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(5) High performance liquid chromatography (HPLC)
After the 1990s, HPLC has become the most widely used method in the detection of β-lactam drug residues
.
Since β-lactam drugs all contain carboxylic acid groups in their structure, ion exchange chromatography (anion exchange column is generally used) was used for separation and determination in the early days
1) UVD detection
Liu Yuan et al.
reported a method for the detection of amoxicillin residues in animal tissues
.
The sample was precipitated with trichloroacetic acid, purified and enriched by solid phase extraction.
2) FLD detection
Luo Daoxu and others determined the ampicillin in milk.
The ampicillin in the extract was heated with formaldehyde under acidic conditions to generate 2-hydroxy-3-phenyl-6-methylpyrazine (a fluorescent derivative of ampicillin), which was treated with phosphoric acid.
Salt buffer-acetonitrile (75+25, v/v) was used as the mobile phase, separated by Supelco Discovery C 8 column (250mm×4.
6mm, 5um), FLD detection, excitation wavelength was 346nm, emission wavelength was 422nm
.
Method LOD is 1μg/L, LOQ is 2μg/L, recovery rate is 70%～110%, intra-assay CV is within 10%, inter-assay CV is less than 15%
Meng et al.
established an HPLC-FLD method for the detection of cephalexin in human plasma
.
Cephalexin was derivatized with fluorene methoxycarbonyl chloride (FMOC-CI) in 5mmoL/L borate solution (pH8.
3) ECD detection
ECD is detected based on electrochemical principles and the electrochemical properties of substances
.
In liquid chromatography, electrochemical detection methods can be used for those substances that have no ultraviolet absorption or fluorescence but are electrically active
Related links: β-lactam drug residue detection (1)
