Development and validation of a gas chromatography-flame ionization detection method for quantifying sucrose in equine serum.
Abstract: A simple and accurate method for quantifying sucrose in equine serum that can be applied to sucrose permeability testing in the horse was developed and validated using gas chromatography with flame ionization detection. The assay provided an acceptable degree of linearity, accuracy, and precision at concentrations of sucrose as low as 2.34 μmol/l and as high as 20.45 μmol/l. Percentage recovery of sucrose from serum ranged from 89% to 102%; repeatability and intermediate precision (relative standard deviation) ranged from 3.6% to 6.7% and 4.1% to 9.3%, respectively. The limit of detection was 0.73 μmol/l. No interfering peaks were observed except lactose, which gave 2 peaks, one of which overlapped partially with sucrose. To evaluate the suitability of the method for quantifying sucrose in serum samples from horses with naturally occurring gastric ulceration, 10 horses with and without naturally occurring gastric ulceration were subjected to sucrose permeability testing. All horses demonstrated an increase in serum sucrose concentration over time following oral administration of sucrose; however, the increase from baseline was significant for horses with gastric ulceration at 45 min (P = 0.0082) and 90 min (P = 0.0082) when compared with healthy horses. It was concluded that gas chromatography with flame ionization detection is a valid method for quantifying sucrose in equine serum and can be applied directly to the analysis of sucrose in equine serum as part of a larger validation study aimed at developing a blood test for the diagnosis of gastric ulcers in horses.
Publication Date: 2014-02-11 PubMed ID: 24518277DOI: 10.1177/1040638713519640Google Scholar: Lookup
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- Journal Article
Summary
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The researchers have developed and validated a technique using gas chromatography to measure the amount of sucrose in horse serum. The method could help identify gastric ulcers in horses through analyzing the presence of sucrose in the blood following an oral dosing.
Method Development and Validation
- To test the permeability of sucrose in horses, a method was developed using gas chromatography with flame ionization detection. Flame ionization was used as it is a simple, accurate and widely used method to detect and quantify various substances.
- The developed method showed satisfactory linearity, accuracy, and precision at sucrose concentrations ranging from 2.34 μmol/l to 20.45 μmol/l.
- The recovery percentage of sucrose from the serum varied between 89% and 102%, indicating the efficiency of the method in extracting sucrose.
- The method showed reliable repeatability and intermediate precision, with a relative standard deviation ranging between 3.6% and 6.7% and 4.1% to 9.3%, respectively.
- The detection limit of the method was found to be 0.73 μmol/l, which implies the lowest amount of sucrose that could be reliably quantified.
- The method did not produce any interfering signals except for lactose, which gave 2 overlapping peaks with sucrose. But this did not affect the overall quantification of sucrose.
Use in Gastric Ulceration Diagnosis
- Researchers tested the developed method on serum samples from horses with and without gastric ulcerations as a case study for its applicability.
- The serum samples were subject to sucrose permeability tests, where the concentration of sucrose was measured following oral administration of sucrose to horses.
- All horses displayed an increase in serum sucrose concentration over time after taking the sucrose dose.
- Horses with gastric ulcers showed a significant increase from baseline in sucrose level at the 45-minute and 90-minute marks compared to healthy horses.
- The study concluded that measuring sucrose levels in horse serum could potentially be used as a diagnostic tool for the identification of gastric ulcers in horses.
Conclusion
- The generated method using gas chromatography with flame ionization detection was successful in quantifying the presence of sucrose in equine serum.
- This validated method may serve a critical role in diagnosing gastric ulcers in horses by monitoring changes in serum sucrose concentration after oral administration of sucrose.
- This validated method paves the way for a larger validation study designed to develop a blood test for the diagnosis of gastric ulcers in horses.
Cite This Article
APA
Hewetson M, Aaltonen K, Tulamo RM, Sankari S.
(2014).
Development and validation of a gas chromatography-flame ionization detection method for quantifying sucrose in equine serum.
J Vet Diagn Invest, 26(2), 232-239.
https://doi.org/10.1177/1040638713519640 Publication
Researcher Affiliations
- 1Michael Hewetson, Faculty of Veterinary Medicine, PO Box 57, 00014, University of Helsinki, Finland. michael.hewetson@helsinki.fi.
MeSH Terms
- Animals
- Chromatography, Gas / methods
- Chromatography, Gas / veterinary
- Flame Ionization / methods
- Flame Ionization / veterinary
- Horses / blood
- Reproducibility of Results
- Sensitivity and Specificity
- Sucrose / blood
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