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High-throughput analysis of testosterone in serum samples by on-line solid phase extraction liquid chromatography-tandem mass spectrometry

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Published/Copyright: July 14, 2011
Clinical Chemistry and Laboratory Medicine
From the journal Clinical Chemistry and Laboratory Medicine Volume 49 Issue 11

Abstract

Background: Liquid chromatography tandem-mass spectrometer (LC-MS/MS) techniques are more and more common in the measurement of testosterone concentrations in biological samples. However, LC-MS/MS methods are more laborious than streamlined automated immunochemistry methods because of the need for tedious pre-purification of the sample before the mass spectrometric analysis. We therefore developed a robust and rapid sample clean-up method to improve the throughput of the whole LC-MS/MS analysis procedure by applying an automated on-line solid-phase extraction (SPE) technique instead of the still widely used conventional liquid-liquid extraction.

Methods: Testosterone was purified by the on-line SPE-column-switching technique after rapid precipitation of serum samples by zinc sulphate/internal standard solution before LC-MS/MS analysis. The results were compared with those of our previous routine LC-MS/MS method using liquid-liquid extraction with tert-butyl methyl ether for the pre-purification of the samples.

Results: The tested on-line SPE-LC-MS/MS method reached the specifications of the previous method with liquid-liquid extraction. The precision of the new method was notably better, especially in the lower concentration range, than with the former method; the total variation was below 10% in the whole quantitation range of 0.25–35 nmol/L. The new method liberates more than 50% of hands-on time of laboratory technicians as well as expensive instrument time for other applications compared with the older method.

Conclusions: The on-line SPE-pre-purification technique tested in long-term use offers a rapid and reliable technique in the LC-MS/MS analysis of serum testosterone and is a valuable tool in the improvement of efficiency in the laborious steroid analytics.

Keywords: testosterone; pre-purification; solid phase extraction; automation; liquid chromatography tandem-mass spectrometer (LC-MS/MS)
Corresponding author: Kari Savolainen, Eastern Finland Laboratory Centre (ISLAB), Puijonlaaksontie 2, 70210 Kuopio, Finland Phone: +358-44-7178741, Fax: +358-17-173200,
Received: 2011-2-1
Accepted: 2011-6-1
Published Online: 2011-07-14
Published in Print: 2011-11-01

©2011 by Walter de Gruyter Berlin Boston

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https://doi.org/10.1515/cclm.2011.663
Keywords for this article
testosterone; pre-purification; solid phase extraction; automation; liquid chromatography tandem-mass spectrometer (LC-MS/MS)

Articles in the same Issue

  1. Editorial
  2. Thank you, indeed!
  3. Reviews
  4. Glomerular filtration rate estimation: performance of serum cystatin C-based prediction equations
  5. Oxidative stress and diabetes mellitus
  6. Human papilloma virus (HPV) molecular diagnostics
  7. Opinion Papers
  8. Urine immunofixation electrophoresis remains important and is complementary to serum free light chain
  9. Permissible performance limits of regression analyses in method comparisons
  10. General Clinical Chemistry and Laboratory Medicine
  11. The value of serum ischemia-modified albumin for assessing liver function in patients with chronic liver disease
  12. Relationship between bilirubin and C-reactive protein
  13. Status of serum-calcium and -albumin measurement in Argentina assessed in 300 representative laboratories with 20 fresh frozen single donation sera
  14. Practical application of Sigma Metrics QC procedures in clinical chemistry
  15. High-throughput analysis of testosterone in serum samples by on-line solid phase extraction liquid chromatography-tandem mass spectrometry
  16. Monitoring of mycophenolic acid and kidney function during combined immunosuppressive therapy
  17. Phenotype of Gc-globulin influences the macrophage activating factor (MAF) levels in serum
  18. Combination of cellular population data and CytoDiff™ analyses for the diagnosis of lymphocytosis
  19. Eosinophiluria is common among patients after ileal conduit surgery
  20. Soluble urokinase plasminogen activator receptor (suPAR) levels in healthy pregnancy and preeclampsia
  21. Uptake of cobalamin and markers of cobalamin status: a longitudinal study of healthy pregnant women
  22. Vitamin B12 absorption judged by measurement of holotranscobalamin, active vitamin B12: evaluation of a commercially available EIA kit
  23. Salivary markers of oxidative stress and Salivette collection systems
  24. Infectious Diseases
  25. Development of a microwell adapted immunoblot system with recombinant antigens for distinguishing human herpesvirus (HHV)6A and HHV6B and detection of human cytomegalovirus
  26. Cardiovascular Disease
  27. High-sensitive troponin T in chronic heart failure correlates with severity of symptoms, left ventricular dysfunction and prognosis independently from N-terminal pro-b-type natriuretic peptide
  28. Plasma symmetric dimethylarginine reference limits from the Framingham offspring cohort
  29. Letters to the Editor
  30. Iron deficiency prevalence is increased among Helicobacter pylori infected Brazilian patients
  31. Helicobacter pylori infection and iron deficiency in a group of Brazilian patients
  32. Multiplex protein assay performance/evaluation and the requirement for precision and correlation to clinical assays
  33. Functional assay or antigen test for protein C and protein S in ischemic stroke: which shows the greatest change?
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