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Prep & Process Columns

Knowledge about large prep and process scale columns

Introduction

Preparative and Industrial scale Solid Phase Extraction, Batch Adsorption, LC, Flash, MPLC,  HPLC and SMB is about  purification and isolation of milligram to kilogram amounts of compounds. Every mode produces different yield and purity of compounds and has different cost structures.

As we move from LC to HPLC operating pressure is increased. As we increase pressure we also increase choice of solvents to be used, plant complexity and equipment costs. We also decrease range of molecules that can be separated, extracted or purified. Under higher pressure some molecules break apart,  protein type of molecules denature. 

Criteria for using large scale columns

The criteria governing preparative and industrial scale separations are market driven.  Economic considerations are most important. They are governed by the following factors.

  • Type of compound to be purified
  • Production out-put
  • Final purity of  target compound
  • Production cost by weight

 

Resolution

Separation Strategy: By optimising the separation between the peak of interest and the nearest contaminant, high sample loads can be achieved without compromising product purity.

Loadability

Loadability is controlled by the silica particles’ pore size and available surface area. The smaller the pore size the larger the surface area. . Application of the smaller pore size silicas is limited by the range of molecular weight materials they can chromatograph.

Chemical Stability

The lifetime of a column is often dependent on the silica’s chemical stability. Conditions of use will be very important.

Physical Stability

The robustness of a silica will determine how many times a material can be repacked into a column. Reversed-phase is the dominant technique used in analytical HPLC. Normal-phase HPLC is still often used in preparative separations due to the high cost of reversed-phase materials and the easier recovery of solute from the organic solvents used.

Two strategies dominate the approach to preparative HPLC. In the ‘scale-up’ approach a method developed for analytical purposes is directly applied to a larger i.d. column. Although typical 3 - 5Ám particles may be replaced with 10 Ám material of identical selectivity, high preparative efficiencies are maintained. Such an approach is particularly suitable for purifying gram quantities of material with low k'-values.

In the alternative ‘overload’ approach resolution is sacrificed by operating the column in an overload situation. Such high loadings maximise column capacity. Separations are poorer but gram to kilogram amounts of material may be purified.

A comparison of scale-up parameters is highlighted in Table 1.

Table 1. Column Capacity

Column Size

Column Internal Diameter (mm)

Relative Flow Rate (ml/min)

Volume of 250mm Length Column (ml)

Weight of Phase1 (g)

Maximum Column Capacity per Injection

Optimum2

Practical3

Analytical

4.6

1.0

4.2

2.5

-

-

Semi-Preparative

10

4.7

20

12

10mg

-

Preparative 1"

20

19

79

47

50mg

2g

Preparative 2"

48

110

450

268

250mg

10g

Preparative 4"

96

440

1800

1072

1g

40g

Production scale  LC columns

100 - 3000

 

 

 

 

 

1 Assumes 250 x 4.6 mm column contains 2.5g material :  2 Scale-up approach,  3 Overload approach 

The technique uses larger particle size silicas, polymers and wider internal diameter columns than in analytical HPLC. Column efficiency can be preserved on scale-up from analytical to preparative separations. However, broader lower efficiency chromatographic peaks are more often observed when the column is used in an overload state. 

Critical Components in a HPLC Column

 

 

Bulk Preparative Materials

We distributes most commercially available preparative HPLC bulk materials. For details please contact us

 

New Style Prep HPLC Columns

SieLC  Prep Column with inbuilt pre-column

The SieLC Prep-column outperforms all previous hardware styles and offers significant benefits to both the column manufacturer and the column user.

New hardware offers:

  • Price savings, since stainless steel has been significantly replaced by less expensive aluminum, yet still offering only stainless steel contact with chromatography liquids.
  • 50 mm x 50 mm , 250 mm x 30 mm, 250 mm x 10 mm, 150 mm x 30 mm , 150 mm x 20mm and 150 by 10 ml.ld,
  • Good heat dissipation and efficient cooling and heating.
  • Lighter weight compared to a stainless steel column of comparable size.
  • Stylish and convenient guard system with simple guard replacement.
  • Optional, direct guard attachment holder
  • Appealing design.
  • Variety of custom aluminum color options.
  • Unified cylindrical body, making it convenient to apply virtually any sized labels to even the shortest column.
  • Simple and convenient packing attachments.
  • After the column is packed, the sliding disconnection of column from packer leaves perfectly cut silica bed.
  • Universal design and style for 10 mm, 20 mm, 30 mm, and 50 mm id columns.
  • Reusable hardware.
  • Patent-pending design

The stationary phase carrying column and Frit is made of SS. The outer body is mande of anodized aluminium. Top and bottom of the column is sealed with PEEK rings.

prep Column

 

Guard-column

UPack -  Self Packing Prep-Column

SIELC Technologies introduces new preparative  chromatography self-packing re-usable column hardware. It is available  in 20 mm, 30 mm, and 50 mm ID format. The hardware allows to pack  columns in a regular chromatography lab, using only a preparative HPLC  pump. The device has a built-in switching valve with position for column packing (fill) and a position for column operation.

No additional equipment is required if you have prep LC system

  • One hardware set provide columns of different length
  • Dynamic and static bed compression
  • Optional air or water heating/cooling
  • Easy packing and repacking
  • Small setup cost, flexible operations
  • Little bench space required
  • Low weight
  • Patent pending design

Simple and inexpensive way of making your own prep columns in 3 steps.

  1. Fill the column with an adsorbent slurry and cap the column with a filter, then switch integrated valve to fill position.
  2. Connect the column to a regular prep pump and setup  pressure limit on the pump to stop at 1000-5000 psi, depending on your  packing material pressure limitation.
  3. Start the pump. When pressure reaches the set limit, switch the integrated valve on a top of the column to operation  position and start you separation.

 

Biochromatography Columns

 

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