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A Practical Guide to Using a New HPLC Column
Introduction Getting a new HPLC column is exciting—but also important. It is a core part of your chromatography system. If you use it the right
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FAQ for HPLC Components
An HPLC column is a device used in High Performance Liquid Chromatography (HPLC) that contains a stationary phase that separates the components of a sample as they flow through the column. The stationary phase can be made up of a variety of materials such as silica, alumina, or polymer, and can be packed with various sizes of particles ranging from 1.7 µm to 10 µm.
Some HPLC columns, known as empty columns, can be refilled or repacked with a new stationary phase. However, many HPLC columns are pre-packed and cannot be refilled. If a column is refillable, it requires special tools and knowledge to ensure that the packing is uniform and tightly packed to prevent column voids.
An HPLC empty column is a type of column that is not pre-packed and requires the user to pack it with their own stationary phase. These columns are typically used for customized applications or for users who prefer to pack their own columns for quality control purposes. Empty columns require special tools and knowledge to ensure that the packing is uniform and tightly packed to prevent column voids.
Packing an HPLC empty column involves selecting the appropriate stationary phase and properly packing the column to ensure uniformity and minimal backpressure. It requires special tools such as a column packing stand and a slurry mixer. The packing process involves mixing the stationary phase in a solvent to form a slurry, which is then poured into the column and allowed to settle. The column is then packed to the recommended pressure and tested to ensure the packing is uniform.
HPLC columns have a finite lifespan that depends on the type of column, the nature of the samples being run, and the operating conditions. In general, most columns can last for several hundred samples. However, if a column is exposed to harsh conditions such as high temperature, high salt concentration or a pH outside its recommended range, its lifespan can be dramatically reduced.
Choosing the right HPLC column involves considering the sample matrix and analyte properties, such as molecular weight, polarity, and functional groups. The choice of stationary phase, particle size, and column dimensions also play a role. For example, if your sample is highly polar, you would choose a stationary phase that is non-polar in nature. Similarly, if your sample contains large molecules, you would choose a column with a larger particle size to prevent clogging.
HPLC hardware consumables are the parts and accessories used in the operation of an HPLC instrument, such as tubing, fittings, filters, and valves. These consumables are critical to the performance and longevity of the instrument and should be replaced regularly to prevent system downtime and chromatographic problems. Common consumables include injector and detector tubing, filters for mobile phase and sample preparation, and check valves.
The frequency of replacement for HPLC hardware consumables depends on usage, sample matrix, and operating conditions. In general, regular maintenance and replacement of worn or damaged parts can improve instrument performance and longevity. Common consumables like injector tubing and filters should be replaced after every few hundred samples or as part of a regular maintenance schedule, while more durable parts like check valves and pumps may only need replacement every few years.
