Evaluation of enzyme immunoassay and radioimmunoassay methods for the measurement of plasma oxytocin
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OBJECTIVE: There is increased interest in measuring peripheral oxytocin levels to better understand the role of this peptide in mammalian behavior, physiology, and disease. The purpose of this study was to compare methods for plasma oxytocin measurement using a commercially available enzyme immunoassay (EIA) and radioimmunoassay (RIA), to evaluate the need for sample extraction, and to assess the immunospecificity of the assays. METHODS: Oxytocin was measured in extracted and unextracted human plasma samples (n = 39). Oxytocin and its degradation products were separated by high-performance liquid chromatography and gel filtration chromatography and then assayed by EIA or RIA to identify oxytocin immunoreactive peaks. RESULTS: Without extraction, plasma measured by EIA was more than 100-fold higher than in extracted plasma, and the correlation between oxytocin levels in extracted and unextracted plasma was minimal (Spearman ρ = -0.10, p = .54). Using the RIA, most samples (>90%) were below the level of detection with or without extraction. After chromatographic fractionation of sample extracts, multiple immunoreactive products were found to be present in addition to oxytocin, which casts doubts on the specificity of the assays. CONCLUSIONS: Changes in oxytocin levels have been reported in social and behavioral challenge studies. This study indicates that sample extraction is necessary to obtain valid assay results. Changes in oxytocin degradation products are likely to contribute to the previously observed responses in circulating oxytocin levels to behavioral and social challenge. There is a critical need for valid and reliable methods to measure oxytocin in biologic samples.
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Three studies that compared ELISA oxytocin measurements of the same samples with and without extraction all found no correlations.
“Three studies have compared ELISA oxytocin measurements of the same samples with and without extraction (69-71): all reported no correlations.”
From Intranasal Oxytocin: Myths and Delusions (2016) · cites this paper as [71]
EIA performed without extraction yielded values two orders of magnitude greater than those in extracted samples and without correlation, due to the presence of multiple adjunctive immunoreactive products besides oxytocin.
“EIA performed without extraction yielded to values that were two orders of magnitude greater than those in extracted samples (Szeto et al., 2011) and without correlation, due to the presence of multiple adjunctive immunoreactive products besides oxytocin (Szeto et al., 2011).”
From Peripheral oxytocin and vasopressin: Biomarkers of psychiatric disorders? A comprehensive systematic review and preliminary meta-analysis (2016) · cites this paper as Szeto et al., 2011
Three studies compared measurements of oxytocin in the same samples by the same ELISA with and without extraction; Robinson et al. found approximately 8 pg/ml in extracted seal plasma versus 300 pg/ml in unextracted heparin samples or 540 pg/ml in unextracted EDTA samples with no correlation, Christensen et al. found radioimmunoassay extracted mean levels of 1.1 pg/ml and spiked levels higher by t
“Three studies have compared measurements of oxytocin in the same samples by the same ELISA, with and without extraction 101, 116, 117. Robinson et al. 116 measured oxytocin levels in seals: in extracted samples, they found plasma levels of approximately 8 pg/ml, compared to 300 pg/ml in unextracted samples collected in heparin vacutainers or 540 pg/ml in unextracted samples collected in ethylenedi”
From Measuring Oxytocin and Vasopressin: Bioassays, Immunoassays and Random Numbers (2016) · cites this paper as [101]
Currently, endogenous oxytocin assessment methods and interpretations of the relation between plasma oxytocin levels and corresponding central nervous system functions are still contested.
“Currently, endogenous OT assessment methods (Szeto et al., 2011) and interpretations of the relation between plasma OT levels and corresponding central nervous system functions are still contested (Meyer-Lindenberg et al., 2011;Churchland and Winkielman, 2012).”
From The effect of oxytocin on cooperation in a prisoner's dilemma depends on the social context and a person's social value orientation (2014) · cites this paper as Szeto et al., 2011
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- Authors
- Szeto, A., McCabe, P. M., Nation, D. A., Tabak, B. A., Rossetti, M. A., McCullough, M. E., Schneiderman, N., & Mendez, A. J.
- Affiliations
- National Institutes of Health (US)
- Corresponding author
- Mendez, Armando J.
- Venue
- Psychosomatic Medicine
- Published
- 2011-06
- Volume
- 73
- Issue
- 5
- Pages
- 393-400
- ISSN
- 0033-3174 (ISSN-L)
- PMC
- PMC3118424
- OpenAlex
- W2060180675
- Cited by
- 330
- References
- 56
- Citations by year
- 2026: 11 · 2025: 12 · 2024: 16 · 2023: 22 · 2022: 23 · 2021: 32 · 2020: 24 · 2019: 26 · 2018: 32 · 2017: 32 · 2016: 29 · 2015: 27 · 2014: 20 · 2013: 15 · 2012: 6
- FWCI
- 19.21
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Psychosomatic Medicine (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Heart, Lung, and Blood Institute (T32HL007426, P01HL036588, HL36588, HL-04726)
- Topics
- Social Sciences › Psychology › Social Psychology › Neuroendocrine regulation and behavior
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuropeptides and Animal Physiology
- Life Sciences › Neuroscience › Behavioral Neuroscience › Stress Responses and Cortisol
- Keywords
- plasma oxytocin, enzyme immunoassay, radioimmunoassay, assay comparison, sample extraction, immunospecificity, immunoreactive peptides, human plasma
- MeSH terms
- Animals, Cattle, Chromatography, Liquid, Arvicolinae, Female, Humans, Immunoenzyme Techniques, Macaca, Male, Oxytocin, Rabbits, Radioimmunoassay, Reagent Kits, Diagnostic, Sensitivity and Specificity, Mice, Rats, Limit of Detection
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