HPLC Analysis of Nitrite and Nitrate Ion on Amaze TH Mixed-Mode Column

Application description

Nitrates and nitrites are polar inorganic ions. Nitrates are mainly used as fertilizers and as oxidizing agents in the production of explosives. Both nitrates and nitrites are used as food additives/components. Both ions are considered toxic and methods for determination* in various samples are very critical for examples in food safety. We have developed a simple, mixed-mode approach to separate and analyze these two ions in one run. ELSD was used for monitoring these two ions during the elution from the Heritage MA mixed-mode HPLC column. This method is also compatible with MS, ELSD and CAD detection, and can be used for the analysis of acidic organic and inorganic ions in one run. Nitrate and nitrite are retained by an ion-exchange mechanism at low organic concentration (below 60% ACN). The retention time is adjusted by the buffer pH and buffer concentration. The ions are retained by the combination of HILIC and anion-exchange when more than 60% of ACN is used. The amount of ACN, buffer pH and buffer concentration will affect elution, retention and resolution of separation

Conditions of Experiment
Column: Amaze TH
Separation Modes: anion-exchange
Column Dimenstions: 4.6x150 mm 5 um, 100A
Mobile Phase: 20% ACN with 20 mM AmAc pH 4
Detection: ELSD
Sample: 1 mg/ml
Injection: 3 uL
Flow rate: 1 ml/min
Class of compounds: Inorganic anion, Organic acid
Nature of compounds: Acidic, Hydrophilic, Polar
Compounds: Nitrite ion, Nitrate ion