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12/04/12

Shodex HPLC Columns

Lesson 3: Partition/Adsorption Chromatography


Lesson 3: Partition/Adsorption Chromatography
1. Reversed-Phase Column
1.1 ODS Column 1.2 Other Silica-Based Column 1.3 Polymer-Based Column

2. Normal-Phase Column 3. Hydrophilic Interaction Chromatography Column 4. Summary of ShodexTM Partition/Adsorption Columns

1. Reversed-Phase Column 1.1 ODS column

The packing material used for reversed-phase column is often made of silica gel modified with functional group. In the sto drink by knowing the bar, but if there is a promoter standing in front of the bar, handing out flyers, more people may stop column. The most often used functional group is octadecyl. Octadecyl functional group is a straight carbon chain of 18. I

It is also often called C18 (C eighteen) column. The name of octadecyl is originated from octa meaning number 8 and de chain is attached on the surface of silica gel. By looking at the structure written above, it may look like a very long an infinite number of chains can be modified on one silica gel. More over these chains are modified even inside of the si

Back to our famous story, some people may pick-up the flyer, but others may not pay any attention. Similarly, some co not. This defines the eluting timing of each component. By using more scientific words, it can be said a separation the sample. This affinity is called partition or adsorption thus this type of separation is also called partition/adsorpti mode is a part of partition/adsorption mode.

The gel made of silica-base and modified with octadecyl functional group is called octadecyl silica (ODS) gel. Also the co column). Among the silica-based column available in the market, about 80% of them are ODS columns.

Ideally, entire surface of ODS gel is modified with C18 functional group; however there will be remaining spaces that are presence of residual silanols can influence the separation. Often end capping is applied to the gels to immobilize the capped, however depending on the type of analyte, presence of silanol may provide better separation results.

1.2 Other Silica-Based Columns

ODS is most popularly used RP column, however since C18 is a long chain, it may retain compounds too much and cons this maybe a case that promotion was too effective making the customer stay at the bar forever, which is not a good m functional group with shorter chain, such as C8, C4, and C3. C8Octyl functional group -CH2CH2CH2CH2CH2CH2CH2CH3 C4Butyl functional group -CH2CH2CH2CH3 C3Trimethyl functional group -CH2CH2CH3 Also there are silica gels modified with phenyl and cyanopropyl functional groups.

1.3 Polymer-Based Columns


Long column life In general, polymer gel is chemically more stable than silica gel and deteriorates slowly. It is difficult to predict a column life, since
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12/04/12

Shodex HPLC Columns

used. Despite that the column life of silica column is about 3 months whereas it is not surprising to see a polymer column provides Good repeatability Gel to gel lot difference of silica gel can be large, whereas that of polymer gel is smaller, since it is relatively easy to control the po performance of a new column is expected to be the same as old column, i.e., negligible level of lot-to-lot difference. Usability under alkaline conditions Silica column cannot be used under alkaline conditions where polymer column can. Some basic samples (often pharmaceutical co thus polymer columns are suitable for such analysis. In another case, columns might be clogged with impurities. Silica columns hav be cleaned with alkaline solvent, it may be possible to remove the impurities and regenerated. Resolution and price General consensus is that polymer columns compared to silica columns are more expensive and provide less resolution. Therefor columns, former is less often used. The price of the polymer column is becoming lower as the technology has been improved. Als polymer column is not extremely more expensive than silica columns. The biggest concern will be the resolution. However, again becoming very comparable to silica columns.

ShodexTM has strength in development of polymer-based RP columns. The most popular Shodex TM polymer column OctaDecyl Polymer. We believe it is easy to remember, as S (silica) in ODS was simply replaced with P (polymer). The gel is made of polymethacrylate. What unique about this gel is that is not modified with any functional group. Instead of u interaction occurs between the sample and the natural characteristics of polymethacrylate. The question about selection many applications on our website to see when ODP/DE-413 is suitable.

2. Normal-Phase Column

It is rather complicated to explain detailed-theoretical differences between RP and normal phase, thus we will keep it sim HPLC analysis have different polarities. Water and oil is a famous example of something does not mix: Water is categor something with small polarity. Oil is a type of carbohydrate, made of carbon and hydrogen; such compound has small pol compound has bigger polarity. Silica gel without modification has large polarity, but when C18 functional group is modifie due to these oxygen and hydrogen; the composition of silica is SiO2, but it has hydroxide group (oxygen + hydrogen) on and mobile phase with large polarity (e.g., water, acetonitrile). Normal-phase mode uses gel with large polarity (e.g. silica chloroform). Instead of using the word small or large polarity, it is also common to use words, hydrophilic or hydrophobic called hydrophilic and something easy to dissolve in oil (i.e., small polarity) is called hydrophobic.

At the very early stage of HPLC development, silica gel without any functional group was only used. Thus, historically no Then the separation mode which uses opposite separation theory to normal phase was developed and named Reverse nowadays, but we cannot change the historical background, and thus they are still called normal and reversed-phase mo

3. Hydrophilic Interaction Chromatography (HILIC) Column

HILIC is a relatively new concept as a member of partition chromatography. It is considered as a part of normal phase b can be either silica or polymer and they may be modified with different types of polar functionalities such as amide, amin phase used for HILIC is very similar to RP mode mobile phase such as mixture of water and acetonitrile. From practical mode separation. Hydrophilic compounds that were too polar to be retained by RP can be analyzed by HILIC using HILIC is popularly used for the separation of carbohydrates, especially saccharide which is hydrophilic. ShodexTM carries a polymer based amino column, named Asahipak NH2P series. NH2 indicates amino functional group polyvinyl alcohol based gel, modified with polyamine. As for RP columns, polymer base compared to silica-base column and good repeatability.

4. Summary of Shodex Partition/Adsorption Columns

Figure 1 is a summary of ShodexTM partition/adsorption type columns. X-axis is the polarity of gel; column on the right sid right side is used for RP mode and left side is for the normal phase mode. Y-axis is the pore size of the gel. For the anal selected and vice versa.

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Shodex HPLC Columns

Figure 1

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