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For many years concentrations of functional steroid hormones or their precursors have been determined mainly in plasma or serum samples of patients, occasionally in saliva or tissues, or as their metabolites, in urine or amniotic fluid [1]. Gas Chromatography/Mass Spectrometry (GC/MS) is well-suited for the identification of a large number of potential steroids and metabolites due to its high chromatographic resolution capacity and reproducible ionization efficiency [2]. Albumin is dominant in the binding, transport, and delivery of a range of endogenous ligands and pharmaceuticals because it accounts for approximately 60% of the total proteins in blood serum with a typical concentration of 3.4–5.0 g/dL [3]. Biblio graphy shows that protein precipitation for human plasma analysis performed by GC/MS can be performed successfully with methanol by centrifugation [4, 5, 6] with acetonitrile [7, 8, 9] with 70% perchloric acid and n-hexane [10], with 5% sulfosalicylic [11], with ethanol [12], with acetone [13], and with trichloroacetic acid [14]. Other methods for plasma deproteinization focus on physical methods as described [15]. These methods include ultrafiltration and centrifugation with varied molecular weight cut-offs, or using other simple methods such as heating the plasma. However, it has not been shown that these deproteinization methods interfere with gaschromatography-ion trap-tandem mass spectrometry (GC/IT-MS/MS) analysis of plasmatic steroids. Hence, the aim of this work was to identify if it is necessary and useful to apply a deproteinization treatment to human plasma before its analysis in order to determine steroids hormones using GC/IT-MS/MS.

Is deproteinization necessary in the determination of human plasmatic steroids by GC/IT-MS/MS analysis?