Stability of ascorbic acid in serum and plasma prior to analysis

Ching, Simon; Prins, Alex; Beilby, John

doi:10.1258/000456302320314566

Cited by 33 publications

(34 citation statements)

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“…With respect to degradation of AA in whole blood stored at room temperature before plasma or serum preparation only scattered data have been reported (Liau et al, 1993;Ching et al, 2002;Rumelin et al, 2003;Terzuoli et al, 2004). Our data suggest that stability is preserved best in heparin and citrate vacutainer tubes.…”

Section: Resultsmentioning

confidence: 53%

“…Similar results were observed when whole blood was stored at the same conditions (data not shown). Previously reported data on AA stability in plasma are scarce, and due to differences in experimental design, the limited data may not easily be compared (Lykkesfeldt et al, 1995;Ching et al, 2002). Our study provides a full elucidation on the degradation of AA in plasma and whole blood under conditions that are highly relevant with respect to experimental studies and clinical practice.…”

Section: Resultsmentioning

confidence: 98%

“…In a few studies data on initial values of AA in different vacutainer tubes have been reported (Ching et al, 2002;Rumelin et al, 2003). Our observation, which is in accordance with these earlier more limited studies, demonstrates that AA is most stable in plasma isolated from heparin tubes.…”

Section: Resultsmentioning

confidence: 99%

“…AA is the most abundant form in plasma and sensitive to oxidation and degradation during blood sampling, handling, storage, and analysis (Washko et al, 1992;Levine et al, 1999). Various factors such as choice of anticoagulant, temperature, light, pH, dissolved oxygen, solvent, ionic strength, trace metals as ferric ions or the presence of oxidizing enzymes have been reported to affect AA stability (Bradley et al, 1973;Lunec and Blake, 1985;Galan et al, 1988;Margolis and Davis, 1988;Comstock et al, 1995;Key et al, 1996;Margolis and Duewer, 1996;Chung et al, 2001;Ching et al, 2002). It is essential to avoid degradation and oxidation of AA during sampling, sample handling and analysis, as this may skew the ratio between DHAA and AA.…”

Section: Introductionmentioning

confidence: 99%

“…To our knowledge, only limited and scattered data have been reported earlier and in general only one of several aspects has been evaluated (Lunec and Blake, 1985;Galan et al, 1988;Margolis and Davis, 1988;Comstock et al, 1995;Key et al, 1996;Margolis and Duewer, 1996;Chung et al, 2001;Ching et al, 2002). Furthermore, the majority of these studies use supplemented plasma or serum, (Margolis and Davis 1988;Margolis et al, 1990Margolis et al, , 1991Margolis and Duewer 1996), which may not reflect the degradation pattern that is expected in fresh samples.…”

Section: Introductionmentioning

confidence: 99%

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Stability of whole blood and plasma ascorbic acid

2007

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The objective of this study was to evaluate the impact of pre-analytical factors on the short and long term stability of ascorbic acid (AA), the main form of vitamin C in whole blood and plasma. The effects of various anticoagulants, acidification, storage temperature and time were tested. A recently developed fast and sensitive HPLC method was used to measure AA levels. AA baseline values observed in heparin plasma were significantly higher than values observed in EDTA, citrate and Stabilyte plasma, as well as in serum. pH and temperature were identified as additional critical pre-analytical factors during the short, medium and long term handling and storage. Thus, assessment of reliable and accurate AA status in biological samples demonstrates to be highly dependent on whether the initial conditions during sample handling are controlled. In conclusion, heparin tubes should be used for blood sample collection. As AA is rapidly degraded, sample collection should be followed by immediate centrifugation and plasma acidification. To avoid further degradation during sample handling, samples should be stored at À701C without delay and analyzed within 80 days.

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Section: Resultsmentioning

confidence: 53%

Section: Resultsmentioning

confidence: 98%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Stability of whole blood and plasma ascorbic acid

2007

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Determination of ascorbic acid in human plasma with a view to stability using HPLC with UV detection

Kanďár¹,

Žáková

2008

J of Separation Science

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A method for the measurement of ascorbic acid using HPLC with UV detection and investigation into the protein precipitation techniques with regard to stability and recovery are described. The effectiveness of various protein precipitants was tested. Stability of ascorbic acid samples for analysis was investigated over 10 h. Ascorbic acid samples extracted with metaphosphoric acid were stable on a cooled autosampler (4 degrees C) for at least 10 h (with a decline of 1.8% for ascorbic acid solution and 2.8% for plasma). Perchloric acid as protein precipitant for ascorbic acid was unsuitable (with a decline of 36.0% for ascorbic acid solution and 7.3% for plasma). Analytical performance of this method is satisfactory. The intra- and interassay coefficients of variation were 2.1% (n = 10) and 5.8% (n = 12), respectively. The calibration curve was linear with the tested range of 2.0-250.0 micromol/L. The recovery was 96.1% with CV = 4.8% (n = 6) and the LOD was 3 micromol/L. The preliminary reference ranges of ascorbic acid in a group of blood donors are 50.8 +/- 22.4 micromol/L. This assay is a highly sensitive and reproducible HPLC method for the determination of ascorbic acid in human plasma.

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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

2016

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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. Graphical Abstract 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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Stability of ascorbic acid in serum and plasma prior to analysis

Abstract: Minimum loss of ascorbic acid is achieved if blood is collected into tubes containing dipotassium EDTA and separated within 2 h, followed by immediate deproteinization and preservation.

Cited by 33 publications

References 1 publication

Stability of whole blood and plasma ascorbic acid

Stability of whole blood and plasma ascorbic acid

Determination of ascorbic acid in human plasma with a view to stability using HPLC with UV detection

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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