Method of packing chromatographic columns

Patent metadata

Field	Value
Publication number	US-9504936-B2
Application number	US-201113639165-A
Country	US
Kind code	B2
Filing date	Apr 5, 2011
Priority date	Apr 5, 2010
Publication date	Nov 29, 2016
Grant date	Nov 29, 2016

Abstract

Improved methods for packing a chromatographic column and columns packed by these methods are provided. The methods include introducing a packing material into a chromatographic column, while simultaneously ultrasonicating and applying pressure to the packing material to form a packed column with radial homogeneity.

First claim

What is claimed:

1. A method for packing a chromatographic column comprising silica particles, the method comprising the steps of
a) introducing the silica particles into the chromatographic column, wherein the chromatographic column has a length of 2 to 10 cm and an inner diameter of 25 μm to 5 cm, and the silica particles have a diameter of 100 nm to 900 nm;
b) ultrasonicating the chromatographic column; and
c) applying pressure to the silica particles,
wherein steps a)-c) are performed simultaneously and wherein a packed chromatographic column with an eddy diffusion term of less than 1 μM is formed, wherein the silica particles are uniformly packed and exhibit no cracks or uninterrupted contact between the inner surface of the column and the silica particles, and wherein the packed chromatographic column is suitable for HPLC, UHPLC, or FPLC.

2. The method of claim 1 , wherein the silica particles have a diameter range from about 150 nm to 900 nm.

3. The method of claim 1 , wherein the silica particles comprise nonporous silica particles, porous silica particles, core-shell silica particles, or combinations thereof.

4. The method of claim 1 , wherein the chromatographic column comprises fused silica, glass, ceramic, stainless steel, aluminum, PEEK, acrylic, or polystyrene.

5. The method of claim 4 , wherein the chromatographic column comprises fused silica.

6. The method of claim 4 , wherein the chromatographic column comprises glass.

7. The method of claim 4 , wherein the chromatographic column comprises stainless steel.

8. The method of claim 1 , wherein a plurality of chromatographic columns are packed simultaneously.

9. The method of claim 1 , wherein the chromatographic column is packed in 6 hours or less.

10. The method of claim 9 , wherein the chromatographic column is packed in 1 hour or less.

11. The method of claim 1 , wherein the pressure is applied within a range from about 200 bar to about 1,000 bar.

12. A packed chromatographic column comprising:
(i) a chromatographic column having a length of 2 to 10 cm and an inner diameter of 25 μm to 5 cm, and
(ii) silica particles inside the chromatographic column, wherein the silica particles have a diameter of 100 nm to 900 nm,
the packed chromatographic column prepared by the steps of
a) introducing the silica particles into the chromatographic column;
b) ultrasonicating the chromatographic column; and
C) applying pressure to the silica particles,
wherein steps a)-c) are performed simultaneously,
wherein the packed chromatographic column has an eddy diffusion term of less than 1 μm;
wherein the silica particles are uniformly packed and exhibit no cracks or uninterrupted contact between the inner surface of the chromatographic column and the silica particles; and
wherein the packed chromatographic column is suitable for HPLC, UHPLC or FPLC.

13. The packed chromatographic column of claim 12 , wherein the chromatographic column comprises a glass capillary column comprising an inner diameter of about 25 μm, a length of about 10 cm, and the silica particles have a diameter within the range of 100 nm to 900 nm wherein the packed chromatographic column is suitable for separating proteins.

14. The packed chromatographic column of claim 12 , wherein the chromatographic column is a stainless steel chromatographic column, a fused silica chromatographic column or a PEEK chromatographic column.

15. The packed chromatographic column of claim 12 , wherein the chromatographic column comprises a fused silica capillary column comprising an inner diameter of about 75 μm, a length of about 5 cm, and the silica particles have a diameter of 750 nm, wherein the packed chromatographic column is suitable for isoelectric focusing of proteins.

16. The packed chromatographic column of claim 12 , wherein the chromatographic column comprises a fused silica capillary column comprising an inner diameter of about 75 μm, a length of about 5 cm, and the silica particles have a diameter of 750 nm, wherein the packed chromatographic column is suitable for hydrophilic interaction chromatography of proteins and peptides.

17. The packed chromatographic column of claim 12 , wherein the chromatographic column comprises a fused silica capillary column comprising an inner diameter of about 75 μm, a length of about 5 cm, and, the silica particles have a diameter of 750 nm, wherein the packed chromatographic column is suitable for reversed phase chromatography of proteins and peptides.

18. The packed chromatographic column of claim 12 , wherein the column has a length of 3 to 5 cm.

19. The packed chromatographic column of claim 12 , wherein the column has an inner diameter of 75 μm to 5 cm.

20. The packed chromatographic column of claim 12 , wherein the column has an inner diameter of 2.1 mm.

21. The packed chromatographic column of claim 12 , wherein the silica particles have a diameter of 150 nm to 900 nm.

22. The packed chromatographic column of claim 12 , wherein the silica particles have a diameter of 300 nm to 900 nm.

23. A packed chromatographic column comprising silica particles inside a column, wherein the silica particles have a diameter of 100 nm to 150 nm, the packed chromatographic column prepared by the steps of
a) introducing the silica particles into the column;
b) ultrasonicating the column; and
c) applying pressure to the silica particles,
wherein steps a)-c) are performed simultaneously,
wherein the packed chromatographic column has an eddy diffusion term of less than 1 μm;
wherein the silica particles are uniformly packed and exhibit no cracks or uninterrupted contact between the inner surface of the column and the silica particles, and
wherein the packed chromatographic column is suitable for HPLC, UHPLC or FPLC.

24. A packed chromatographic column comprising silica particles inside a column, the packed chromatographic column prepared by the steps of
a) introducing the silica particles into the column;
b) ultrasonicating the column; and
c) applying pressure to the silica particles,
wherein steps a)-c) are performed simultaneously,
wherein the packed chromatographic column has an eddy diffusion term of less than 100nm;
wherein the silica particles are uniformly packed and exhibit no cracks or uninterrupted contact between the inner surface of the column and the silica particles, and
wherein the packed chromatographic column is suitable for HPLC, UHPLC or FPLC.

Assignees

• Wirth Mary Jean
• Kodithuwakkuge Sampath Ranasinghe
• Yerneni Charu
• Birdsall Robert
• Ranasinghe Kodituwakkuge Tharanga
• Purdue Research Foundation

Inventors

• Wirth Mary Jean
• Kodithuwakkuge Sampath Ranasinghe
• Yerneni Charu
• Birdsall Robert

Classifications

• G01N2030/565

  slurry packing · CPC title

• B01J20/283

  based on silica · CPC title

• G01N2030/8831

  involving peptides or proteins · CPC title

• B01D15/168

  pH gradient or chromatofocusing, i.e. separation according to the isoelectric point pI · CPC title

• B01D15/305

  Hydrophilic interaction chromatography [HILIC] · CPC title