)2064 427-270
Fax: +49 (0)2064 427-222