L-Histidine hydrochloride monohydrate can be analyzed by this reverse phase (RP) HPLC method with simple conditions. The mobile phase contains an acetonitrile (MeCN), water, and phosphoric acid. For Mass-Spec (MS) compatible applications the phosphoric acid needs to be replaced with formic acid. Smaller 3 µm particles columns available for fast UPLC applications. This liquid chromatography method is scalable and can be used for isolation impurities in preparative separation. It also suitable for pharmacokinetics.
High Performance Liquid Chromatography (HPLC) Method for Analysis of Histidine and Histidine Methyl Ester Dihydrochloride
Histidine and Histidine Methyl Ester Dyhydrochloride are related compounds with interesting uses. Histidine is used in biosynthesis to create proteins, while histidine methyl ester dihydrochloride (a derivative of histidine) is used to create various other compounds.
This amino acid and its derivative can be detected in the low UV regime. Using a Primesep 100 mixed-mode column and a mobile phase consisting of water and acetonitrile (MeCN) with a sulfuric acid (H2SO4) buffer, Histidine and Histidine Methyl Ester Dyhydrochloride can be retained, separated, and analyzed. This analysis method can be UV detected at 200 nm.