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Assessment of the Stability of Supraphysiological Ascorbate in Human Blood: Appropriate Handling of Samples from Clinical Trials for Measurements of Pharmacological Ascorbate
Journal article   Open access   Peer reviewed

Assessment of the Stability of Supraphysiological Ascorbate in Human Blood: Appropriate Handling of Samples from Clinical Trials for Measurements of Pharmacological Ascorbate

Michael S Petronek, Brett A Wagner, Nancy J Hollenbeck, Joseph M Caster, Douglas R Spitz, Joseph J Cullen, Garry R Buettner and Bryan G Allen
Radiation research, Vol.191(6), pp.491-496
06/2019
DOI: 10.1667/RR15328.1
PMCID: PMC6608566
PMID: 31039080
url
https://www.ncbi.nlm.nih.gov/pmc/articles/6608566View
Open Access

Abstract

Based on encouraging results from several early-phase clinical trials, there is renewed interest in the use of pharmacological ascorbate (i.e., intravenous administration resulting in >≈10 m plasma ascorbate concentrations) in combination with standard-of-care cancer treatments including radiation and/or chemotherapy. Under normal, healthy physiological conditions, humans maintain plasma ascorbate concentrations in the range of 40-80 l . However, antitumor activity requires supraphysiological plasma concentrations on the order of ≈20 m . The stability of ascorbate in whole blood has been well studied. The goal of this work was to determine the appropriate handling methods of blood samples, after treatment with pharmacological ascorbate, which allow for the optimal measurement of ascorbate in plasma for dosing verification. Our findings indicate that ascorbate concentrations (m ) are relatively stable in whole blood collected in sodium heparin tubes and stored on ice (or at 4°C) for up to 24 h. After 24 h, ascorbate levels in plasma are relatively stable at 4°C for up to 72 h. At -20°C, plasma concentrations are relatively stable for 2-3 weeks, while at -80°C, ascorbate concentrations in plasma are stable for at least one month. In contrast, patient samples showed better stability when stored as whole blood compared to plasma at 4°C but increasing hemolysis over time may significantly skew ascorbate measurements. Additionally, patient samples can be reliably stored as plasma at -20°C for up to three weeks in either a frost-containing or frost-free environment. This information can guide the collection, processing and storage of clinical samples after pharmacological ascorbate infusions amenable to multi-center clinical trials.
Cold Temperature Time Factors Humans Female Male Ascorbic Acid - blood Clinical Trials as Topic Blood Specimen Collection - methods

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