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Rapid and harmonized analytical workflow for the determination of peptidic and non-peptidic doping agents in dried and liquid blood matrices
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DOI：10.1039/D6AN00455E.png)
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Abstract
En 中文
Recently; methods for detecting small peptides in dried blood spots have been published. These procedures typically involve multiple sample preparation steps; resulting in labor-intensive and costly workflows. In the present study; we report a fast; streamlined; and harmonized analytical workflow to detect 54 prohibited peptidic and non-peptidic compounds in dried blood spots; serum; and plasma. Sample preparation is based on a single microextraction step using 500 µL of a methanol/water (8:2; v/v) mixture. Detection was performed using liquid chromatography coupled with high-resolution mass spectrometry. The validation results showed satisfactory performance with respect to selectivity (no interferences were detected at the retention times of the analytes); detection limits (0.05-1.25 ng/mL); carry-over (no signals in the negative sample injected after the positive sample); matrix effect (5-33%); extraction yield (15-80%); and extract stability (the target analytes were stable for at least 72 h in the autosampler at 10 °C). The method was successfully applied to samples containing sub-ng levels of ibutamoren; confirming that the analytical procedure presented in this study is fit for purpose within the doping-control framework. Stability studies showed that all compounds were stable (variation lower than 15%) for at least two months at −20 °C in all the blood matrices considered. At 4 and 22 °C; alexamorelin; AOD9604; buserelin; hGH 176–191; kisspeptin-10 and LHRH were extensively degraded after one week in serum and plasma; whereas BPC-157; TB500; vasopressin; lypressin; and terlipressin showed complete degradation only in serum. In contrast; in dried matrices all compounds remained detectable throughout the entire duration of the study; indicating that samples can be transported and stored under non-refrigerated conditions; thereby reducing costs.
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