How to effectively extend the service life of high performance liquid chromatography system through filtration
High performance liquid chromatography (HPLC) is an analytical method developed on the basis of classical liquid chromatography. Because of its fast sample analysis, it is called high performance liquid chromatography. At present, it has been widely used in many fields such as biology, medicine, organic chemistry, food and beverage, and environmental monitoring.
High-performance liquid chromatography analyzers generally consist of components such as an injector, a degasser, a pump, a column, a column oven, and a detector, as shown in Figure 1. The principle is that the analyte to be injected into the column with the mobile phase is separated in the column due to the different interaction forces between the different types of analyte and the stationary phase, and finally different peak signals are obtained through the detector.
Figure 1 high performance liquid chromatography analyzer diagram
High-performance liquid chromatography is a sophisticated and expensive analytical system. It is especially important to standardize the operation and maintain it so that each component can reach its maximum life. Here I recommend microfiltration to extend the life of each component.
(1) The mobile phase of the filtered mobile phase acts as a carrier for the substance to be tested, which in turn passes through a degasser, a high pressure infusion pump, a sampler, a chromatography column, and finally enters the detector. The quality of the mobile phase directly affects the service life of the relevant components. If the mobile phase is not filtered, the internal particles will enter the system, causing blockage or damage to the small-hole pipe, column, and high-pressure infusion pump.
Anno filtration suggests that the mobile phase be filtered prior to use with a 0.45 μm or 0.22 μm pore size Φ47 mm filter-matched glass filter. Filtration can also be carried out using a Φ33 mm needle filter of the same pore size. In addition to removing fine particles, mobile phase filtration can also remove small bubbles, reduce the burden on the degasser, and reduce fluctuations during the analysis.
(2) Filtration before sample injection High-performance liquid chromatography has a wide variety of samples to be tested, and the particle content and particle size distribution of different samples also vary greatly. If the sample is injected without filtration, the column will be blocked in a short time, resulting in increased column pressure and a significant decrease in service life. See Figure 2.
Therefore, before the injection, Anno recommends filtering the sample with a 0.45 μm or 0.22 μm pore size filter. Generally, a needle filter of a suitable specification can be selected according to the volume of the sample to be processed. Φ5mm filtration volume is the smallest (<1ml), and the retention volume is also the smallest (<15μl); Φ13mm filtration volume is larger than the former (<10ml); Φ25mm filtration volume is the largest (<80ml). For difficult-to-filter samples with large particle loading, a needle filter containing a glass or PP pre-filter layer can be selected.
Figure 2 Relationship between column pressure and filtration
Filtration of the mobile phase and the sample to be tested according to the above recommendations can greatly guarantee the service life of high performance liquid chromatography.
Filtering considerations:
(1) Please select a suitable filter according to the chemical compatibility table.
(2) Please pay attention to the hydrophobicity of the filter. For example, hydrophobic PTFE is recommended to filter organic solvents due to its inherently strong hydrophobicity but it is not recommended to filter aqueous solutions.
(3) Since the filter or the membrane itself may cause the dissolution of the interference material of UV absorption, and the chromatographic peak generated by the dissolution of the interference substance is too large, the analysis and judgment of the experimental data may be disturbed. Therefore, it is strongly recommended that the user prefer a manufacturer that has verified the dissolution of the interference of the UV absorption of the filter or filter, such as ANOW, Millipore, Whatman, and the like.
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