Agilent hplc column

Types of Agilent HPLC column

Agilent HPLC columns are manufactured in different types. Below are some of the most common types.

C18 columns

C18 columns are the most commonly used of all Agilent HPLC columns. Usually, they contain 72 percent carbon and 12 percent hydrogen. The carbon atoms of the C18 chain are bonded to oxygen-saturated silicon spheres. The columns are employed because of their versatility in function, especially for reversing phase chromatography.

Sec columns

SEC columns are used for size exclusion chromatography. The SEC common packs contain porous silica. These pores give columns the ability to separate samples based on their molecular sizes. Larger molecules travel through columns quickly than smaller ones.

Amino columns

Amino columns come with a polar amino functional group (C1H4N) attached to a silica particle. In reversed chromatography, the polar amino group increases the column's polarity. This allows polar, non-ionized, or amine-containing compounds to elute selectively during separation.

Phenyl columns

Columns, such as phenyl-hexyl and phenyl ether columns, include a phenyl group with a non-polar carbon chain bonded to the silica surface. The phenyl moiety interacts with aromatic compounds, allowing them to separate efficiently during liquid chromatography.

Cyano columns

Cyano columns include a cyanide functional group consisting of a carbon and nitrogen atom. These columns are used for both polar and non-polar separation. Their versatility makes them ideal for separating mixtures containing both polar and non-polar components.

Nitro columns

Nitro columns contain nitro functional groups. They are used as a polar end for selective affinity or normal-phase chromatography. Nitro groups can cause strong electronic interaction with analytes containing electron-rich functional groups. That is why they are good at ligand exchange chromatography.

Industry applications of Agilent HPLC column

Agilent HPLC columns are applicable in the following industries:

Pharmaceutical industry

Pharmaceutical companies use Agilent 1200 series HPLC columns in drug formulation and analysis to ensure the efficacy and safety of drugs. Agilent columns are highly sensitive and can be used to analyze the purity and potency of active pharmaceutical ingredients and excipients in drugs.

Environmental testing laboratories

Natural disturbances such as floods, storms, and drought affect human systems' ecological and economic build resilience. Environmental testing laboratories analyze pollutants and contaminants in water, soil, and air samples with Agilent columns.

Food and beverage industry

Agilent columns separate and analyze vitamins, amino acids, fatty acids, and carbohydrates in food and beverages. The columns help identify and quantify pesticides, additives, and contaminants to ensure safety and compliance with regulations.

Biotechnology and life sciences research laboratories

In biomolecule analysis and purification, such as proteins, peptides, nucleotides, and carbohydrates, the bio-separation HPLC columns from Agilent are frequently used. The columns help characterize complex biological samples in proteomics and metabolomics studies.

Agricultural chemistry and petrochemical analysis

Agilent columns are suitable for analyzing and characterizing polymers, petrochemicals, and agricultural chemicals. The versatility of the columns allows for separation of a wide range of compounds present in these complex mixtures while providing reliable and consistent chromatography.

Product specifications and features of Agilent HPLC column

The specification and features include:

Technical specifications

• Column type: Reversed-phase, normal phase, ion-exchange, SEC.
• Particle size: 1.8, 3.5, 5, 7.5 micrometer.
• Column dimension: 30 mm to 300 mm by 2.1 mm to 30 mm.
• Pore size: 80 to 300 nanometer.
• Temperature range: Maximum 60 and 100 degrees centigrade, depending on the column type.
• Material: Silica-based media.

How to use

Remove the HPLC column cover and ensure the guard column is fixed and the solvent has filled the system. Fix the analytical column and attach it to the outlet of the guard column. Ensure the column adapter is tightened to avoid leaking. Turn on the HPLC system to equilibrate the columns.

At this point, the mobile phase will pass through both columns without any sample. Set the detector to the right wavelength and prepare mobile phase and sample. Turn on the HPLC system and inject the sample. After that, record and analyze the data.

Maintenance and repair

• Equilibrate the column monthly with 10 column volumes of strong solvent.
• Use a column reconditioning kit and follow the manufacturer's instructions.
• Store the column in the recommended solvent for the column type.
• Ensure proper mobile phase filtration to prevent particulate contamination.
• Regularly inspect and replace damaged or worn parts within the HPLC system.
• Minimize temperature and pressure fluctuations for consistent performance.
• Add Agilent 1100 HPLC column in the harsh environment periodically.

Agilent HPLC column quality and safety considerations

Here are some of the quality and safety considerations for Agilent LCMS HPLC columns:

• Only store and use Agilent HPLC columns within the specified temperature and pressure range to avoid damage and maintain optimal performance.
• Use column protection devices, such such as inline filters and guard columns. They prevent particulate matter and contaminants from entering the analytical column, ensuring longer column life and improved performance.
• Always handle columns with care, and wear gloves to avoid direct skin contact with the column to prevent contamination and damage to sensitive column media.
• To achieve good separation and accurate results, always ensure the mobile phase composition is correct and premixed properly before use.
• Use only recommended solvents and chemicals to prevent damage to the column packing material and ensure the integrity of the results.
• Check for leaks in the column connections and throughout the instrument regularly. Agilent HPLC columns have high-pressure operation, making them prone to leaks. In addition, leaks can affect results and pose safety risks.

Here are some of the safety considerations to observe:

• Use properly labeled and SDS (safety data sheet)-compliant chemicals to reduce the risk of exposure to hazardous materials.
• Properly dispose of waste solvent in compliance with local environmental regulations to reduce the impact on the environment and prevent contamination.
• Have contingency plans for solvent spills and exposure. Keep absorbent materials nearby to quickly address spills and minimize risk.

Q&A

Q1. What is the lifespan of an Agilent HPLC column?

A1. An Agilent HPLC column can last for one to five years. Factors that affect the column lifespan include the type of analysis done, the sample processed, and the column type used. Also, regular maintenance and proper usage can extend the column lifespan.

Q2. Can I use a Agilent HPLC column in different chromatography types?

A2. Sections 2 and 3 of this post discuss the types, applications, and benefits of care HPLC columns. Some of the columns are designed for specific chromatography types. For example, only SEC columns can be used in size exclusion chromatography. Others, like C18 and amino columns, are versatile and can be used in different chromatography types.

Q3. Are there any guidelines for regularly replacing Agilent HPLC columns?

A3. Users should put into practice quality monitoring, control, and assessment guidelines for replacing the columns. Age, column efficiency, resolution, peak shape, and retention time repro must also be considered.

Q4. Are Agilent columns suitable for high-temperature chromatography?

A4. HPLC columns are generally manufactured to operate at a maximum temperature of 60 degrees centigrade. Nonetheless, some columns can be used at higher temperatures. Check the user manual for restrictions and limitations on temperature properties.