Ultra sensitive quantitation of endogenous oxytocin in rat and human plasma using a two-dimensional liquid chromatography-tandem mass spectrometry assay
Abstract
Oxytocin (OT) is a neuropeptide with an extremely low endogenous level (low pg/ml) in human plasma. It is very challenging to develop a highly sensitive assay to measure endogenous OT, including radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA). Electrospray ionization (ESI) liquid chromatography-tandem mass spectrometry (LC-MS/MS) can provide high-throughput and selective methods for quantification of peptides in biological samples. A novel and highly sensitive two-dimensional LC-MS/MS (2D-LC-MS/MS) assay combining solid-phase extraction (SPE) has been developed and validated for the determination of endogenous OT in both human and rat plasma. The lower limit of quantification (LLOQ) was 1.00 pg/ml for human and 50.0 pg/ml for rat. Human plasma diluted with water (1:6, v/v) was successfully optimized as a surrogate matrix for human to prepare standard curves without endogenous interference. The extraction efficiency and absolute recovery were above 65.8% using the HLB SPE procedure, and matrix effects were lower than 12%. The method was validated in the range of 1.00-250 pg/ml for human plasma and 50.0-10,000 pg/ml for rat plasma with precision less than 12.7% and accuracy less than 7%.
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Validated radioimmunoassays, confirmed by combined liquid chromatography/mass spectrometry, indicate that basal circulating oxytocin and vasopressin levels in humans are 1-10 pg/ml.
“Validated radioimmunoassays have long converged on the conclusion that basal circulating levels of oxytocin and vasopressin in man are in the range 1-10pg/ml, confirmed recently by combined LC/mass spectrometry (57).”
From Intranasal Oxytocin: Myths and Delusions (2016) · cites this paper as [57]
Plasma oxytocin was measured using human plasma diluted with water (1:6) as a surrogate matrix, and basal levels of 2.4 pg/ml for human but 298 pg/ml for rat were reported; these high levels in the rat may indicate that human plasma is not a suitable surrogate matrix for measurements in rat.
“Plasma oxytocin was measured using human plasma diluted with water (1 : 6) as a surrogate matrix, and basal levels of 2.4 pg/ml for human but 298 pg/ml for rat were reported; these high levels in the rat may indicate that human plasma is not a suitable surrogate matrix for measurements in rat.”
From Measuring Oxytocin and Vasopressin: Bioassays, Immunoassays and Random Numbers (2016) · cites this paper as [125]
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- Authors
- Zhang, G., Zhang, Y., Fast, D. M., Lin, Z., & Steenwyk, R.
- Affiliations
- Pfizer (United States) (US)
- Corresponding author
- Zhang, Guodong
- ORCID
- Zhang, Guodong
- Venue
- Analytical Biochemistry
- Publisher
- Elsevier BV
- Published
- 2011-09
- Volume
- 416
- Issue
- 1
- Pages
- 45-52
- ISSN
- 0003-2697 (ISSN-L)
- OpenAlex
- W2081826142
- Cited by
- 79
- References
- 40
- Citations by year
- 2026: 3 · 2025: 3 · 2024: 5 · 2023: 5 · 2022: 5 · 2021: 4 · 2020: 2 · 2019: 3 · 2018: 9 · 2017: 8 · 2016: 8 · 2015: 8 · 2014: 7 · 2013: 5 · 2012: 3
- FWCI
- 6.96
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Analytical Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Social Sciences › Psychology › Social Psychology › Neuroendocrine regulation and behavior
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical Methods in Pharmaceuticals
- Keywords
- oxytocin, two-dimensional liquid chromatography, tandem mass spectrometry, solid-phase extraction, human plasma, rat plasma, matrix effects, HLB extraction
- MeSH terms
- Animals, Chromatography, High Pressure Liquid, Humans, Oxytocin, Sensitivity and Specificity, Rats, Solid Phase Extraction, Tandem Mass Spectrometry
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