Development of a liquid chromatography coupled to tandem mass spectrometry method for the quantification of Arginine Vasopressin in human plasma.
A new LC-MS/MS method for measuring Arginine Vasopressin in plasma has been developed, showing high sensitivity and accuracy.
Where it sits
this study against the rest of the lypressin corpusSummary and findings
A highly sensitive LC-MS/MS method was developed for quantifying Arginine Vasopressin (AVP) in human plasma. The method was validated over a concentration range from 0.2 ng/L to 60 ng/L. Precision was below 10%, and accuracy ranged from 96% to 109%.
Abstract
Arginine Vasopressin (AVP) is a nonapeptide synthesized in the hypothalamus and released by the neurohypophysis. It plays a central role in water and electrolyte homeostasis, urine concentration and blood pressure regulation. Dysregulation of AVP secretion is associated with several pathological conditions, including central diabetes insipidus and syndrome of inappropriate antidiuretic hormone secretion. Currently, AVP quantification mainly relies on immunoassays which suffers from limitations such as cross reactivity, limited sensitivity and complex sample handling. Given the very low endogenous concentrations of AVP in human plasma (low ng/L range), a highly sensitive LC-MS/MS method was developed and validated for plasma AVP quantification. AVP was extracted using Oasis WCX solid-phase extraction prior to LC-MS/MS analysis. PBS containing 0.1% BSA was used as surrogate matrix for calibration and validation samples to avoid endogenous interferences. Method validation was performed according to the Clinical Laboratory Standards Institute guidelines. Precision, accuracy, and measurement uncertainty were evaluated using single-nested analysis of variation and e.noval software. Stability in whole blood and plasma samples was also investigated. The method was validated over a concentration range from 0.2 ng/L to 60 ng/L, with precision (coefficient of variation) below 10%, accuracy ranging from 96% and 109%, measurement uncertainty below 15%, and a lower limit of quantification of 0.2 ng/L. EDTA blood samples remained stable up to 3 h at room temperature before centrifugation. In conclusion, we have developed and validated a highly sensitive LC-MS/MS method for the quantification of AVP in plasma. Our novel methodology is well-suited for clinical applications.
Background
This paper addresses the need for accurate quantification of Arginine Vasopressin (AVP) in human plasma, which is crucial for understanding various pathological conditions. Previous methods, primarily immunoassays, have limitations such as cross-reactivity and sensitivity issues. The development of a reliable LC-MS/MS method is significant for clinical applications where precise AVP measurement is necessary.
Methods
The study describes a method development and validation process for quantifying AVP in human plasma using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The method involved solid-phase extraction using Oasis WCX and calibration with PBS containing 0.1% BSA. Validation was performed according to Clinical Laboratory Standards Institute guidelines, assessing precision, accuracy, and measurement uncertainty.
Results
The method was validated over a concentration range from 0.2 ng/L to 60 ng/L, with precision below 10% and accuracy ranging from 96% to 109%. The measurement uncertainty was reported to be below 15%, with a lower limit of quantification of 0.2 ng/L.
Interpretation
The findings suggest that the developed LC-MS/MS method offers improved sensitivity and reliability compared to traditional immunoassays for AVP quantification. While the precision and accuracy are statistically significant, the clinical relevance of these findings depends on their application in diagnosing conditions related to AVP dysregulation. Limitations such as the lack of detailed clinical validation and potential confounding factors in sample handling may affect the broader applicability of the results.
Key findings
- Validated over a concentration range from 0.2 ng/L to 60 ng/L.
- Precision (coefficient of variation) below 10%.
- Accuracy ranging from 96% to 109%.
- Measurement uncertainty below 15%.
- Lower limit of quantification of 0.2 ng/L.
Limitations
- Not reported in abstract.
- Lack of detailed clinical validation.
- Potential confounding factors in sample handling.