HPLC Analysis of Ascorbic and Citric Acids on Amaze TH Mixed-Mode Column

HPLC Analysis of Ascorbic and Citric Acids on Amaze TH Mixed-Mode Column chromatogram
Conditions of Experiment
Amaze TH
Separation Modes:
HILIC, anion-exchange
Column Dimenstions: 4.6 x 150 mm, 5 um, 100A,
Mobile Phase: 75% ACN with 25 mM AmAc pH 4.2
Detection: ELSD
Sample: 0.3 mg/ml
Injection: 3 uL
Flow rate: 1 ml/min
1. Ascorbic acid
2. Citric acid

Class of compounds: Aromatic acid, Organic acid, Preservative, Supplement, Vitamin
Nature of compounds: Acidic, Hydrophilic, Polar

Application description

Ascorbic acid is a vitamin and preservative. Citric acid is a preservative. Both are widely used in food industry. Both acids are polar and acidic in nature. Retention on a regular C18 column can be achieved with addition of ion-pairing reagent. We have developed a simple and robust method for separation of ascorbic and citric acid in HILIC/anion-exchange mode on Amaze TH HPLC column. Method is using volatile mobile phase which is compatible with MS spectrometry. Method can be used for analysis of ascorbic and citric acids in various sample matrices including biofluids and complex food food matrices. Amaze TH column can retain compounds by combination of HILIC, cation- and anion-exchange. Amount of ACN, buffer concentration and buffer pH will affect retention of polar and ionizable compounds.

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