Document Detail

Validation of radioimmunoassay for estradiol-17 beta by isotope dilution-mass spectrometry and by a test of radiochemical purity.
MedLine Citation:
PMID:  6360429     Owner:  NLM     Status:  MEDLINE    
Isotope dilution-mass spectrometry (ID-MS) was used as a reference method to determine the concentration of estradiol-17 beta (E2) in five different plasma pools (concentrations ranging from 0.040 to 65 nmol/l). The same plasma pools were also subjected to radioimmunoassay (RIA) using five different antisera of largely varying specificity. With the best antiserum (E), a direct RIA apparently gave accurate results (i.e. results statistically indistinguishable from those obtained by ID-MS) at all levels except the lowest one (0.040 nmol/l). It was shown, however, that the apparent accuracy of this RIA to some extent could be due to a lowering effect of lipids in the serum masking a lack of specificity of the antibodies. With the least specific antiserum (A), accurate results were obtained only after chromatography. However, in the assay of the lowest concentration of E2 with this antiserum there was a significant overestimation, even after chromatography. The other three antisera (B, C, D) of average quality gave accurate results in assays of plasma diethyl ether extracts in various numbers of the plasma pools tested, depending on their intrinsic specificity. This specificity was not correlated with the cross-reaction reported for individual antisera. ID-MS is difficult to use in most laboratories. We demonstrate here that the validity of a RIA may in this case be assessed by a relatively simple method, the test of radiochemical purity (RP-test). This test is based on the measurements of specific activity (e.g. dpm/pg) in small consecutive fractions of the chromatographic zone which is usually employed for the RIA.
S Xing; S Z Cekan; U Diczfalusy; O Falk; S A Gustafsson; E Akerlöf; I Björkhem
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Clinica chimica acta; international journal of clinical chemistry     Volume:  135     ISSN:  0009-8981     ISO Abbreviation:  Clin. Chim. Acta     Publication Date:  1983 Dec 
Date Detail:
Created Date:  1984-02-14     Completed Date:  1984-02-14     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  1302422     Medline TA:  Clin Chim Acta     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  189-201     Citation Subset:  IM    
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MeSH Terms
Estradiol / blood*
Evaluation Studies as Topic
Indicator Dilution Techniques
Lipids / blood
Mass Spectrometry
Reg. No./Substance:
0/Lipids; 50-28-2/Estradiol

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