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The effect of pH on thrombin generation–An unrecognized potential source of variation

BACKGROUND: When CO(2) escapes from plasma, the pH of the plasma increases. In samples left open or kept in long‐term storage, the pH may increase considerably. Assays in which the ratio of plasma sample relative to the total volume including reagents is high may be sensitive to the pH of the plasma...

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Autores principales: Kristensen, Søren Risom, Nybo, Jette, Pedersen, Shona
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7040540/
https://www.ncbi.nlm.nih.gov/pubmed/32110752
http://dx.doi.org/10.1002/rth2.12289
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author Kristensen, Søren Risom
Nybo, Jette
Pedersen, Shona
author_facet Kristensen, Søren Risom
Nybo, Jette
Pedersen, Shona
author_sort Kristensen, Søren Risom
collection PubMed
description BACKGROUND: When CO(2) escapes from plasma, the pH of the plasma increases. In samples left open or kept in long‐term storage, the pH may increase considerably. Assays in which the ratio of plasma sample relative to the total volume including reagents is high may be sensitive to the pH of the plasma sample. OBJECTIVE: The aim was to investigate the effect of the pH of plasma samples used in the calibrated automated thrombin generation (CAT) assay in which the ratio (plasma sample) / (total volume) is high. METHODS: Plasma pH was increased by allowing CO(2) to escape in open beakers before the CAT analysis. The effect of pH was also investigated by mixing plasma with buffers with different pH levels. RESULTS: At a pH close to 8.0, endogenous thrombin potential (ETP) and peak decreased considerably, whereas lagtime and time‐to‐peak were modestly increased. Mixtures of plasma and buffer with pH levels between 7 and 8 showed that ETP and peak decreased at alkaline pH; lagtime and time‐to‐peak were higher at acidic pH levels but were shortened, partly in contrast to first results, at alkaline pH levels. The addition of 4‐(2‐hydroxyethyl)‐1‐piperazineethanesulfonic acid buffer to plasma with a high pH attenuated the effects; however, the effect was most significant if added before the CO(2) escaped. CONCLUSION: Modifications of plasma pH can significantly alter thrombin generation. In alkaline samples, for example, after lengthy storage in a freezer where pH can increase considerably, thrombin generation is lowered. To minimize this effect, plasma should be stored in tubes filled to the maximum volume.
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spelling pubmed-70405402020-02-27 The effect of pH on thrombin generation–An unrecognized potential source of variation Kristensen, Søren Risom Nybo, Jette Pedersen, Shona Res Pract Thromb Haemost Original Articles: Thrombosis BACKGROUND: When CO(2) escapes from plasma, the pH of the plasma increases. In samples left open or kept in long‐term storage, the pH may increase considerably. Assays in which the ratio of plasma sample relative to the total volume including reagents is high may be sensitive to the pH of the plasma sample. OBJECTIVE: The aim was to investigate the effect of the pH of plasma samples used in the calibrated automated thrombin generation (CAT) assay in which the ratio (plasma sample) / (total volume) is high. METHODS: Plasma pH was increased by allowing CO(2) to escape in open beakers before the CAT analysis. The effect of pH was also investigated by mixing plasma with buffers with different pH levels. RESULTS: At a pH close to 8.0, endogenous thrombin potential (ETP) and peak decreased considerably, whereas lagtime and time‐to‐peak were modestly increased. Mixtures of plasma and buffer with pH levels between 7 and 8 showed that ETP and peak decreased at alkaline pH; lagtime and time‐to‐peak were higher at acidic pH levels but were shortened, partly in contrast to first results, at alkaline pH levels. The addition of 4‐(2‐hydroxyethyl)‐1‐piperazineethanesulfonic acid buffer to plasma with a high pH attenuated the effects; however, the effect was most significant if added before the CO(2) escaped. CONCLUSION: Modifications of plasma pH can significantly alter thrombin generation. In alkaline samples, for example, after lengthy storage in a freezer where pH can increase considerably, thrombin generation is lowered. To minimize this effect, plasma should be stored in tubes filled to the maximum volume. John Wiley and Sons Inc. 2020-01-17 /pmc/articles/PMC7040540/ /pubmed/32110752 http://dx.doi.org/10.1002/rth2.12289 Text en © 2020 The Authors. Research and Practice in Thrombosis and Haemostasis published by Wiley Periodicals, Inc on behalf of International Society on Thrombosis and Haemostasis. This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.
spellingShingle Original Articles: Thrombosis
Kristensen, Søren Risom
Nybo, Jette
Pedersen, Shona
The effect of pH on thrombin generation–An unrecognized potential source of variation
title The effect of pH on thrombin generation–An unrecognized potential source of variation
title_full The effect of pH on thrombin generation–An unrecognized potential source of variation
title_fullStr The effect of pH on thrombin generation–An unrecognized potential source of variation
title_full_unstemmed The effect of pH on thrombin generation–An unrecognized potential source of variation
title_short The effect of pH on thrombin generation–An unrecognized potential source of variation
title_sort effect of ph on thrombin generation–an unrecognized potential source of variation
topic Original Articles: Thrombosis
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7040540/
https://www.ncbi.nlm.nih.gov/pubmed/32110752
http://dx.doi.org/10.1002/rth2.12289
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