Related Experiment Video
Updated: Jul 5, 2026
10:22
Improved UPLC-UV Method for the Quantification of Vitamin C in Lettuce Varieties (Lactuca sativa L.) and Crop Wild Relatives (Lactuca spp.)
Published on: June 30, 2020
Hydrophilic interaction liquid chromatography method for the determination of ascorbic acid
Lucie Nováková1, Dagmar Solichová, Sona Pavlovicová
1Department of Metabolic Care and Gerontology, Charles University, Faculty of Medicine and University Hospital in Hradec Králové, Hradec Králové, Czech Republic. novakoval@faf.cuni.cz
Journal of Separation Science
|April 23, 2008
Summary
A new hydrophilic interaction liquid chromatography (HILIC) method rapidly determines ascorbic acid stability. Low temperatures and specific stabilizing agents significantly enhance ascorbic acid
Area of Science:
- Analytical Chemistry
- Biochemistry
- Pharmaceutical Analysis
Background:
- Ascorbic acid (vitamin C) is a vital nutrient with known instability issues.
- Accurate determination and stability assessment are crucial for pharmaceutical and food applications.
- Existing methods for ascorbic acid analysis can be time-consuming or lack comprehensive stability data.
Purpose of the Study:
- To develop a rapid and reliable hydrophilic interaction liquid chromatography (HILIC) method for ascorbic acid determination.
- To conduct an extensive stability study of ascorbic acid under various conditions.
- To identify key factors influencing ascorbic acid stability for improved preservation.
Main Methods:
- Development of a HILIC method utilizing a ZIC HILIC column and a mobile phase of acetonitrile and ammonium acetate buffer.
- Detection of ascorbic acid at its maximum absorbance wavelength (268 nm) using diode array detection.
- Systematic investigation of ascorbic acid stability concerning temperature, stabilizing agents (o-phosphoric acid, oxalic acid), and oxygen levels.
Main Results:
- The developed HILIC method is fast and simple, enabling efficient ascorbic acid quantification.
- Ascorbic acid stability is significantly influenced by temperature, stabilizing agents, and oxygen presence.
- Low temperatures and the addition of o-phosphoric acid or oxalic acid were identified as critical for enhancing ascorbic acid stability.
Conclusions:
- The HILIC method provides a robust analytical tool for ascorbic acid determination and stability studies.
- Optimal storage and formulation strategies for ascorbic acid should incorporate low temperatures and specific stabilizing agents.
- This research offers valuable insights for preserving ascorbic acid integrity in various products.
Related Concept Videos
03:38Polyprotic Acids
Acids are classified by the number of protons per molecule that they can give up in a reaction. Acids such as HCl, HNO3, and HCN that contain one ionizable hydrogen atom in each molecule are called monoprotic acids. Their reactions with water are:
01:11High-Performance Liquid Chromatography: Introduction
High-performance liquid chromatography(HPLC), formerly referred to as High-pressure liquid chromatography, is a powerful technique used to separate, identify, and quantify components in complex mixtures. The term "high pressure" refers to using high pressure to push the liquid mobile phase through the tightly packed columns.
In HPLC, two phases play a critical role in the separation process:
In HPLC, two phases play a critical role in the separation process:
00:57High-Performance Liquid Chromatography: Instrumentation
High-performance liquid chromatography, or HPLC, is an analytical technique that separates liquid samples under high pressures. An HPLC instrument consists of glass bottles for storing solvents called mobile phase reservoirs. HPLC-grade solvents are used to maintain high purity, and the dissolved gases are removed using a degasser, such as a vacuum pumping system or sparging with helium. The solvents are then pumped into the analytical column using a screw-driven syringe or reciprocating pumps.
01:05High-Performance Liquid Chromatography: Elution Process
In High-Performance Liquid Chromatography (HPLC), the elution process is critical to the separation of analytes and the quality of chromatographic results. Elution describes how compounds move through the column and separate based on their interactions with the mobile and stationary phases. This process determines the resolution, peak shape, and retention times in the chromatogram, which are essential for identifying and quantifying components in complex mixtures. Understanding the elution...