The effect of pH on thrombin generation–An unrecognized potential source of variation
Abstract Background When CO2 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...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2020-02-01
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| Series: | Research and Practice in Thrombosis and Haemostasis |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/rth2.12289 |
| _version_ | 1827850479395143680 |
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| author | Søren Risom Kristensen Jette Nybo Shona Pedersen |
| author_facet | Søren Risom Kristensen Jette Nybo Shona Pedersen |
| author_sort | Søren Risom Kristensen |
| collection | DOAJ |
| description | Abstract Background When CO2 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 CO2 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 CO2 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. |
| first_indexed | 2024-03-12T10:17:56Z |
| format | Article |
| id | doaj.art-a78b5390e19a4427a148207d6303d0b0 |
| institution | Directory Open Access Journal |
| issn | 2475-0379 |
| language | English |
| last_indexed | 2024-03-12T10:17:56Z |
| publishDate | 2020-02-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Research and Practice in Thrombosis and Haemostasis |
| spelling | doaj.art-a78b5390e19a4427a148207d6303d0b02023-09-02T10:20:10ZengElsevierResearch and Practice in Thrombosis and Haemostasis2475-03792020-02-014222422910.1002/rth2.12289The effect of pH on thrombin generation–An unrecognized potential source of variationSøren Risom Kristensen0Jette Nybo1Shona Pedersen2Department of Clinical Biochemistry Aalborg University Hospital Aalborg DenmarkDepartment of Clinical Biochemistry Aalborg University Hospital Aalborg DenmarkDepartment of Clinical Biochemistry Aalborg University Hospital Aalborg DenmarkAbstract Background When CO2 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 CO2 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 CO2 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.https://doi.org/10.1002/rth2.12289alkaline plasmacarbon dioxideendogenous thrombin potentialpHpreanalytical variation |
| spellingShingle | Søren Risom Kristensen Jette Nybo Shona Pedersen The effect of pH on thrombin generation–An unrecognized potential source of variation Research and Practice in Thrombosis and Haemostasis alkaline plasma carbon dioxide endogenous thrombin potential pH preanalytical 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 | alkaline plasma carbon dioxide endogenous thrombin potential pH preanalytical variation |
| url | https://doi.org/10.1002/rth2.12289 |
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