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Setup and validation of a convenient sampling procedure to promptly and effectively stabilize vitamin C in blood and plasma specimens stored at routine temperatures

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Abstract

Vitamin C (ascorbic acid, AA) is very labile in nature and decays quickly after blood withdrawal. To ensure AA stability, the current procedure prescribes immediate plasma acidification followed by sample storage at ultra-low temperature. The aim of this study was to set up a pre-analytical procedure to promptly stabilize AA at routine temperatures while minimizing both specimen manipulation and instrumental requirement. Blood from healthy subjects was collected in lithium-heparin gel separator tubes containing or not different reducing agents (dithioerythritol, tris(2-carboxyethyl)phosphine, n-acetylcysteine and sodium thiosulfate). Plasma AA stability during blood and plasma storage at routine temperatures was evaluated. Plasma AA concentration was assayed by RP-HPLC-UV under ion suppression conditions. Each of the reductants tested was able to slow down the ex vivo degradation of plasma AA; dithioerythritol was the most effective. Five to 10 mmol/L dithioerythritol did not interfere with blood separation and allowed plasma AA to be stabilized up to 6 h, 24 h and 60 days at room temperature, +4 °C and −25 °C, respectively. The method worked well even in case of delayed blood separation and/or incomplete vacutainer filling. The procedure is feasible and reliable. Of special usefulness in clinical and epidemiological studies, prompt plasma manipulation after blood withdrawal or special storage equipments are not required.

Collecting blood in tubes containing a reducing agent is a feasible method to promptly and effectively stabilize plasma vitamin C at routine temperature

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Acknowledgments

The authors are indebted with Dr. Catia Valdarchi and Dr. Davide Francomano for their friendly collaboration and assistance in donor recruitment and blood sampling.

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Authors and Affiliations

  1. Department of Hematology, Oncology and Molecular Medicine, Istituto Superiore di Sanità, Viale Regina Elena, 299, 00161, Rome, Italy

    Barbara Rossi, Francesca Tittone & Simonetta Palleschi

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  1. Barbara Rossi
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  2. Francesca Tittone
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  3. Simonetta Palleschi
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Correspondence to Simonetta Palleschi.

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The authors declare that they have no conflict of interest

Human and animal rights and informed consent

All procedures performed in the present study were in accordance with the ethical standards of our institutional research committee and with the Helsinki declaration as amended in October 2013. Informed, written consent was signed by all the participants included in the study.

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Rossi, B., Tittone, F. & Palleschi, S. Setup and validation of a convenient sampling procedure to promptly and effectively stabilize vitamin C in blood and plasma specimens stored at routine temperatures. Anal Bioanal Chem 408, 4723–4731 (2016). https://doi.org/10.1007/s00216-016-9560-6

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  • DOI: https://doi.org/10.1007/s00216-016-9560-6

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