Evaluation of a one-step sample preparation protocol for analysis of total serum testosterone by LC–MS/MS

madman

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4. Summary

This study compared our one-step procedure to a recently reported two-step sample preparation extraction procedure for the accurate and efficient measurement of testo in human serum (e.g., postmenopausal, premenopausal, male and prepubescent) in terms of specificity, sensitivity, data accuracy, recovery and lipid removal. We found the one step procedure simplified sample preparation and did not compromise data reliability; there was no discernable impact of the matrix effect on data accuracy. The difference in accuracy between the two procedures is less than 5% for adult sera, while it is less than 10% for prepubescent sera. The specificity was monitored by the ion ratio technique (i.e., the ratio of the response of transition 289 > 97 to that of 289 > 109) and no interference in the testosterone peak was observed for samples prepared by either the one-step or two-step procedure. However, recovery in unaltered serum using the one-step procedure was approximately 15% greater across the low to high concentration range than that for the two-step procedure,while lipids were removed efficiently in the two-step procedure as verified by monitoring the typical phospholipid MRM transition of m/z 496 > 184. A further benefit of the simplicity of the one-step procedure is a reduction in the potential for human error.
 

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Estradiol (E2)

A form of estrogen produced from testosterone. Important for bone health, mood, and libido. Too high can cause side effects; too low can affect well-being.

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Dihydrotestosterone is a potent androgen derived from testosterone. Affects hair growth, prostate health, and masculinization effects.

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The biologically active form of testosterone not bound to proteins. Directly available for cellular uptake and biological effects.

Scientific Reference

Lakshman KM, Kaplan B, Travison TG, Basaria S, Knapp PE, Singh AB, LaValley MP, Mazer NA, Bhasin S. The effects of injected testosterone dose and age on the conversion of testosterone to estradiol and dihydrotestosterone in young and older men. J Clin Endocrinol Metab. 2010 Aug;95(8):3955-64.

DOI: 10.1210/jc.2010-0102 | PMID: 20534765 | PMCID: PMC2913038

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