Basic Knowledge Of HPLC Column
HLPC means High-Performance Liquid Chromatography, which is also known as “High-pressure Liquid Chromatography”, “High-speed Liquid Chromatography”, “High separation Liquid Chromatography”, “modern column Chromatography”, etc.
HPLC is an important branch of chromatography, using the high-pressure infusion system to put the single solvents with different polarity or different portion of mixed solvents and mobile phases into a stationary phase chromatographic column, in which each component is separated and then entered the detector for testing, so as to realize the analysis of the sample.
This is normally shown as column length * inside diameter. For example: 150*4.6, 50 * 2.1, etc. Generally, 4.6 of diameter is used on the conventional detector and 2.1 is used with MS detector.
Types of HAWACH HPLC Column
There are 2 kinds of HAWACH HPLC columns:
Xcheoma series: with many types of fixed phase; with complete varieties and high quality. C18-Aqueous; C18-universal;C18-low pH; C18-Alkaline; C8-universal; C8-Low pH; C8-Fluorine; C4; C30; C1;Phenyl; PAH; etc
Echroma series: less column type, slightly cheaper. C18: 5um; C8: 5um.
Characteristics of HPLC Column
1. High column quality recognition; imported chromatographic filler with high purity and goof uniformity
2. High column efficiency and high peak capacity.
3. Excellent batch stability.
4. Multiple types of chromatographic columns, there are many different types of C18 column and series.
Complete column specifications, particularly, X series column has various fillers, different varieties of column length and internal specifications, which can meet the requirements of the majority of analysis experiments.
Application of HPLC Column
(1)food nutrition analysis: proteins, amino acids, sugars, pigments, vitamins, spices, minerals, etc
(2)food additive analysis: sweeteners, preservatives, colorants, antioxidants, etc
(3)food contamination analysis: mycotoxin, trace elements, polycyclic aromatic diameter, etc
2.Environmental analysis: such as pesticide residue
3.Life Sciences: (1)essential for the study of life sciences, genetic engineering, clinical chemistry and molecular biology
(2) Low molecular weight substance: separation and determination of amino acids,organic acids, organic amines, steroids, porphyrins, carbohydrates, vitamins, etc
(3)High molecular weight substance: purification, isolation and determination of peptides, RNA, proteins and enzymes
4.(1)Medical examination: determination of metabolites in body fluids; Pharmacokinetic study; clinical drug testing.
(2)Synthetic drugs: antibiotics, anti-depressants, sulfonamides
(3)Natural medicine: alkaloids
5.Inorganic analysis: analysis of yang and anion
HPLC columns are prone to clogging and damage, so some laboratories would use a protective column in front of the column to extend the service life of the HPLC column.
The protection column generally includes: card sleeve+ column core + peek joint.
Regeneration of HPLC Column
It is recommended to flush in the direction of the arrow of the column, and try not to recoil as far as possible. The reference solvent sequence used for flushing is water, methanol, chloroform, isopropanol. And it is better to flush the column with a solvent equivalent to 20 times of the column volume in the following order.