Hydrophilic interaction liquid chromatography method for the determination of ascorbic acid
Abstract
Hydrophilic interaction liquid chromatography (HILIC) method using internal standard for the determination and stability study of ascorbic acid was developed. HILIC method was very fast and simple using the following analytical conditions: ZIC HILIC (150 x 2.1 mm, 3.5 microm) chromatographic column and mobile phase composed of ACN and 50 mM ammonium acetate buffer pH 6.8 (78:22 v/v). Diode array detection was performed and chromatograms were processed at 268 nm, the maximum wavelength of absorbance of ascorbic acid. An extensive stability study of ascorbic acid as a function of various factors including temperature, stabilizing agents, oxygen presence and its concentration in solution was performed in order to gain information about the quantitative influence of individual stability factors. Low temperature and stabilizing agents (o-phosphoric acid and oxalic acid) were found to be key factors enabling substantial enhancement of the stability of ascorbic acid.
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Internal standard quantification of L-ascorbic acid in biological material is difficult because it is difficult to find a compound suitable for both extraction and analysis steps similar to L-ascorbic acid.
“An internal standard is not routinely used to determine L-ascorbic acid in biological material because it is difficult to find a compound that is suitable for both extraction and analysis steps similar to L-ascorbic acid.”
From Vitamin C-Sources, Physiological Role, Kinetics, Deficiency, Use, Toxicity, and Determination (2021) · cites this paper as [213]
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- Authors
- Nováková, L., Solichová, D., Pavlovičová, S., & Solich, P.
- Affiliations
- Charles University (CZ)
- Venue
- Journal of Separation Science
- Publisher
- Wiley
- Published
- 2008-05
- Volume
- 31
- Issue
- 9
- Pages
- 1634-1644
- ISSN
- 1615-9306 (ISSN-L)
- 1615-9314
- OpenAlex
- W2156576576
- Cited by
- 50
- References
- 38
- Citations by year
- 2026: 1 · 2025: 3 · 2024: 3 · 2023: 4 · 2022: 5 · 2021: 3 · 2020: 2 · 2019: 1 · 2018: 1 · 2017: 1 · 2016: 4 · 2015: 2 · 2014: 3 · 2013: 4 · 2012: 1
- FWCI
- 4.01
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Separation Science (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical Methods in Pharmaceuticals
- Physical Sciences › Chemistry › Spectroscopy › Analytical Chemistry and Chromatography
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical chemistry methods development
- Keywords
- vitamin C, hydrophilic interaction liquid chromatography, internal standard, stability studies, diode array detection, oxalic acid, temperature effects, oxygen concentration, stabilizers
- MeSH terms
- Ascorbic Acid, Chlorogenic Acid, Chromatography, High Pressure Liquid, Chromatography, Liquid, Drug Stability, Humans, Hydrogen-Ion Concentration, Indicators and Reagents, Oxygen, Reference Standards, Solvents, Spectrophotometry, Ultraviolet, Temperature, Phase Transition
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