Streamlined three step total vitamin C analysis by HILIC-UV for laboratory testing

Michael Fitzpatrick; Paul Bonnitcha; Van Long Nguyen

doi:10.1515/cclm-2021-0614

Article

Streamlined three step total vitamin C analysis by HILIC-UV for laboratory testing

Michael Fitzpatrick , Paul Bonnitcha and Van Long Nguyen

Published/Copyright: September 3, 2021

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From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 59 Issue 12

Abstract

Objectives

In the clinical setting, the analysis and quantification of vitamin C (ascorbic acid) poses several challenges including analyte instability and poor retention by reverse phase HPLC systems. In this article we describe a rapid hydrophilic interaction chromatography ultraviolet method for the measurement of total vitamin C in plasma which overcomes these issues.

Methods

Ascorbic acid and the internal standard were separated under isocratic conditions using a Waters BEH-Amide column and a mobile phase containing 0.005 M potassium phosphate in 80% acetonitrile.

Results

The proposed method was validated and showed good precision (coefficient of variation <5%), accuracy (>99%), and analyte stability after extraction (>24 h).

Conclusions

The simple sample preparation allows full automation and rapid analytical run times of the assay and is therefore suitable for a high-throughput clinical chromatography laboratory.

Keywords: ascorbic acid; high performance liquid chromato-graphy-diode array detection (HPLC-DAD); high performance liquid chromatography-ultraviolet detection (HPLC-UV); hydrophilic interaction chromatography (HILIC); plasma; vitamin C

Corresponding author: Michael Fitzpatrick, Department of Chemical Pathology, NSW Health Pathology, Royal Prince Alfred Hospital, Camperdown, NSW 2050, Australia; and RCPAQAP-Australasian Association of Clinical Biochemists, Vitamins Advisory Committee, Alexandria, NSW, Australia, Phone: +61 (2) 95154631, E-mail: michael.fitzpatrick1@health.nsw.gov.au

Acknowledgments

The authors would like to acknowledge the RCPAQAP for use of their proficiency testing data. The authors would also like to acknowledge the support of the staff and Professor David Sullivan of the New South Wales Health Pathology, Chemical Pathology Department at Royal Prince Alfred Hospital.

Research funding: None declared.
Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.
Competing interests: Authors state no conflict of interest.
Informed consent: Not applicable.
Ethical approval: Not applicable.

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Received: 2021-05-24

Accepted: 2021-08-13

Published Online: 2021-09-03

Published in Print: 2021-11-25

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Articles in the same Issue

https://doi.org/10.1515/cclm-2021-0614

Keywords for this article

ascorbic acid; high performance liquid chromato-graphy-diode array detection (HPLC-DAD); high performance liquid chromatography-ultraviolet detection (HPLC-UV); hydrophilic interaction chromatography (HILIC); plasma; vitamin C