USP Methods for Chloramphenicol using a Legacy L1 Column



Application Notes: Chloramphenicol is a common broad spectrum antibiotic developed in the 1940’s. According to the USP methods, chloramphenicol should contain no less than 97.0% and no more than 103% of chloramphenicol calculated on a dried basis. The USP HPLC method for the analysis of chloramphenicol was developed on Legacy L1 column according to the US Pharmacopeia methodology. L1 classification is assigned to reversed-phase HPLC column containing C18 ligand. Support for the material is spherical silica gel with particles size 3-10 um and pore size of 100-120A.

Application Columns: Legacy L1 C18 HPLC column

Application compounds: Chloramphenicol

Mobile phase: MeOH/H2O/AcOH 45/55/0.1

Detection technique: UV

Reference: USP 30: NF35


Column Legacy L1, 4.6×150 mm, 5 µm, 100A
Mobile Phase MeOH/H2O/AcOH 45/55/0.1
Buffer AcOH
Flow Rate 1.0 ml/min
Detection UV, 270 nm



Class of Compounds
Drug,  Hydrophobic, Ionizable
Analyzing Compounds Chloramphenicol


Application Column

SIELC Legacy L1 HPLC column

Legacy L1

SIELC's family of Legacy columns is based on the United States Pharmacopeia's (USP) published chromatographic methods and procedures. Numerous brands have columns used in USP reference standards and methods. USP has created various designations to group together columns with similar types of packing and properties in the solid phase. SIELC's Legacy columns adhere to these strict requirements and properties, allowing you to easily replace older columns that are no longer available without needing to significantly modify your method or SOPs.

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Application Analytes:

Application Detection:
UV Detection
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to so we can ensure you get optimal results for your compound/s of interest.