USP Methods for the Analysis of Amitriptyline using a Legacy L1 Column

Application Notes: Amitriptyline is a tricyclic antidepressant. While it is a older drug, it is still just as effective as newer SSRI’s. According to the USP methods, amitriptyline hydrochloride conatin no less than 99% and no more than 100.5% of amitriptyline. The USP HPLC method for the separation of amitriptyline 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. Resolution between critical pairs corresponds to rules and specifications of USP. Application Columns: Legacy L1 C18 HPLC column Application compounds: Amitriptyline Mobile phase: MeCN/92mM monobasic sodium Detection technique: UV Reference: USP35: NF30


Column Legacy L1, 4.6x150 mm, 5 µm, 100A
Mobile Phase MeCN/92 mM NaH2PO4  pH 2.5 (42/58)
Buffer NaH2PO4
Flow Rate 1.0 ml/min
Detection UV, 250 nm


Class of Compounds Drug, Nerve pain medication and antidepressant, Hydrophobic, Ionizable
Analyzing Compounds Amitriptyline
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Application Analytes:

Application Detection:
UV Detection