Determination of vitamin C (ascorbic acid) using high performance liquid chromatography coupled with electrochemical detection
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Vitamin C (ascorbic acid, ascorbate, AA) is a water soluble organic compound that participates in many biological processes. The main aim of this paper was to utilize two electrochemical detectors (amperometric - Coulouchem III and coulometric - CoulArray) coupled with flow injection analysis for the detection of ascorbic acid. Primarily, we optimized the experimental conditions. The optimized conditions were as follows: detector potential 100 mV, temperature 25 °C, mobile phase 0.09% TFA:ACN, 3:97 (v/v) and flow rate 0.13 mL·min-1. The tangents of the calibration curves were 0.3788 for the coulometric method and 0.0136 for the amperometric one. The tangent of the calibration curve measured by the coulometric detector was almost 30 times higher than the tangent measured by the amperometric detector. Consequently, we coupled a CoulArray electrochemical detector with high performance liquid chromatography and estimated the detection limit for AA as 90 nM (450 fmol per 5 μL injection). The method was used for the determination of vitamin C in a pharmaceutical preparations (98 ± 2 mg per tablet), in oranges (Citrus aurantium) (varied from 30 to 56 mg/100 g fresh weight), in apples (Malus sp.) (varied from 11 to 19 mg/100 g fresh weight), and in human blood serum (varied from 38 to 78 μM). The recoveries were also determined.
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L-ascorbic acid and dehydroascorbic acid can degrade during sample preparation for HPLC/CE analysis.
“Since the target analytes can degrade during the sample preparation process, stabilizers and/or chelators need to be added to the sample [223,226,228].”
From Vitamin C-Sources, Physiological Role, Kinetics, Deficiency, Use, Toxicity, and Determination (2021) · cites this paper as [228]
The Methods section (‘Chemicals, material and pH measurements’; page unavailable) says oxygen oxidizes ascorbic acid in samples to dehydroascorbic acid and describes using a reducing agent to prevent oxidation. It does not say that dehydroascorbic acid itself degrades during preparation or that chelators need to be added. (LLM, conf=0.84)
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- Authors
- Gazdik, Z., Zitka, O., Petrlova, J., Adam, V., Zehnalek, J., Horna, A., Reznicek, V., Beklova, M., & Kizek, R.
- Affiliations
- Department of Breeding and Propagation of Horticultural Plants, Faculty of Horticulture, Valticka 337, CZ-691 44 Lednice, Faculty of Agronomy, Zemedelska 1, CZ-613 00 Brno, Mendel University of Agriculture and Forestry, Czech Republic
- Department of Chemistry and Biochemistry, Faculty of Agronomy, Zemedelska 1, CZ-613 00 Brno, Mendel University of Agriculture and Forestry, Czech Republic
- Tomas Bata University, T.G. Masaryka 275, CZ-762 72 Zlin, Czech Republic
- Department of Veterinary Ecology and Environmental Protection, Faculty of Veterinary Hygiene and Ecology, University of Veterinary and Pharmaceutical Sciences, Palackeho 1-3, CZ-612 42 Brno, Czech Republic
- Venue
- Sensors
- Publisher
- MDPI AG
- Published
- 2008-11-07
- Volume
- 8
- Issue
- 11
- Pages
- 7097-7112
- ISSN
- 1424-8220 (ISSN-L)
- PMC
- PMC3787433
- OpenAlex
- W2116929359
- Cited by
- 170
- References
- 70
- Citations by year
- 2026: 10 · 2025: 17 · 2024: 24 · 2023: 14 · 2022: 17 · 2021: 14 · 2020: 8 · 2019: 8 · 2018: 8 · 2017: 8 · 2016: 10 · 2015: 5 · 2014: 5 · 2013: 4 · 2012: 7
- FWCI
- 2.06
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Sensors (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Repository of TBU publications (Univerzita Tomase Bati ze Zline) (submittedVersion, cc-by)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Topics
- Physical Sciences › Chemistry › Electrochemistry › Electrochemical Analysis and Applications
- Physical Sciences › Engineering › Electrical and Electronic Engineering › Electrochemical sensors and biosensors
- Physical Sciences › Chemistry › Spectroscopy › Analytical Chemistry and Chromatography
- Keywords
- vitamin C, electrochemical detection, amperometric detection, coulometric detection, HPLC-ELSD, flow injection analysis, detection limit, tablets, citrus fruit, apple, serum, calibration curve
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